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首页> 外文期刊>Vaccine >A modified surface killing assay (MSKA) as a functional in vitro assay for identifying protective antibodies against pneumococcal surface protein A (PspA)
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A modified surface killing assay (MSKA) as a functional in vitro assay for identifying protective antibodies against pneumococcal surface protein A (PspA)

机译：改良的表面杀伤测定法（MSKA）作为功能性体外测定法，用于鉴定针对肺炎球菌表面蛋白A（PspA）的保护性抗体

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Streptococcus pneumoniae causes otitis media, meningitis and pneumonia in patients worldwide; predominantly affecting young children, the elderly, and the immune compromised. Current vaccines against invasive pneumococcal disease are based on the polysaccharide capsules of the most clinically relevant serotypes. Due to serotype replacement, non-vaccine serotypes of S. pneumoniae have become more clinically relevant and as a result pneumococcal vaccines are becoming increasingly complex. These events emphasize the need to evaluate the potential for pneumococcal cross-reactive proteins to contribute to future vaccines. Antibody elicited by the immunization of humans with pneumococcal surface protein A (PspA) can passively protect mice from infection. However, robust in vitro functional assays for antibody to PspA are not available to predict the protective capacity of immune serum. For polysaccharide based vaccines, a standardized opsonophagocytosis killing assay (OPKA) is used. Antibody to PspA, however, does not work well in the standard OPKA. The present studies take advantage of past observations that phagocytosis is more efficient on tissue surfaces than in solution. In a modified surface killing assay (MSKA), monoclonal antibody to PspA, in the presence of complement, opsonized pneumococci for killing by phagocytes on an agar surface. Five monoclonal antibodies to PspA were tested; three demonstrated increased amounts of killing compared to the diluent control and protected mice by passive protection against type 3 pneumococci. The two antibodies that were not functional in the MSKA also failed to protect mice. Thus, an MSKA might be useful as a functional assay for immunity to PspA
机译：肺炎链球菌在全球范围内引起中耳炎，脑膜炎和肺炎。主要影响幼儿，老年人和免疫功能低下的人。当前针对侵袭性肺炎球菌疾病的疫苗是基于临床上最相关血清型的多糖胶囊。由于血清型的替代，肺炎链球菌的非疫苗血清型在临床上变得越来越重要，结果肺炎球菌疫苗变得越来越复杂。这些事件强调需要评估肺炎球菌交叉反应蛋白对未来疫苗做出贡献的潜力。用肺炎球菌表面蛋白A（PspA）免疫人类后产生的抗体可以被动保护小鼠免受感染。但是，目前尚无针对PspA抗体的强大的体外功能测定方法来预测免疫血清的保护能力。对于基于多糖的疫苗，使用标准化的调理吞噬作用杀灭测定法（OPKA）。但是，PspA抗体在标准OPKA中不能很好地发挥作用。本研究利用过去的观察结果，即吞噬作用在组织表面上比在溶液中更有效。在改良的表面杀伤试验（MSKA）中，抗PspA的单克隆抗体在补体存在下调理了肺炎球菌，以被琼脂表面的吞噬细胞杀死。测试了五种针对PspA的单克隆抗体;与稀释剂对照和通过对3型肺炎球菌的被动保护而得到保护的小鼠相比，三只动物的杀伤量增加。在MSKA中不起作用的两种抗体也未能保护小鼠。因此，MSKA可能可用作针对PspA免疫的功能测定

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