Lin Jisheng, Smith Mark A, Benjamin William H, Kaminski Robert W, Wenzel Heather, Nahm Moon H
Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama, USA.
Subunit Enteric Vaccines and Immunology, Walter Reed Army Institute of Research, Washington, DC, USA.
Clin Vaccine Immunol. 2016 Aug 5;23(8):681-8. doi: 10.1128/CVI.00148-16. Print 2016 Aug.
There is a significant need for an effective multivalent Shigella vaccine that targets the most prevalent serotypes. Most Shigella vaccines under development utilize serotype-specific lipopolysaccharides (LPSs) as a major component based on protection and epidemiological data. As vaccine formulations advance from monovalent to multivalent, assays and reagents need to be developed to accurately and reproducibly quantitate the amount of LPSs from multiple serotypes in the final product. To facilitate this effort, we produced 36 hybridomas that secrete monoclonal antibodies (MAbs) against the O antigen on the LPS from Shigella flexneri 2a, Shigella flexneri 3a, and Shigella sonnei We used six of these monoclonal antibodies for an inhibition enzyme-linked immunosorbent assay (iELISA), measuring LPSs with high sensitivity and specificity. It was also demonstrated that the Shigella serotype-specific MAbs were useful for bacterial surface staining detected by flow cytometry. These MAbs are also useful for standardizing the serum bactericidal assay (SBA) for Shigella Functional assays, such as the in vitro bactericidal assay, are necessary for vaccine evaluation and may serve as immunological correlates of immunity. An S. flexneri 2a-specific monoclonal antibody killed S. flexneri 2b isolates, suggesting that S. flexneri 2a LPS may induce cross-protection against S. flexneri 2b. Overall, the Shigella LPS-specific MAbs described have potential utility to the vaccine development community for assessing multivalent vaccine composition and as a reliable control for multiple immunoassays used to assess vaccine potency.
迫切需要一种有效的多价志贺氏菌疫苗，该疫苗能够针对最常见的血清型。根据保护作用和流行病学数据，大多数正在研发的志贺氏菌疫苗都将血清型特异性脂多糖（LPS）作为主要成分。随着疫苗配方从单价发展到多价，需要开发检测方法和试剂，以准确且可重复地定量最终产品中多种血清型的LPS含量。为推动这项工作，我们制备了36种杂交瘤，这些杂交瘤分泌针对福氏志贺氏菌2a、福氏志贺氏菌3a和宋内志贺氏菌LPS上O抗原的单克隆抗体（MAb）。我们使用其中六种单克隆抗体进行抑制酶联免疫吸附测定（iELISA），以高灵敏度和特异性测量LPS。还证明了志贺氏菌血清型特异性MAb可用于通过流式细胞术检测细菌表面染色。这些MAb对于标准化志贺氏菌的血清杀菌试验（SBA）也很有用。功能测定，如体外杀菌试验，对于疫苗评估是必要的，并且可以作为免疫的免疫学相关指标。一种福氏志贺氏菌2a特异性单克隆抗体可杀死福氏志贺氏菌2b分离株，这表明福氏志贺氏菌2a LPS可能诱导对福氏志贺氏菌2b的交叉保护。总体而言，所描述的志贺氏菌LPS特异性MAb对疫苗开发领域具有潜在用途，可用于评估多价疫苗组成，并作为用于评估疫苗效力的多种免疫测定的可靠对照。
