Cullen C M, Bonventre P F, Heeg H, Bluethmann H, Mountz J D, Edwards C K
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, OH 45267, USA.
Pathobiology. 1995;63(6):293-304. doi: 10.1159/000163965.
Recombinant toxic shock syndrome toxin-1 (rTSST-1) administered to MRL-lpr/lpr TCR V beta 8.2 transgenic mice at doses of 0.1 microgram/mouse resulted in 100% mortality. This was an unexpected finding since TSST-1 does not activate V beta 8.2 T cells. In contrast, control mice heterozygous at the lpr locus and also for the transgene (MRL-lpr/+; V beta 8.2/0) survived doses of superantigen 100 times higher. The transgenic mice which succumbed to rTSST-1 challenge exhibited histopathology of the liver consistent with toxic shock (generalized inflammation and hepatocellular necrosis) as well as substantially elevated serum TNF-alpha, IL-2, and IL-6 cytokine levels. Splenic T cells derived from transgenic mice stimulated with rTSST-1 in vitro did not undergo detectable proliferation as measured in a standard mitogen assay. However, PCR amplification of cDNA prepared from the V beta 8.2 splenocytes revealed the presence of minor populations of TSST-1-reactive V beta elements (i.e. V beta 3 and V beta 15). Furthermore, an expansion of the V beta 3 and V beta 15 T-cell families was detected by PCR assay of spleen cell cultures stimulated with rTSST-1. These results suggested that the exquisite sensitivity of the MRL-lpr/lpr V beta 8.2 transgenic animals to rTSST-1 was not dependent exclusively on T-cell proliferation but was augmented by the influence of a defective fas antigen receptor expressed in homozygous lpr mice. To test this hypothesis more directly, we compared the sensitivity of MRL-lpr/lpr mice (not carrying the V beta 8.2 transgene) to MRL-+/+ mice. The MRL-lpr/lpr fas antigen-defective mice were substantially more susceptible to rTSST-1 challenge. Mice carrying the lpr mutation on another genetic background (C57BL/6.C3H-lpr/lpr) were also more sensitive to rTSST-1 challenge than were C57BL/6.C3H-+/+ mice. Although induction of toxic shock is clearly associated with T-cell proliferation, defects in fas antigen receptor or ligand may also contribute substantively to superantigen-mediated lethal shock by still undefined mechanisms.
以0.1微克/只的剂量给MRL-lpr/lpr TCR Vβ8.2转基因小鼠注射重组中毒性休克综合征毒素-1(rTSST-1),导致100%的死亡率。这是一个意外发现,因为TSST-1不会激活Vβ8.2 T细胞。相比之下,lpr位点以及转基因均为杂合的对照小鼠(MRL-lpr/+; Vβ8.2/0)在剂量高出100倍的超抗原作用下存活了下来。死于rTSST-1攻击的转基因小鼠表现出与中毒性休克相符的肝脏组织病理学变化(全身性炎症和肝细胞坏死),同时血清TNF-α、IL-2和IL-6细胞因子水平大幅升高。在标准促有丝分裂原试验中,用rTSST-1体外刺激转基因小鼠来源的脾T细胞,未检测到可察觉的增殖。然而,对从Vβ8.2脾细胞制备的cDNA进行PCR扩增,发现存在少量对TSST-1有反应的Vβ元件群体(即Vβ3和Vβ15)。此外,通过对用rTSST-1刺激的脾细胞培养物进行PCR分析,检测到Vβ3和Vβ15 T细胞家族有所扩增。这些结果表明,MRL-lpr/lpr Vβ8.2转基因动物对rTSST-1的高度敏感性并非仅取决于T细胞增殖,而是因纯合lpr小鼠中表达的缺陷型fas抗原受体的影响而增强。为了更直接地验证这一假设,我们比较了MRL-lpr/lpr小鼠(不携带Vβ8.2转基因)与MRL-+/+小鼠的敏感性。MRL-lpr/lpr fas抗原缺陷小鼠对rTSST-1攻击的敏感性明显更高。在另一种遗传背景(C57BL/6.C3H-lpr/lpr)上携带lpr突变的小鼠对rTSST-1攻击也比C57BL/6.C3H-+/+小鼠更敏感。虽然中毒性休克的诱导显然与T细胞增殖有关,但fas抗原受体或配体的缺陷也可能通过仍未明确的机制,对超抗原介导的致死性休克有实质性影响。