PO.IM01.03 · 免疫学
推进新型疫苗：在HLA-A2.1/HLA-DRB1*1.1人源化小鼠中的临床前评价
Advancing novel vaccines: Preclinical evaluation in HLA-A2.1/HLA-DRB1*1.1 humanized mice
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
人白细胞抗原（HLA）系统在协调适应性免疫应答中发挥核心作用。虽然现有的单等位基因人源化模型颇具价值，但越来越需要能够更充分地重现人HLA I类和II类限制性免疫的临床前平台，以评估复杂的免疫疗法。为此，我们开发了一种新型MHC I/II双人源化小鼠模型，即B-HLA-A2.1/HLA-DRB1*1.1小鼠。B-HLA-A2.1/HLA-DRB1*1.1模型通过将人HLA-A02:01和HLA-DRB1*01:01基因共整合到C57BL/6背景中生成。通过流式细胞术在脾脏、外周血和骨髓白细胞上确认人HLA-A2.1和HLA-DRB1蛋白表达。进行了全面的免疫细胞图谱分析以表征T和B细胞群。该模型的功能性在使用人表位靶向疫苗的治疗性肿瘤接种研究中得到验证。流式细胞术分析确认纯合子小鼠中HLA-A2.1和HLA-DRB1蛋白均稳定而强健的细胞表面表达。脾脏、血液和淋巴结中CD8+ T细胞的频率显著降低，而CD4+ T细胞的频率显著增加，表明HLA-A02:01和HLA-DRB1*01:01的引入影响了T细胞的发育。包括血液学、血清生化以及主要器官组织病理学检查（H&E染色）在内的其他分析表明无自发性病理，确认了该模型的整体健康和生理稳定性。在疫苗疗效研究中，与对照相比，用人抗原特异性疫苗免疫显著抑制双人源化小鼠中的肿瘤生长，证明了该模型产生功能性HLA限制性免疫应答的能力。我们已成功开发并初步验证了新型B-HLA-A2.1/HLA-DRB1*1.1双人源化小鼠模型。它代表了一种强大且具有生理相关性的临床前工具，用于研究包括疫苗和免疫检查点抑制剂在内的联合免疫疗法，并在更完整的类人背景下研究抗肿瘤和感染性疾病免疫。
查看英文原文 English abstract
The human leukocyte antigen (HLA) system plays a central role in orchestrating adaptive immune responses. While existing single-allele humanized models have been valuable, there is a growing need for preclinical platforms that more fully recapitulate human HLA class I and II restricted immunity to evaluate complex immunotherapies. To address this, we developed a novel MHC I/II dual-humanized mouse model, the B-HLA-A2.1/HLA-DRB1*1.1 mouse.The B-HLA-A2.1/HLA-DRB1*1.1 model was generated by co-integrating the human HLA-A02:01 and HLA-DRB1*01:01 genes into the C57BL/6 background. Human HLA-A2.1 and HLA-DRB1 protein expression was confirmed by flow cytometry on spleen, peripheral blood, and bone marrow leukocytes. Comprehensive immune cell profiling was performed to characterize T and B cell populations. The model's functionality was validated in a therapeutic tumor vaccination study using a human epitope-targeting vaccine.Flow cytometric analysis confirmed stable and robust cell surface expression of both HLA-A2.1 and HLA-DRB1 proteins in homozygous mice. The frequency of CD8+ T cells in the spleen, blood, and lymph node was significantly decreased, while the frequency of CD4+ T cells was significantly increased, demonstrating that the introduction of HLA-A02:01 and HLA-DRB1*01:01 affected the development of T cells. Additional analyses, including hematology, serum biochemistry, and histopathological examination (H&E staining) of major organs, indicated no spontaneous pathology, confirming the overall health and physiological stability of the model. In a vaccine efficacy study, immunization with a human antigen-specific vaccine significantly inhibited tumor growth in the dual-humanized mice compared to controls, demonstrating the model's capability to mount a functional HLA-restricted immune response.We have successfully developed and preliminarily validated the novel B-HLA-A2.1/HLA-DRB1*1.1 dual-humanized mouse model. It represents a powerful and physiologically relevant preclinical tool for investigating combination immunotherapies, including vaccines and immune checkpoint inhibitors, and for studying anti-tumor and infectious disease immunity within a more complete human-like context.
利益披露 Disclosure
D. Emerson, None..
Z. Niu, None..
S. Zhang, None..
X. Zhou, None.