Tambourgi D V, Kipnis T L, da Silva W D, Joiner K A, Sher A, Heath S, Hall B F, Ogden G B
Departamento de Imunologia, Universidade de São Paulo, Brazil.
Infect Immun. 1993 Sep;61(9):3656-63. doi: 10.1128/iai.61.9.3656-3663.1993.
Resistance to complement-mediated lysis in Trypanosoma cruzi is due to the expression of complement-regulatory factors by the virulent developmental forms of this protozoan parasite. An 87- to 93-kDa molecule, which we have termed T-DAF (trypomastigote decay-accelerating factor), is present on the surface of the parasite and inhibits complement activation in a manner functionally similar to the mammalian complement regulatory component, decay-accelerating factor. In this report, we characterized monospecific polyclonal and monoclonal antibodies which were obtained from mice and rabbits immunized with fast protein liquid chromatography-purified T-DAF. These polyclonal antibodies were shown to inhibit T-DAF activity and were capable of inducing lysis of the parasites. Both the polyclonal and monoclonal antibodies were used to screen a cDNA expression library prepared from T. cruzi trypomastigote mRNA. From this library, we obtained a partial lambda gt11 cDNA clone which showed genetic and functional similarity to the human C3 convertase inhibitor DAF (A. Nicholson-Weller, J. Burge, D. T. Fearon, P. F. Weller, and K. F. Austen, J. Immunol. 129:184-189, 1982).
克氏锥虫对补体介导的溶解作用具有抗性,这归因于这种原生动物寄生虫的致病发育形式表达补体调节因子。一种87至93千道尔顿的分子,我们将其命名为T-DAF(锥鞭毛体衰变加速因子),存在于寄生虫表面,并以一种功能上类似于哺乳动物补体调节成分衰变加速因子的方式抑制补体激活。在本报告中,我们对用快速蛋白质液相色谱纯化的T-DAF免疫的小鼠和兔子获得的单特异性多克隆抗体和单克隆抗体进行了表征。这些多克隆抗体被证明可抑制T-DAF活性,并能够诱导寄生虫裂解。多克隆抗体和单克隆抗体都用于筛选从克氏锥虫锥鞭毛体mRNA制备的cDNA表达文库。从该文库中,我们获得了一个部分λgt11 cDNA克隆,该克隆显示出与人类C3转化酶抑制剂DAF(A. Nicholson-Weller、J. Burge、D. T. Fearon、P. F. Weller和K. F. Austen,《免疫学杂志》129:184 - 189,1982)在遗传和功能上的相似性。