Research of dendritic cells pulsed with HPV peptide inducing specific CTL in vitro / 中华微生物学和免疫学杂志

ABSTRACT

Objective To explore the potential of autologous dendritic cells (DC) pulsed with HLA-A201-binding peptide E613-21(KLPDLCTEL) and E786-94(TLGIVCPI)in inducing specific T cells respouse in vitro.Methods Cervical carcinoma patients with positive HLA-A201 were enrolled and their monocytes isolated and induced into dendritic cells and pulsed with HLA-A201-binding peptide E613-21 and E786-94.PBLs were primed by DCs every week for thee times.The cytokine level of supernatant of CTLs was tested by ELISA.The percentage of special CTLs was tested by flow cytometry.The specific killing effect of CTLs was tested by MTT.Results the numbers of DCs of eleven cervical carcinoma patients were (10.79±0.88) ×106(100 ml peripheral blood).CDllc+HLA-DR+(97.15±2.41)％,CD80+(84.28+5.39)％,CD83 +(85.17±5.06) ％,CD86 + (97.74+0.87) ％.Proliferation index of PBLs primed by DCs three times was 15.4± 1.5.Cytokine levels including IL-2,IL-12,IFN-γ and TNF-α were obviously higher than nonpriming PBLs[(2551.9+195.3) pg/ml,(554.9±64.0) pg/ml,(2416.9±281.7) pg/ml,(632.4 +71.1)pg/ml,respectively] (P＜0.05),but IL-10 was no significant difference between priming CTLs and nonpriming CTLs.The average percentage of special CTLs was obviously higher than control group[(6.32±1.54)％,P＜0.05].The killing effect of CTLs was obviously higher than control group(P＜0.05).Conclusion Dendritic cells pulsed with peptide E613-21 and E786-94 can induce special CTLs in vitro and stimulate CTLs secret cytokines.This will provide science basis for research of therapeutic HPV vaccine.