A method for immortalizing swine monoclonal B cells secreting anti-PRRSV antibodies.

Jian Wang; Jing Zhang; Kun LI; Pu Sun; Guoxiu LI; Jiaoyang LI; Yimei Cao; Zhixun Zhao; Hong Yuan; Yuanfang FU; Pinghua LI; Dong LI; Zaixin Liu; Zengjun LU

Return

A method for immortalizing swine monoclonal B cells secreting anti-PRRSV antibodies.

Author: Jian WANG ¹ ; Jing ZHANG ¹ ; Kun LI ¹ ; Pu SUN ¹ ; Guoxiu LI ¹ ; Jiaoyang LI ¹ ; Yimei CAO ¹ ; Zhixun ZHAO ¹ ; Hong YUAN ¹ ; Yuanfang FU ¹ ; Pinghua LI ¹ ; Dong LI ¹ ; Zaixin LIU ¹ ; Zengjun LU ¹
Author Information

1. State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, Gansu, China.
Publication Type:Journal Article
Keywords: B cell immortalization; PRRSV; magnetic bead method; monoclonal antibody
MeSH: Animals; Antibodies, Monoclonal; Antibodies, Neutralizing; Antibodies, Viral; Humans; Porcine Reproductive and Respiratory Syndrome/prevention & control*; Porcine respiratory and reproductive syndrome virus/genetics*; Swine
From: Chinese Journal of Biotechnology 2022;38(8):2872-2882
CountryChina
Language:Chinese
Abstract: Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which causes great economic losses. At the moment, no effective neutralizing antibody is available for scientific research and treatment. Therefore, developing a method for screening the neutralizing monoclonal antibodies is of great significance for the prevention and treatment of PRRSV and the screening of antigen sites. Monoclonal antibodies have been widely used in the treatment and diagnosis of many human and animal diseases. Therefore, screening effective neutralizing antibodies for different pathogens is an urgent task. Among the methods for monoclonal antibody screening, B cell immortalization is an effective method to obtain neutralizing monoclonal antibody. Specifically, in this study, the bcl-6 and bcl-xl genes were connected by f2a and then the yielded product was ligated to a vector for retrovirus packaging. The swine lymphocytes immunized with PRRSV were infected the yielded mature viruses and cultured in the complete medium containing CD40L and IL21 cytokines. Then, CD21 was used as the marker to screen B cells with the magnetic bead method. Finally, monoclonal B cells were obtained and the secretion of antibodies was tested. The results showed that the plasmid, either being transfected alone or with the packaged plasmids, could be expressed, and that the packaged retrovirus could infect the cells. Moreover, the infected lymphocytes secreted antibodies, so did the screened B cells. Therefore, the method for screening monoclonal antibody against PRRSV was successfully established.