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[Identification of immunological effector cells after autologous cytokine-induced killer cells treatment and its clinical implication in hepatocellular carcinoma patients].

OBJECTIVE
To investigate the alteration of the cellular profiles of T lymphocyte subsets and dendritic cell subsets in peripheral blood of primary hepatocellular carcinoma (HCC) patients after being transfused with autologous cytokine-induced killer cells (CIK) in patients, then to evaluate the clinical efficacy of the immune therapeutic strategy.


METHODS
Peripheral blood mononuclear cells (PBMCs) from 13 patients with primary were collected using blood cell separator, and expanded in the fresh AIM-V medium in the presence of cytokine cocktail including interferon-gamma (IFN-gamma), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). The phenotypic patterns of CIK cells were longitudinally characterized by flow cytometry on day 0, 4, 7, 10,13 and 15 during the incubation period. PBMCs obtained from HCC patients before or after CIK cells transfusion into bodies to assay the changes of proportion of DC1 or DC2 in peripheral blood.


RESULTS
After in vitro incubation for 14 or 15 days, a large of CD3(+)CD56(+) cells were produced from their progenitors and the percentages of CD3(+)CD8(+), CD3(+)CD56(+), CD25(+) cells significantly increased from 33.5% +/- 10.1%, 7.7% +/- 2.8%, and 12.3% +/- 4.5% at the beginning to 36.6% +/- 9.0% (P < 0.05), 18.9% +/- 6.9% (P < 0.01), and 16.4% +/- 5.9% (P < 0.05) at the day 15, respectively. In contrast, the percentages of CD3(+)CD4(+) and NK cells displayed no significant difference. The percentages of CD3(+), CD3(+)CD8(+) cells was held at a higher level during the whole incubation period, however those of the CD25(+), and CD3(+)CD56(+) cells began decreasing on day 7 and day 13, respectively. The proportion of type I of dendritic cells (DC1) and type II of dendritic cells (DC2) subsets increased from 0.59% +/- 0.23% and 0.26% +/- 0.12% before CIK cell transfusion to 0.85% +/- 0.27% and 0.43% +/- 0.20% (all P < 0.01) after CIK cell transfusion. The symptom of HCC patients receiving the CIK cell therapy was markedly ameliorated, and not side effect was seen in the treatment.


CONCLUSION
Our results indicated that autologous CIK cells is able to boost the cellular immunological function in HCC patients, which probably provide a potent immune therapeutic strategy for HCC patients.