通过原位杂交对B细胞集落进行基因分型分析。成年C57BL/6和BALB/c小鼠中三个VH家族的化学计量表达。
Genotypic analysis of B cell colonies by in situ hybridization. Stoichiometric expression of three VH families in adult C57BL/6 and BALB/c mice.
作者信息
Schulze D H, Kelsoe G
出版信息
J Exp Med. 1987 Jul 1;166(1):163-72. doi: 10.1084/jem.166.1.163.
Abstract
The filter paper disc method for cloning inducible lymphocytes was used to census the splenic B cell population of C57BL/6 and BALB/c mice for the expression of three VH gene-families, VH X-24, -Q52, and -J558. B cell colonies, arising from single founder lymphocytes, were identified by in situ hybridization with VH family- and C mu-specific cDNA probes. Some 6.7 X 10(4) C mu+ colonies were screened. Among C57BL/6- or BALB/c-derived colonies, approximately 3% were VH X-24+, approximately 19% were VH Q52+, and approximately 54% were VH J558+. These frequencies are consistent with a process of equiprobable expression for individual VH segments, and provide direct evidence that normal splenic B lymphocytes use a process of random genetic combinatorics to generate the antibody repertoire.
摘要
采用滤纸圆盘法克隆诱导性淋巴细胞，以普查C57BL/6和BALB/c小鼠脾脏B细胞群体中三个VH基因家族（VH X-24、-Q52和-J558）的表达情况。通过与VH家族和Cμ特异性cDNA探针进行原位杂交，鉴定来自单个起始淋巴细胞的B细胞集落。共筛选了约6.7×10⁴个Cμ⁺集落。在源自C57BL/6或BALB/c的集落中，约3%为VH X-24⁺，约19%为VH Q52⁺，约54%为VH J558⁺。这些频率与单个VH区段等概率表达的过程一致，并提供了直接证据，表明正常脾脏B淋巴细胞利用随机基因组合过程来产生抗体库。
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