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Journal of Biomedicine and Biotechnology
Volume 2010, Article ID 596432, 13 pages
Strategies for Cancer Vaccine Development
Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA
Received 13 January 2010; Accepted 17 May 2010
Academic Editor: Zhengguo Xiao
Copyright © 2010 Matteo Vergati et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Treating cancer with vaccines has been a challenging field of investigation since the 1950s. Over the years, the lack of effective active immunotherapies has led to the development of numerous novel strategies. However, the use of therapeutic cancer vaccines may be on the verge of becoming an effective modality. Recent phase II/III clinical trials have achieved hopeful results in terms of overall survival. Yet despite these encouraging successes, in general, very little is known about the basic immunological mechanisms involved in vaccine immunotherapy. Gaining a better understanding of the mechanisms that govern the specific immune responses (i.e., cytotoxic T lymphocytes, CD4 T helper cells, T regulatory cells, cells of innate immunity, tumor escape mechanisms) elicited by each of the various vaccine platforms should be a concern of cancer vaccine clinical trials, along with clinical benefits. This review focuses on current strategies employed by recent clinical trials of therapeutic cancer vaccines and analyzes them both clinically and immunologically.
1. Introduction
Cancer is the second leading cause of death in the United States, exceeded only by heart disease (23.1% versus 26.0% of total deaths, resp.). Currently, 1 in 4 deaths in the United States is due to cancer. According to American Cancer Society statistics, an estimated 1,479,350 new cases and 562,340 deaths from cancer are expected during 2009, with a slightly higher incidence and death rate in the male population. Prostate, lung, and colorectal cancers are the most common types of cancer in men; breast, lung, and colorectal cancers are most common among women. Altogether, lung, breast, prostate, and colorectal cancers account for 49% of cancer-related deaths in the U.S. population [1]. Overall, except for lung cancer in women, incidence and mortality rates have steadily decreased for all 4 types of cancer in both men and women, probably due to both an increase in early diagnosis and improvements in therapy and combination therapies (surgery, radiotherapy, chemotherapy, and, lately, targeted therapy). But despite these encouraging advances, cancer is still a major public health problem worldwide, requiring new strategies and treatment modalities to optimize patient outcomes.
In this context, immunotherapy has always been an attractive and potentially efficient treatment for cancer patients [19]. Tumor immunotherapy can generally be classified as (a) passive (or adaptive), consisting of administration of cells or antibodies ex vivo, and (b) active, represented by vaccines, aimed at eliciting a specific immune response against tumor-specific antigens (TSAs) and tumor-associated antigens (TAAs). Prophylactic and therapeutic vaccines represent one of the most intriguing approaches in the multidisciplinary treatment of cancer patients. Compared to all other standard modalities (surgery, chemotherapy, radiotherapy, and adaptive immunotherapy), an effective vaccine-based immune response against tumor may be the only cancer treatment with the potential to last a lifetime. Theoretically, vaccinated patients could mount an immune response able to either cure tumor or keep it under constant restraint (i.e., immune surveillance), delaying tumor recurrence and prolonging survival.
One of the major problems in developing an efficient cancer vaccine is the lack of TSAs and the weakness of immune responses against TAAs, usually recognized by the immune system as self-antigens. During the last decades, various strategies for therapeutic cancer vaccines have been proposed to overcome this weak immune response against TAAs, including cell-based vaccines, DNA- or RNA-based vaccines, protein- or peptide-based vaccines, and vector-based vaccines [20]. The common rationale for all these modalities is the activation of antigen-presenting cells (APCs) and the stimulation of an antigen-specific cytotoxic T lymphocyte-(CTL-) mediated immune response. Dendritic cells (DCs) are the most potent APCs, and various strategies have been used to enhance their ability to activate T cells. This review focuses on the state of the art of these modalities and analyzes the most promising phase II/III clinical trials, emphasizing vaccines directed against carcinomas (Table 1). Despite recent achievements, one criticism of some of these clinical trials has been the lack of immunological data supporting the significant improvements in time to progression and overall survival (OS) observed. An effort should be made to define the specific components of each immune response as a consequence of anticancer vaccination. In this context, both the specificity and the identification of potential escape mechanisms (i.e., increase of Treg number or function, balance between positive and negative regulators of antitumor responses, such as CD28, cytotoxic T lymphocyte-associated antigen 4 (CTLA-4), programmed death-1 molecule (PD-1) and its ligands PD-L1 and PD-L2) should be investigated. Increasing our understanding of how these modalities modulate the CTL response is vital to developing novel and effective antitumor vaccines.
The goal of therapeutic cancer vaccines is to "teach" the patient's own immune system to specifically recognize and eliminate tumor cells. The potential target for the immune response can be either TSAs (antigens present only on tumor cells) or TAAs (antigens present mostly on tumor cells but also on some normal cells). Theoretically, TSAs are the ideal target for cancer immunotherapy because of their specificity. They are largely composed of mutant proteins caused by somatic mutations in the original sequence of the protein. A major advantage of targeting TSAs is that many of these proteins have been demonstrated to be essential for tumorigenesis and cancer progression [21]. On the other hand, a major drawback of targeting TSAs is the fact that most of the mutations identified are unique to each tumor, potentially requiring the development of personalized immunotherapy for individual patients. In contrast, TAAs are commonly expressed on tumors with the same histology and are shared among tumors of different origin. A major limitation of targeting TAAs is that they are weakly immunogenic due to the tolerance for self-antigens acquired by the immune system in its developmental stages [22].
In the last decades, several different mechanisms have been proposed to "instruct" DCs, the most potent APCs known, to induce Th and CTL responses against tumor antigens, thus breaking immune tolerance. Antigen-loading techniques include (a) infecting DCs with viral, bacterial, or yeast vectors, (b) pulsing DCs with proteins or peptides, (c) loading DCs with tumor cells or tumor-cell lysates, and (d) transfecting DCs with DNA or RNA (Figure 1) [20].
Encouraged by positive preclinical and clinical data [23–27], further studies are currently ongoing to evaluate the possibility to enhance vaccine-induced immunity by combining vaccines with low doses of chemotherapeutic agents (i.e., cyclophosphamide, doxorubicin, docetaxel) or radiation therapy, that showed synergistic immunotherapeutic effects when given in proper sequence.
2. Vaccines with Viral, Bacterial, or Yeast Vectors
As mentioned above, one of the major difficulties in cancer immunotherapy is to develop a strategy to overwhelm the characteristically weak immune response of the host against TAAs. Several vectors can be used to deliver recombinant genes (including genes expressing TAAs, costimulatory molecules, or cytokines) into APCs. Recombinant vector-based vaccines may induce the immune system to generate a strong inflammatory response, directed mainly towards vector proteins. In turn, this inflammatory response may lead to an increased immune response against the genes of interest that have been inserted into the vector. One advantage of using vectors as vehicles for TAAs is that this type of delivery of a recombinant protein is much more immunogenic than administering the protein with adjuvants [28, 29].
Vectors used in cancer immunotherapy include viral, bacterial, and yeast vectors. The choice of vector can have important consequences for the subsequent immune response against TAAs because each vector has its own characteristics and is potentially able to uniquely stimulate the host immune system. A further factor that must be taken into account in the development of an efficient vector-based vaccine strategy is the balance between the stimulation of innate versus adaptive responses, Th1 versus Th2 responses, or the preferred activation of subsets of cells mainly committed to regulatory (Tregs, Tr1, and Th3) or proinflammatory functions (Th17).
Poxviral vectors are among the most heavily exploited in vaccine development. The prototype is vaccinia virus, which was used successfully to eradicate smallpox [30]. The poxvirus family is composed of double-stranded DNA viruses that replicate within the cytoplasm of infected cells. This feature is important for the safe use of poxviruses as recombinant vaccines, since no genetic sequence from the virus will be inserted into the host cell genome. However, owing to concerns about the use of replicating vectors in potentially immunocompromised patients and immune responses generated against the vector by immunocompetent patients, developing safe, nonreplicating viral vectors has been the focus of extensive research. Other attenuated poxviruses have been identified and are currently available for clinical use. Fowlpox, an avipoxvirus, can infect mammalian cells abortively, but recombinant-encoded genes are transcribed [31]. The drawback is that recombinant fowlpox usually generates a weaker immune response in humans than vaccinia and is thus often used for booster vaccinations after a primary vaccination with recombinant vaccinia. Modified vaccinia Ankara (MVA) is a highly attenuated strain of vaccinia that was developed by hundreds of passages of vaccinia virus in chick embryo fibroblasts. MVA can infect mammalian cells and undergo DNA replication in them but has lost the ability to produce infective viral particles [32]. Preclinical and clinical studies have demonstrated the superiority of a priming vaccination with recombinant vaccinia followed by multiple boosts with recombinant fowlpox, over different dosing schedules or the continuous use of either vector alone [33–36].
The large genome of poxviruses (approximately 130 kb for mammalian poxviruses and 300 kb for avian poxviruses) allows for insertion of more than 10 kb of foreign DNA.
Moreover, gene products are usually expressed at high levels, resulting in a potent cellular immune response. As mentioned previously, poxviruses can also be modified to express one or more T-cell costimulatory molecules along with the transgene for a TAA, or cytokines such as GM-CSF. Tumor recognition by CTLs is a complex mechanism that requires several different signals. DCs provide T cells with antigenic signal 1 through the specific interaction between the peptide-MHC I complex and the T-cell receptor. A costimulatory signal 2 is needed for the activation and expansion of T cells. Finally, DCs provide an additional polarizing signal 3 through the release of different cytokines, driving the immune response toward type-1 or type-2 immunity. Therefore, costimulatory molecules are critical in the generation of potent T-cell responses, particularly toward weak antigens such as TAAs. The most studied costimulatory signals involve the interaction between B7.1 (CD80) expressed on APCs and CD28 or CTLA-4 on T cells, between intercellular adhesion molecule-1 (ICAM-1 or CD54) on APCs and leukocyte function-associated antigen-1 (LFA-1) on T cells, and between LFA-3 (CD58) on APCs and CD2 on T cells [37].
PSA-TRICOM vaccine (prostate-specific antigen plus a TRIad of COstimulatory Molecules; PROSTVAC) consists of a priming vaccination with recombinant vaccinia- (rV-) PSA-TRICOM and booster vaccinations with recombinant fowlpox- (rF-) PSA-TRICOM. Each vaccine consists of the transgenes for PSA, including an agonist epitope [38], and 3 immune costimulatory molecules (B7.1, ICAM-1, and LFA3; designated TRICOM). The efficacy of PSA-TRICOM has been evaluated in 2 phase II clinical trials in patients with metastatic hormone-refractory prostate cancer (mHRPC). In the first multicenter clinical trial, 122 patients with Gleason scores of 7 were randomized 2 : 1 to receive PSA-TRICOM plus GM-CSF () versus an empty-vector placebo (). A vaccinia-based vector was used as prime, followed by 6 boosts with a fowlpox-based vector. Vaccinated patients had a greater 3-year OS compared to the placebo arm (30% versus 17%, resp.) and an improvement in median OS of 8.5 months (24.5 months versus 16 months, resp.; ). T-cell responses to vaccine or vector were not evaluated in this trial [2–4].
In a concurrent phase II clinical trial at the National Cancer Institute employing the identical PSA-TRICOM vaccine, 32 patients (representing all Gleason scores) were randomized to 1 of 4 cohorts. Cohort 1 received no immune adjuvant; cohort 2 received recombinant human GM-CSF protein; cohort 3 received 107 plaque-forming units (pfu) rF-GM-CSF; cohort 4 received 108 pfu rF-GM-CSF. All patients were primed with rV-PSA-TRICOM s.c. on day 1 and then received monthly boosts of rF-PSA-TRICOM until progression. Patients who remained on-study after 12 months had booster vaccinations every 3 months. With a median follow-up of 44.6 months, the median OS for all 32 patients on-study was 26.6 months, compared to a median Halabi nomogram-predicted survival of 17.4 months (an improvement of 9.2 months) [5]. No major differences were observed among the 4 cohorts. The subanalysis of patients with a Halabi-predicted survival (HPS) of 18 months showed a minimal difference between actual OS and HPS. However, patients with HPS 18 months had a significant increase in actual OS (37.3 months, median not reached, with 8 of 15 patients still alive at 44.6 months) compared to HPS (20.9 months). PBMCs from patients pre and post vaccination were analyzed by ELISPOT assay to evaluate the specific immune response against the HLA-A2 PSA peptide PSA-3 [39]. Thirteen of 29 patients analyzed had an enhanced (2-fold) PSA-3-specific T-cell immune response post vaccination. Furthermore, patients with a postvaccination ELISPOT response to PSA epitope 6-fold seemed to live longer, compared to patients with a postvaccination ELISPOT response to PSA epitope 6-fold (). We also analyzed Treg function pre and post vaccination. Among patients who survived longer than predicted, Treg suppressive function decreased in 10/13 (77%) after 3 vaccinations versus pre vaccination. In contrast, among patients whose survival was less than predicted, Treg function increased in 6/8 (75%) after 3 vaccinations versus pre vaccination. These data strongly suggest that Tregs play a significant role in the modulation of antitumor immune response [40].
PANVAC-VF, another poxviral-based vaccine, consists of a priming vaccination with rV encoding CEA(6D), MUC1(L93), and TRICOM plus booster vaccinations with rF expressing the identical transgenes. CEA(6D) and MUC1(L93) represent carcinoembryonic antigen and mucin 1 glycoprotein, respectively, with a single amino acid substitution designed to enhance their immunogenicity [41, 42]. A phase III study in patients with advanced pancreatic cancer treated with PANVAC-VF as second-line therapy showed no improvement in survival [6]. The vaccine is currently under evaluation in several different types of CEA- or MUC1-expressing carcinomas and in patients with a life expectancy 3 months. In our experience, PANVAC-VF was well tolerated in a pilot study enrolling 25 patients with metastatic carcinomas. After vaccination, CAP1(6D)-specific CD8 immune responses were detected in 3/8 patients by ELISPOT, CAP1(6D)-tetramer, and intracellular IFN-γ staining. We also evaluated CD4 immune responses in 15 patients included in the study, using CEA protein as antigen. Six of 15 patients with undetectable levels of IFN- pre vaccination showed measurable levels in response to CEA protein. Four of 14 patients were positive for the generation of MUC1-specific T cells post vaccination [43].
The rationale for the use of microbes such as yeast as delivery vehicles for TAAs is based on the ability of these agents to activate a proinflammatory response through the interaction of pathogen-associated molecular patterns with pattern-recognition receptors, such as Toll-like receptors, expressed on APCs. These interactions play a central role in the activation of innate and adaptive immunity [44]. Over the years, several different bacterial and yeast vectors, such as Escherichia coli, Salmonella, Shigella, Yersinia, Listeria monocytogenes, and Saccharomyces cerevisiae, have been investigated for use as vaccine vectors.
The development of genetic engineering technology and efficient fermentation procedures has made large-scale, cost-effective production of these vectors possible and is one of the major advantages of their use in antitumor vaccines. Unfortunately, development of yeast-based vaccines has lagged behind that of cell-, protein-, and viral-based vaccines, and clinical experience has been limited to phase I/II studies [45]. One such vector currently being evaluated is a whole, heat-killed, recombinant S. cerevisiae yeast (Tarmogens GI-4000, GlobeImmune) intended to generate a T-cell response to eliminate tumor cells expressing the 7 most common mutations in the ras oncogene product. A randomized, double-blind, placebo-controlled phase IIa clinical trial has enrolled 100 patients with resected pancreatic cancer, with half receiving adjuvant gemcitabine plus placebo and half receiving adjuvant gemcitabine plus GI-4000 [46].
3. Vaccines with Proteins or Peptides
The use of proteins or peptides to stimulate a specific immune response against cancer has long been investigated and covers a broad spectrum of possibilities employing single agents or combinations of proteins, heat-shock proteins (HSPs) [47], peptides and agonist peptides [48–51], anti-idiotype antibodies [52, 53], and fusion proteins [54]. These protein- or epitope-based vaccines have 2 main advantages over the use of tumor cells or lysates: (a) production, storage, and distribution are faster and more cost-effective, and (b) the identification and administration of TSAs is preferable since tumor-cell preparations mostly contain self-proteins with no therapeutic benefit and are potentially capable of generating an autoimmune response. On the other hand, this approach has certain drawbacks: (a) first is the weak immunogenicity of a single protein or, especially, a single epitope; (b) tumors can easily escape immune recognition through antigen mutation or loss; (c) their use is HLA-restricted (mainly for epitope-based vaccines) and limited to a subset of patients (usually HLA-A); (d) they have a poor ability to induce balanced activation of CD4 and CD8 subsets, which is thought to be essential for effective, long-lasting antitumor immunity. To date, in fact, most epitope-based vaccines induce HLA-A2-restricted responses that efficiently kill tumor cells but are characterized by a limited lifespan in the absence of CD4 helper T cells. Protein-based vaccines are capable of generating stronger CD4 responses (MHC class II-restricted), but at the cost of less effective induction of CTLs [55, 56]. Most of the issues described above could be easily overcome by the use of longer peptides or the combination of several different epitopes in the same vaccine, while the relatively poor immunogenicity of peptides could necessitate that they be administered with adjuvants or loaded onto DCs [57, 58].
The use of specific proteins or peptides as targets for immunotherapy clearly requires a careful choice of the targeted TAAs and their epitopes, involving knowledge of their structural and functional characteristics. Single-peptide epitopes composed of 8 to 10 amino acids are able to induce a CTL response by binding to MHC class I molecules expressed on APCs. Each epitope is composed of conserved anchor residues (mostly at position 2 and the C-terminal position) needed to bind to the cleft of MHC I molecules and residues that are specific for T-cell recognition. Theoretically, changes in the former do not affect the specificity of the latter, and they have been used as a strategy to increase the immunogenicity of several different epitopes (agonist epitopes) [38, 41, 42, 50, 59]. Furthermore, the ideal TAA should be widely expressed in different tumor types and also play a central role in oncogenic processes or in cancer cell survival, to avoid immune escape by mutations or loss of antigens by tumor cells.
Identification of novel TAAs can be achieved through 2 experimental processes: direct immunology (starting from patient-derived autologous tumor-specific CTL clones specific for an unknown epitope) and reverse immunology (starting from a predicted epitope). The former has been used since the discovery of the first tumor-specific CTL epitope, MAGE-1 [60]. Direct immunology is further subdivided into genetic or biochemical approaches. Briefly, in the genetic approach, a patient-derived CTL clone is screened by using target cells transfected with tumor-derived cDNA libraries. Subsequently, the increased release of cytokines in the supernatant due to the recognition by the tumor-specific CTL clone allows one to select the cells that contain the antigen-encoding cDNA; these are then subcloned and rescreened to finally identify the cDNA that encodes the specific antigen. The biochemical approach consists of the purification of peptides eluted from MHC class I molecules of antigen-expressing cells by high-performance liquid chromatography fractionation. Antigen-negative target cells expressing the appropriate HLA molecule are used to load these peptides and tested for CTL recognition. Positive fractions are analyzed by mass spectrometry to identify the amino acid sequence of the epitope recognized by CTLs [61]. The need for expensive specialized equipment, plus the labor-intensive method, probably accounts for the increasing use of reverse immunology. Over the years, a growing understanding of HLA-specific peptide-binding motifs has led to the development of several computer algorithms for amino acid sequences with predicted binding capacity. Reverse immunology consists of two different phases: in the epitope prediction phase, proteins are analyzed for the presence of potential epitopes by the use of prediction algorithms. Subsequently, in the epitope validation phase, the candidate peptides are tested by binding and stability assays in vitro. Nevertheless, differences between the processing machinery in normal and tumor cells might be liable for the lack of activity against tumor cells of several CTLs raised against high-affinity binding TAAs [62]. Nowadays, indeed, the most recent algorithms also take into account the proteasomal processing and transporters associated with antigen processing- (TAP-)translocation, 2 other fundamental processes in the antigen-presentation pathway. Despite many efforts, the use of epitope-based vaccines has not advanced beyond phase I or II clinical trials, probably due to the drawbacks described above. To date, the best results have been achieved with the use of fusion protein- or HSP-based vaccines.
Provenge (sipuleucel-T, Dendreon Corporation) is in late-stage development for the treatment of mHRPC. Sipuleucel-T is an immunotherapy product designed to stimulate T-cell immunity against prostatic acid phosphatase (PAP). It consists of autologous APCs isolated by leukapheresis, cultured with a PAP-GM-CSF fusion protein, and reinfused into the patient. The time from apheresis to infusion of final product is approximately 48 hours. The efficacy of Provenge was evaluated in 2 randomized, double-blind, placebo-controlled phase III clinical trials (D9901 and D9902A) [7, 8]. D9901 enrolled 127 patients with asymptomatic mHRPC, who were randomly assigned 2 : 1 to receive 3 infusions of Provenge () or placebo () every 2 weeks. Enrollment in D9902A was stopped at 98 patients after D9901 showed encouraging results in terms of disease progression, and the study was amended to become D9902B (IMPACT), enrolling 512 patients with OS as the primary endpoint. An integrated analysis of 225 patients in D9901 and D9902A (147 in the vaccine arm and 78 in the placebo arm) demonstrated a survival benefit for patients treated with Provenge versus placebo (23.2 months versus 18.9 months, resp.), with a 33% reduction in the risk of death. The only immunological data to emerge from these studies are limited to the correlation between the upregulation of CD54 molecules on the cell surface of sipuleucel-T-treated APCs and OS, whereas no data are available about a specific immune response against PAP. At the American Urological Association 2009 Annual Meeting, Dendreon Corporation announced that the phase III IMPACT clinical trial had met its primary endpoint of significantly improving OS by 4.1 months compared to placebo 25.8 months versus 21.7 months, respectively, , HR = 0.775 [95% CI: 0.614, 0.979]. The U.S. Food and Drug Administration (FDA) will respond to the existing Dendreon's amended Biologics License Application (BLA) for the licensing of Provenge in men with metastatic castrate-resistant prostate cancer (CRPC) by May 2010. If approved, Provenge will be the first active cellular immunotherapy to decisively demonstrate a survival benefit for cancer patients.
Oncophage (vitespen, Antigenics), an autologous tumor-derived HSP gp96 peptide complex, has been evaluated in 2 phase III clinical trials in stage IV melanoma patients and in renal cell carcinoma (RCC) patients at high risk of recurrence after nephrectomy [9, 10]. Oncophage consists of a purified preparation of the HSP gp96 from tumor. HSPs are noncovalently bound to peptides derived from self- and tumor-specific proteins. Immunization with gp96 peptide complexes leads to their uptake by DCs through CD91 (an HSP receptor) and stimulation of cognate T cells. In the first phase III clinical trial, 322 patients with stage IV melanoma were randomized 2 : 1 to receive Oncophage or a treatment of the physician's choice. The first 4 injections were administered weekly and subsequent injections were given every other week. Results from this trial suggested a survival benefit in the subpopulation of patients with M1a or M1b disease who were able to receive 10 or more doses of vaccine. In the second phase III trial of 818 patients with postnephrectomy RCC, no difference in recurrence-free survival or OS was observed between patients receiving Oncophage versus no treatment, although a trend toward a decrease in recurrence-free survival was reported in stage I or II disease in the experimental arm.
In a prospective randomized multicenter phase III trial, 185 patients with locally advanced stage III or stage IV melanoma were randomized to receive high-dose (HD) IL-2 alone (94 patients) or a synthetic peptide from the gp100 melanoma-associated antigen [gp100 : 209-217(210M)] plus an adjuvant (Montanide ISA) followed by HD IL-2 (91 patients). Overall response rate (RR, 22.1% versus 9.7%, ) and progression-free survival (PFS) (2.9 months versus 1.6, ) were significantly improved in the experimental arm compared with the HD IL-2 arm, respectively. Median OS was 17.6 months in the HD IL-2 + vaccine arm versus 12.8 in the HD IL-2 alone arm () [11].
Stimuvax (BLP25 liposome vaccine, L-BLP25, Oncothyreon partnered with Merck KGaA) is a cancer vaccine designed to induce an immune response against the extracellular core peptide of MUC1, a type I membrane glycoprotein widely expressed on many tumors (i.e., lung cancer, breast cancer, prostate cancer, and colorectal cancer). Stimuvax consists of MUC1 lipopeptide BLP25 [STAPPAHGVTSAPDTRPAPGSTAPPK(Pal)G], an immuno-adjuvant monophosphoryl lipid A, and three lipids (cholesterol, dimyristoyl phosphatidylglycerol, and dipalmitoyl phosphatidylcholine), capable of enhancing the delivery of the vaccine to APCs.
A randomized phase IIB clinical trial evaluated the effect of Stimuvax on survival and toxicity in 171 patients (88 in the L-BLP25 arm and 83 in the best supportive care arm (BSC)) with stage IIIB and IV nonsmall-cell lung cancer (NSCLC), after stable disease or response to a first-line chemotherapy [12]. Median OS was 17.4 months in the L-BLP25 arm and 13.0 months in the BSC arm, respectively, with a nonsignificant improvement of 4.4 months in the experimental arm (). T-cell proliferation assays were conducted on 78 of 88 patients enrolled in the L-BLP25 group, before and after immunization. Sixteen patients showed a MUC1-specific T-cell response (only two with a locoregional stage IIIB disease). No severe toxicities were reported. After a median follow-up of 53 months, updated survival data reported a median OS of 30.6 months in the Stimuvax arm versus 13.3 months in the BSC arm, in the subgroup of patients with locoregional stage IIIB (65 patients, of whom 35 were randomized to the vaccine arm and 30 were randomized to the BSC arm) () [63]. Although nonsignificant, considering the magnitude of the difference and the prolonged follow-up, these results suggest a signal of efficacy for the vaccine.
Based on these data, Merck is currently conducting three large phase III clinical trials of Stimuvax. START (Stimulating Targeted Antigenic Responses To NSCLC) is a double-blind, placebo-controlled, randomized, multicenter phase III clinical trial that will enroll patients with unresectable stage IIIA or IIIB NSCLC, after stable disease or response to a platinum-based chemo-radiotherapy. This study will involve more than 1,300 patients.
The INSPIRE study (Stimuvax trial In Asian NSCLC Patients: stimulating Immune REsponse) will enroll approximately 420 patients with unresectable stage III NSCLC across China, Hong Kong, South Korea, Singapore, and Taiwan. STRIDE (STimulating immune Response In aDvanced brEast cancer) is a randomized, double-blind, controlled, multicenter Phase III study designed to evaluate the efficacy of Stimuvax, in combination with hormonal therapy, in patients with inoperable, locally advanced, recurrent, or metastatic breast cancer.
4. Vaccine with Tumor Cells or Tumor-Cell Lysates
Theoretically, tumor-cell vaccines have at least 3 advantages over the single-target approaches discussed above in terms of eliciting an immune response: (a) different and unknown antigens can be targeted at the same time; (b) the immune response is not HLA-restricted; (c) the variety of both MHC class I and class II epitopes processed is likely to be able to stimulate both an innate (NK cells, macrophages, and eosinophils) and adaptive (CD8+ and CD4+ T cells) response.
The first important distinction is between vaccines using autologous (patient-specific) or allogeneic (nonpatient-specific) tumor cells. Second, these cells may be unmodified, modified for expression of MHC, costimulatory molecules, or cytokines, or used in combination with adjuvants such as GM-CSF and Bacille Calmette-Guerin (BCG). Third, these cells can be used in the form of tumor-cell lysates [64].
The mechanism for priming naïve T cells in response to whole-cell or lysate vaccination is still unclear. Tumor antigens are probably phagocytosed by DCs and cross-presented to CD8+ cells by MHC class I molecules. In some models, a CD4+ response seems to be required for effective tumor rejection [65, 66]. A mesothelin-specific CD8+ T cell response has been shown in a clinical trial employing vaccination with GM-CSF-secreting pancreatic cancer cell lines. The results of this study provide the first direct evidence that a cross-priming mechanism mediated by professional APCs is involved in a postvaccination induction of CD8+ T cell response [51].
In the past 20 years, several different vaccines derived from whole tumor cells or tumor-cell lysates have been evaluated in preclinical models and clinical trials. OncoVAX (Vaccinogen) is composed of autologous irradiated tumor cells, with or without BCG as an adjuvant. In a multicenter phase III clinical trial, 254 patients with stage II and III colon cancer were randomly assigned, after curative resection for primary tumor, to receive OncoVAX or no adjuvant treatment [13]. The 5.8-year median follow-up showed a 20.4% reduction in risk of disease progression in patients receiving OncoVAX compared to the control group. Analysis by stage showed no significant benefit of OncoVAX in stage III disease, whereas a statistically significant improvement in recurrence-free survival in stage II was reported, with a 41.4% reduction in relative risk of disease progression () in the OncoVAX arm. The OS rate for the OncoVAX-treated group was higher compared to control, with an 11.1% and a 33.3% relative risk reduction in all patients and stage II patients, respectively [14]. Besides the clinical data and a prospective study of medical and economic benefits, the only immunological mechanism proposed by the authors was the presence of a significant delayed cutaneous hypersensitivity response to tumor cells after the third and fourth OncoVAX treatments (which lack BCG), as a measure of the immugenicity of the treatment, potentially correlated with long-term survival [15].
Reniale (LipoNova) is a vaccine designed to treat RCC. It is based on a lysate of autologous tumor cells, preincubated with IFN- to increase the antigenicity of these cells, and tocopherol acetate to protect cell membranes during the incubation process. A randomized, open-label, multicenter phase III clinical trial compared adjuvant treatment with Reniale after radical nephrectomy versus radical nephrectomy alone in nonmetastatic RCC (pT2-3b, pN0-3, M0) [16]. Prior to surgery, 558 patients at 55 institutions in Germany were enrolled in the trial and were randomized to receive 6 s.c. vaccinations at 4-week intervals, or no adjuvant therapy (control group). The intention-to-treat (ITT) population consisted of 379 patients in the primary analysis (177 patients in the vaccine group and 202 patients in the control group). Progression-free survival at 5 years for patients at all tumor stages was 77.4% in the Reniale group and 67.8% in the control group (). Interestingly, patients with a higher risk (T3 subgroup) showed greater benefit from adjuvant treatment with Reniale, with a 5-year PFS of 67.5% in the vaccine group and 49.7% in the control group. A secondary ITT analysis on 477 patients (233 patients in the Reniale group and 244 patients in the control group) showed a statistically significant advantage in the experimental arm in terms of PFS (); there was no statistically significant difference in OS between the 2 arms (). However, a per-protocol analysis of 352 patients revealed a statistically significant increase in PFS () and OS () in the vaccine arm [17]. No immunological data from this study have been reported.
GVAX for prostate cancer (Cell Genesys) is an allogeneic vaccine composed of 2 irradiated human prostate cancer cell lines, LNCaP and PC-3, modified by ex vivo transduction with an adenoassociated viral vector encoding the human GM-CSF gene. A preclinical study has demonstrated that s.c. administration of these cells invokes a local immune response, characterized by a local infiltration of neutrophils, CD4+ T cells, and apoptotic cells. The irradiated tumor cells persist and secrete high levels of GM-CSF at the injection site for 21 days. Theoretically, secretion of GM-CSF by allogeneic tumor cells improves the antigen presentation of TSAs and TAAs through recruitment and maturation of DCs at the site of immunization. DCs then migrate to draining lymph nodes and activate antigen-specific CD4+ T cells, characterized by the production of both Th1 and Th2 cytokines. Moreover, DCs may efficiently capture apoptotic tumor cells and cross-present multiple TAAs on MHC class I molecules for recognition by host CD8+ T cells, as demonstrated by the ability of GM-CSF-secreting tumor cells to generate T-cell responses to multiple TAAs capable of targeting antigenically related but distinct tumors [67]. Based on encouraging clinical and immunological responses in 5 phase I/II clinical trials with nearly 200 prostate cancer patients [68–70], 2 phase III trials were initiated. VITAL-1 completed patient accrual in 2007, enrolling 626 patients with mHRPC randomized to receive GVAX as monotherapy for up to 6 months or standard docetaxel chemotherapy. The primary endpoint of the trial was improvement in OS. In 2008, Cell Genesys terminated the trial based on the results of a futility analysis conducted by the study's Independent Data Monitoring Committee (IDMC), which indicated a 30% chance of meeting the primary endpoint. VITAL-2 was a phase III trial designed to compare GVAX plus docetaxel versus docetaxel alone in mHRPC. The primary endpoint of VITAL-2 was also improved in OS. The trial was initiated in 2005 and enrolled 408 patients. In 2008, Cell Genesys announced its decision to terminate VITAL-2, as recommended by a safety review in which the IDMC reported an imbalance in deaths between the 2 treatment arms (67/114 deaths in the GVAX plus docetaxel arm and 47/114 deaths in the docetaxel-alone arm). In this case, despite encouraging preclinical and immunological data, GVAX failed to meet the defined endpoints of both phase III clinical trials [6].
Further clinical trials, employing GVAX cancer immunotherapies, are underway and include pancreatic and breast cancers. A randomized three-arm clinical trial is currently evaluating the efficacy and toxicity of GVAX for pancreatic cancer (GM-CSF secreting allogeneic pancreatic cancer vaccine) administered either alone or in combination with either a single intravenous dose or daily metronomic oral doses of cyclophosphamide for the treatment of patients undergoing chemotherapy and radiation therapy for stage I or II disease, surgically resectable.
Recently, studies of combination therapies of GVAX vaccine and CTLA-4-blocking antibodies have shown activity in melanoma and ovarian carcinoma, representing a potential new strategy to enhance vaccine-mediated antitumor effects [71].
5. DNA and RNA Vaccines
DNA-based vaccines are a recently developed strategy that has proven capable of activating strong immunity against weak TAAs. Several approaches have been developed and evaluated for enhancing the potency of DNA-based vaccines, including improved delivery systems (Gene Gun, cationic liposomes) [72, 73], simultaneous administration of cytokines (GM-CSF or IL2) [74], and the use of separate plasmids encoding nonself-antigens (i.e., hepatitis B surface antigen) [75]. The immunogenicity of DNA-based vaccines can also be enhanced by various modifications of plasmid-encoded antigens [76, 77].
Recently, several phase I/II clinical trials employing DNA-based vaccines targeting different TAAs (i.e., PSA, PAP, gp100, CEA, hsp65) have been conducted in patients with prostate cancer [78, 79], melanoma [80, 81], colorectal cancer [75], and head and neck carcinomas [82]. In all these trials, DNA-based vaccines were administered either as monotherapy or in association with different delivery systems and adjuvants. In terms of immune response, most of these trials showed a low immunogenicity of TAAs. The small sample size of these phase I/II studies precludes achieving a statistical correlation between development of an immune response and clinical outcomes in vaccinated patients. Evidence of clinical benefit must be evaluated in larger studies.
mRNA-based gene transfer vaccines are another attractive immunotherapeutic approach to cancer treatment [83, 84]. This method, based primarily on transient transfection of nondividing cells, is regarded as pharmaceutically safe because the transfected mRNA does not integrate into the host genome [85]. In addition, high transfection efficiency can be achieved by electroporation [86, 87]. mRNA, which can be effectively overexpressed in target cells, is generated by in vitro transcription from a bacteriophage promoter-equipped plasmid DNA. It is composed of a cap structure at the end, the coding RNA for target antigen, and a tail of poly-adenosine (polyA tail) [88]. The target antigen used can be a single peptide PSA [89] or CEA [90], allogeneic cancer cell lines [18, 91, 92], or autologous tumor mRNA [93]. The mRNA-based vaccine containing the mRNA-coding TAA is transfected into DCs and translated into proteins. After protein processing, the antigen can be loaded on MHC molecules for antigen presentation, thus activating an antigen-specific CTL response [94].
Clinical trials have been performed employing mRNA-transfected DCs or injecting mRNA directly into patients with prostate cancer [18, 89, 95], RCC [96], ovarian cancer [97], lung cancer, breast cancer [90], pediatric brain cancer [98], neuroblastoma [99], and melanoma [100, 101]. A phase I clinical trial was performed using PSA-mRNA-transfected DCs in patients with metastatic prostate cancer [89]. When the effects of repeated vaccinations with PSA-mRNA-transfected DCs were examined, the results demonstrated that the vaccine was able to increase PSA-specific CTL responses.
In a phase I/II clinical trial in androgen-resistant prostate cancer, patients were vaccinated with DCs transfected with mRNA from 3 allogeneic prostate cancer cell lines (DU145, LNCaP, and PC-3) [18]. Twelve of 19 patients showed specific T-cell responses; 10 of those 12 had a positive response in IFN- by ELISPOT assay and 9 had a specific T-cell proliferation response. Two CD8+ CTL clones were generated from a patient who showed a positive response in both the ELISPOT and proliferation assay. The CTL clones demonstrated specific killing of tumor mRNA-transfected DCs and PC-3 cells. Of the 19 patients on-study, 11 showed stable disease and 10 developed specific T-cell responses; only 2 of 8 patients with disease progression showed T-cell responses. These results demonstrate a correlation between immune response and clinical response.
In another clinical trial, patients with metastatic RCC received a vaccine consisting of DCs transfected with total RNA extracted from clear cell carcinoma, with or without IL2 prevaccination [97]. The results showed a significant increase in the frequency of tumor-specific CD4+ and CD8+ T cells as well as a decrease in Treg frequency. This trial demonstrated that mRNA-transfected DCs can increase immune response, and that this immune response in combination with depletion of Tregs can have a synergistic effect on antitumor immunity.
6. Conclusions
The promising results of recent phase II/III clinical trials may herald a new era for cancer vaccine immunotherapy. However, in spite of exciting improvements in the activity and efficacy of various vaccine platforms, including objective response, disease-free survival, progression-free survival, and overall survival, there is still much to learn about the immunological mechanisms by which these results can be improved. Further research is required to improve our understanding of CTL antigen-specific activation, decreased Treg numbers and functionality, NK activation, antigen cascade, and the impact of tumor escape.
A paradigm shift is necessary in order to improve the design of immuno-oriented clinical trials, increase understanding of the balance between proinflammatory and immunosuppressive responses in antitumor immunity, and define new criteria for the immunological evaluation of antitumor activity and clinical outcomes. Such knowledge would not only improve the efficacy of cancer vaccines but would help to guide decisions regarding patient selection, vaccine scheduling, and the combination of vaccines and other treatment modalities such as surgery, radiotherapy, chemotherapy, and targeted therapy.
The authors thank Bonnie Casey and Debra Weingarten for their editorial assistance in preparation of this manuscript.
- A. Jemal, R. Siegel, E. Ward, Y. Hao, J. Xu, and M. J. Thun, "Cancer statistics, 2009," CA Cancer Journal for Clinicians, vol. 59, no. 4, pp. 225–249, 2009. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- P. Kantoff, L. Glode, S. Tannenbaum, D. Bilhartz, W. Pittman, and T. Schuetz, "Randomized, double-blind, vector-controlled study of targeted immunotherapy in patients (pts) with hormone-refractory prostate cancer (HRPC) [abstract]," Journal of Clinical Oncology, vol. 24, no. 18S, abstract 2501, 2006. View at Google Scholar
- P. Kantoff, T. Schuetz, B. Blumenstein et al., "Overall survival (OS) analysis of a phase II randomized controlled trial (RCT) of a poxviral-based PSA targeted immunotherapy in metastatic castration-resistant prostate cancer (mCRPC) [abstract]," Journal of Clinical Oncology, vol. 27, no. 15S, abstract 5013, 2009. View at Google Scholar
- P. Kantoff, T. Schuetz, B. Blumenstein et al., "Overall survival (OS) analysis of a phase II randomized controlled trial (RCT) of a poxviral-based PSA targeted immunotherapy in metastatic castration-resistant prostate cancer (mCRPC)," Journal of Clinical Oncology, vol. 28, no. 7, pp. 1099–1105, 2010. View at Google Scholar
- S. Halabi, E. J. Small, and E. J. Small, "Prognostic model for predicting survival in men with hormone-refractory metastatic prostate cancer," Journal of Clinical Oncology, vol. 21, no. 7, pp. 1232–1237, 2003. View at Publisher · View at Google Scholar
- B. Goldman and L. DeFrancesco, "The cancer vaccine roller coaster," Nature Biotechnology, vol. 27, no. 2, pp. 129–139, 2009. View at Publisher · View at Google Scholar · View at PubMed
- E. J. Small, P. F. Schellhammer, and P. F. Schellhammer, "Placebo-controlled phase III trial of immunologic therapy with Sipuleucel-T (APC8015) in patients with metastatic, asymptomatic hormone refractory prostate cancer," Journal of Clinical Oncology, vol. 24, no. 19, pp. 3089–3094, 2006. View at Publisher · View at Google Scholar · View at PubMed
- C. S. Higano, P. F. Schellhammer, and P. F. Schellhammer, "Integrated data from 2 randomized, double-blind, placebo-controlled, phase 3 trials of active cellular immunotherapy with sipuleucel-T in advanced prostate cancer," Cancer, vol. 115, no. 16, pp. 3670–3679, 2009. View at Publisher · View at Google Scholar · View at PubMed
- A. Testori, J. Richards, and J. Richards, "Phase III comparison of vitespen, an autologous tumor-derived heat shock protein gp96 peptide complex vaccine, with physician's choice of treatment for stage IV melanoma: the C-100-21 study group," Journal of Clinical Oncology, vol. 26, no. 6, pp. 955–962, 2008. View at Publisher · View at Google Scholar · View at PubMed
- C. Wood, P. Srivastava, and P. Srivastava, "An adjuvant autologous therapeutic vaccine (HSPPC-96; vitespen) versus observation alone for patients at high risk of recurrence after nephrectomy for renal cell carcinoma: a multicentre, open-label, randomised phase III trial," The Lancet, vol. 372, no. 9633, pp. 145–154, 2008. View at Publisher · View at Google Scholar · View at PubMed
- D. J. Schwartzentruber, D. Lawson, and J. Richards, "A phase III multi-institutional randomized study of immunization with the gp100:209-217(210M) peptide followed by high-dose IL-2 compared with high-dose IL-2 alone in patients with metastatic melanoma," Journal of Clinical Oncology, vol. 27, no. 18S, abstract CRA9011, 2009. View at Google Scholar
- C. Butts, N. Murray, and N. Murray, "Randomized phase IIB trial of BLP25 liposome vaccine in stage IIIB and IV non-small-cell lung cancer," Journal of Clinical Oncology, vol. 23, no. 27, pp. 6674–6681, 2005. View at Publisher · View at Google Scholar · View at PubMed
- J. B. Vermorken, A. M. E. Claessen, and A. M. E. Claessen, "Active specific immunotherapy for stage II and stage III human colon cancer: a randomised trial," Lancet, vol. 353, no. 9150, pp. 345–350, 1999. View at Publisher · View at Google Scholar
- C. A. Uyl-de Groot, J. B. Vermorken, and J. B. Vermorken , "Immunotherapy with autologous tumor cell-BCG vaccine in patients with colon cancer: a prospective study of medical and economic benefits," Vaccine, vol. 23, no. 17-18, pp. 2379–2387, 2005. View at Publisher · View at Google Scholar · View at PubMed
- H. C. Hoover Jr., M. Surdyke, and R. B. Dangel, "Delayed cutaneous hypersensitivity to autologous tumor cells in colorectal cancer patients immunized with an autologous tumor cell: Bacillus Calmette-Guérin vaccine," Cancer Research, vol. 44, no. 4, pp. 1671–1676, 1984. View at Google Scholar
- D. Jocham, A. Richter, and A. Richter, "Adjuvant autologous renal tumour cell vaccine and risk of tumour progression in patients with renal-cell carcinoma after radical nephrectomy: phase III, randomised controlled trial," Lancet, vol. 363, no. 9409, pp. 594–599, 2004. View at Publisher · View at Google Scholar · View at PubMed
- C. Doehn, A. Richter, R. Theodor, W. Lehmacher, and D. Jocham, "Prolongation of progression-free and overall survival following an adjuvant vaccination with Reniale in patients with non-metastatic renal cell carcinoma: secondary analysis of a multicenter phase-III trial [abstract]," in Proceedings of the 27th German Cancer Congress Program and Abstracts, vol. 395, 2006.
- L. J. Mu, J. A. Kyte, and J. A. Kyte, "Immunotherapy with allotumour mRNA-transfected dendritic cells in androgen-resistant prostate cancer patients," British Journal of Cancer, vol. 93, no. 7, pp. 749–756, 2005. View at Publisher · View at Google Scholar · View at PubMed
- M. Dougan and G. Dranoff, "Immune therapy for cancer," Annual Review of Immunology, vol. 27, pp. 83–117, 2009. View at Publisher · View at Google Scholar · View at PubMed
- C. Palena, S. I. Abrams, J. Schlom, and J. W. Hodge, "Cancer vaccines: preclinical studies and novel strategies," Advances in Cancer Research, vol. 95, pp. 115–145, 2006. View at Publisher · View at Google Scholar · View at PubMed
- G. M. Bendle, A. Holler, A.-M. Downs, S.-A. Xue, and H. J. Stauss, "Broadly expressed tumour-associated proteins as targets for cytotoxic T lymphocyte-based cancer immunotherapy," Expert Opinion on Biological Therapy, vol. 5, no. 9, pp. 1183–1192, 2005. View at Publisher · View at Google Scholar · View at PubMed
- A. Schietinger, M. Philip, and H. Schreiber, "Specificity in cancer immunotherapy," Seminars in Immunology, vol. 20, no. 5, pp. 276–285, 2008. View at Publisher · View at Google Scholar · View at PubMed
- L. A. Emens, J. M. Asquith, J. M. Leatherman et al., "Timed sequential treatment with cyclophosphamide, doxorubicin, and an allogeneic granulocyte-macrophage colony-stimulating factor-secreting breast tumor vaccine: a chemotherapy dose-ranging factorial study of safety and immune activation," Journal of Clinical Oncology, vol. 27, no. 35, pp. 5911–5918, 2009. View at Publisher · View at Google Scholar · View at PubMed
- D. Laheru, E. Lutz, and E. Lutz, "Allogeneic granulocyte macrophage colony-stimulating factor-secreting tumor immunotherapy alone or in sequence with cyclophosphamide for metastatic pancreatic cancer: a pilot study of safety, feasibility, and immune activation," Clinical Cancer Research, vol. 14, no. 5, pp. 1455–1463, 2008. View at Publisher · View at Google Scholar · View at PubMed
- C. T. Garnett, J. Schlom, and J. W. Hodge, "Combination of docetaxel and recombinant vaccine enhances T-cell responses and antitumor activity: effects of docetaxel on immune enhancement," Clinical Cancer Research, vol. 14, no. 11, pp. 3536–3544, 2008. View at Publisher · View at Google Scholar · View at PubMed
- M. E. C. Lutsiak, R. T. Semnani, R. De Pascalis, S. V. S. Kashmiri, J. Schlom, and H. Sabzevari, "Inhibition of T regulatory cell function implicated in enhanced immune response by low-dose cyclophosphamide," Blood, vol. 105, no. 7, pp. 2862–2868, 2005. View at Publisher · View at Google Scholar · View at PubMed
- R. J. Lechleider, P. M. Arlen, and P. M. Arlen, "Safety and immunologic response of a viral vaccine to prostate- specific antigen in combination with radiation therapy when metronomic-dose interleukin 2 is used as an adjuvant," Clinical Cancer Research, vol. 14, no. 16, pp. 5284–5291, 2008. View at Publisher · View at Google Scholar · View at PubMed
- J. Kantor, S. Abrams, K. Irvine, P. Snoy, H. Kaufman, and J. Schlom, "Specific immunotherapy using a recombinant vaccinia virus expressing human carcinoembryonic antigen," Annals of the New York Academy of Sciences, vol. 690, pp. 370–373, 1993. View at Google Scholar
- E. Kass, J. Schlom, J. Thompson, F. Guadagni, P. Graziano, and J. W. Greiner, "Induction of protective host immunity to carcinoembryonic antigen (CEA), a self-antigen in CEA transgenic mice, by immunizing with a recombinant vaccinia-CEA virus," Cancer Research, vol. 59, no. 3, pp. 676–683, 1999. View at Google Scholar
- B. Acres and J.-Y. Bonnefoy, "Clinical development of MVA-based therapeutic cancer vaccines," Expert Review of Vaccines, vol. 7, no. 7, pp. 889–893, 2008. View at Publisher · View at Google Scholar · View at PubMed
- J. Taylor and E. Paoletti, "Fowlpox virus as a vector in a non-avian species," Vaccine, vol. 6, no. 6, pp. 466–468, 1988. View at Google Scholar
- H. Stickl, V. Hochstein-Mintzel, A. Mayr, H. C. Huber, H. Schäfer, and A. Holzner, "MVA vaccination against smallpox: clinical tests with an attenuated live vaccinia virus strain (MVA) (author's transl)," Deutsche Medizinische Wochenschrift, vol. 99, no. 47, pp. 2386–2392, 1974. View at Google Scholar
- J. W. Hodge, J. P. McLaughlin, J. A. Kantor, and J. Schlom, "Diversified prime and boost protocols using recombinant vaccinia virus and recombinant non-replicating avian pox virus to enhance T-cell immunity and antitumor responses," Vaccine, vol. 15, no. 6-7, pp. 759–768, 1997. View at Publisher · View at Google Scholar
- D. W. Grosenbach, J. C. Barrientos, J. Schlom, and J. W. Hodge, "Synergy of vaccine strategies to amplify antigen-specific immune responses and antitumor effects," Cancer Research, vol. 61, no. 11, pp. 4497–4505, 2001. View at Google Scholar
- R. Slack, L. Ley, and P. Chang, "Association between CEA-specific T cell responses (TCR) following treatment with vaccinia-CEA (V) and Alvac-CEA (A) and survival in patients with CEA-bearing cancers [abstract]," WProceedings of the American Society of Clinical Oncology, vol. 20, no. 1086, 2001. View at Google Scholar
- H. L. Kaufman, W. Wang, and W. Wang, "Phase II randomized study of vaccine treatment of advanced prostate cancer (E7897): a trial of the Eastern Cooperative Oncology Group," Journal of Clinical Oncology, vol. 22, no. 11, pp. 2122–2132, 2004. View at Publisher · View at Google Scholar · View at PubMed
- J. W. Hodge and J. Schlom, "Costimulatory molecules in vaccine design," Ernst Schering Research Foundation Workshop, no. 30, pp. 23–52, 2000. View at Google Scholar
- H. Terasawa, K.-Y. Tsang, J. Gulley, P. Arlen, and J. Schlom, "Identification and characterization of a human agonist cytotoxic T-lymphocyte epitope of human prostate-specific antigen," Clinical Cancer Research, vol. 8, no. 1, pp. 41–53, 2002. View at Google Scholar
- J. Gulley, A. P. Chen, and A. P. Chen, "Phase I study of a vaccine using recombinant vaccinia virus expressing PSA (rV-PSA) in patients with metastatic androgen-independent prostate cancer," Prostate, vol. 53, no. 2, pp. 109–117, 2002. View at Publisher · View at Google Scholar · View at PubMed
- J. L. Gulley, P. M. Arlen, and P. M. Arlen, "Immunologic and prognostic factors associated with overall survival employing a poxviral-based PSA vaccine in metastatic castrate-resistant prostate cancer," Cancer Immunology, Immunotherapy, vol. 59, no. 5, pp. 1–12, 2010. View at Publisher · View at Google Scholar · View at PubMed
- S. Zaremba, E. Barzaga, M. Zhu, N. Soares, K.-Y. Tsang, and J. Schlom, "Identification of an enhancer agonist cytotoxic T lymphocyte peptide from human carcinoembryonic antigen," Cancer Research, vol. 57, no. 20, pp. 4570–4577, 1997. View at Google Scholar
- K.-Y. Tsang, C. Palena, J. Gulley, P. Arlen, and J. Schlom, "A human cytotoxic T-lymphocyte epitope and its agonist epitope from the nonvariable number of tandem repeat sequence of MUC-1," Clinical Cancer Research, vol. 10, no. 6, pp. 2139–2149, 2004. View at Publisher · View at Google Scholar
- J. L. Gulley, P. M. Arlen, and P. M. Arlen, "Pilot study of vaccination with recombinant CEA-MUC-1-TRICOM poxviral-based vaccines in patients with metastatic carcinoma," Clinical Cancer Research, vol. 14, no. 10, pp. 3060–3069, 2008. View at Publisher · View at Google Scholar · View at PubMed
- G. Trinchieri and A. Sher, "Cooperation of Toll-like receptor signals in innate immune defence," Nature Reviews Immunology, vol. 7, no. 3, pp. 179–190, 2007. View at Publisher · View at Google Scholar · View at PubMed
- D. G. Brockstedt and T. W. Dubensky Jr., "Promises and challenges for the development of Listeria monocytogenes-based immunotherapies," Expert Review of Vaccines, vol. 7, no. 7, pp. 1069–1084, 2008. View at Publisher · View at Google Scholar · View at PubMed
- A. Franzusoff, R. C. Duke, T. H. King, Y. Lu, and T. C. Rodell, "Yeasts encoding tumour antigens in cancer immunotherapy," Expert Opinion on Biological Therapy, vol. 5, no. 4, pp. 565–575, 2005. View at Publisher · View at Google Scholar · View at PubMed
- A. Bolhassani and S. Rafati, "Heat-shock proteins as powerful weapons in vaccine development," Expert Review of Vaccines, vol. 7, no. 8, pp. 1185–1199, 2008. View at Publisher · View at Google Scholar · View at PubMed
- V. Cereda, D. J. Poole, and D. J. Poole, "New gene expressed in prostate: a potential target for T cell-mediated prostate cancer immunotherapy," Cancer Immunology, Immunotherapy, vol. 59, no. 1, pp. 63–71, 2010. View at Publisher · View at Google Scholar · View at PubMed
- C. Palena, D. E. Polev, and D. E. Polev, "The human T-box mesodermal transcription factor Brachyury is a candidate target for T-cell–mediated cancer immunotherapy," Clinical Cancer Research, vol. 13, no. 8, pp. 2471–2478, 2007. View at Publisher · View at Google Scholar · View at PubMed
- J. Yokokawa, T. K. Bera, and T. K. Bera, "Identification of cytotoxic T-lymphocyte epitope(s) and its agonist epitope(s) of a novel target for vaccine therapy (PAGE4)," International Journal of Cancer, vol. 121, no. 3, pp. 595–605, 2007. View at Publisher · View at Google Scholar · View at PubMed
- A. M. Thomas, L. M. Santarsiero, and L. M. Santarsiero, "Mesothelin-specific CD T cell responses provide evidence of in vivo cross-priming by antigen-presenting cells in vaccinated pancreatic cancer patients," Journal of Experimental Medicine, vol. 200, no. 3, pp. 297–306, 2004. View at Publisher · View at Google Scholar · View at PubMed
- W. Z. Ai, R. Tibshirani, B. Taidi, D. Czerwinski, and R. Levy, "Anti-idiotype antibody response after vaccination correlates with better overall survival in follicular lymphoma," Blood, vol. 113, no. 23, pp. 5743–5746, 2009. View at Publisher · View at Google Scholar · View at PubMed
- A. M. Hernández, D. Toledo, and D. Toledo, "Characterization of the antibody response against NeuGcGM3 ganglioside elicited in non-small cell lung cancer patients immunized with an anti-idiotype antibody," Journal of Immunology, vol. 181, no. 9, pp. 6625–6634, 2008. View at Google Scholar
- R. So-Rosillo and E. J. Small, "Sipuleucel-T (APC8015) for prostate cancer," Expert Review of Anticancer Therapy, vol. 6, no. 9, pp. 1163–1167, 2006. View at Publisher · View at Google Scholar · View at PubMed
- S. Ostrand-Rosenberg, "Immune surveillance: a balance between protumor and antitumor immunity," Current Opinion in Genetics and Development, vol. 18, no. 1, pp. 11–18, 2008. View at Publisher · View at Google Scholar · View at PubMed
- P. Kalinski, J. Urban, R. Narang, E. Berk, E. Wieckowski, and R. Muthuswamy, "Dendritic cell-based therapeutic cancer vaccines: what we have and what we need," Future Oncology, vol. 5, no. 3, pp. 379–390, 2009. View at Publisher · View at Google Scholar · View at PubMed
- V. Cerundolo, I. F. Hermans, and M. Salio, "Dendritic cells: a journey from laboratory to clinic," Nature Immunology, vol. 5, no. 1, pp. 7–10, 2004. View at Publisher · View at Google Scholar · View at PubMed
- W. J. Lesterhuis, I. J. M. de Vries, G. J. Adema, and C. J. A. Punt, "Dendritic cell-based vaccines in cancer immunotherapy: an update on clinical and immunological results," Annals of Oncology, vol. 15, supplement 4, pp. iv145–iv151, 2004. View at Publisher · View at Google Scholar · View at PubMed
- J. Yokokawa, C. Palena, and C. Palena, "Identification of novel human CTL epitopes and their agonist epitopes of mesothelin," Clinical Cancer Research, vol. 11, no. 17, pp. 6342–6351, 2005. View at Publisher · View at Google Scholar · View at PubMed
- P. van der Bruggen, C. Traversari, and C. Traversari, "A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma," Science, vol. 254, no. 5038, pp. 1643–1647, 1991. View at Google Scholar
- J. H. Kessler and C. J. M. Melief, "Identification of T-cell epitopes for cancer immunotherapy," Leukemia, vol. 21, no. 9, pp. 1859–1874, 2007. View at Publisher · View at Google Scholar · View at PubMed
- M. Campoli and S. Ferrone, "HLA antigen changes in malignant cells: epigenetic mechanisms and biologic significance," Oncogene, vol. 27, no. 45, pp. 5869–5885, 2008. View at Publisher · View at Google Scholar · View at PubMed
- C. M. Butts, A. Maksymiuk, and G. Goss, "A multicentre phase IIB randomized controlled study of BLP25 liposome vaccine (L-BLP25 or Stimuvax) for active specific immunotherapy of non-small cell lung cancer (NSCLC): updated survival analysis: B1–01 [abstract]," Journal of Thoracic Oncology, vol. 2, pp. S332–S333, 2007. View at Google Scholar
- J. Copier and A. Dalgleish, "Overview of tumor cell-based vaccines," International Reviews of Immunology, vol. 25, no. 5-6, pp. 297–319, 2006. View at Publisher · View at Google Scholar · View at PubMed
- S. E. Hamilton, M. C. Wolkers, S. P. Schoenberger, and S. C. Jameson, "The generation of protective memory-like CD T cells during homeostatic proliferation requires CD T cells," Nature Immunology, vol. 7, no. 5, pp. 475–481, 2006. View at Publisher · View at Google Scholar · View at PubMed
- E. M. Janssen, E. E. Lemmens, T. Wolfe, U. Christen, M. G. von Herrath, and S. P. Schoenberger, "CD T cells are required for secondary expansion and memory in CD T lymphocytes," Nature, vol. 421, no. 6925, pp. 852–856, 2003. View at Publisher · View at Google Scholar · View at PubMed
- A. D. Simmons, B. Li, and B. Li, "GM-CSF-secreting cancer immunotherapies: preclinical analysis of the mechanism of action," Cancer Immunology, Immunotherapy, vol. 56, no. 10, pp. 1653–1665, 2007. View at Publisher · View at Google Scholar · View at PubMed
- J. W. Simons, M. A. Carducci, and M. A. Carducci, "Phase I/II trial of an allogeneic cellular immunotherapy in hormone-naïve prostate cancer," Clinical Cancer Research, vol. 12, no. 11, part 1, pp. 3394–3401, 2006. View at Publisher · View at Google Scholar · View at PubMed
- E. J. Small, N. Sacks, and N. Sacks, "Granulocyte macrophage colony-stimulating factor-secreting allogeneic cellular immunotherapy for hormone-refractory prostate cancer," Clinical Cancer Research, vol. 13, no. 13, pp. 3883–3891, 2007. View at Publisher · View at Google Scholar · View at PubMed
- C. S. Higano, J. M. Corman, and J. M. Corman, "Phase 1/2 dose-escalation study of a GM-CSF-secreting, allogeneic, cellular immunotherapy for metastatic hormone-refractory prostate cancer," Cancer, vol. 113, no. 5, pp. 975–984, 2008. View at Publisher · View at Google Scholar · View at PubMed
- F. S. Hodi, M. Butler, and M. Butler, "Immunologic and clinical effects of antibody blockade of cytotoxic T lymphocyte-associated antigen 4 in previously vaccinated cancer patients," Proceedings of the National Academy of Sciences of the United States of America, vol. 105, no. 8, pp. 3005–3010, 2008. View at Publisher · View at Google Scholar · View at PubMed
- S. R. Best, S. Peng, and S. Peng, "Administration of HPV DNA vaccine via electroporation elicits the strongest CD8+ T cell immune responses compared to intramuscular injection and intradermal gene gun delivery," Vaccine, vol. 27, no. 40, pp. 5450–5459, 2009. View at Publisher · View at Google Scholar · View at PubMed
- L. U'Ren, R. Kedl, and S. W. Dow, "Vaccination with liposome-DNA complexes elicits enhanced antitumor immunity," Cancer Gene Therapy, vol. 13, no. 11, pp. 1033–1044, 2006. View at Publisher · View at Google Scholar · View at PubMed
- S. Pasquini, Z. Xiang, Y. Wang, Z. He, H. Deng, M. Blaszczyk-Thurin, and H. C. J. Ertl, "Cytokines and costimulatory molecules as genetic adjuvants," Immunology and Cell Biology, vol. 75, no. 4, pp. 397–401, 1997. View at Google Scholar
- R. M. Conry, D. T. Curiel, and D. T. Curiel, "Safety and immunogenicity of a DNA vaccine encoding carcinoembryonic antigen and hepatitis B surface antigen in colorectal carcinoma patients," Clinical Cancer Research, vol. 8, no. 9, pp. 2782–2787, 2002. View at Google Scholar
- R. J. Binder and P. K. Srivastava, "Peptides chaperoned by heat-shock proteins are a necessary and sufficient source of antigen in the cross-priming of T cells," Nature Immunology, vol. 6, no. 6, pp. 593–599, 2005. View at Publisher · View at Google Scholar · View at PubMed
- B. H. Segal, X.-Y. Wang, C. G. Dennis, R. Youn, E. A. Repasky, M. H. Manjili, and J. R. Subjeck, "Heat shock proteins as vaccine adjuvants in infections and cancer," Drug Discovery Today, vol. 11, no. 11-12, pp. 534–540, 2006. View at Publisher · View at Google Scholar · View at PubMed
- M. Pavlenko, A.-K. Roos, and A.-K. Roos, "A phase I trial of DNA vaccination with a plasmid expressing prostate-specific antigen in patients with hormone-refractory prostate cancer," British Journal of Cancer, vol. 91, no. 4, pp. 688–694, 2004. View at Google Scholar
- D. G. McNeel, E. J. Dunphy, and E. J. Dunphy, "Safety and immunological efficacy of a DNA vaccine encoding prostatic acid phosphatase in patients with stage D0 prostate cancer," Journal of Clinical Oncology, vol. 27, no. 25, pp. 4047–4054, 2009. View at Publisher · View at Google Scholar · View at PubMed
- J. Yuan, G. Y. Ku, and G. Y. Ku, "Safety and immunogenicity of a human and mouse gp100 DNA vaccine in a phase I trial of patients with melanoma," Cancer Immunity, vol. 9, no. 5, 2009. View at Google Scholar
- R. D. Cassaday, P. M. Sondel, and P. M. Sondel, "A phase I study of immunization using particle-mediated epidermal delivery of genes for gp100 and GM-CSF into uninvolved skin of melanoma patients," Clinical Cancer Research, vol. 13, no. 2, pp. 540–549, 2007. View at Publisher · View at Google Scholar · View at PubMed
- P. Michaluart, K. A. Abdallah, and K. A. Abdallah, "Phase I trial of DNA-hsp65 immunotherapy for advanced squamous cell carcinoma of the head and neck," Cancer Gene Therapy, vol. 15, no. 10, pp. 676–684, 2008. View at Publisher · View at Google Scholar · View at PubMed
- J. A. Kyte and G. Gaudernack, "Immuno-gene therapy of cancer with tumour-mRNA transfected dendritic cells," Cancer Immunology, Immunotherapy, vol. 55, no. 11, pp. 1432–1442, 2006. View at Publisher · View at Google Scholar · View at PubMed
- B. Weide, C. Garbe, H.-G. Rammensee, and S. Pascolo, "Plasmid DNA- and messenger RNA-based anti-cancer vaccination," Immunology Letters, vol. 115, no. 1, pp. 33–42, 2008. View at Publisher · View at Google Scholar · View at PubMed
- A. Yamamoto, M. Kormann, J. Rosenecker, and C. Rudolph, "Current prospects for mRNA gene delivery," European Journal of Pharmaceutics and Biopharmaceutics, vol. 71, no. 3, pp. 484–489, 2009. View at Publisher · View at Google Scholar · View at PubMed
- V. F. I. Van Tendeloo, P. Ponsaerts, and P. Ponsaerts, "Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells," Blood, vol. 98, no. 1, pp. 49–56, 2001. View at Publisher · View at Google Scholar
- P. Ponsaerts, S. van der Sar, V. F. I. Van Tendeloo, P. G. Jorens, Z. N. Berneman, and P. B. Singh, "Highly efficient mRNA-based gene transfer in feeder-free cultured H9 human embryonic stem cells," Cloning and Stem Cells, vol. 6, no. 3, pp. 211–216, 2004. View at Publisher · View at Google Scholar · View at PubMed
- P. A. Krieg and D. A. Melton, "Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs," Nucleic Acids Research, vol. 12, no. 18, pp. 7057–7070, 1984. View at Google Scholar
- A. Heiser, D. Coleman, and D. Coleman, "Autologous dendritic cells transfected with prostate-specific antigen RNA stimulate CTL responses against metastatic prostate tumors," Journal of Clinical Investigation, vol. 109, no. 3, pp. 409–417, 2002. View at Publisher · View at Google Scholar
- M. A. Morse, S. K. Nair, and S. K. Nair, "Immunotherapy with autologous, human dendritic cells transfected with carcinoembryonic antigen mRNA," Cancer Investigation, vol. 21, no. 3, pp. 341–349, 2003. View at Publisher · View at Google Scholar
- E. A. Hirschowitz, T. Foody, R. Kryscio, L. Dickson, J. Sturgill, and J. Yannelli, "Autologous dendritic cell vaccines for non-small-cell lung cancer," Journal of Clinical Oncology, vol. 22, no. 14, pp. 2808–2815, 2004. View at Publisher · View at Google Scholar · View at PubMed
- H. S. Pandha, R. J. John, J. Hutchinson, N. James, M. Whelan, C. Corbishley, and A. G. Dalgleish, "Dendritic cell immunotherapy for urological cancers using cryopreserved allogeneic tumour lysate-pulsed cells a phase I/II study," BJU International, vol. 94, no. 3, pp. 412–418, 2004. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- J. A. Kyte, G. Kvalheim, S. Aamdal, S. Sæbøe-Larssen, and G. Gaudernack, "Preclinical full-scale evaluation of dendritic cells transfected with autologous tumor-mRNA for melanoma vaccination," Cancer Gene Therapy, vol. 12, no. 6, pp. 579–591, 2005. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- S. Pascolo, "Messenger RNA-based vaccines," Expert Opinion on Biological Therapy, vol. 4, no. 8, pp. 1285–1294, 2004. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- Z. Su, J. Dannull, and J. Dannull, "Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD and CD T cell responses in patients with metastatic prostate cancer," Journal of Immunology, vol. 174, no. 6, pp. 3798–3807, 2005. View at Google Scholar · View at Scopus
- Z. Su, J. Dannull, and J. Dannull, "Immunological and clinical responses in metastatic renal cancer patients vaccinated with tumor RNA-transfected dendritic cells," Cancer Research, vol. 63, no. 9, pp. 2127–2133, 2003. View at Google Scholar · View at Scopus
- J. Dannull, Z. Su, and Z. Su, "Enhancement of vaccine-mediated antitumor immunity in cancer patients after depletion of regulatory T cells," Journal of Clinical Investigation, vol. 115, no. 12, pp. 3623–3633, 2005. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- D. A. Caruso, L. M. Orme, and L. M. Orme, "Results of a phase 1 study utilizing monocyte-derived dendritic cells pulsed with tumor RNA in children and young adults with brain cancer," Neuro-Oncology, vol. 6, no. 3, pp. 236–246, 2004. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- D. A. Caruso, L. M. Orme, and L. M. Orme, "Results of a phase I study utilizing monocyte-derived dendritic cells pulsed with tumor RNA in children with stage 4 neuroblastoma," Cancer, vol. 103, no. 6, pp. 1280–1291, 2005. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- J. A. Kyte, L. Mu, and L. Mu, "Phase I/II trial of melanoma therapy with dendritic cells transfected with autologous tumor-mRNA," Cancer Gene Therapy, vol. 13, no. 10, pp. 905–918, 2006. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus
- B. Weide, S. Pascolo, and S. Pascolo, "Direct injection of protamine-protected mRNA: results of a phase 1/2 vaccination trial in metastatic melanoma patients," Journal of Immunotherapy, vol. 32, no. 5, pp. 498–507, 2009. View at Publisher · View at Google Scholar · View at PubMed · View at Scopus


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Now that the dust has settled from this past weekend, when Floyd Mayweather easily dispatched Robert Guerrero over 12 rounds, the question is just when and who does "Money" face next? After signing his groundbreaking, six-fight, 30-month deal with Showtime, he made it very clear that he absolutely intended to fulfill this contract.
At the post-fight press conference on Saturday night, the victorious Mayweather mentioned some discomfort in his right hand.
But on Monday afternoon, Golden Boy Promotions CEO Richard Schaefer told Maxboxing, "I've been told that the hand is OK. It was bruised. So as Floyd stated in the post-fight press conference, he fully intends to fight on September the 14th."
As you probably know by now, Top Rank has put in a request with the Nevada State Athletic Commission to stage a show on that same day at the Thomas and Mack Center between Juan Manuel Marquez and Tim Bradley. But that's on the assumption that Mayweather doesn't actually perform at the MGM Grand that same evening.
So where does this leave unified junior middleweight champion Saul "Canelo" Alvarez, who is coming off a big victory over Austin Trout on April 20th at the Alamodome in San Antonio?
"Well, hopefully in a fight with Floyd," said Schaefer, chuckling. "So I'll be meeting this week with Floyd's adviser [Al Haymon] and we will start having conversations. I know that Floyd is the kind of fighter who doesn't turn down anyone. He's always fought anyone and everyone and I know that with 'Canelo,' that's the fight he really wants. So we'll just have to see if we put it together. But I'll be working on that. That's my number one priority."
Being that September 14th is Mexican Independence Day weekend, you'd figure it would make sense that Mayweather face a Mexican boxer. Currently, there is no more popular fighter from that country than 'Canelo,' who drew nearly 40,000 in his last fight and does huge TV ratings in Mexico.
Just looking at the chess pieces on the board, it's pretty obvious that those in the Mayweather Sweepstakes will come from the pool of talent affiliated with Golden Boy and/or Haymon. It's clear that during the duration of this deal, it will be Golden Boy's job to develop and cultivate opponents for Mayweather.
One name thrown around last week was Amir Khan, who struggled mightily against Julio Diaz on April 27th in the U.K.
"Amir's going to be doing one of two things once he's back, which will be in late November, early December. We're actually holding December 7th for Amir," said Schaefer, who explained that with Khan's upcoming wedding and observance of Ramadan, he will be out of commission for a stretch. "The idea would be that he would be either fighting the winner of Danny Garcia [vs.] the winner of [Lucas] Matthysse and [Lamont] Peterson, who are going to be fighting in a couple of weeks. We'd like to match the winner of Matthysse and Peterson against Danny Garcia on September the 7th, which would be a week before our pay-per-view.
"It would be a very strong lead-in for the pay-per-view and then the winner of that September 7th card, the idea was to have them fight against Amir Khan on December 7th. Now, that's one direction. The other direction could be is if Devon Alexander is successful in his mandatory [IBF welterweight] title defense against Lee Purdy, which is going to take place on May 18th. There could be a possibility that we could do a fight between Amir and Devon for his welterweight title and basically get Amir up to 147 and see how he does there. Or give Devon a big win at '47 and so that's another scenario.
"It was never the scenario that Floyd would fight Amir next."
But Schaefer didn't rule out that a Mayweather-Khan match-up could happen in 2014.
"I think it's a possibility. You obviously have Floyd who's made it very clear that he would like to fight in the U.K. He loves the U.K., the U.K. fan-base he has there, which is substantial. But there are some things that would have to be worked out," said Schaefer, alluding to the taxes that Mayweather would be subject to for performing in jolly ol' England. Schaefer mentioned that track superstar Usain Bolt negotiated a special arrangement that allowed him to run in Britain.
Down the line, perhaps whoever comes out of the Golden Boy 140-pound tournament would be an attractive opponent for Mayweather. And Schaefer said that the fact Matthysse signed with Haymon (who just happens to rep Garcia and Mayweather among many others) would be no impediment to the hard-hitting Argentinean facing Garcia, should he come out victorious on May 18th in Atlantic City.
Schaefer says, "Matthysse wants the fight. Danny wants the fight. If Peterson should win, there's no obstacle to that either. So I really don't foresee any issues of the winner of that fight, fighting Danny Garcia September the 7th."
There's a lot of speculation that the "May Day" event did under one million buys. On Tuesday, esteemed colleagues, Dan Rafael and Kevin Iole, openly speculated on that possibility on Twitter. And yeah, I've heard the same things (hey, we all basically have the same sources and talk to the same group of people) but no official numbers have been released by anyone associated with the event.
The number being thrown around the most is 900,000 buys. And if that's accurate, let's do the math; at those amount of buys at $70 each (assuming everyone got the high-definition telecast), that's $63 million dollars of revenue of which about 55 percent goes to the cable and satellite operators for their participation in the event. So that's right around $31 million left for the promotion. Also you include what the closed-circuit did and the international rights fees. According to the NSAC, Mayweather's purse was $32.5 million. Add in the costs of the undercard, marketing and other expenses related to the event, well, you see there might be a significant deficit here (the site fee or ticket money from the MGM Grand most likely goes to the promotion, not the network).
Industry sources say Showtime needed between 1.1 or 1.2 to break even on this venture.
It's widely known that in this deal with Showtime, Mayweather has his money guaranteed throughout the length of this pact, meaning whether his fights garner a thousand pay-per-view sales or two million, he's protected financially. Some might say the results from the past weekend will serve as an impetus for Mayweather to face better opposition that would theoretically sell better to the public. Well, the reality is no matter what happens, Mayweather has a very lucrative safety net regardless of how his future fights perform on pay-per-view.
The question is, does Showtime have the right to (dis)approve future opponents or is there merely a list of already pre-approved opponents available (many of whom have already been mentioned in this column)? The way this deal is structured may have actually de-incentivized Mayweather from actually facing real challenges and everyone knows how important retiring undefeated is to him.
There are several factors that might have hurt this promotion. First, it was announced relatively late as there were a lot of moving parts (such as Mayweather moving away from HBO to Showtime). With that, there wasn't the customary press tour that accompanies this caliber of events. And again, with Floyd having a financial guarantee, from what I was told, he didn't do much to extend himself to the press like he had before in the past. More than a few of the established boxing scribes who were used to getting a certain type of access to Mayweather groused that they were shut down this time around for whatever reason.
Also, the preview shows and documentaries played to much smaller audiences on Showtime than they have in the past on HBO for whatever reason (HBO's subscriber base is still larger than Showtime's).
Perhaps the biggest factor in all this was that Guerrero simply wasn't an ideal B-side. While a solid fighter, unlike a Miguel Cotto or Ricky Hatton, he simply didn't have his own constituency to draw upon. This should be put into perspective; a very small percentage of pay-per-view events ever crack the half-million mark and this point should be overlooked. This promotion still did a significant amount of buys and it can be argued that no other boxer could have put up these numbers with a dance partner like "The Ghost." But again, there is that $32.5 million nut.
Even a past pay-per-view titan like Oscar De la Hoya didn't routinely put up a million pay-per-view buys and his tallies were affected by who he faced. Don't believe me? Look up the fluctuation in numbers when he faced the likes of Yori Boy Campas and the likes of Fernando Vargas and Felix Trinidad. The opponent absolutely matters.
Mayweather isn't immune to this.
The fight most clamored for now is Mayweather against "Canelo" but that comes with its own set of problems. Alvarez is a rock-solid junior middleweight. It's not clear if Mayweather is willing to move up to 154 pounds anymore. Also, there are the economics. Unlike a Guerrero, who received $3 million this past weekend, you can bet the redhead from Mexico commands at least three times that much.
While some will state Mayweather needs that fight for his legacy, it can be argued that it's really Showtime who needs it for its bottom line.
© 2000 - 2011 Knockout Entertainment Ltd & SecondsOut.com

subset: commoncrawl


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Jim Tressel reading daughter's poetry at UA showcase in Merriman Valley11/04/2013
Jim Tressel is stepping out of his usual role as The University of Akron's vice president for student success to share another love: poetry.
Jim Tressel and Carlee Tressel Alson.
Tressel plans to read works by his daughter — poet, essayist and fiction writer Carlee Tressel Alson — during "Poetry Night in the Valley" on Tuesday, Nov. 12, at 7 p.m. at Max McQ's Sports Bar and Grill, 1562 Akron Peninsula Road, Akron.
Free and open to the public, the event will showcase the creative writing program of UA's Department of English.
Also during the event, published authors who teach in UA's creative writing program will read their works, sign books and share tips about publishing. They include Mary Biddinger, Julie Drew, David Giffels, James Renner and Eric Wasserman.
Biddinger, UA associate professor of English and author of five collections of poetry, will read from "Small Enterprise," her soon-to-be published full-length collection of poems. She will be joined by UA student writers Sarah Dravec of Boardman, Nathan Kemp of Mansfield and James Phillips of Medina.
An editor, writer and the assistant chair of the UA Department of English, Biddinger teaches literature and creative writing with a focus on poetry. Her work has appeared in many poetry journals as well as her four books: "Prairie Fever" (2007), "Saint Monica" (2011), "O Holy Insurgency" (2013) and the upcoming "A Sunny Place with Adequate Water" (2014).
The event will include a meet-and-greet with the authors from 5:30 to 7 p.m. The poetry reading at 7 p.m. will be followed at 8:30 p.m. by an open mic for community poets so that attendees can share their own creative works. For more information, call 330-972-7472.
"I look forward to hearing Jim give voice to his daughter's poetry," said William Thelin, UA English professor and department chair.
"Poetry and writing really are for everyone," added Thelin. "Because of the vitality of our creative writing program and the strength of our faculty, The University of Akron has much to offer to aspiring writers."
Media contact: Cyndee Snider, 330-972-5196 or email@example.com.


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In the wake of the unthinkable tragedy in Aurora, Colo., some good news came in the form of a new baby named Hugo Medley. Katie Medley gave birth to her son, NBC News reports, at 7:11 a.m. Tuesday morning after narrowly escaping the attacks in the Colorado theater shooting. Though she escaped the attacks unharmed, her husband, Caleb Medley, lies in the same hospital in a medically induced coma, unaware that his son was born.
Plus: Woman Gives Birth After Being Shot and Set on Fire
The two high school sweethearts were celebrating one final date night before the birth of their son, which was scheduled to take place on July 23. Unfortunately, the evening took a tragic turn as Caleb was shot in the face. He has lost his right eye and suffered brain damage from the attack.
Plus: Stories of Miracle Babies
The young couple does not have health insurance and friends have rallied to help them collect money for what is sure to be hefty hospital bills. They have organized a website to collect monetary donations and share Caleb, Katie, and now baby Hugo's inspirational story.
Watch an interview with a friend of Caleb Medley:
We send the Medley family best wishes for a healthy, speedy recovery together!


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College Savings Plans of Maryland, an independent Maryland state agency, offers two tax-advantaged 529 plans to pay college expenses
Both the Maryland Prepaid College Trust and the Maryland College Investment Plan can be used at most federally accredited schools across the country, including many trade and technical schools.
Earnings in the plans are tax free as long as they are used for qualified higher education expenses.
For information, go to http://www.CollegeSavingsMD.org/Webinar.
An enrollment kit that describes the investment objectives, risks, expenses and other important information that should considered is available online or by calling 1-888-4MD-GRAD.


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Featured News - Current News - Archived News - News Categories
The Grand Island Fire Co. will host its annual open house on Thursday, Oct. 10, at GIFC headquarters, 2275 Baseline Road.
This year's event features a live firefighting demonstration, a fire engine ride raffle, emergency medical service cardiac lifesaving fire prevention tips, a display of fire and rescue equipment, a basket raffle and refreshments.
The open house runs from 5 until 8 p.m.
In the video below, check out Larry Austin's photos of last year's GIFC open house live fire demonstration.


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FENDER PROMO! Buy Fender American Standard Get FREE Fender Champion 600
Buy a Fender American Standard guitar from Dolphin Music and win a FREE Fender Champion 600 valve amp worth £140!!! Offer ends 30th September, so get yours today!*
* this offer is valid only for UK customers!
The Fender Champion 600 is a 5W tube amp with a 2-tone retro styling. Its vintage 'TV' front makes it look as though it's from the pioneering days of the late 1940s, when Leo Fender was turning out small guitar amps from his radio repair shop, laying down the benchmark for all future guitar amp designs.
Of course, the circuits have been updated to incorporate the hi-gain and overdriven tone required by today's guitar player. The Champion 600 features high or low gain inputs and an extra speaker jack enabling the connection of an external speaker cab.
The Fender Champion 600 is a great vintage-style amp, and if you can't wait to try one of them, here's the great news: Fender is giving away a Fender Champion 600 to anyone who purchases a Fender American Standard guitar - but offer ends 30th September.
How to get a FREE Fender Champion 600 Combo Amp
- Download your unique, serialised Champion 600 voucher here
- Purchase any qualifying Fender American Standard Guitar (see below) between 18th July and 30th September 2009 from Dolphin Music.
- Present your voucher in-store and receive your Fender Champion 600, absolutely FREE!
Qualifying Fender Guitars:
Also qualifying but not currently in stock:
- Fender American Standard Stratocaster Left-handed Rosewood Fretboard
- Fender American Standard Stratocaster Left-handed Maple Fretboard
- Fender American Standard Stratocaster HSS
- Fender American Standard Stratocaster HSS
- Fender American Standard Telecaster Left-handed Maple Fretboard
- call 0844 815 0888 for availability or email us at email@example.com
All colour options qualify for offer. For terms & conditions, please click here
Contact Dolphin Music for the best deals on Fender American Standard guitars and claim your FREE Fender Champion 600 guitar amp.


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HIV/AIDS: Practice Interventions
Kosta N. Kalogerogiannis, Richard Hibbert, Lydia M. Franco, Taiwanna Messam, and Mary M. McKay
For over 20 years, social workers have been involved in service delivery for HIV and AIDS infected and affected individuals. It is estimated that more than 1 million people are living with HIV or AIDS in the United States. The rates of HIV infections continue to rise, with more than 40,000 individuals being diagnosed each year in the United States. This entry explores the current trends in HIV primary prevention, secondary prevention, and counseling and psychotherapy services for people living with or affected by HIV/AIDS.
Men: Health and Mental Health Care
Darrell P. Wheeler
Reproductive Health
Marjorie R. Sable and Patricia J. Kelly
Reproductive health includes family planning, prenatal care, and the broader scope of primary care. Because a woman's health status at conception is as important as prenatal care, genetic screening and 20th century medical technology, reproductive health includes "the preconceptual and interconceptual periods and the menopause, and finally, not only reproductive tract problems but the wide range of risk factors that influence a woman's health in general." Quantitative indicators of reproductive outcomes are useful for summarizing progress in reproductive health. Important indicators are discussed and reveal significant racial disparities.
Sex Workers and HIV/AIDS in India
Sunny Sinha
The risk of HIV infection looms large among male, female, and transgender sex workers in India. Several individual, sociocultural, and structural-environmental factors enhance the risk of HIV infection among sex workers by restricting their ability to engage in safer sexual practices with clients and/or intimate partners. While most HIV prevention programs and research focus on visible groups of women sex workers operating from brothels (Pardasani, 2005) and traditional sex workers, for example, Devadasis (Orchard, 2007); there is a whole subgroup of the sex worker population that remains invisible within HIV prevention programs, such as the male, female, and transgender sex workers operating from non-brothel-based settings. This paper provides an overview of the different types and contexts of sex work prevalent in Indian society, discusses the factors that increase a sex worker's risk of HIV infection, describes the varied approaches to HIV prevention adopted by the existing HIV prevention programs for sex workers, discusses the limitations of the HIV prevention programs, and concludes with implications for social work practice and education.
Women: Health Care
Marian A. Aguilar
This entry provides an abbreviated version of the status of women's health in the United States, highlighting health care utilization, health care expenditures, policy issues, barriers to health care, and the impact on populations at risk. The findings accentuate the importance of moving the women's health care agenda forward because of the persistence of health disparities not just among women of color but among women with disabilities, adolescents, women in violent relationships, women with AIDS, women who are incarcerated, women who are homeless, older low-income women, women on welfare, and lesbian women.
Women's Health
Vimla Nadkarni and Roopashri Sinha
The entry outlines a historical and global overview of women's health in the context of human rights and public health activism. It unravels social myths, traditional norms, and stereotypes impacting women's health because social workers must understand the diverse factors affecting women's health in a continually changing and globalized world. There is need for more inclusive feminist and human rights models to study and advocate women's health. There is as much scope for working with women in a more holistic manner as there is for researching challenging issues and environments shaping women's health.


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Featured News - Current News - Archived News - News Categories
Due to Wednesday's rain delay of game three of the National League Championship Series, the conclusion of the "Judges' Homes" round of "The X Factor" will air in its entirety on Tuesday, Oct. 23 (8 to 10 p.m.), on FOX. The competition kicks into high gear on Tuesday, when the judges reveal which four acts in each of the four categories - Teens, Young Adults, Groups and Over 25s - continue and which two acts are eliminated.
The remainder of "The X Factor" schedule remains intact, with the first live episodes airing Thursday, Nov. 1 (8 to 10 p.m.), and Sunday, Nov. 4 (8 until 10 p.m.).
Visitwww.TheXFactorusa.com for up-to-the-minute news, videos and information. Become a fan of "The X Factor" on Facebook at http://www.facebook.com/TheXFactorUSA and follow the series on Twitter @TheXFactorUSA.
The "Raising Hope," "Ben And Kate," "New Girl" and "The Mindy Project" episodes originally scheduled for Tuesday, Oct. 23, are pre-empted and will be scheduled at a later date.


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Ableton Live 8 upgrade
Live and Suite users can now upgrade their software directly from Ableton
Ableton Live 8 and Suite 8 are being reminded to upgrade their software to Ableton Live 8.0.5, with the new data now available for download at the Ableton website. This latest update is free and fixes a number of gripes that have been discovered since the version 8 software was launched in April this year, thus the developer recommends that users download and install it.
Users can click on the 'Check for Updates' command from Live's help menu or can simply go to Abletons latest version area.


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Auto Body Shops
Beyer Collision Center
Alexandria, VA 22309703-780-7200
|Checkbook's Top Rating ()
- 40 Ratings
- 98% Superior Overall quality
- Prices 10% higher than average
- Compare 132 Auto Body Shops nearby
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Auto Body Shops Articles
How to Find a Good Auto Body Shop
While even the best body shops can't undo an accident, our ratings reveal that many shops will get your car back on the road without a hitch. But our ratings also reveal that some shops may compound your misery with lousy work.
Ask About Source of Replacement Parts
In addition to checking how shops were rated by their customers, judging how well shops communicate, and certifications, ask about the source of any parts that will be used. There is some concern over the quality of used and aftermarket parts.
How to Avoid Overpaying for Auto Body Work
Since insurance companies pay for most repairs, for most auto body repair customers price is less important than quality. But for work paid for by the customer, price is an important consideration.
Advice for Dealing with Your Insurer and Auto Body Shop
Choosing a good shop is an important step toward obtaining quality service, but certainly not the only one. How you deal with your shop and, when you're involved in accidents, with your insurer, are equally important.