Romano T J, Lerman S P, Thorbecke G J
Eur J Immunol. 1976 Jun;6(6):434-42. doi: 10.1002/eji.1830060611.
Incubation of trinitrophenylated hemocyanin (TNP-KLH)-primed spleen cells with microgram amounts of 2,4-dinitrophenyl (DNP) or 2,4,6-trinitrophenyl (TNP) conjugates of pneumococcal polysaccharide type 3 (SIII) for as little as 5 min at 4 degrees C results in a specific "block" of the 19 S and 7 S adoptive memory response to TNP-KLH. This hapten-SIII-induced block of anti-hapten memory B cell responsiveness seems to be an example of specific receptor blockade. The block is specific and can be prevented by simultaneous incubation of the primed cells with hapten-protein conjugates which presumably compete with the hapten-polysaccharide for attachment to the B cell surface via anti-hapten Ig receptors. Removal via capping of these Ig receptors by exposure of TNP-KLH-primed memory cells to rabbit anti-mouse Fab serum for 45 min at 37 degrees C renders these cells refractory to the blocking effect of hapten-SIII. Once the hapten-SIII has attached to the memory cells, these blocked cells can be "rescued" (i.e. returned to a state of responsiveness) by incubating these cells with either mouse anti-SIII at 37 degrees C or rabbit anti-DNP serum at 4 degrees C. Since a papain digest of the IgG fraction of rabbit anti-DNP did not rescue the cells while the intact IgG did, a capping off of the TNP-SIII was proposed as the mechanims for this return to responsiveness of the hitherto blocked cells. A rescue was not seen by treatment of recipient mice with such B cell mitogens as dextran sulfate, endotoxin or purified protein derivative of tuberculin.
用微克量的3型肺炎球菌多糖(SIII)的2,4 - 二硝基苯基(DNP)或2,4,6 - 三硝基苯基(TNP)缀合物在4℃下与三硝基苯基化血蓝蛋白(TNP - KLH)致敏的脾细胞孵育仅5分钟,就会导致对TNP - KLH的19S和7S过继性记忆反应出现特异性“阻断”。这种半抗原 - SIII诱导的抗半抗原记忆B细胞反应性阻断似乎是特异性受体阻断的一个例子。该阻断是特异性的,并且可以通过将致敏细胞与半抗原 - 蛋白质缀合物同时孵育来预防,推测这些半抗原 - 蛋白质缀合物通过抗半抗原Ig受体与半抗原 - 多糖竞争附着于B细胞表面。通过将TNP - KLH致敏的记忆细胞在37℃下暴露于兔抗小鼠Fab血清45分钟,通过封帽去除这些Ig受体,使这些细胞对半抗原 - SIII的阻断作用产生抗性。一旦半抗原 - SIII附着于记忆细胞,通过在37℃下用小鼠抗SIII或在4℃下用兔抗DNP血清孵育这些细胞,可以“拯救”这些被阻断的细胞(即恢复到反应性状态)。由于兔抗DNP的IgG部分的木瓜蛋白酶消化产物不能拯救细胞,而完整的IgG可以,因此提出TNP - SIII的封帽是使迄今被阻断的细胞恢复反应性的机制。用诸如硫酸葡聚糖、内毒素或结核菌素纯化蛋白衍生物等B细胞有丝分裂原处理受体小鼠未观察到拯救现象。