Characterization of a neutralizing monoclonal antibody directed at the lipopolysaccharide of Chlamydia pneumoniae.
Characterization of a neutralizing monoclonal antibody directed at the lipopolysaccharide of Chlamydia pneumoniae.
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针对肺炎衣原体脂多糖的中和单克隆抗体的表征。
DOI：
10.1128/iai.66.8.3848-3855.1998
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发表时间：
1998
影响因子：
3.1
通讯作者：
Brade,H
中科院分区：
文献类型：
--
作者：
Peterson,EM;delaMaza,LM;Brade,L;Brade,H
识别保护性表位是发展亚单位疫苗的第一步。实现这一点的一种方法是通过使用能够在体外和体内减弱感染性的单抗(MAb)来识别结构或表位。到目前为止，对肺炎衣原体使用这种方法的尝试都失败了。本报告首次报道了一种针对衣原体脂多糖的单抗，它在体内和体外都能中和衣原体的感染性。肺炎。单抗CP-33是一种免疫球蛋白G2B(IgG2b)，由C免疫的小鼠脾细胞融合而成。肺炎TW-183。通过Western印迹分析，单抗CP-33具有属特异性反应活性，因为它识别LPS OFC。肺炎、沙眼衣原体和鹦鹉热衣原体。单抗CP-33与15种革兰氏阴性和革兰氏阳性细菌及白色念珠菌均不反应。利用明尼苏达州肠炎沙门氏菌再突变株的分离内毒素和3-脱氧-d-甘露-OCT-2-磺酸(KDO)转移酶基因KdtAofC的重组子。沙眼衣原体单抗CP-33需要与属特异性三糖表位αKdo(2→8)αKdo(2→4)αKdo结合。通过使用人工合成的寡糖和新糖偶联物进行酶免疫分析(EIA)和EIA抑制，进一步表明单抗CP-33不同于广泛研究的衣原体单抗S25-23。最有可能的是，单抗CP-33识别一个构象表位，其中αKdo(2→8)αKdo(2→4)αKdo三糖是必需的结构成分。在体外中和试验中，单抗CP-33的中和效价为8 ng/ml。肺炎TW-183。然而，该单抗并不能中和其他C。肺炎衣原体、沙眼衣原体、ORC。鹦鹉。肺炎衣原体TW-183用单抗CP-33或对照免疫球蛋白免疫，然后通过呼吸道途径接种小鼠。接种5天后，对照组和单抗CP-33处理组的小鼠感染数和肺培养回收的包涵体单位数差异有统计学意义(P<；0.05)。综上所述，衣原体特异性脂多糖单抗能够在体外中和FC的感染性。肺炎TW-183。
Identification of protective epitopes is one of the first steps in the development of a subunit vaccine. One approach to accomplishing this is to identify structures or epitopes by using monoclonal antibodies (MAb) that can attenuate infectivity in vitro and in vivo. To date attempts to use this approach withChlamydia pneumoniaehave failed. This report is the first description of a MAb directed to the lipopolysaccharide (LPS) ofChlamydiathat neutralizes both in vitro and in vivo the infectivity ofC. pneumoniae. MAb CP-33, an immunoglobulin G2b (IgG2b), was identified from a fusion using splenocytes from mice immunized withC. pneumoniaeTW-183. By Western blot analysis, MAb CP-33 exhibited genus-specific reactivity in that it recognized the LPSs ofC. pneumoniae,Chlamydia trachomatis, andChlamydia psittaci. MAb CP-33 did not react with 15 genera of gram-negative and gram-positive bacteria andCandida albicans. By using isolated LPS of Re mutants ofEscherichia coli,Salmonella entericaserovar Minnesota, and recombinants expressing the 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) transferase genekdtAofC. trachomatis, MAb CP-33 was shown to require for binding the presence of the genus-specific trisaccharide epitope αKdo(2→8)αKdo(2→4)αKdo. By employing synthetic oligosaccharides and neoglycoconjugates in an enzyme immunoassay (EIA) and EIA inhibition, it was further shown that MAb CP-33 differed from the extensively investigated prototype chlamydial LPS MAb S25-23. Most likely, MAb CP-33 recognizes a conformational epitope in which the αKdo(2→8)αKdo(2→4)αKdo trisaccharide is an essential structural component. When tested in an in vitro neutralization assay, MAb CP-33 gave a 50% neutralization titer of 8 ng/ml againstC. pneumoniaeTW-183. However, this MAb did not neutralize otherC. pneumoniaestrains,C. trachomatis, orC. psittaci. C. pneumoniaeTW-183 was treated with either MAb CP-33 or a control IgG and then used to inoculate mice by the respiratory route. Five days after inoculation, there was a difference between the mice inoculated with the control IgG-treated inoculum and those inoculated with the MAb CP-33-treated organisms as to the number of mice infected as well as the number of inclusion-forming units recovered from lung cultures (P< 0.05). In summary, aChlamydia-specific LPS MAb was able to neutralize in vitro the infectivity ofC. pneumoniaeTW-183.