Schook L B, Gutmann D H, Marlin L E, Niederhuber J E
Transplantation. 1984 Jun;37(6):585-90. doi: 10.1097/00007890-198406000-00012.
Cultures of bone marrow stem cells, grown in the presence of L-cell-conditioned medium, were harvested on successive days to determine the expression of Ia antigens and the acquisition of Ir4 gene-regulated antigen presentation. I-A subregion antigen expression was detected by an indirect radiobinding assay (RBA) as early as day 3 and reached maximal binding at day 7, before declining with additional time in culture. Indirect immunofluorescence (IIF) demonstrated 20% Ia+ cells on day 3 of culture and peaked at 60% on day 7 before declining with continued incubation. A mixed lymphocyte reaction (MLR) across an I region difference was used to assess the kinetics of bone-marrow-derived macrophage ( BMDM ) Ia expression. Maximal stimulation occurred with BMDM stimulator cells obtained from 5-9-day cultures. To investigate the acquisition of Ir gene function, BMDM harvested after various days in culture were used to reconstitute the T cell proliferative response to poly Glu60Ala30Tyr10 (GAT). The ability of BMDM to present GAT was detected after day 5, increased to maximal levels on day 7, and then declined with weak proliferative responses obtained using day 12 BMDM . The presentation of GAT by BMDM was inhibited by monoclonal anti-Ia. 17 antibody. Thus, the acquisition of Ir gene function by BMDM was found to parallel the expression of Ia molecules. Additional experiments were performed to determine whether treatment of BMDM at day 7 with lymphokines or beta-interferon could extend Ia antigen expression. Whereas treatment of day 7 BMDM cultures with Con-A-stimulated rat splenocyte supernatants extended and augmented Ia expression for an additional three days, beta-interferon treatment did not result in augmentation or extension of Ia antigen expression.
在L细胞条件培养基存在下培养的骨髓干细胞，连续数天收获，以确定Ia抗原的表达以及Ir4基因调控的抗原呈递的获得情况。早在第3天通过间接放射结合测定（RBA）检测到I-A亚区抗原表达，并在第7天达到最大结合，之后随着培养时间延长而下降。间接免疫荧光（IIF）显示培养第3天有20%的Ia+细胞，第7天达到峰值60%，之后随着继续孵育而下降。通过跨I区差异的混合淋巴细胞反应（MLR）来评估骨髓来源巨噬细胞（BMDM）Ia表达的动力学。用从5 - 9天培养物中获得的BMDM刺激细胞时发生最大刺激。为了研究Ir基因功能的获得情况，使用培养不同天数后收获的BMDM来重建T细胞对聚谷氨酸60丙氨酸30酪氨酸10（GAT）的增殖反应。BMDM呈递GAT的能力在第5天后被检测到，在第7天增加到最大水平，然后随着使用第12天BMDM获得的微弱增殖反应而下降。BMDM对GAT的呈递被单克隆抗Ia.17抗体抑制。因此，发现BMDM对Ir基因功能的获得与Ia分子的表达平行。进行了额外的实验以确定在第7天用淋巴因子或β干扰素处理BMDM是否能延长Ia抗原表达。虽然用Con - A刺激的大鼠脾细胞上清液处理第7天的BMDM培养物可使Ia表达延长并增强另外三天，但β干扰素处理并未导致Ia抗原表达的增强或延长。
