Verma Shailendra Kumar, Kumar Subodh, Gupta Nimesh, Vedi Satish, Bhattacharya Shailja Misra, Lakshmana Rao P V
Microbiology Division, Defense Research and Development Establishment, Jhansi Road, Gwalior-474002, India.
Vaccine. 2009 Nov 16;27(49):6905-9. doi: 10.1016/j.vaccine.2009.09.006. Epub 2009 Sep 15.
Japanese encephalitis is a major cause of encephalitis in Asia. Cases occur largely in rural areas of the South and East Asian region resulting in significant morbidity and mortality. Multiple vaccines exist to control Japanese encephalitis, but all suffer from problems. Envelope protein domain III of Japanese encephalitis virus is involved in binding to host receptors and it contains specific epitopes that elicit virus-neutralizing antibodies. Earlier, the protective efficacy of domain III has been evaluated in mice by some researchers, but these studies are lacking in explanation of humoral and cellular immune responses. We have earlier reported cloning, expression, purification and in vitro refolding of Japanese encephalitis virus envelope protein domain III (rJEV-DIII). Ninety percent JEV is neutralized when the serum against refolded rJEV-DIII is used at a dilution of 1:80. In the present study, we have evaluated the immunomodulatory potential of refolded rJEV-DIII protein in BALB/c mice with Freunds complete/incomplete adjuvants. Mice were tested for humoral immune response by ELISA. Cell-mediated immune response was tested by lymphocyte proliferation assay and cytokine profiling. The rJEV-DIII generated high IgG antibody and its isotypes (IgG2a and IgG3) and induced significant expression of INF-gamma and IL-2 cytokines. The rJEV-DIII induced significant lymphoproliferation of splenocytes. In conclusion rJEV-DIII induced Th1 type of immune response which plays an important role in protection for intracellular pathogens.
日本脑炎是亚洲脑炎的主要病因。病例主要发生在南亚和东亚地区的农村,导致了显著的发病率和死亡率。有多种疫苗可用于控制日本脑炎,但都存在问题。日本脑炎病毒的包膜蛋白结构域III参与与宿主受体的结合,并且包含能引发病毒中和抗体的特定表位。此前,一些研究人员已在小鼠中评估了结构域III的保护效力,但这些研究缺乏对体液免疫和细胞免疫反应的解释。我们之前报道过日本脑炎病毒包膜蛋白结构域III(rJEV-DIII)的克隆、表达、纯化及体外重折叠。当使用以1:80稀释的抗重折叠rJEV-DIII血清时,90%的日本脑炎病毒被中和。在本研究中,我们评估了重折叠的rJEV-DIII蛋白在弗氏完全/不完全佐剂存在的情况下对BALB/c小鼠的免疫调节潜力。通过ELISA检测小鼠的体液免疫反应。通过淋巴细胞增殖试验和细胞因子分析检测细胞介导的免疫反应。rJEV-DIII产生了高IgG抗体及其亚型(IgG2a和IgG3),并诱导了INF-γ和IL-2细胞因子的显著表达。rJEV-DIII诱导了脾细胞的显著淋巴细胞增殖。总之,rJEV-DIII诱导了Th1型免疫反应,这在针对细胞内病原体的保护中起重要作用。