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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Use of Highly Encapsulated Streptococcus pneumoniae Strains in a Flow-Cytometric Assay for Assessment of the Phagocytic Capacity of Serotype-Specific Antibodies
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Use of Highly Encapsulated Streptococcus pneumoniae Strains in a Flow-Cytometric Assay for Assessment of the Phagocytic Capacity of Serotype-Specific Antibodies

机译：流式细胞术中高度封装的肺炎链球菌菌株在评估血清型特异性抗体吞噬能力中的应用

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A phagocytosis assay for Streptococcus pneumoniae based on flow cytometry (FACS) with human polymorphonuclear cells and human complement was developed for the study of human vaccination antisera. Human prevaccination sera already contain high levels of C-polysaccharide (C-PS) antibodies, which are not protective in humans but which might give false positive results in a flow-cytometry-based assay. Cultures of S. pneumoniae grown to log phase on three consecutive days, followed by heat inactivation, yielded stable and highly encapsulated strains for serotypes 6A, 6B, 14, 19F, and 23F. As a result, only serotype-specific antibodies were able to facilitate phagocytosis of these strains, whereas no phagocytosis was observed with antibodies against C-PS or pneumococcal surface proteins. No, or weak, phagocytosis was observed with human prevaccination sera, whereas in general, postvaccination antisera facilitated phagocytosis. A highly significant correlation was observed between enzyme-linked immunosorbent assay titers and FACS phagocytosis titers (r = 0.98, P < 0.001) for serotype 23F pneumococci with human vaccination antisera. For all serotypes, interassay variation was below 10%. Major advantages of this assay over the classical killing assay are that (i) limited amounts of sera are required (10 μl per titration curve), (ii) 600 samples can be processed in one day by one person, and (iii) cells can be fixed and measurement of the samples can be performed up to 1 week later.
机译：基于人多形核细胞和人补体的流式细胞术（FACS），开发了一种肺炎链球菌的吞噬作用试验方法，用于研究人接种疫苗的抗血清。人疫苗接种前血清中已经含有高水平的C-多糖（C-PS）抗体，该抗体对人没有保护作用，但在基于流式细胞仪的测定中可能会产生假阳性结果。 S的文化。肺炎连续三天生长到对数期，然后热灭活，产生了稳定的且高度包被的血清型6A，6B，14、19F和23F菌株。结果，仅血清型特异性抗体能够促进这些菌株的吞噬作用，而针对C-PS或肺炎球菌表面蛋白的抗体则未观察到吞噬作用。在人疫苗接种前血清中未观察到吞噬作用或弱于吞噬作用，而一般而言，疫苗接种后抗血清可促进吞噬作用。血清型23F肺炎球菌与人源疫苗接种抗血清的酶联免疫吸附滴度与FACS吞噬作用滴度（ r = 0.98， P <0.001）之间观察到高度相关。对于所有血清型，批间变异均低于10％。与传统的杀灭测定相比，该测定的主要优势在于：（i）需要的血清量有限（每滴定曲线10μl）;（ii）一个人一天可以处理600个样品;（iii）细胞可以固定并可以在1周后进行样品测量。

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