Atkin C L, Cole B C, Sullivan G J, Washburn L R, Wiley B B
J Immunol. 1986 Sep 1;137(5):1581-9.
Previous studies established that Mycoplasma arthritidis produces a soluble T cell mitogen (MAM), and that response of murine T cells to MAM is genetically restricted. MAM appeared predominantly in the supernatants of senescent cultures, but was not extracted in significant amounts from whole cells. A quantitative assay of MAM activity was devised. MAM formed noncovalent complexes with nucleic acids and uncharacterized high m.w. constituents of sera and of complex media. Partially purified MAM was adsorbed or denatured by glass and plastic surfaces. MAM was protease-labile, had pI greater than or equal to 9, and had Mr ca 15,000 according to gel filtration experiments. MAM was a very minor component of culture supernatant proteins, and even after 200- to estimated 5 X 10(4)-fold purification was not identified as a stainable or ultraviolet-absorbing entity in electrophoretigrams or chromatograms. It was estimated that MAM was half-optimally active at less than 1000th the half-optimal concentration of concanavalin A or phytohemagglutinin. Culture supernatants and highly purified MAM exhibited the same haplotype specificity (H-2k-dependent response) for stimulated proliferation of lymphocytes and for induction of interferon in vitro.
先前的研究证实,关节炎支原体可产生一种可溶性T细胞促有丝分裂原(MAM),并且小鼠T细胞对MAM的反应受遗传限制。MAM主要出现在衰老培养物的上清液中,但从完整细胞中提取的量并不多。设计了一种MAM活性的定量测定方法。MAM与核酸以及血清和复合培养基中未鉴定的高分子量成分形成非共价复合物。部分纯化的MAM会被玻璃和塑料表面吸附或变性。MAM对蛋白酶敏感,其pI大于或等于9,根据凝胶过滤实验,其相对分子质量约为15,000。MAM是培养上清液蛋白中的一种非常次要的成分,即使经过200倍至估计5×10⁴倍的纯化后,在电泳图或色谱图中也未被鉴定为可染色或有紫外线吸收的物质。据估计,MAM的半最佳活性浓度不到伴刀豆球蛋白A或植物血凝素半最佳浓度的千分之一。培养上清液和高度纯化的MAM在体外刺激淋巴细胞增殖和诱导干扰素方面表现出相同的单倍型特异性(依赖H-2k的反应)。