Characterization of a neutralizing monoclonal antibody directed at variable domain I of the major outer membrane protein of Chlamydia trachomatis C-complex serovars.
Characterization of a neutralizing monoclonal antibody directed at variable domain I of the major outer membrane protein of Chlamydia trachomatis C-complex serovars.
复制标题
针对沙眼衣原体 C 复合物血清型主要外膜蛋白可变结构域 I 的中和单克隆抗体的表征。
DOI：
10.1128/iai.61.4.1365-1370.1993
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发表时间：
1993
影响因子：
3.1
通讯作者：
Peterson,EM
中科院分区：
文献类型：
--
作者：
Qu,Z;Cheng,X;delaMaza,LM;Peterson,EM
鉴定出一种单克隆抗体 (MAb) C10，它可以在体外中和沙眼衣原体血清型 C、I、J 和 L3（C 和 C 相关复合物的成员）的感染性。在斑点印迹测定中，在 15 个主要血清型和沙眼衣原体、鹦鹉热衣原体和肺炎衣原体的小鼠肺炎菌株中，它们被用作未处理和热处理（56 摄氏度，30 分钟）的抗原，只有血清型 C、I、J 和 L3 被天然和处理的抗原识别。蛋白质印迹（免疫印迹）结果显示 MAb C10 识别这四种血清型的主要外膜蛋白。合成了与沙眼衣原体血清型 C 主要外膜蛋白可变结构域 (VD) I、II、III 和 IV 相对应的重叠六聚肽，肽筛选显示 MAb C10 映射到 VD I 氨基酸序列 VAGLQNDPT。体外中和测定的结果与间接免疫荧光测定、蛋白质印迹和斑点印迹测定的结果相关，因为只有血清型 C、I、J 和 L3 被 MAb C10 中和。在体外竞争性中和实验中，使用代表血清型 C 的 VD I 的肽与沙眼衣原体血清型 C 竞争 MAb C10 结合，结果表明 MAb C10 的血清学活性和中和活性均被 VD I 肽抑制。在使用经 MAb C10 预处理的血清型 L3 进行的体内毒性/感染性测定中，感染致死剂量的小鼠在 48 小时内存活率为 100%。相比之下，对照组由注射了相同剂量 L3 且用不识别 L3 的 MAb 预处理的小鼠组成，在 48 小时观察期内没有幸存者。总之，由于 MAb C10 识别的表面暴露的连续表位结合对 C 和 C 相关复合物亚种特异性的中和抗体，因此应考虑将其纳入衣原体疫苗的开发中。
A monoclonal antibody (MAb), C10, that neutralized in vitro the infectivity of serovars C, I, J, and L3 (members of the C and C-related complexes) of Chlamydia trachomatis was identified. Of the 15 major serovars and the mouse pneumonitis strain of C. trachomatis, Chlamydia psittaci, and Chlamydia pneumoniae, which were used as nontreated and heat-treated (56 degrees C, 30 min) antigens in a dot blot assay, only serovars C, I, J, and L3 were recognized with both the native and treated antigens. Western blot (immunoblot) results showed that MAb C10 recognized the major outer membrane protein of these four serovars. Overlapping hexameric peptides corresponding to variable domains (VDs) I, II, III, and IV of the major outer membrane protein of C. trachomatis serovar C were synthesized, and peptide screening showed that MAb C10 mapped to the VD I amino acid sequence VAGLQNDPT. Results of an in vitro neutralization assay correlated with those of the indirect immunofluorescence assay, Western blot, and dot blot assay in that only serovars C, I, J, and L3 were neutralized by MAb C10. In vitro competitive neutralization experiments, using a peptide representing VD I of serovar C to compete with C. trachomatis serovar C for MAb C10 binding, revealed that both serological and neutralizing activities of MAb C10 were inhibited by the VD I peptide. In an in vivo toxicity/infectivity assay using serovar L3 pretreated with MAb C10, there was 100% survival of mice infected with a lethal dose at 48 h. In contrast, the control group, consisting of mice injected with the same dose of L3 pretreated with a MAb that does not recognize L3, had no survivors during a 48-h observation period. In summary, since the surface-exposed contiguous epitope recognized by MAb C10 binds neutralizing antibodies that are subspecies specific for the C and C-related complexes, it should be considered for inclusion in the development of a chlamydial vaccine.