Muñoz M L, Lamoyi E, León G, Tovar R, Pérez-García J, De La Torre M, Murueta E, Bernal R M
Department of Genetics and Molecular Biology, Centro de Investigación y de Estudios Avanzados del Instituto, Politécnico Nacional, Mexico, D.F.
J Clin Microbiol. 1990 Nov;28(11):2418-24. doi: 10.1128/jcm.28.11.2418-2424.1990.
In vitro interaction of Entamoeba histolytica with collagen induces intracellular formation and release of electron-dense granules (EDG) and stimulation of collagenolytic activity. Purified EDG contain 1.66 U of collagenase per mg of protein. Thus, EDG may participate in tissue destruction during invasive amebiasis. Monoclonal antibodies (MAbs) L1.1 and L7.1 reacted specifically with EDG in enzyme-linked immunosorbent assay (ELISA) and immunofluorescence and immunoelectron microscopy. MAb L7.1 immunoprecipitated three polypeptides with molecular weights of 95,000, 68,000, and 28,000 from lysates of biosynthetically labeled E. histolytica. Both MAbs recognized the pathogenic E. histolytica axenic strains HM1:IMSS, HM38:IMSS, and HK-9 but failed to react in ELISA with Entamoeba moshkovskii, Entamoeba invadens, and E. histolytica-like Laredo. In addition, MAb L7.1 reacted with one E. histolytica isolate from a symptomatic patient but did not react with four of five isolates from asymptomatic patients. EDG antigens were detected by a MAb L7.1-based ELISA in E. histolytica-containing fecal samples from symptomatic, but not asymptomatic, individuals. These results suggest that the EDG antigen detected with MAb L7.1 may be differentially expressed in pathogenic and nonpathogenic E. histolytica.
溶组织内阿米巴与胶原蛋白的体外相互作用可诱导电子致密颗粒(EDG)在细胞内形成并释放,同时刺激胶原酶活性。纯化后的EDG每毫克蛋白质含有1.66单位的胶原酶。因此,EDG可能参与侵袭性阿米巴病期间的组织破坏。单克隆抗体(MAb)L1.1和L7.1在酶联免疫吸附测定(ELISA)、免疫荧光和免疫电子显微镜中与EDG发生特异性反应。MAb L7.1从生物合成标记的溶组织内阿米巴裂解物中免疫沉淀出三种分子量分别为95,000、68,000和28,000的多肽。两种单克隆抗体均识别致病性溶组织内阿米巴无菌株HM1:IMSS、HM38:IMSS和HK-9,但在ELISA中不与莫氏内阿米巴、侵袭内阿米巴和溶组织内阿米巴样拉雷多反应。此外,MAb L7.1与一名有症状患者的一株溶组织内阿米巴分离株发生反应,但不与五株无症状患者分离株中的四株发生反应。通过基于MAb L7.1的ELISA在有症状个体而非无症状个体的含溶组织内阿米巴粪便样本中检测到EDG抗原。这些结果表明,用MAb L7.1检测到的EDG抗原可能在致病性和非致病性溶组织内阿米巴中差异表达。