School of Clinical Veterinary Science, University of Bristol, Langford House, Langford, Bristol BS40 5DU, UK.
Vet Microbiol. 2010 Aug 26;144(3-4):415-21. doi: 10.1016/j.vetmic.2010.02.021. Epub 2010 Feb 18.
Chlamydophila felis is a common cause of conjunctivitis in cats. Greater understanding of C. felis infection and immunity and identification of protective antigens will facilitate improved vaccine design. Chlamydial polymorphic membrane proteins (Pmps) represent a family of homologous proteins of likely importance in chlamydial infection and immunity. To identify immunogenic C. felis Pmps, we generated recombinant C. felis Pmps (rPmps) and used these to detect serum antibody reactivity against Pmps arising during C. felis infection in cats. Sequencing of Pmp genes 1, 7, 13, 18, 19 and 20 from 3 laboratory strains of C. felis (K2487, 1497V and Cello) and alignment with the Fe/C-56 genome revealed high genetic identity in Pmp genes between strains. PCR products lacking the predicted N-terminal signal sequence peptide and C-terminal domain were generated, cloned and expressed in Escherichia coli prior to purification by nickel-agarose affinity chromatography. Serum samples from 4 cats collected up to 55 days post-inoculation with C. felis (K2487) were analysed by western blotting and rPmp-specific ELISAs for evidence of serum antibody reactivity against each rPmp. Strong serum antibody reactivity against rPmps 1 and 7, and weak heterogeneous serum immunoreactivity against rPmps 13, 19 and 20, were detected from 14 to 21 days post-infection (dpi), peaking at 28-35 dpi and tending to plateau thereafter. No significant serum antibody reactivity was detected against rPmp18. This study provides the first evidence that C. felis Pmps 1 and 7 are likely to represent immunodominant proteins and recommends investigation of their potential as serodiagnostic antigens and novel vaccine candidates.
猫衣原体是猫结膜炎的常见病因。深入了解猫衣原体感染和免疫以及鉴定保护性抗原将有助于改进疫苗设计。衣原体多态性膜蛋白(Pmps)是一组同源蛋白,可能在衣原体感染和免疫中具有重要作用。为了鉴定具有免疫原性的猫衣原体 Pmps,我们生成了重组猫衣原体 Pmps(rPmps),并使用这些 rPmps 来检测猫衣原体感染期间血清中针对 Pmps 的抗体反应性。对 3 株实验室分离株(K2487、1497V 和 Cello)的 Pmp1、7、13、18、19 和 20 基因进行测序,并与 Fe/C-56 基因组进行比对,显示了菌株间 Pmp 基因的高度遗传同一性。生成了缺少预测的 N 端信号序列肽和 C 端结构域的 PCR 产物,克隆并在大肠杆菌中表达,然后通过镍琼脂糖亲和层析进行纯化。通过 Western blot 和 rPmp 特异性 ELISA 分析了 4 只猫在感染衣原体(K2487)后 55 天内采集的血清样本,以检测针对每个 rPmp 的血清抗体反应性。从感染后 14 至 21 天(dpi)检测到针对 rPmps1 和 7 的强烈血清抗体反应性,对 rPmps13、19 和 20 则呈现出较弱的异质血清免疫反应性,在 28-35dpi 时达到高峰,此后趋于稳定。对 rPmp18 未检测到显著的血清抗体反应性。本研究首次证明猫衣原体 Pmps1 和 7 可能代表免疫显性蛋白,并建议对其作为血清学诊断抗原和新型疫苗候选物的潜力进行研究。