Tarkowski A, Moldoveanu Z, Koopman W J, Radl J, Haaijman J J, Mestecky J
Department of Microbiology, University of Alabama, Birmingham 35294.
Clin Exp Immunol. 1991 Aug;85(2):341-8. doi: 10.1111/j.1365-2249.1991.tb05730.x.
Using modified ELISA and spot-ELISA, which permit the parallel determination of heavy chain subclass and the presence of covalently linked J chain, we analysed IgA found in cell culture supernatants or secreted by individual cells from peripheral blood, spleen, bone marrow, gingiva and synovial tissue, with respect to its polymeric or monomeric IgA form (pIgA, mIgA) and IgA1 or IgA2 subclass. The ELISA for determination of J chain in tissue culture supernatants was specific and highly sensitive (detection limit in pg). The results demonstrated that IgA1-producing cells predominated in the tissues examined, and that J chain could be detected in association with the majority of IgA1 and IgA2 secreted by individual cells. With respect to the frequency of cells secreting polymeric, J chain-containing IgA, only 20-30% of cells from the bone marrow were engaged in the synthesis of PIgA. In other tissues the frequency of cells secreting pIgA1 and pIgA2 was considerably higher. Peripheral blood mononuclear cells secreting pIgA2 were easily inducible during stimulation with T cell-dependent pokeweed mitogen, whereas Epstein-Barr virus-transformed cells secreted preferentially mIgA1. When the frequencies of pIgA-, pIgA1- or pIgA2-secreting cells (determined by spot-ELISA technique) from different tissues were correlated with the proportion of pIgA to mIgA (and IgA subclasses) secreted in tissue culture supernatants, data obtained suggest that many individual IgA-producing cells could be engaged in simultaneous secretion of mIgA and pIgA.
使用改良酶联免疫吸附测定法(ELISA)和斑点ELISA,这些方法可同时测定重链亚类以及共价连接的J链的存在情况,我们分析了细胞培养上清液中或外周血、脾脏、骨髓、牙龈和滑膜组织单个细胞分泌的IgA,涉及聚合或单体IgA形式(pIgA、mIgA)以及IgA1或IgA2亚类。用于测定组织培养上清液中J链的ELISA具有特异性且高度灵敏(检测限为皮克)。结果表明,在所检测的组织中,产生IgA1的细胞占主导,并且可以检测到与单个细胞分泌的大多数IgA1和IgA2相关联的J链。关于分泌含J链的聚合IgA的细胞频率,骨髓中只有20% - 30%的细胞参与pIgA的合成。在其他组织中,分泌pIgA1和pIgA2的细胞频率要高得多。在用T细胞依赖性商陆有丝分裂原刺激期间,分泌pIgA2的外周血单核细胞很容易被诱导,而爱泼斯坦 - 巴尔病毒转化细胞优先分泌mIgA1。当通过斑点ELISA技术确定来自不同组织的分泌pIgA、pIgA1或pIgA2的细胞频率与组织培养上清液中分泌的pIgA与mIgA(以及IgA亚类)的比例相关时,所获得的数据表明,许多单个产生IgA的细胞可能同时参与mIgA和pIgA的分泌。