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.

概括：鼠MHC II类分子存在于两个不同的构象状态中，由差分配对形成 II类跨膜（TM）域GXXXG二聚体基序（即M1和M2配对II类），并且这些两个构象异构体与不同的免疫学功能有关。此外，我们最近报告了：1） HLA II类HLA-DR，DP和DQ分子镜鼠标II类具有三个TM域GXXXG 主题，2）在计算机分子建模中表明，HLA-DR和DQ TM域可以在能量上采用有利的M1和M2配对状态，以及3）与HLA-DR或DQ TM域的混合II类分子可以同时采用M1和M2构象。在此提案中，我们证明了商业上可用的反HLA II类MAB（H0002）表现出对野生型与HLA-DR M1 GXXXG> VXXXXV突变体的反应性差异分子（将改变构象异构态）。这些发现强烈暗示着喜欢鼠标 II类，人类II类分子也存在于两个不同的构象状态（M1和M2配对II类）中。除了发表了关于M1与M2配对II类的差分信号的发表工作。细胞，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B细胞。在此，我们表明，尽管M1 II类选择性参与抑制B细胞淋巴瘤增殖，M1和M2 II类的共同结合可以阻止这种效果。这很重要，因为三个抗HLA-DR mAb（Immu-114，1D09C3和HU1D10）已接受临床试验作为B细胞的治疗白血病/淋巴瘤伴有混合结果。由于这三个抗Dr mab的构象异构体特异性是尚不清楚，这增加了抗DR MAB治疗功效与MAB构象反应性有关的可能性。这些发现导致了以下假设：像小鼠II类一样，人类HLA II类分子存在于两个中基于跨膜结构域GXXXG二聚体的替代配对的不同构象状态主题和这两个不同的HLA II类构象异构体与不同的临床B细胞有关信号通路。为了检验这一核心假设，我们将； 1）定义大型的HLA-DR构象异构体特异性可用的抗HLA II类MAB的面板，包括接受临床试验作为治疗疗法的抗HLA MAB B细胞白血病/淋巴瘤和2）开发了一组新的单克隆抗体，以识别 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类构象异构体范式带到HLA领域，提供深入了解临床与临床相关的抗Dr MAB的构象特异性之间是否存在相关性并表现出治疗作用，并为该领域提供一套新的工具（即，构象异构的抗 - HLA-DR和抗HLA-DQ mABS）进一步研究M1和M2 HLA II类在人类健康中的作用和疾病。