PARTIAL PROTECTION AGAINST GENITAL REINFECTION BY IMMUNIZATION OF GUINEA-PIGS WITH ISOLATED OUTER-MEMBRANE PROTEINS OF THE CHLAMYDIAL AGENT OF GUINEA-PIG INCLUSION CONJUNCTIVITIS
PARTIAL PROTECTION AGAINST GENITAL REINFECTION BY IMMUNIZATION OF GUINEA-PIGS WITH ISOLATED OUTER-MEMBRANE PROTEINS OF THE CHLAMYDIAL AGENT OF GUINEA-PIG INCLUSION CONJUNCTIVITIS
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DOI：
10.1099/00221287-139-12-2965
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发表时间：
1993-12-01
期刊：
影响因子：
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通讯作者：
SODERBERG, LSF
中科院分区：
文献类型：
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作者：
BATTEIGER, BE;RANK, RG;SODERBERG, LSF
由于针对再感染的部分保护是通过生殖器衣原体感染的豚鼠模型中的实验性感染诱导的，因此我们试图通过免疫来诱导免疫力。用从 SDS 聚丙烯酰胺凝胶（SDS-MOMP、SDS-61 kDa）洗脱的豚鼠包涵性结膜炎剂（GPIC）的主要外膜蛋白（MOMP）和富含半胱氨酸的 61 kDa 外膜蛋白（61 kDa）对雌性豚鼠进行皮下免疫。通过免疫印迹测量，免疫后血清和分泌物含有高滴度的 SDS 纯化蛋白抗体，而使用整个原体作为抗原的酶免疫测定 (EIA) 显示滴度显着较低 (P < 0.001)。同样，外周单核细胞对 GPIC 基体的胚细胞反应也很弱。用 SDS-MOMP 和 SDS-61 kDa 免疫的动物对阴道内攻击完全敏感，用不含蛋白质的缓冲液免疫的对照动物也是如此。另一组动物用通过提取衣原体外膜复合物与辛基P-D-吡喃葡萄糖苷（OGP）和二硫苏糖醇制备的材料进行免疫，该复合物主要由MOMP（OGP-MOMP）组成。与SDS-MOMP组相比，OGP-MOMP组的血清和分泌物在EIA中显示出高滴度，并且通过免疫印迹显示出高滴度的MOMP抗体；然而，大多数动物也具有 61 kDa、72 kDa 和 ca 的抗体。 84 kDa 外膜蛋白。尽管通过培养物分离测量的感染持续时间与对照动物相似，但与对照动物相比，OGP-MOMP 动物的纳入分数较低，这证明了 OGP-MOMP 动物受到了部分保护，免受生殖器攻击。使用含有 MOMP 四个可变结构域的重组融合肽对来自免疫动物和一组感染后免疫动物的血清进行免疫印迹分析。当比较受保护和非受保护动物的血清时，没有观察到反应模式的一致差异。因此，高度精制的外膜制剂能够对生殖器感染产生部分免疫力。需要进一步研究以确定这种保护作用是由于 MOMP 本身还是由于 OGP-MOMP 免疫原中少量发现的其他外膜蛋白。结果表明，不连续的 MOMP 表位可能在豚鼠模型中诱导保护性免疫反应中发挥作用，这一概念需要进一步评估。
Because partial protection against reinfection is induced by experimental infection in the guinea-pig model of genital chlamydial infection, we sought to induce immunity by immunization. Female guinea-pigs were immunized subcutaneously with the major outer-membrane protein (MOMP) and the 61 kDa cysteine-rich outer-membrane protein (61 kDa) of the agent of guinea-pig inclusion conjunctivitis (GPIC) eluted from SDS-polyacrylamide gels (SDS-MOMP, SDS-61 kDa). Post-immunization sera and secretions contained antibodies to the SDS-purified proteins at high titre as measured by immunoblotting, whereas enzyme inmunoassays (EIA) using whole elementary bodies as antigen showed significantly lower titres (P < 0.001). Likewise, blastogenic responses of peripheral mononuclear cells to GPIC elementary bodies were weak. Animals immunized with SDS-MOMP and SDS-61 kDa were fully susceptible to intravaginal challenge, as were control animals immunized with buffer without protein. Another group of animals were immunized with material prepared by extraction of chlamydial outer-membrane complexes with octyl P-D-glucopyranoside (OGP) and dithiothreitol, which consisted largely of MOMP (OGP-MOMP). In contrast to the SDS-MOMP group, sera and secretions in the OGP-MOMP group showed high titres in EIA, and high titre antibodies to MOMP by immunoblot; however, most animals also had antibodies to 61 kDa, 72 kDa and ca. 84 kDa outer-membrane proteins. OGP-MOMP animals were partially protected against genital challenge as evidenced by low inclusion scores compared to control animals, although duration of infection measured by culture isolation was similar to controls. Immunoblot analysis of sera from immunized animals and from a group of immune animals post-infection was performed using recombinant fusion peptides containing the four variable domains of MOMP. No consistent differences in reaction patterns were observed when sera from protected and non-protected animals were compared. Thus, a highly refined outer-membrane preparation is capable of producing partial immunity to genital infection. Further study is required to determine whether the protection is due to MOMP itself or to other outer-membrane proteins found in small amounts in the OGP-MOMP immunogen. The results suggest the possibility that discontinuous MOMP epitopes could play a role in inducing a protective immune response in the guinea-pig model, a concept that requires further evaluation.