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Development of Conformer-Specific Anti-HLA Class II mAbs

构建体特异性抗 HLA II 类单克隆抗体的开发

基本信息

批准号：
10330612
负责人：
James R Drake
金额：
$ 8.15万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-27 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10330612
关键词：
Adopted Alleles B-Cell Leukemia B-Cell Lymphomas B-Lymphocytes Binding CD4 Positive T Lymphocytes Calcium Signaling Cell Communication Cells Cholesterol Clinical Clinical Trials Data Dendritic Cells Development Dimerization Disease Distal Epitopes Exhibits Extracellular Domain Follow-Up Studies Genetic Polymorphism Goals HLA-DQ Antigens HLA-DR Antigens Health Histocompatibility Antigens Class II Hu1D10 Human Hybridomas Hybrids Immune Immunize Immunobiology Immunologics In Vitro Lead Ligation Link Liquid substance Lymphoma MHC Class II Genes Mediating Membrane Membrane Microdomains Molecular Molecular Conformation Monoclonal Antibodies Mus Mutation Pathway interactions Peptides Phase Protocols documentation Publishing Reagent Reporting Research Research Methodology Rest Role Scientist Shapes Signal Pathway Signal Transduction Site Sodium Chloride Specificity T-Lymphocyte Technology Testing Therapeutic Therapeutic Effect Therapeutic Monoclonal Antibodies Therapeutic Uses Transmembrane Domain Treatment Efficacy Work antigen processing base clinically relevant conformer crosslink human leukocyte antigen testing immune function in silico in vivo insight leukemia/lymphoma macrophage molecular modeling mutant neoplastic research and development response tool

项目摘要

Summary: Murine MHC class II molecules exist in two distinct conformational states formed by differential pairing of class II transmembrane (TM) domain GxxxG dimerization motifs (i.e., M1 and M2 paired class II), and these two conformers are linked to different immunological functions. Moreover, we have recently reported that: 1) HLA class II HLA-DR, DP and DQ molecules mirror mouse class II in possessing three TM domain GxxxG motifs, 2) in silico molecular modeling reveals that HLA-DR and DQ TM domains can adopt energetically favorable M1 and M2 paired states, and 3) hybrid class II molecules with an HLA-DR or DQ TM domain can adopt both an M1 and M2 conformation. In this proposal, we show that a commercially-available anti-HLA class II mAb (H0002) exhibits differential reactivity to wild type vs. an HLA-DR M1 GxxxG>VxxxV mutant molecule (which would have an altered conformer state). These findings strongly suggesting that like mouse class II, human class II molecules also exist in two distinct conformational states (M1 and M2 paired class II). In addition to published work on the differential signaling of M1 vs. M2 paired class II in normal B murine cells, our preliminary studies show that M1 and M2 class II are linked to distinct signaling pathways in neoplastic B cells. Herein, we show that while selective engagement of M1 class II inhibits B cell lymphoma proliferation, co-ligation of M1 and M2 class II blocks this effect. This is important because three anti-HLA-DR mAbs (IMMU-114, 1D09C3 and Hu1D10) have undergone clinical trials as treatment for B cell leukemia/lymphoma with mixed results. Since the conformer specificity of these three anti-DR mAbs is unclear, it raises the possibility that anti-DR mAb therapeutic efficacy is linked to mAb conformer reactivity. These findings lead to the hypothesis that like mouse class II, human HLA class II molecules exist in two distinct conformational states based on alternative pairing of transmembrane domain GxxxG dimerization motifs and that these two distinct HLA class II conformers are linked to different clinically-relevant B cell signaling pathways. To test this core hypothesis, we will; 1) Define the HLA-DR conformer specificity of a large panel of available anti-HLA class II mAbs, including those that have undergone clinical trials as therapeutics for B cell leukemia/lymphoma, and 2) Develop a panel of new monoclonal antibodies that recognize the conformational differences between M1 and M2 paired human HLA-DR and HLA-DQ class II molecules. Completion of these aims will help bring the M1/M2 class II conformer paradigm to the HLA field, provide insight into whether there is a correlation between the conformer-specificity of clinically-relevant anti-DR mAbs and demonstrated therapeutic effect, and provide the field with a new set of tools (i.e., conformer-specific anti- HLA-DR and anti-HLA-DQ mAbs) to further investigate the role of M1 and M2 HLA class II in human health and disease.

总结：小鼠MHCII类分子存在于两种不同的构象状态中，这两种构象状态是由MHCII类分子的差异配对形成的。 II类跨膜（TM）结构域GxxxG二聚基序（即，M1和M2配对II类），这些两种构象异构体与不同的免疫功能有关。此外，我们最近报告说：1） HLA-DR、DP和DQ分子反映了具有三个TM结构域GxxxG的小鼠II类 2）计算机分子模拟显示HLA-DR和DQ TM结构域可以在能量上采用有利的M1和M2配对状态，和3）具有HLA-DR或DQ TM结构域的杂合II类分子可以采用M1和M2构象。在这个建议中，我们表明，商业上可获得的抗HLA II类mAb（H 0002）对野生型与HLA-DR M1 GxxxG>VxxxV突变体显示出不同的反应性分子（其将具有改变的构象状态）。这些发现有力地表明，像老鼠一样，人II类分子也以两种不同的构象状态存在（M1和M2成对的II类分子）。除了已发表的关于正常B小鼠中M1与M2配对II类的差异信号传导的工作外，我们的初步研究表明，M1和M2 II类与不同的信号通路有关，肿瘤性B细胞。在此，我们表明，虽然选择性参与M1 II类抑制B细胞淋巴瘤，增殖，II类M1和M2的共连接阻断了这种作用。这一点很重要，因为三种抗HLA-DR mAb（IMMU-114、1D 09 C3和Hu 1D 10）已经经历了作为B细胞治疗的临床试验白血病/淋巴瘤，结果混杂。由于这三种抗DR mAb的构象特异性是尚不清楚，这提出了抗DR mAb治疗功效与mAb构象反应性有关的可能性。这些发现导致了这样的假设，即像小鼠II类分子一样，人类HLA II类分子存在于两个细胞中。基于跨膜结构域GxxxG二聚化的交替配对的不同构象状态这两种不同的HLA II类构象异构体与不同的临床相关B细胞相关，信号通路为了验证这一核心假设，我们将：1）定义HLA-DR构象特异性的大的一组可用的抗HLA II类mAb，包括那些已经进行临床试验作为治疗 B细胞白血病/淋巴瘤，和2）开发一组新的单克隆抗体， M1和M2配对的人HLA-DR和HLA-DQ II类分子之间的构象差异。这些目标的完成将有助于将M1/M2 II类构象异构体范式引入HLA领域，深入了解临床相关抗DR mAb的一致性特异性之间是否存在相关性并证明了治疗效果，并为该领域提供了一套新的工具（即，适形剂特异性抗 HLA-DR和抗HLA-DQ单克隆抗体）进一步研究M1和M2 HLA II类在人类健康中的作用和疾病