Pichugin A V, Khaidukov S V, Moroz A M, Apt A S
Laboratory for Immunogenetics, Central Institute for Tuberculosis, Moscow, Russia.
Clin Exp Immunol. 1998 Feb;111(2):316-24. doi: 10.1046/j.1365-2249.1998.00498.x.
It is firmly established that the allelic composition of the H-2 complex has a prominent impact on the course of tuberculosis (TB) infection in mice, including granuloma formation, mycobacterial spread in the lungs, and the dynamics of mortality. Although intuitively obvious, the role of long-term specific T cell responses in the expression of corresponding phenotypes is poorly understood. In this study we have compared polyclonal lymph node cell response (cell yield, proliferation, surface markers, IL-4/interferon-gamma (IFN-gamma) production) to Mycobacterium tuberculosis H37Rv sonicate in repeated 10-day cycles of stimulation/rest between H-2 congenic IE-negative mouse strains, categorized on the basis of mortality following lethal challenge as TB-susceptible (C57B1/6), TB-resistant (4R) and BCG non-protected (B10.M). The capacity to retain specific responsiveness to repeated stimulation by mycobacterial antigens depended upon both the H-2 haplotype of the host and the immunizing dose of the antigen. 4R lymph node cells following either 50 microg/mouse or 100 microg/mouse immunization constantly responded to sonicate, increased in numbers, and after the third stimulation/rest cycle developed into a stable CD3+CD4+ cell line. B6 cells following either 50 microg/mouse or 100 microg/mouse immunization, and B10.M cells following 100 microg/mouse (but not 50 microg/mouse) immunization, lost the capacity to incorporate methyl-3H-thymidine during the second cycle, and died. Analogous results were obtained in the in vivo experiments, when the dynamics of the response over 12 weeks following a single immunization with the antigen was studied. In response to the antigen, cells from all three mouse strains produced significant amounts of IL-2 and IFN-gamma, but not IL-4, indicating that they belong predominantly to the Th1-like subset. Among noteworthy differences between the mouse strains was a clear deficiency of CD8+ T cells in B6 cultures, and an unusually high proportion of CD3+CD4-CD8- (double-negative) T cells in B10.M cultures following a high-dose immunization.
H-2复合体的等位基因组成对小鼠结核病(TB)感染进程有显著影响,这一点已得到确证,包括肉芽肿形成、肺部分枝杆菌传播以及死亡率动态变化。尽管直观上显而易见,但长期特异性T细胞反应在相应表型表达中的作用却知之甚少。在本研究中,我们比较了H-2同类基因IE阴性小鼠品系在重复的10天刺激/休息周期中,针对结核分枝杆菌H37Rv超声裂解物的多克隆淋巴结细胞反应(细胞产量、增殖、表面标志物、IL-4/干扰素-γ(IFN-γ)产生)。这些小鼠品系根据致死性攻击后的死亡率分为TB易感(C57B1/6)、TB抗性(4R)和卡介苗无保护作用(B10.M)。对分枝杆菌抗原重复刺激保持特异性反应的能力既取决于宿主的H-2单倍型,也取决于抗原的免疫剂量。用50微克/小鼠或100微克/小鼠免疫后的4R淋巴结细胞持续对超声裂解物产生反应,数量增加,在第三个刺激/休息周期后发展成为稳定的CD3+CD4+细胞系。用50微克/小鼠或100微克/小鼠免疫后的B6细胞,以及用100微克/小鼠(而非50微克/小鼠)免疫后的B10.M细胞,在第二个周期失去了掺入甲基-3H-胸腺嘧啶核苷的能力并死亡。在体内实验中,当研究单次免疫抗原后12周内的反应动态时,也得到了类似结果。针对该抗原,所有三个小鼠品系的细胞都产生了大量的IL-2和IFN-γ,但未产生IL-4,表明它们主要属于Th1样亚群。小鼠品系之间值得注意的差异包括B6培养物中CD8+T细胞明显缺乏,以及高剂量免疫后B10.M培养物中CD3+CD4-CD8-(双阴性)T细胞比例异常高。