Antigen-specific immunoglobulin A antibodies secreted from circulating B cells are an effective marker for recent local immune responses in patients with cholera: Comparison to antibody-secreting cell responses and other immunological markers
Antigen-specific immunoglobulin A antibodies secreted from circulating B cells are an effective marker for recent local immune responses in patients with cholera: Comparison to antibody-secreting cell responses and other immunological markers
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DOI：
10.1128/iai.71.8.4808-4814.2003
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发表时间：
2003-08-01
影响因子：
3.1
通讯作者：
Calderwood, SB
中科院分区：
文献类型：
--
作者：
Qadri, F;Ryan, ET;Calderwood, SB
肠源性淋巴细胞短暂迁移通过外周循环，然后返回粘膜部位，可以使用基于 ELISPOT 的抗体分泌细胞 (ASC) 测定进行检测。或者，可以在体外培养瞬时循环淋巴细胞，并可以测定培养物上清液的抗原特异性反应（淋巴细胞上清液中的抗体[ALS]测定）。 ALS 测定尚未在自然粘膜感染中得到广泛验证，也未将 ALS 反应与 ASC 测定和其他霍乱特异性免疫反应进行比较。因此，我们检查了孟加拉国 30 名急性霍乱成年患者的免疫反应，并与 10 名健康对照者进行比较，测量了 ALS-免疫球蛋白 A (IgA)、ASC-IgA，以及对两种强效霍乱弧菌免疫原（即无毒的霍乱毒素 B 亚基 (CtxB) 和脂多糖 (LPS) 以及较弱的霍乱弧菌免疫原，即霍乱弧菌免疫原）的血清和粪便 IgA 反应。甘露糖敏感血凝素（MSHA）。我们使用 ALS 测定发现霍乱发病后 7 天培养的淋巴细胞上清液中抗 CtxB、抗 LPS 和抗 MSHA IgA 显着增加。我们发现 ALS 和 ASC 反应相关性极好；两者都具有与杀弧菌反应相当的敏感性，并且两种方法都比粪便 IgA 测量更敏感。 ALS 测定用于研究粘膜免疫反应的一个优点是能够冷冻上清液中的抗体以进行后续评估；与 ASC 检测一样，ALS 检测可以区分近期和远期粘膜感染，而在流行环境中使用其他程序可能很难做出这种区分。
Gut-derived lymphocytes transiently migrate through the peripheral circulation before homing back to mucosal sites and can be detected using an ELISPOT-based antibody secreting cell (ASC) assay. Alternatively, transiently circulating lymphocytes may be cultured in vitro, and culture supernatants may be assayed for antigen-specific responses (antibody in lymphocyte supernatant [ALS] assay). The ALS assay has not been validated extensively in natural mucosal infection, nor has the ALS response been compared to the ASC assay and other cholera-specific immunological responses. Accordingly, we examined immune responses in 30 adult patients with acute cholera in Bangladesh, compared with 10 healthy controls, measuring ALS-immunoglobulin A (IgA), ASC-IgA, and serum and fecal IgA responses to two potent Vibrio cholerae immunogens, the nontoxic B subunit of cholera toxin (CtxB) and lipopolysaccharide (LPS) and a weaker V. cholerae immunogen, the mannose-sensitive hemagglutinin (MSHA). We found significant increases of anti-CtxB, anti-LPS, and anti-MSHA IgA in supernatants of lymphocytes cultured 7 days after onset of cholera using the ALS assay. We found that ALS and ASC responses correlated extremely well; both had comparable sensitivities as the vibriocidal responses, and both procedures were more sensitive than fecal IgA measurements. An advantage of the ALS assay for studying mucosal immune responses is the ability to freeze antibodies in supernatants for subsequent evaluation; like the ASC assay, the ALS assay can distinguish recent from remote mucosal infection, a distinction that may be difficult to make in endemic settings using other procedures.