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首页> 外文期刊>Infection and immunity >Protection Elicited by Native Outer Membrane Protein Oms66 (p66) against Host-Adapted Borrelia burgdorferi: Conformational Nature of Bactericidal Epitopes
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Protection Elicited by Native Outer Membrane Protein Oms66 (p66) against Host-Adapted Borrelia burgdorferi: Conformational Nature of Bactericidal Epitopes

机译：天然的外膜蛋白Oms66（p66）对宿主适应性疏螺旋体伯氏疏螺旋体的保护作用：杀菌表位的构象性质

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Oms66 is a Borrelia burgdorferi outer membrane porin protein whose role in Lyme disease pathogenesis and immunity has not been well established. Oms66 was solubilized from whole-cell lysates of strain B313 (which is derived from B31 but lacks OspA, -B, -C, and -D) and purified to homogeneity by fast-protein liquid chromatography. Purified native Oms66 (nOms66), which retained the ability to form large channels in a planar lipid bilayer model membrane system, and denatured Oms66 (hOms66) were used to immunize New Zealand White rabbits. The resulting Oms66 antisera were tested in a complement-dependent borreliacidal assay in parallel with basal serum and with serum from rabbits immune to reinfection with B. burgdorferi (IRS). IRS showed high-titer complement-dependent killing of both strains B31 and B313. Sera from animals immunized with nOms66 showed high-titer complement-dependent killing activity against strain B313 but exhibited no killing of B31. By comparison, serum generated from immunizations with hOms66 showed no killing activity against either strain. Following adsorption of antiserum to nOms66 with recombinant Oms66 (rOms66), the serum antibodies no longer bound to rOms66 or to nOms66 that had been denatured with 8 M urea. However, the antibodies still bound to nOms66 and killing activity against B313 was retained, thus suggesting that native, conformational epitopes are targets of this bactericidal activity. Six C3H HeJ mice were immunized with nOms66 and were challenged using “host-adapted” B. burgdorferi B31 by skin implantation of infected mouse ear tissue. Four of the six mice were protected against both localized and disseminated infection. These findings indicate that native Oms66 can elicit potent bactericidal activity and significant protective immunity against host-adapted organisms.
机译：Oms66是伯氏疏螺旋体外膜孔蛋白，其在莱姆病的发病机理和免疫中的作用尚未完全确立。从菌株B313（衍生自B31，但缺少OspA，-B，-C和-D）的全细胞裂解物中溶解Oms66，并通过快速蛋白质液相色谱法纯化至均质。纯化的天然Oms66（nOms66）保留了在平面脂质双层模型膜系统中形成大通道的能力，并使用变性的Oms66（hOms66）免疫新西兰白兔。在补体依赖性硼酸测定法中，与基础血清以及来自对伯氏疏螺旋体（IRS）进行再感染的兔血清平行进行了补体依赖性硼酸测定，测试了所得的Oms66抗血清。 IRS对B31和B313菌株均显示出高滴度的补体依赖性杀伤作用。用nOms66免疫的动物的血清对菌株B313表现出高滴度的依赖补体的杀伤活性，但对B31没有杀伤力。相比之下，用hOms66免疫产生的血清对任何一种菌株均无杀伤活性。用重组Oms66（rOms66）将抗血清吸附到nOms66之后，血清抗体不再与rOms66或已被8 M尿素变性的nOms66结合。但是，抗体仍然与nOms66结合，并且保留了针对B313的杀伤活性，因此表明天然的构象表位是该杀菌活性的目标。用nOms66免疫六只C3H HeJ小鼠，并通过皮肤移植感染的小鼠耳朵组织，使用“宿主适应型” B. burgdorferi B31对其进行攻击。保护六只小鼠中的四只免受局部和传播感染。这些发现表明，天然Oms66可以引发有效的杀菌活性，并具有针对宿主适应性生物的显着保护性免疫力。

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