Zhu Weiyan, Thomas Christopher E, Chen Ching-Ju, Van Dam Cornelius N, Johnston Robert E, Davis Nancy L, Sparling P Frederick
University of North Carolina at Chapel Hill, Department of Medicine, Division of Infectious Disease Research, 8341 Medical Biomolecular Research Bldg., 103 Mason Farm Road, CB# 7031, Chapel Hill, NC 27599, USA.
Infect Immun. 2005 Nov;73(11):7558-68. doi: 10.1128/IAI.73.11.7558-7568.2005.
Porin (PorB) is a major outer membrane protein produced by all Neisseria gonorrhoeae strains and has been a focus of intense interest as a vaccine candidate. In this study, the immunogenicity of PorB in mice was investigated after several immunization regimens. Outer membrane vesicles (OMV), recombinant renatured PorB (rrPorB), and PorB-expressing Venezuelan equine encephalitis (VEE) virus replicon particles (PorB VRP) were delivered intranasally (i.n.) or subcutaneously (s.c.) into the dorsal area or the hind footpad in three-dose schedules; the PorB VRP-immunized mice were given a single additional booster dose of rrPorB in Ribi adjuvant. Different delivery systems and administration routes induced different immune responses. Mice immunized s.c. with rrPorB in Ribi had the highest levels of PorB-specific serum immunoglobulin G (IgG) by enzyme-linked immunosorbent assay. Surprisingly, there was an apparent Th1 bias, based on IgG1/IgG2a ratios, after immunization with rrPorB in Ribi in the footpad while the same vaccine given in the dorsal area gave a strongly Th2-biased response. PorB VRP-immunized mice produced a consistent Th1 response with a high gamma interferon response in stimulated splenic lymphocytes and very low IgG1/IgG2a ratios. Immunization by OMV delivered i.n. was the only regimen that resulted in a serum bactericidal response, and it generated an excellent mucosal IgA response. Serum from mice immunized with rrPorB preferentially recognized the surface of whole gonococci expressing a homologous PorB, whereas serum from PorB VRP-immunized mice had relatively low whole-cell binding activity but recognized both heterologous and homologous PorB equally. The data resulting from this direct comparison suggested that important aspects of the immune response can be manipulated by altering the form of the antigen and its delivery. This information coupled with an understanding of protective antigonococcal immune responses will enable the design of the optimal vaccine for N. gonorrhoeae.
孔蛋白（PorB）是所有淋病奈瑟菌菌株产生的一种主要外膜蛋白，作为候选疫苗一直备受关注。在本研究中，研究了几种免疫方案后PorB在小鼠中的免疫原性。将外膜囊泡（OMV）、重组复性PorB（rrPorB）和表达PorB的委内瑞拉马脑炎（VEE）病毒复制子颗粒（PorB VRP）以三剂量方案经鼻内（i.n.）或皮下（s.c.）注射到背部区域或后足垫；用PorB VRP免疫的小鼠在Ribi佐剂中额外单次注射rrPorB加强剂量。不同的递送系统和给药途径诱导了不同的免疫反应。通过酶联免疫吸附测定，在Ribi中用rrPorB皮下免疫的小鼠具有最高水平的PorB特异性血清免疫球蛋白G（IgG）。令人惊讶的是，基于IgG1/IgG2a比值，在足垫中用Ribi中的rrPorB免疫后出现明显偏向Th1的反应，而在背部区域给予相同疫苗则产生强烈偏向Th2的反应。用PorB VRP免疫的小鼠在刺激的脾淋巴细胞中产生一致的Th1反应，γ干扰素反应高，IgG1/IgG2a比值非常低。经鼻内递送OMV进行免疫是唯一导致血清杀菌反应的方案，并且产生了出色的黏膜IgA反应。用rrPorB免疫的小鼠血清优先识别表达同源PorB的全淋球菌表面，而用PorB VRP免疫的小鼠血清全细胞结合活性相对较低，但对异源和同源PorB的识别相同。这种直接比较产生的数据表明，免疫反应的重要方面可以通过改变抗原的形式及其递送方式来操控。这些信息加上对保护性抗淋球菌免疫反应的理解将有助于设计针对淋病奈瑟菌的最佳疫苗。
