ENHANCED POLYCLONAL B CELL ACTIVATION BY PMN LYSATES

牙周炎病变的组织学特征是致密的 中性粒细胞（PMNs）、T细胞和异常大的 活化的B细胞的积累。 越来越多的证据表明， 这些病变中大量的B细胞是非特异性的结果， 龈下菌斑诱导的多克隆B细胞活化 细菌 具核梭杆菌（FN）和其它革兰氏阴性杆菌的细胞壁 经常从牙周炎病变中分离出的阴性细菌非常 在体外对人PBA产生强刺激作用。 在研究FN诱导的PBA（FN-PBA）的细胞调节过程中， 发现将超声处理的中性粒细胞（PMN）裂解物加入淋巴细胞中， 培养物使FN细胞壁诱导的PBA增加2 - 5倍。 裂解物 对FN-PBA无促进作用。 本提案旨在隔离和识别PMN因素 其在体外增强FN-PBA。 具体而言，PMN将被细分 分为1）细胞质，2）核团，3）颗粒（溶酶体），和4） 膜馏分通过常规程序。 每个分数将是 测试在人淋巴细胞培养物中增强FN-PBA的能力。 这些实验的数据将使我们能够详细说明 PMN因子对B细胞活化的作用。 多克隆B细胞 活化与几种慢性炎症性疾病有关， 其中1）微生物诱导的PBA被推测参与疾病， 2）PMN和活化的B细胞紧密并列。 PMN 因此，因子可能参与增强的非特异性激活， 在牙周炎以及其他慢性炎症中观察到的B细胞 疾病

Periodontitis lesions are histologically characterized by a dense infiltration of neutrophils (PMNs), T cells, and an unusually large accumulation of activated B cells. There is increasing evidence that the large number of B cells in these lesions is the result of nonspecific, polyclonal B cell activation (PBA) induced by the subgingival plaque bacteria. The cell walls of Fusobacterium nucleatum (FN) and other gram negative bacteria frequently isolated from periodontitis lesions are very potent stimulants of human PBA in vitro. In the course of studying cellular regulation of FN-induced PBA (FN-PBA) we found that the addition of sonicated neutrophil (PMN) lysates to lymphocyte cultures increased PBA induced by FN cell walls two- to five-fold. Lysates of sonicated lymphocytes did not enhance FN-PBA. The purpose of this proposal is to isolate and identify the PMN factor(s) which enhance FN-PBA in vitro. Specifically, PMNs will be fractionated into 1) cytoplasm, 2) nuclear pellets, 3) granules (lysosomes), and 4) membrane fractions through routine procedures. Each fraction will then be tested for the capacity to enhance FN-PBA in human lymphocyte cultures. The data from these experiments will enable us to detail the mechanism of action of the PMN factor(s) on B cell activation. Polyclonal B cell activation is associated with several chronic inflammatory diseases in which 1) microbial-induced PBA is speculated to be involved in the disease, and 2) PMNs and activated B cells are seen in close juxtaposition. PMN factors may therefore be involved in the enhanced nonspecific activation of B cells observed in periodontitis as well as other chronic inflammatory diseases.