CD40-DEFICIENT MICE GENERATED BY RECOMBINATION-ACTIVATING GENE-2-DEFICIENT BLASTOCYST COMPLEMENTATION
CD40-DEFICIENT MICE GENERATED BY RECOMBINATION-ACTIVATING GENE-2-DEFICIENT BLASTOCYST COMPLEMENTATION
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DOI：
10.1073/pnas.91.25.12135
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发表时间：
1994-12-06
影响因子：
11.1
通讯作者：
GEHA, RS
中科院分区：
文献类型：
--
作者：
CASTIGLI, E;ALT, FW;GEHA, RS
为了研究B细胞抗原CD 40在免疫应答中的作用，将其中编码CD 40的基因的两个拷贝都已通过同源重组被破坏的小鼠胚胎干（ES）细胞注射到RAG-2（重组激活基因-2）缺陷型胚泡中，以产生嵌合体，其中所有成熟淋巴细胞均来自CD 40缺陷型ES细胞。在CD 40（-/-）嵌合体中T和B细胞数量和表型正常。然而，B细胞未能增殖和进行同种型转换在体外响应可溶性CD 40配体（sCD 40 L）与白细胞介素4（IL-4），但正常响应脂多糖（LPS）与IL-4。CD 40（-/-）嵌合体完全未能安装抗原特异性抗体反应或发展后，免疫与T细胞依赖性（TD）抗原钥孔血蓝蛋白的生发中心。相反，CD 40（-/-）突变小鼠对T细胞非依赖性（TI）抗原2，4，6-三硝基苯基（TNP）-LPS和TNP-Ficoll反应正常。幼龄（6-8周龄）CD 40（-/-）动物血清免疫球蛋白水平最显著的变化是IgE缺失和IgG 1和IgG 2a严重降低。这些结果证实了CD 40-CD 40 L相互作用在对TD抗原的抗体应答和同种型转换中的重要作用。
To study the role of the B-cell antigen CD40 in immune responses, mouse embryonic stem (ES) cells in which both copies of the gene encoding CD40 had been disrupted by homologous recombination were injected in RAG-2 (recombination-activating gene-2)-deficient blastocysts to generate chimeras in which all mature lymphocytes are derived from the CD40-deficient ES cells. T- and B-cell number and phenotype were normal in the CD40(-/-) chimeras. However, B cells failed to proliferate and undergo isotype switching in vitro in response to soluble CD40 ligand (sCD40L) with interleukin 4 (IL-4) but responded normally to lipopolysaccharide (LPS) with IL-4. CD40(-/-) chimeras completely failed to mount an antigen-specific antibody response or to develop germinal centers following immunization with the T cell-dependent (TD) antigen keyhole limpet hemocyanin. In contrast, CD40(-/-) mutant mice responded normally to the T cell-independent (TI) antigens 2,4,6-trinitrophenyl (TNP)-LPS and TNP-Ficoll. The most noticeable alteration in the serum immunoglobulin levels of young (6-8 weeks old) CD40(-/-) animals was absence of IgE and severe decrease of IgG1 and IgG2a. These results confirm the essential role of CD40-CD40L interactions in the antibody response to TD antigens and in isotype switching.