Lukehart S A, Baker-Zander S A, Gubish E R
J Immunol. 1982 Aug;129(2):833-8.
The major antigens of Treponema pallidum, Nichols strain were examined by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting techniques. Organisms were extracted from rabbit testes, purified by differential centrifugation, washed, and disrupted by sonication before solubilization in SDS buffer, SDS-PAGE analysis on 12.5% gels revealed at least 35 distinguishable polypeptides with molecular weights between 14,000 and 100,000. Proteins were electrophoretically transferred from gels to nitrocellulose paper and were stained with pooled rabbit anti-T. pallidum serum (IRS) and 125I-protein A. At least eight distinct antigenic molecules were revealed, with the major component residing at 48,000 daltons. Extensive washing of T. pallidum before solubilization did not alter the antigenic profile. Five molecules with common treponemal antigenic determinants were identified both by staining blots of T. phagedenis, biotype Reiter with IRS and by staining blots of T. pallidum with rabbit anti-Reiter antiserum. Three putative T. pallidum-specific antigens were identified by staining T. pallidum with IRS that had been extensively adsorbed with Reiter treponemes.
采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和蛋白质印迹技术对梅毒螺旋体Nichols株的主要抗原进行了检测。从兔睾丸中提取梅毒螺旋体，通过差速离心法进行纯化，洗涤后经超声破碎，然后溶解于SDS缓冲液中。在12.5%的凝胶上进行SDS-PAGE分析，结果显示至少有35种可区分的多肽，其分子量在14000至100000之间。蛋白质通过电泳从凝胶转移至硝酸纤维素膜上，并用兔抗梅毒螺旋体混合血清（IRS）和125I-蛋白A进行染色。结果显示至少有8种不同的抗原分子，其中主要成分位于48000道尔顿处。在溶解前对梅毒螺旋体进行充分洗涤，并未改变其抗原谱。通过用IRS对致病变种梅毒螺旋体生物型Reiter的印迹进行染色，以及用兔抗Reiter抗血清对梅毒螺旋体的印迹进行染色，鉴定出了5种具有共同梅毒螺旋体抗原决定簇的分子。通过用已被Reiter梅毒螺旋体广泛吸附的IRS对梅毒螺旋体进行染色，鉴定出了3种推测的梅毒螺旋体特异性抗原。
