Peterson J D, Karpus W J, Clatch R J, Miller S D
Department of Microbiology-Immunology, Northwestern University Medical School, Chicago, IL 60611.
Eur J Immunol. 1993 Jan;23(1):46-55. doi: 10.1002/eji.1830230109.
Theiler's murine encephalomyelitis virus (TMEV) produces a chronic, inflammatory demyelinating disease in susceptible mouse strains that is used as a model for multiple sclerosis. Because disease susceptibility correlates temporally with the development of virus-specific delayed-type hypersensitivity (DTH) responses, we studied methods and mechanisms by which virus-specific DTH could be specifically inhibited. The intravenous injection of UV-inactivated TMEV coupled to syngeneic splenocytes via a carbodiimide linkage (TMEV-SP), prior to immunization, induced a significant degree of tolerance in virus-specific helper (Th) cells as determined by decreased DTH and T cell proliferative responses, and decreased interleukin (IL)-2 and interferon (IFN)-gamma protein and mRNA levels. In contrast to the reduced levels of Th1-specific lymphokine mRNA levels, IL-4-specific mRNA levels in response to virus stimulation were not affected in tolerant mice. Surprisingly, the total anti-TMEV antibody response in DTH tolerant mice was enhanced 20-100-fold over sham-tolerized controls and was composed of reduced levels of anti-virus IgG2a, but dramatically increased levels of anti-virus IgG1. The "split-tolerance" was antigen specific, dependent on the concentrations of TMEV and carbodiimide used in the coupling procedure, and varied with the number of coupled syngeneic splenocytes administered. The fixative effects of carbodiimide on antigen-presenting function were necessary for the induction of DTH tolerance with TMEV-SP, since intravenous administration of virus coupled to splenocytes via a biotin-avidin linkage led to enhanced virus-specific antibody responses, but was unable to inhibit DTH unless concomitantly fixed with carbodiimide. Collectively, the data indicate that Th1 cells (mediating DTH, IL-2 and IFN-gamma production, and helper function for IgG2a production) were specifically anergized, with concomitant stimulation of Th2 cells (producing IL-4 and mediating helper function for IgG1 antibody production).
泰勒氏鼠脑脊髓炎病毒（TMEV）在易感小鼠品系中引发一种慢性炎症性脱髓鞘疾病，该疾病被用作多发性硬化症的模型。由于疾病易感性在时间上与病毒特异性迟发型超敏反应（DTH）的发展相关，我们研究了特异性抑制病毒特异性DTH的方法和机制。在免疫前，静脉注射通过碳二亚胺连接与同基因脾细胞偶联的紫外线灭活TMEV（TMEV-SP），通过降低的DTH和T细胞增殖反应以及降低的白细胞介素（IL）-2和干扰素（IFN）-γ蛋白及mRNA水平来确定，诱导了病毒特异性辅助（Th）细胞的显著程度的耐受。与Th1特异性淋巴因子mRNA水平降低相反，耐受小鼠中对病毒刺激的IL-4特异性mRNA水平不受影响。令人惊讶的是，DTH耐受小鼠中的总抗TMEV抗体反应比假耐受对照增强了20至100倍，并且由降低水平的抗病毒IgG2a组成，但抗病毒IgG1水平显著增加。这种“分裂耐受”是抗原特异性的，取决于偶联过程中使用的TMEV和碳二亚胺的浓度，并随给予的偶联同基因脾细胞数量而变化。碳二亚胺对抗抗原呈递功能的固定作用对于用TMEV-SP诱导DTH耐受是必要的，因为通过生物素-抗生物素蛋白连接与脾细胞偶联的病毒静脉内给药导致增强的病毒特异性抗体反应，但除非同时用碳二亚胺固定，否则无法抑制DTH。总体而言，数据表明Th1细胞（介导DTH、IL-2和IFN-γ产生以及IgG2a产生的辅助功能）被特异性无反应，同时刺激Th2细胞（产生IL-4并介导IgG1抗体产生的辅助功能）。
