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转基因小鼠在B细胞上选择性缺乏MHCII类 (I-E) 抗原表达:一种体内方法来研究Ia基因功能
G Widera1, L C Burkly, C A Pinkert
1Biogen Research Corporation, Cambridge, Massachusetts 02142.
Cell
|October 23, 1987
概括
表达EαMHCII类基因的转基因小鼠表现出受限的表达,特别是在B淋巴细胞中缺席. 这表明B细胞表达的关键调节序列位于5'侧边区域的-1.4kb和-2kb之间.
科学领域:
- 免疫遗传学 免疫遗传学
- 分子生物学分子生物学
- 转基因动物模型
背景情况:
- Eα基因是MHCII类分子的关键组成部分,对于T细胞激活至关重要.
- 了解控制Eα基因表达的调节元件对于免疫系统研究至关重要.
- 之前的研究已经确定了MHCII类基因的5'侧翼序列内的调节区域.
研究的目的:
- 研究5'侧翼序列在Eα基因的组织特异性表达中的作用.
- 确定B淋巴细胞中Eα表达所需的最小调节元素.
- 分析转基因结构修改对Eα基因表达模式和免疫耐受性的影响.
主要方法:
- 携带EαMHCII类基因的转基因小鼠的生成,具有定义的5'和3'侧翼序列.
- 使用分子技术,分析各种组织 (甲状腺,脏,腹排泄细胞) 中的转基因表达.
- 通过混合白细胞反应实验对转基因小鼠免疫耐受性诱导的评估.
主要成果:
- 转基因表达在胸膜组织和附着的脏细胞中被观察到,并且可通过玛干扰因子在腹膜液细胞中被诱导.
- 与内源基因不同的是,B淋巴细胞中显著缺少Eα转基因表达.
- 在-1.4kb到2kb之间删除5'侧边序列似乎会使B细胞特异性调节元件失活.
- 与转基因相关的pBR327DNA的存在抑制了外周表达,导致只有胆髓表达,并诱导了对IE的耐受性.
结论:
- Eα基因的5'侧边区域包含B淋巴细胞特异性表达的关键调节元素.
- 在5'侧翼序列内的特定删除可以消除B细胞中的Eα表达.
- Eα转基因结构及其相关元素影响基因表达模式和诱导免疫耐受性.
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