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Augments Chemotherapy-Induced Apoptosis Cells Facilitates CTL Responses and Activation of CD40 in Cervical Carcinoma

In this study, we describe the expression and function of CD40, a TNF receptor family member, in cervical carcinomas. CD40 was present at very low levels in normal cervical epithelium but was overexpressed in human papillomavirus-infected lesions and advanced squamous carcinomas of the cervix. The stimulation of CD40-positive cervical carcinoma cell lines with soluble CD40L (CD154) resulted in activation of the NF- (cid:1) B and MAPK signaling pathways and up-regulation of cell surface markers and intracellular molecules associated with Ag processing and presentation. Concomitantly, the CD154-induced activation of CD40 in carcinoma cells was found to directly inﬂuence susceptibility to CTL-mediated killing. Thus, CD40 stimulation in cervical carcinoma cell lines expressing a TAP-dependent human papillomavirus 16 E6 Ag epitope resulted in their enhanced killing by speciﬁc CTLs. However, CD154 treatment of carcinoma cells expressing proteasome-dependent but TAP-independent Ags from the EBV-encoded BRLF1 and BMLF1 failed to increase tumor cell lysis by speciﬁc CTLs. Moreover, we demonstrate that chemotherapeutic agents that suppress protein synthesis and reverse the CD40-mediated dissociation of the translational repressor eukaryotic initiation factor 4E-binding protein from the initiation factor eukaryotic initiation factor 4E, such as 5-ﬂuorouracil, etoposide, and quercetin, dramatically increase the susceptibility of cervical carcinoma cells to CD40L-induced apoptosis. Taken together, these observations demonstrate the functional SD) and from a representative experiment performed in MS751-CD40 cells, where the SD of percent lysis was (cid:4) 9% of the mean of triplicate determinations. C CD40 ligation does not increase CTL responses in HPV16 E6 peptide- pulsed cervical carcinoma cells. CaSki cells were stimulated with CD40L for 24–30 h ( (cid:1) L) or left untreated ( (cid:7) L) and then pulsed for 1 h with 1 (cid:1) M HPV16 E6 peptide TIHDIILECV before being analyzed for susceptibility to peptide-speciﬁc CTL lysis by 51 Cr release assays.