用 RT-PCR 对沙鼠-鼠杂种瘤细胞进行同型分析。

Isotype analysis of gerbil-mouse heterohybridomas by RT-PCR.

机构信息

Department of Animal Science and Resources, College of Bioresource Sciences, Nihon University, Kameino 1866, Fujisawa 252-0880, Japan.

出版信息

J Immunol Methods. 2012 Dec 14;386(1-2):108-11. doi: 10.1016/j.jim.2012.09.001. Epub 2012 Sep 10.

DOI:10.1016/j.jim.2012.09.001

PMID:22974835

Abstract

We designed primer sets specific to the immunoglobulin (Ig) heavy-chain constant region (IGHC) genes in Mongolian gerbil (Meriones unguiculatus) to amplify five gerbil IGHC cDNA sequences, Cμ, Cγ1, Cγ2, Cε, and Cα. Five gerbil-mouse heterohybridomas B11D2(C2), B11E2(D5).M, B5-3, D5, and C11 respectively expressed Cγ1, Cμ, Cγ2, Cγ2, and Cγ1. In contrast, a commercial isotyping kit for mouse Igs identified Cγ1, Cμ, Cγ3, Cγ3, and Cγ1, respectively, misidentifying gerbil IgG2 as IgG3 by cross-reactivity with anti-mouse IgG3 polyclonal antibody. These primer sets will allow the accurate estimation of gerbil Ig classes and IgG subclasses. These results from three gerbil strains indicate that the primer sets can be used for isotype analysis of gerbil mAbs and for evaluation of humoral immunity.

摘要

我们设计了针对蒙古沙鼠（Meriones unguiculatus）免疫球蛋白（Ig）重链恒定区（IGHC）基因的引物对，以扩增五个沙鼠 IGHC cDNA 序列，Cμ、Cγ1、Cγ2、Cε和 Cα。五个沙鼠-小鼠异源杂交瘤 B11D2(C2)、B11E2(D5).M、B5-3、D5 和 C11 分别表达 Cγ1、Cμ、Cγ2、Cγ2 和 Cγ1。相比之下，用于小鼠 Igs 的商业分型试剂盒分别鉴定出 Cγ1、Cμ、Cγ3、Cγ3 和 Cγ1，通过与抗小鼠 IgG3 多克隆抗体的交叉反应错误地将沙鼠 IgG2 鉴定为 IgG3。这些引物对将允许准确估计沙鼠 Ig 类和 IgG 亚类。来自三个沙鼠品系的这些结果表明，这些引物对可用于沙鼠 mAbs 的同种型分析以及体液免疫的评估。