B-CELL DYSFUNCTION FOLLOWING BONE MARROW TRANSPLANTATION

骨髓移植后 B 细胞功能障碍

• 批准号: 6236433
• 负责人: Tucker W LeBien
• 金额: $ 2.48万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-01-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6236433
• 关键词: B lymphocyte; autologous transplantation; bone marrow; bone marrow transplantation; cell adhesion; cell differentiation; gene expression; genetic library; genetic mapping; histochemistry /cytochemistry; homologous transplantation; human tissue; immunoconjugates; immunofluorescence technique; immunotoxicity; interleukin 1; interleukin 4; interleukin 7; lymphoblast; lymphocyte proliferation; nucleic acid sequence; polymerase chain reaction; tissue /cell culture; transforming growth factors

Immune reconstitution following allogeneic (allo) or autologous (auto) bone marrow transplantation (BMT) has been, and continues to be, a serious clinical problem. In the early post-BMT period deficiencies in humoral immunity have been attributed to B cell dysfunction. Unfortunately, no consensus has emerged that can explain the mechanism underlying the early B cell dysfunction in most transplant patients; nor has there been an analysis of the emerging B cell repertoires. During the next grant period, new experimental approaches to address the problem of B cell dysfunction post-BMT will be employed. We will test the hypothesis that B cell dysfunction post-BMT reflects a defect in BM microenvironmental adherent cell/B-cell precursor interactions required for normal self-renewal and differentiation. Adherent cells established from the BM of post-BMT patients will be characterized for patterns of gene expression using histochemistry, immunofluorescence and the polymerase chain reaction (PCR). Adherent cells deriVed from normal donor BM will be similarly characterized. The post-BMT and normal donor adherent cells will then be compared for their ability to support adhesion, IL-7 driven proliferation in long-term culture, and differentiation of BCP. We will test the hypothesis that an ordered re-establishment of antibody repertoires occurs post-BMT which may contribute to early, restricted B cell responses. Peripheral blood B cells taken at various time points from post-BMT recipients will be used to generate heavy and light chain Ig repertoire DNA and RNA libraries by PCR amplification. Libraries will be screened with heavy and light chain Ig probes specific for V gene families to evaluate expression frequency. Restriction mapping and DNA sequencing will be used to define specific V gene usage. A chronologic analysis of individual V gene repertoires post-BMT will be compared among patients, as well as with repertoire patterns existing in normal individuals. A better understanding of B cell differentiation post-BMT may stimulate the development of corrective strategies that could reverse or inhibit humoral immunodeficiency in these patients.

同种异体（allo）或自体（auto）骨后的免疫重建 骨髓移植（BMT）一直是，并将继续是，一个严重的， 临床问题 在骨髓移植后早期， 免疫性归因于B细胞功能障碍。 可惜没有 人们已经达成共识，可以解释早期B的机制 大多数移植患者的细胞功能障碍;也没有一个 新出现的B细胞库的分析。 在下一个资助期， 解决B细胞功能障碍问题的新实验方法 将采用后BMT。 我们将检验B细胞 骨髓移植后功能障碍反映了骨髓微环境粘附缺陷 正常自我更新所需的细胞/B细胞前体相互作用， 分化 从骨髓移植后的骨髓中建立贴壁细胞 患者将被表征为基因表达模式， 组织化学、免疫荧光和聚合酶链反应（PCR）。 来源于正常供体骨髓的贴壁细胞将类似地 表征了 然后将BMT后和正常供体贴壁细胞进行体外培养。 与支持粘附的能力相比，IL-7驱动的增殖 在长期培养中，BCP的分化。 我们将测试 假设抗体库发生有序重建 这可能有助于早期限制性B细胞应答。 BMT后不同时间点采集的外周血B细胞 受体将用于产生重链和轻链IG库DNA 和通过PCR扩增的RNA文库。 将筛选文库， 特异于V基因家族的重链和轻链IG探针，以评估 表达频率 将使用限制性酶切图谱和DNA测序 来定义特定的V基因用法 个体V的年代学分析 BMT后的基因库将在患者之间进行比较，以及与 存在于正常个体中的所有模式。 更好地理解 BMT后B细胞分化的增加可能会刺激 纠正策略，可以逆转或抑制体液 这些患者的免疫缺陷。