Peeling R W, Brunham R C
Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada.
Infect Immun. 1991 Aug;59(8):2624-30. doi: 10.1128/iai.59.8.2624-2630.1991.
Monoclonal antibodies to the major outer membrane protein of Chlamydia trachomatis were used to neutralize C. trachomatis infectivity in HeLa 229 cells and to determine the kinetics and stoichiometry of the reaction. In vitro neutralization of C. trachomatis infectivity proceeded as a first-order reaction and required an activation energy of approximately 20 kcal/mol (ca. 84 kJ/mol). The rate of neutralization was linear with respect to antibody concentration and reaction temperature. The efficiency of neutralization decreased exponentially as the ratio of noninfective to infective chlamydiae increased in the antigen preparation. The neutralization assay was also significantly affected by reaction parameters such as the reaction volume and the duration of incubation. Stoichiometric calculations showed that an average ratio of 10(3) and 10(4) immunoglobulin molecules per chlamydial particle was required to yield 50% neutralization by monoclonal antibodies specifying serovar-specific and species-specific epitopes, respectively. The implications of these findings for vaccine design and for the role of the major outer membrane protein in the pathogenesis of chlamydial infections are discussed.
利用针对沙眼衣原体主要外膜蛋白的单克隆抗体来中和HeLa 229细胞中沙眼衣原体的感染性,并确定该反应的动力学和化学计量关系。体外对沙眼衣原体感染性的中和反应呈一级反应,需要约20千卡/摩尔(约84千焦/摩尔)的活化能。中和速率与抗体浓度和反应温度呈线性关系。随着抗原制剂中无感染性衣原体与感染性衣原体的比例增加,中和效率呈指数下降。中和试验也受到反应体积和孵育时间等反应参数的显著影响。化学计量计算表明,对于分别识别血清型特异性和种特异性表位的单克隆抗体,要产生50%的中和效果,每个衣原体颗粒平均需要10³和10⁴个免疫球蛋白分子。讨论了这些发现对疫苗设计以及主要外膜蛋白在衣原体感染发病机制中的作用的意义。