A major cell surface antigen of Coccidioides immitis which elicits both humoral and cellular immune responses
A major cell surface antigen of Coccidioides immitis which elicits both humoral and cellular immune responses
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DOI：
10.1128/iai.68.2.584-593.2000
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发表时间：
2000-02-01
影响因子：
3.1
通讯作者：
Cole, GT
中科院分区：
文献类型：
--
作者：
Hung, CY;Ampel, NM;Cole, GT
粗球孢子菌分离物的多核寄生细胞（小球）在体外产生膜状外壁成分（SOW），据报道，该成分可与球孢子菌感染患者的抗体发生反应，引发小鼠免疫 T 细胞的有效增殖反应，并在球孢子菌病小鼠模型中具有免疫保护能力。为了鉴定 SOW 的抗原成分，首先对粗壁材料进行 Triton 处理X-114 提取物以及由该处理获得的母液可溶性级分在体液和细胞免疫测定中检查了蛋白质组成和反应性。蛋白质电泳显示三种不同的 C.immitis 分离株的水性级分各自含有一种或两种主要糖蛋白 (SOWgps)，其分子大小范围为 58 至 82 kDa。然而，SOWgps 显示相同的 N 端氨基酸序列，并且每种都被来自 C 型细丝炎感染患者的血清。针对 Silveira 分离物的纯化 58-kDa 糖蛋白 (SOWgp58) 产生的抗体用于蛋白质印迹和免疫定位分析。 SOWgp的表达被证明是寄生期特异性的，并且抗原定位于膜SOW。测试SOW的水溶性级分和纯化的SOWgp58刺激人外周单核细胞(PBMC)增殖的能力。后者是从对球蛋白（粗线念珠菌的一种寄生期抗原）有阳性皮试反应的健康志愿者和对同一抗原没有表现出皮试反应的志愿者中获得的。 SOW制剂刺激皮试阳性供体的PBMC增殖，但不刺激皮试阴性供体的PBMC增殖，并且激活的细胞分泌γ干扰素，这表明免疫反应的T辅助1途径。这项研究的结果表明，SOWgp 是一种主要的寄生细胞表面表达的抗原，可在球虫感染患者中引发体液和细胞免疫反应。
Multinucleate parasitic cells (spherules) of Coccidioides immitis isolates produce a membranous outer wall component (SOW) in vitro which has been reported to be reactive with antibody from patients with coccidioidal infection, elicits a potent proliferative response of murine immune T cells, and has immunoprotective capacity in a murine model of coccidioidomycosis, To identify the antigenic components of SOW, the crude wall material was first subjected to Triton X-114 extraction, and a mater-soluble fraction derived from this treatment was examined for protein composition and reactivity in humoral and cellular immunoassays, Protein electrophoresis revealed that the aqueous fraction of three different isolates of C. immitis each contained one or two major glycoproteins (SOWgps), distinguished by their molecular sizes, which ranged from 58 to 82 kDa, The SOWgps, however, showed identical N-terminal amino acid sequences, and each was recognized by sera from patients with C, immitis infection. Antibody raised against the purified 58-kDa glycoprotein (SOWgp58) of the Silveira isolate was used for Western blot and immunolocalization analyses. Expression of SOWgp was shown to be parasitic phase specific, and the antigen was Localized to the membranous SOW, The water-soluble fraction of SOW and the purified SOWgp58 were tested for the ability to stimulate proliferation of human peripheral monocytic cells (PBMC). The latter were obtained from healthy volunteers with positive skin test reaction to spherulin, a parasitic-phase antigen of C. immitis, and from volunteers who showed no skin test reaction to the same antigen. The SOW preparations stimulated proliferation of PBMC from skin test-positive but not skin test-negative donors, and the activated cells secreted gamma interferon, which is indicative of a T helper 1 pathway of immune response. Results of this study suggest that SOWgp is a major parasitic cell surface-expressed antigen that elicits both humoral and cellular immune responses in patients with coccidioidal infection.