C D4＋T 细胞剔除对复制型重组痘苗病毒免疫原性及毒力的影响  

Study of the immunogenicity and virulence of replication-competent recombinant vaccinia virus in CD4＋T cell-depleted mice

作　　者：刘颖[1] 王书晖[1] 刘莹[1] 彭虹[1] 刘畅[1] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心传染病预防控制国家重点实验室,北京102206

出　　处：《中华微生物学和免疫学杂志》2014年第11期811-816,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家科技重大专项（2012zx10001-D08）;国家自然科学基金国际合作项目（8102008030）

摘　　要：目的：探讨CD4＋T细胞剔除对复制型重组痘苗病毒免疫原性及毒力的影响。方法BALB／c小鼠经腹腔注射CD4单克隆抗体剔除CD4＋T细胞后，尾部划痕接种重组痘苗病毒rTV或天坛株VTT，同时设正常小鼠对照。免疫后定期观察小鼠体重变化和接种部位痘斑变化，并检测病毒排出量和CD4＋T细胞比例。免疫后第28天处死小鼠，检测小鼠体液和细胞免疫，取卵巢组织检测病毒滴度。结果 CD4＋T细胞剔除小鼠和正常小鼠在接种rTV或VTT后均出现典型的种痘后局部反应，但均未出现次级或卫星病损。 CD4＋T细胞剔除小鼠的痘斑愈合时间、病毒排出时间及体重恢复时间均长于正常小鼠。 CD4＋T细胞剔除小鼠和正常小鼠的卵巢中均未检测出痘苗病毒。 CD4＋T细胞剔除小鼠的HIV抗体和痘苗病毒抗体平均A值分别为0．119和0．168，显著低于正常小鼠的平均A值（P＜0．05，P＜0．01）。 McAb／rTV组的痘苗病毒中和抗体滴度几何均值为1∶321，显著低于rTV组1∶1286（P＜0．05）。 McAb／rTV组诱导的痘苗病毒特异性IFN-γ＋／CD4＋T效应细胞百分比均值为0．654％，显著低于rTV组（P＝0．0004），但IFN-γ＋／CD8＋T效应细胞百分比与rTV组的差异无统计学意义。结论剔除CD4＋T细胞的小鼠能有效控制重组痘苗病毒的复制和播散。 CD4＋T细胞的剔除显著降低了体液免疫应答和CD4＋T细胞免疫应答，对CD8＋T细胞免疫应答无显著影响。Objective To investigate the effects of CD4＋T cell depletion in BALB/c mice on the immunogenicity and virulence of replication-competent recombinant vaccinia virus. Methods Twelve BALB/c mice were inoculated with recombinant Tiantan vaccinia （ rTV, n=8 ） or vaccinia virus Tiantan strain （ VTT, n=4） by tail scarification after the depletion of CD4＋T cells with anti-CD4 monoclonal anti-body（ McAb） injected intraperitoneally for three days before virus inoculation.A control group without anti-body treatment was set up accordingly.Several parameters including the body weight, pocks on tail, viral shedding and the percentage of CD4＋T cells were monitored.In the fourth week after virus infection, ovaries were collected from mice and viral loads in the tissue were titrated by plaque forming assay on chick embryo fibroblast （ CEF） cells.The specific cellular immune responses against vaccinia virus and HIV induced by inoculation of VTT and rTV respectively were detected by intracellular cytokine staining （ ICS） assay.ELISA was used to detect antibodies against vaccinia virus and HIV-1 gp120.Results All mice with or without CD4 McAb treatment showed typical poxvirus pocks on tails after inoculation of vaccinia viruses, but none of them developed secondary or satellite lesions.It took a longer time for CD4＋T cell-depleted mice to heal from lesions, to regain body weights and to release viruses than the mice in control group.No vaccinia virus was detected in the ovaries of CD4＋T cell-depleted mice or mice in control group.The mean absorbance（ A） values for the detection of HIV-specific and vaccinia virus-specific antibodies in CD4＋T cell-depleted mice with ELISA were respectively 0.119 and 0.168, which were significantly lower than those in mice of control group.The titers of neutralizing antibodies against vaccinia virus in McAb/rTV treated mice （1 ∶321） were lower than those in rTV treated mice （1 ∶1286） （P〈0.05）.The percentages of CD4＋T cells secreting IFN-γ（0.654

关 键 词：复制型重组痘苗病毒 CD4+T细胞 CD8+T细胞 病毒清除 免疫原性 

分 类 号：R392[医药卫生—免疫学]