丙型肝炎患者血清HSP70水平的检测及HSP70-HCV肽诱导CTL作用

Detection of serum heat shock protein 70 level in patients with hepatitis C infection and its role in cytotoxicity of CTLs induced by HSP70-HCV peptide complex

目的 :检测丙型肝炎患者及正常人血清中HSP70的水平 ,探讨HSP70 HCV肽体外诱导特异性CTL的作用。方法 :先用ELISA法筛选出丙型肝炎患者及正常人血清 ,再用双夹心ELISA法检测血清HSP70 ,并分析抗HCV抗体和HSP70之间的关系。用HSP70 HCV肽激活外周血单个核细胞后 ,通过51Cr释放试验测定CTL的杀伤活性。结果 :76份血清 (抗HCV抗体阳性 2 8例 ,正常人血清 4 8例 )中 ,抗HCV抗体阳性患者HSP70的检出率为 82 .1% ,正常人血清中HSP70的检出率为18.8%。HSP70的检出率与HCV感染呈显著相关 ( χ2 =2 8.77,P <0 .0 1)。HCV感染者血清HSP70的含量显著高于正常人。HSP70与HCVC区肽 (DLMGYIPAV)的混合物 ,可诱导 1例患者的CTL对自体HCV肽致敏靶细胞的杀伤率达 37.8% ,而用HCV肽单独作用则不显著。结论 :HCV感染可刺激机体过表达HSP70 。

AIM: To detect serum HSP70 in patients with hepatitis C and evaluate its role in cytotoxicity of specific CTLs induced by HSP70 HCV peptide complex. METHODS: The serum HSP70 level in patients infected with HCV and normal individuals were detected by ELISA. The relationship between anti HCV antibody and HSP70 was evaluated. Peripheral blood mononuclear cells (PBMCs) were activated by HSP70 HCV peptide complex and then 4 hour 51 Cr release assay were used to detect the killer activity of CTLs. RESULTS: Detection rates of HSP70 were 82.1%(22/28) and 18.8% (9/48) in patients with anti HCV antibody and in normal individuals, respectively. There was significant correlation between the detection rate of HSP70 and HCV infection ( χ 2=28.77, P <0.01). The level of serum HSP70 in patients infected with HCV was markedly more higher than that in normal individuals. CTLs from one patient induced by HSP70 HCV C region peptide (DLMGYIPAV) complex could lyse autologous B lymphoblastoid cell lines (BLcLs) with killer rate of 37.8%, whereas CTLs induced by peptide alone could not lyse autologous BLcLs cells. CONCLUSION: HCV infection can induce over expression of HSP70. HSP70 may have the action of enhancing presentation of HCV epitope peptide and promoting clearance cells infected with HCV.