用 Sputnik V 或 Comirnaty 强化一个月后,从体外刺激的记忆 b 细胞中获得的病毒特异性抗体的热敏性不会改变

E. Astakhova
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摘要

接种 COVID-19 疫苗后,长期免疫记忆的保护特性表现为血清抗体和记忆 B 细胞在反复接触抗原后分泌的抗体的中和活性。记忆 B 细胞免疫球蛋白基因中发生的体细胞高突变是提高抗体亲和力的机制之一。目前,人们对用载体疫苗加强接种 COVID-19 对记忆 B 细胞成熟的影响仍知之甚少。这项工作的目的是确定 COVID-19 强化疫苗如何影响记忆 B 细胞分泌的 RBD 特异性 IgG 抗体的亲和力。从使用 Sputnik V 或 Comirnaty 重新接种 COVID-19 的志愿者的外周单核血细胞中分离出 B 淋巴细胞。在体外用 A549 饲养细胞表面表达的 CD40L 和 IL-21 刺激 B 细胞。使用离心浓缩器将上清浓缩 8 倍。通过酶联免疫吸附试验(ELISA)测定从刺激记忆 B 细胞获得的上清中野生型 RBD 特异性 IgG 抗体的水平。为确定亲和性指数,提供了 7M 尿素酶联免疫吸附试验。结果表明,尽管从受刺激的记忆 B 细胞中获得的抗原特异性 IgG 抗体的数量普遍增加,但两组供体在强化一个月后这些抗体的嗜性没有变化。所获得的结果有助于了解接种 COVID-19 强化疫苗后记忆 B 细胞成熟的机制,并可能有助于决定强化接种的策略。
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The avidity of virus-specific antibodies obtained from in vitro stimulated memory b cells does not change one month after booster with Sputnik V or Comirnaty
The protective properties of long-term immunological memory after vaccination against COVID-19 are characterized by the neutralizing activity of serum antibodies and antibodies secreted by memory B cells upon repeated encounter with the antigen. Somatic hypermutations occurring in the immunoglobulin genes of memory B cells are one of the mechanisms for increasing the affinity of antibodies. At the moment, the effect of booster vaccination against COVID-19 with vector vaccines, on the maturation of memory B cells remains poorly understood. The purpose of this work was to determine how COVID-19 booster affects the affinity of RBD-specific IgG antibodies secreted by memory B cells. B lymphocytes were isolated from peripheral mononuclear blood cells of volunteers who had been revaccinated against COVID-19 with Sputnik V or Comirnaty. B cells were stimulated in vitro with CD40L expressed on the surface of A549 feeder cells and IL-21. Supernatants were concentrated 8-fold using centrifugal concentrators. In the obtained supernatants from stimulated memory B cells, the level of IgG antibodies specific to wild-type RBD was determined by enzyme-linked immunosorbent assay (ELISA). To determine the avidity index, ELISA with 7M urea was provided. It was shown that despite a general increase in the amount of antigen-specific IgG antibodies obtained from stimulated memory B cells, there was no change in the avidity of these antibodies one month after booster in both groups of donors. The obtained results contribute to the understanding of the mechanisms of memory B cell maturation after booster vaccinations against COVID-19 and may be useful for deciding on the strategy of booster vaccination.
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