Baker-Zander S A, Sell S, Lukehart S A
Infect Immun. 1982 Aug;37(2):568-78. doi: 10.1128/iai.37.2.568-578.1982.
Sera from rabbits with early experimental syphilis were tested for their effect on in vitro lymphocyte transformation responses to related specific antigens (sonicated T. pallidum), unrelated specific antigens (sheep erythrocytes), and the T cell mitogen, concanavalin A. Results were compared with responses in preinfection sera and in sera from sham-infected rabbits. Titration experiments in which normal serum was used indicated that optimal lymphocyte responsiveness is obtained with a final serum concentration of 1%. Under these conditions, no differences in concanavalin A stimulation were observed in cultures with syphilitic sera. Responses to sonicated T. pallidum were inhibited, but only by 17 to 25% when compared with the response in preinfection sera. In cultures containing 10% serum, inhibition of lymphocyte proliferation to sonicated T. pallidum antigens was evident with sera from all syphilitic animals from day 10 (55% inhibition) through day 31 (80% inhibition) of infection. Responses to concanavalin A and sheep erythrocytes were significantly inhibited by day 10 sera; only 20% of the sera tested demonstrated substantial nonspecific inhibitory capacity. No differences were evident among sera from any of the sham-infected animals or among the preinfection sera from either group. Pooled serum with high inhibitory activity was fractionated by ammonium sulfate precipitation, DEAE ion exchange chromatography, and Sephadex G-200 gel filtration. Two separate inhibitors were identified: (i) a low-molecular-weight, ammonium sulfate-soluble, nonspecific inhibitory fraction containing albumin and alpha-globulins with the capacity to inhibit both antigen and mitogen responses and (ii) a high-molecular-weight, ammonium sulfate-precipitable, inhibitory fraction containing alpha-globulin and FTA-ABS-reactive immunoglobulin M which affected only the antigen-specific response to sonicated T. pallidum. Immunodiffusion failed to detect immunoglobulin or T. pallidum antigens in either fraction. DEAE-purified immunoglobulin G from immune serum was not inhibitory.
检测早期实验性梅毒兔血清对体外淋巴细胞转化反应的影响，该反应针对相关特异性抗原（超声处理的梅毒螺旋体）、非相关特异性抗原（绵羊红细胞）以及T细胞有丝分裂原刀豆球蛋白A。将结果与感染前血清以及假感染兔血清中的反应进行比较。使用正常血清的滴定实验表明，最终血清浓度为1%时可获得最佳淋巴细胞反应性。在这些条件下，用梅毒血清培养时，刀豆球蛋白A刺激未见差异。对超声处理的梅毒螺旋体的反应受到抑制，但与感染前血清中的反应相比，仅抑制了17%至25%。在含有10%血清的培养物中，从感染第10天（抑制55%）至第31天（抑制80%），所有梅毒动物的血清对超声处理的梅毒螺旋体抗原的淋巴细胞增殖抑制明显。到第10天血清时，对刀豆球蛋白A和绵羊红细胞的反应受到显著抑制；所检测的血清中只有20%表现出显著的非特异性抑制能力。假感染动物的任何血清之间或两组的感染前血清之间均无明显差异。通过硫酸铵沉淀、DEAE离子交换色谱和Sephadex G - 200凝胶过滤对具有高抑制活性的混合血清进行分级分离。鉴定出两种不同的抑制剂：(i) 一种低分子量、硫酸铵可溶的非特异性抑制级分，含有白蛋白和α球蛋白，能够抑制抗原和有丝分裂原反应；(ii) 一种高分子量、硫酸铵可沉淀的抑制级分，含有α球蛋白和FTA - ABS反应性免疫球蛋白M，仅影响对超声处理的梅毒螺旋体的抗原特异性反应。免疫扩散未在任何一个级分中检测到免疫球蛋白或梅毒螺旋体抗原。免疫血清经DEAE纯化的免疫球蛋白G无抑制作用。
