Résumé
Objective To explore the HLA-A2 restriction and immunogenicity of 5 previously identified HCV-speeific CTL epitopes. Methods Based on T2 cell, to explore the HLA-A2 restriction of previously identified HCV-specific CTL epitopes by MHC-peptide complex stabilization assay;To detect pep-tide-specific CTL in HLA-A2~+ PBMC stimulated by HLA-A2-restricted peptides by intracellular cytokine staining(ICS) and ELISPOT; To explore the cytotoxicity of peptide-specific CTL to same peptide-loaded T2 cells (target cells) by CTL cytotoxicity test. Results Among 5 previously identified CTL epitopes NS4b_78 (SMMAFSAAL) and NS5a_367 (TVSSALAEL) have high-affinity for HLA-A2 molecules(FI 1) ;ELISPOT results shown that NS4b_78(SMMAFSAAL) and NSSa_367(TVSSALAEL) induced high levels of IFN-γ-se-creting cells [(60±6) SFC/10~4 PBMC vs (4±1 ) SFC/10~4 PBMC, P < 0.01 ; (10 ± 3 ) SFC/10~4 PBMC vs (2±1 ) SFC/10~4 PBMC, P <0.01, respectively] ;ICS results indicated that there were high percentages of CD8~+ IFN-γ~+ T cells in total CD8~+T cells stimulated by these peptides [(2.33 ±0.22 ) % vs (0.05±0.01)%, P <0.001 ; (0.36±0.06)% vs (0.03±0.01)%, P <0.001, respectively]. Furthermore,peptide-specific CTL could effectively kill same peptide-loadcd T2 cells. Conclusion NS4b_78 (SMMAF-SAAL) and NSSa_367 (TVSSALAEL) were identified as HLA-A2-restricted CTL epitopes which could in-duce immune response in vitro.
Résumé
Objective To detect the serum antibody to HPV6 in patients with condylomata acuminata (CA) by use of human papilloma virus(HPV)6bL 1 virus-like particles (VLPS), and to assess the significance of detecting HPV infection. Methods HPV6bL 1 VLPS were produced by a HPV6bL 1 recombinant baculovirus in insect cells,purified by CsCl density gradients centrifugation,and used for ELISA to screen 136 human serum samples in three groups. Results The prevalence of antibody to HPV6bL 1 VLPs was much higher in patients with codylomata acuminata (75%Ab+;mean abs 0.111?0.094) than that in patients with cervical cancer (14.3%Ab+;mean Abs 0.029?0.022)and that in control groups (2.9%Ab+;mean Abs 0.012?0.024). The differences among three groups are significant(P