Shen H D, Choo K B, Yu K W, Ling W L, Chang F C, Han S H
Department of Medical Research, Veterans General Hospital, Taipei, Taiwan.
Int Arch Allergy Appl Immunol. 1991;96(2):142-8. doi: 10.1159/000235485.
A 41-kD component of Candida albicans was identified to be the major antigen radioimmunoprecipitated by antibodies with increased titers in the sera of patients with invasive candidiasis. A mouse monoclonal antibody (RJ5) was generated which, by immunoblotting, showed positive reactivity to the immunoprecipitated 41-kD component. By two-dimensional gel electrophoresis and immunoblotting, MoAb RJ5 was shown to react with different isoforms of the 41-kD component with pI values from 6.1 to 6.9. Furthermore, MoAb RJ5 showed positive reactivity to cytoplasmic antigens of C. albicans by frozen section and immunoperoxidase staining. By SDS-polyacrylamide gel electrophoresis and immunoblotting, MoAb RJ5 showed no cross-reactivity to antigens of Candida tropicalis and Candida parapsilosis. The epitope of the 41-kD molecule recognized by MoAb RJ5 was susceptible to treatment of proteinase K at concentrations of greater than or equal to 5 micrograms/ml, and was relatively resistant to periodate oxidation with concentration of NaIO4 up to 20 mM. This MoAb may be useful in the purification and characterization of the immunodominant 41-kD antigen of C. albicans, and as a probe in the detection of Candida antigens in the sera of patients with invasive candidiasis.
白色念珠菌的一种41-kD成分被鉴定为侵袭性念珠菌病患者血清中滴度升高的抗体进行放射免疫沉淀的主要抗原。制备了一种小鼠单克隆抗体(RJ5),通过免疫印迹法,该抗体对免疫沉淀的41-kD成分显示出阳性反应。通过二维凝胶电泳和免疫印迹法,发现单克隆抗体RJ5与41-kD成分的不同同工型发生反应,其等电点值在6.1至6.9之间。此外,通过冰冻切片和免疫过氧化物酶染色,单克隆抗体RJ5对白色念珠菌的细胞质抗原有阳性反应。通过SDS-聚丙烯酰胺凝胶电泳和免疫印迹法,单克隆抗体RJ5对热带念珠菌和近平滑念珠菌的抗原无交叉反应。单克隆抗体RJ5识别的41-kD分子的表位对浓度大于或等于5微克/毫升的蛋白酶K处理敏感,对浓度高达20毫摩尔的高碘酸钠氧化相对耐受。这种单克隆抗体可能有助于白色念珠菌免疫显性41-kD抗原的纯化和特性鉴定,并可作为检测侵袭性念珠菌病患者血清中念珠菌抗原的探针。