In Vitro Immue Response Of Cytotoxic T Lymphocyte Induced By Dendritic Cells Pulsed With Tumor Associated Antigen MUC4

Objective: To explore the in vitro immune response to tumor associated antigen MUC4-derived cytotoxic T lymphocyte (CTL) epitope-pulsed dendritic cell (DC).Methods: DCs was induced from the HLA-A*0201-positive healthy individuals' peripheral blood mononuclear cells (PBMC). DCs' phenotype was identified by flow cytometry. CD8~+ T cells were isolated and purified through MACS (magnetic activated cell sorting). Mature DCs were pulsed with each of the five synthesized peptides which were selected as possible CTL epitopes by software analysis previously. Autologous CD8~+ T cells from the HLA-A* 0201 healthy donor were stimulated with the peptide-pulsed DCs as CTL. CTL activity was assessed by lactate dehydrogenase (LDH) release assay and IFN-γreleased by enzyme-linked immuno spot assay (ELISPOT).Results: Mature DCs could be induced from PBMCs through several cytokines' action. CTL induced by peptide P01204 could lyse T2 cells pulsed with peptide P01204 (9.03%±0.24%,19.62%±0.44%, 27.93%±2.22% at ratios of 5:1,10:1,20:1) and HCT-116 cells (MUC4~+, HLA-A2~+) (7.26%±0.18%,16.83%±0.40%,25.23%±1.35% at ratios of 5:1,10:1,20:1). The number of CTL induced by peptide P01204 which could secret IFN-γ(130.33±6.58) was obviously higher than the negative group.Conclusion: Tumor associated antigen MUC4-derived HLA-A*0201-restrictive cytotoxic T lymphocyte (CTL) epitope P01204 could induce PBMCs' CTL reaction. The CTL could secret immunologic active material to induce the specific target cells' lysis.