Monedero Marta, Infantes-Lorenzo José A, Moreno Inmaculada, Martín Ana Belén, Macias Rocio I R, Toraño Alfredo, Domínguez Mercedes
Unidad de Inmunología Microbiana, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Madrid, Spain; Experimental Hepatology and Drug Targeting (HEVEFARM), University of Salamanca, IBSAL, CIBERehd, Salamanca, Spain.
VISAVET Health Surveillance Centre, Universidad Complutense de Madrid, Madrid, Spain.
Comp Immunol Microbiol Infect Dis. 2019 Feb;62:7-12. doi: 10.1016/j.cimid.2018.11.008. Epub 2018 Nov 24.
To study complement function in mammalian leishmanioses, we developed mouse monoclonal antibodies to the human complement system components C1q, C4, factor D, factor H, factor B, properdin, C5 and C9. Antibody specificity was determined by indirect and capture ELISA and by Western blot. In flow cytometry analysis, seven antibodies recognized the cognate component on human serum-opsonized Leishmania promastigotes. Antibody reactivity was screened against promastigotes opsonized with sera of nine mammalian genera: pig, guinea pig, goat, rabbit, cat, dog, hamster, jird and rat. No antibody recognized jird epitopes on promastigotes. Anti-C4, -properdin, and -C5b reacted with the orthologous protein of all other mammals tested except cat (anti-properdin) and hamster (anti-C5b); anti-C9 only recognized the rabbit ortholog, and anti-C1q, -factor B and -factor H did not react with any of the nine orthologs. Such interspecies crossreactive antibodies can be valuable tools for analysis of mammalian complement function in infectious diseases.
为研究补体在哺乳动物利什曼病中的功能，我们制备了针对人补体系统成分C1q、C4、D因子、H因子、B因子、备解素、C5和C9的小鼠单克隆抗体。通过间接ELISA、捕获ELISA和蛋白质印迹法确定抗体特异性。在流式细胞术分析中，七种抗体识别经人血清调理的利什曼原鞭毛体上的同源成分。针对用九个哺乳动物属（猪、豚鼠、山羊、兔、猫、狗、仓鼠、沙鼠和大鼠）血清调理的原鞭毛体筛选抗体反应性。没有抗体识别原鞭毛体上的沙鼠表位。抗C4、抗备解素和抗C5b与除猫（抗备解素）和仓鼠（抗C5b）外的所有其他受试哺乳动物的直系同源蛋白发生反应；抗C9仅识别兔直系同源物，而抗C1q、抗B因子和抗H因子不与九个直系同源物中的任何一个发生反应。这种种间交叉反应性抗体可能是分析传染病中哺乳动物补体功能的有价值工具。
