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目的:观察热休克蛋白65的真核表达载体(pHSP65)对HBV DNA疫苗诱导BALB/c小鼠(H-2d)免疫应答的调节作用。方法:肌内注射空载体pcDNA3,HBV DNA疫苗加HSP65佐剂(pHBVS2S+pHSP65)或不加佐剂(pHBVS2S);ELISA法测定血清抗HBs抗体;ELISPOT检测分泌IFN-γ的脾淋巴细胞;4 h51Cr释放法检测小鼠脾细胞CTLs活性。结果:HBV DNA佐剂组免疫小鼠抗HBsAg抗体滴度虽高于不加佐剂组,但无显著性差异;其IgG1/IgG2 a的比例不同于多肽免疫组,二者分别为0.395与10。佐剂组小鼠分泌IFN-γ的脾淋巴细胞量是不加佐剂组的2~3倍。CTLs细胞杀伤活性(E∶T=100)佐剂组与不加佐剂组分别为:(53.68±7.5)%、(42.81±7.7)%,差异显著(P<0.05)。结论:HBV DNA疫苗具有较强的免疫原性,能够诱导机体产生特异性的抗体及CTLs反应;HSP65佐剂能够有效提高小鼠对DNA疫苗的细胞免疫应答,有望成为DNA疫苗的免疫佐剂。
AIM: To investigate the regulatory effect of heat shock protein 65 eukaryotic expression vector (pHSP65) on immune response induced by HBV DNA vaccine in BALB / c mice (H-2d). Methods: The empty vector pcDNA3, HBV DNA vaccine plus HSP65 adjuvant (pHBVS2S + pHSP65) or without adjuvant (pHBVS2S) were injected intramuscularly. The anti-HBs antibodies were detected by ELISA. The spleen lymphocytes secreting IFN- The release assay was used to detect the CTLs activity of mouse spleen cells. Results: The titer of anti-HBsAg antibody in HBV DNA adjuvant group was significantly higher than that in non-adjuvant group, but there was no significant difference between the two groups. The IgG1 / IgG2-a ratio was different from that of peptide-immunized group (0.395 and 10 respectively). The amount of splenic lymphocytes secreting IFN-γ in the adjuvant group was 2-3 times higher than that in the adjuvant-free group. The cytotoxicity of CTLs (E: T = 100) were (53.68 ± 7.5)% and (42.81 ± 7.7)%, respectively, with significant difference (P <0.05). Conclusion: HBV DNA vaccine has strong immunogenicity and can induce specific antibodies and CTLs responses. HSP65 adjuvant can effectively improve the cellular immune response to DNA vaccine in mice and is expected to be an immune adjuvant for DNA vaccine.