Clark-Curtiss J E, Thole J E, Sathish M, Bosecker B A, Sela S, de Carvalho E F, Esser R E
Department of Molecular Microbiology, Washington University, St. Louis, MO 63130.
Res Microbiol. 1990 Sep-Oct;141(7-8):859-71. doi: 10.1016/0923-2508(90)90122-7.
Protein antigens of Mycobacterium leprae have been identified by screening the lambda gt11, pYA626 and pHC79::M. leprae genomic libraries with pooled sera from leprosy patients and with antiserum to M. leprae cell wall protein (CWP) aggregate. Immunological screening of the lambda gt11 library with pooled sera from 21 lepromatous (LL) leprosy patients resulted in the identification of 19 antigens that are apparently different from previously identified M. leprae antigens. Five additional antigens were identified by screening the lambda gt11 library with pooled sera from 30 borderline tuberculoid or tuberculoid patients. Four other antigens were identified by screening the lambda gt11 library with anti-CWP. Two groups of recombinant cosmids were identified by screening the pHC79 library with LL patients' sera: one group specified proteins that reacted with monoclonal antibodies (mAb) against the 65-kDa protein and against the 18-kDa protein; the other group specified a 15-kDa protein that did not react with any of the mAb that were tested. One pYA626 clone also specified a 15-kDa protein that reacted with LL patients' sera, but did not react with any mAb. Genes specifying several of these antigens have been subcloned into the Asd+ plasmid vector pYA292 and have been introduced into a delta cya delta crp delta asd Salmonella typhimurium strain to evaluate the ability of individual M. leprae proteins to elicit immune responses against M. leprae infection.
通过用麻风病人的混合血清以及抗麻风杆菌细胞壁蛋白(CWP)聚集体的抗血清筛选λgt11、pYA626和pHC79::麻风杆菌基因组文库,已鉴定出麻风杆菌的蛋白质抗原。用21例瘤型(LL)麻风病人的混合血清对λgt11文库进行免疫筛选,结果鉴定出19种抗原,这些抗原显然与先前鉴定的麻风杆菌抗原不同。用30例界线类偏结核样型或结核样型病人的混合血清筛选λgt11文库,又鉴定出另外5种抗原。用抗CWP筛选λgt11文库,鉴定出另外4种抗原。用LL病人的血清筛选pHC79文库,鉴定出两组重组黏粒:一组编码与抗65 kDa蛋白和抗18 kDa蛋白的单克隆抗体(mAb)发生反应的蛋白质;另一组编码一种不与所测试的任何mAb发生反应的15 kDa蛋白。一个pYA626克隆也编码一种与LL病人血清发生反应,但不与任何mAb发生反应的15 kDa蛋白。已将编码其中几种抗原的基因亚克隆到Asd + 质粒载体pYA292中,并导入cya缺失、crp缺失、asd缺失的鼠伤寒沙门氏菌菌株,以评估单个麻风杆菌蛋白引发针对麻风杆菌感染的免疫反应的能力。