Chromatin modifications in immunoglobulin switch recombination

免疫球蛋白开关重组中的染色质修饰

• 批准号: 7734113
• 负责人: MARTIN F. GELLERT
• 金额: $ 41.97万
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7734113
• 关键词: Acetylation; Allergic; Antibodies; B-Lymphocytes; Cell Proliferation; Cell surface; Cells; Chromatin; Cytidine Deaminase; Cytokine Activation; DNA Methylation; Deoxycytidine; Event; Genetic Transcription; Histone H4; Histones; Human; IL4 gene; IgE; Immunoglobulin Class Switching; Immunoglobulin Genes; Immunoglobulin Switch Recombination; Immunoglobulins; Interleukin-4; Maps; Mediating; Modification; Nucleosomes; Pattern; RNA Splicing; Relative (related person); Reporting; Resolution; TNFRSF5 gene; Transcript; Variant; Work; constant region gene; promoter; receptor; response

Switching to IgE can be activated by a combination of IL4, which induces transcription in that region, and activation of the cell surface CD40 receptor, which induces cell proliferation. Activation can be produced by anti-CD40 antibody or by a trimeric form of CD40 ligand.Our studies have focused on the human CL-01 line of B cells, which has been reported to undergo switch recombination. We have shown that IL4 causes a 100X increase in transcript, both unspliced and spliced, over the IgE switch region. We also observed a sizable (5X) increase in transcription of AID, the cytidine deaminase that is responsible for initiating CSR. A high resolution study of histone modifications in uninduced and induced CL-01 cells has been carried out, focusing on the region between the I-epsilon promoter and the 3 end of the IgE switch region, the most important region associated with this CSR event. The largest increases were found for acetyl-H3K9/14 (or acetyl-H3K9 alone) and for trimethyl-H3 K4, while other modifications including histone H4 tetra-acetylation (K5/8/12/16) and H3 K27 trimethylation were not greatly altered following IL4 induction. This pattern is consistent with broad activation of the region. Furthermore, the distribution of a given modification was in most cases non-uniform: for example, dimethyl-H3K4, tetraacetyl-H4, and acetyl-H3 K9/14 all showed relative peaks over the I-epsilon promoter. To interpret these results the abundance of nucleosomes was mapped across the same region, and a relative depletion over I-epsilon was observed, without any further change on induction. We are now extending these observations to studying the distribution of the histone variants H3.3 and H2A.Z. In addition we are studying DNA methylation in the same region. We have shown that treatment of the cells with 5-aza-deoxycytidine, a known demethylating agent, leads to an additional increase in IL4-stimulated transcription.

转换到IgE可以通过IL4和细胞表面CD40受体的结合来激活，IL4可以诱导该区域的转录，而CD40受体的激活可以诱导细胞增殖。激活可以通过抗CD40抗体或CD40配体的三聚体形式产生。我们的研究主要集中在人B细胞CL-01系，据报道，它经历了开关重组。我们已经证明，IL4在IgE开关区域导致转录物增加100倍，无论是未剪接的还是剪接的。我们还观察到，负责启动CSR的胞苷脱氨酶AID的转录显著增加（5倍）。