Department of Clinical Immunology and Allergy, Medical Faculty of Masaryk University, University Centre for Primary Immunodeficiencies, St. Anne's University Hospital, Pekarska 53, CZ-656 91 Brno, Czech Republic.
Vaccine. 2011 May 31;29(24):4142-50. doi: 10.1016/j.vaccine.2011.03.087. Epub 2011 Apr 5.
Common variable immunodeficiency (CVID), the most frequent primary antibody disorder, is characterized by hypogammaglobulinaemia and impaired antibody production. Poor vaccination response is essential for the diagnosis of CVID. Their under laying defects remain to be elucidated. Routine determination of antibody production in serum from CVID patients after vaccination and investigation of B cell function in vivo is complicated due to substitution therapy. Therefore we investigated antibody production on the B-cell level by ELISPOT and characterized changes in B-cell subpopulations in CVID patients, including plasmablasts, in peripheral blood by flow cytometry after vaccination for specification of the diagnosis. Thirty-seven CVID patients and eighty healthy volunteers were immunized with tetanus toxoid and pneumococcal polysaccharide vaccines. Specific antibody levels and B cell subpopulations were measured before vaccination and on day 7 after vaccination by ELISPOT assay and flow cytometry respectively. Of the thirty-seven well defined CVID patients studied, thirty lacked detectable spot forming cells producing specific IgG, IgA or IgM antibodies against employed vaccines and seven had only weak responses compared to controls. In the control group, an increase in circulating plasmablasts on day 7 post immunization corresponded with the appearance of antibody forming cells. In contrast, CVID patients failed to increase plasmablasts significantly in peripheral blood after antigen challenge. Our findings indicate that CVID patients have a block in terminal B-cell differentiation and that flow based assessment of plasmablasts in peripheral blood after vaccination serves as a surrogate diagnostic marker for assessing in vivo antibody responses in patients suspected to have CVID.
常见变异性免疫缺陷症（CVID）是最常见的原发性抗体疾病，其特征为低丙种球蛋白血症和抗体产生受损。接种疫苗后不良的疫苗反应对 CVID 的诊断至关重要。但其潜在缺陷仍有待阐明。由于替代疗法，常规测定 CVID 患者接种疫苗后血清中的抗体产生并调查体内 B 细胞功能变得复杂。因此，我们通过 ELISPOT 研究了接种疫苗后 B 细胞水平的抗体产生，并通过流式细胞术在 CVID 患者外周血中表征了浆母细胞等 B 细胞亚群的变化，以明确诊断。我们对 37 例 CVID 患者和 80 名健康志愿者用破伤风类毒素和肺炎球菌多糖疫苗进行了免疫接种。通过 ELISPOT 检测和流式细胞术分别在接种前和接种后第 7 天测定特异性抗体水平和 B 细胞亚群。在 37 例明确诊断的 CVID 患者中，有 30 例未能检测到针对所用疫苗产生特异性 IgG、IgA 或 IgM 抗体的可检测的斑点形成细胞，而 7 例患者与对照组相比仅有较弱的反应。在对照组中，接种后第 7 天循环浆母细胞的增加与抗体形成细胞的出现相对应。相比之下，CVID 患者在抗原刺激后未能使外周血中的浆母细胞显著增加。我们的研究结果表明，CVID 患者存在末端 B 细胞分化障碍，并且接种疫苗后外周血中浆母细胞的流式细胞术评估可作为评估疑似 CVID 患者体内抗体反应的替代诊断标志物。
