Analysis of Antibody Induction by Macrophages Treated Ex Vivo with Human Proteins in Mice.

IF 1.6 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Reports of Biochemistry and Molecular Biology Pub Date : 2023-01-01 DOI:10.52547/rbmb.11.4.694
Nurtleu Malika, Adish Zhansaya, Mukanov Kasym, Tursunov Kanat, Ramankulov Yerlan, Mukantayev Kanatbek
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Abstract

Background: Macrophages are essential cellular components in various body tissues and tumor microenvironments. The high infiltration of macrophages into the tumor microenvironment determines the importance of ex vivo treatment of personalized macrophages with recombinant cytotoxic T-lymphocyte-associated protein 4 (rCTLA-4), programmed death-ligand 1 (rPD-L1), and programmed cell death protein 1 (rPD-1) proteins to block immune checkpoints.

Methods: We investigated the development of humoral immunity against CTLA-4, PD-L1, and PD-1 receptors by introducing macrophages treated ex vivo with the corresponding proteins into mice. Peritoneal macrophages from BALB/c mice were cultured in medium containing recombinant human CTLA-4, PD-L1, and PD-1 proteins. Macrophages processing recombinant proteins were analyzed via immunofluorescence staining using antibodies against CTLA-4, PD-L1, and PD-1. The treated macrophages were administered intraperitoneally to mice to induce anti-CTLA-4, anti-PD-L1, and anti-PD-1 antibodies. The antibody titer in vaccinated mice was determined via enzyme-linked immunosorbent assays, followed by statistical analysis of the results. The specificity of the antibodies was determined via immunofluorescence staining in MCF7 cells.

Results: The ex vivo treatment of macrophages with rCTLA-4, rPD-L1, and rPD-1 induced the formation of specific antibodies in vaccinated mice. The various rPD-L1 and rPD-1 concentrations used to treat macrophages had no significant effect on the specific antibody titers, while the anti-rCTLA-4 titer was dependent on the protein concentration in the culture medium. Immunofluorescence analysis revealed that anti-CTLA-4 and PD-L1 antibodies reacted with MCF7 cells.

Conclusion: The ex vivo treatment of macrophages with rCTLA-4, rPD-L1, and rPD-1 can help induce humoral immunity and develop new approaches for cancer immunotherapy.

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人蛋白体外处理小鼠巨噬细胞诱导抗体的研究。
背景:巨噬细胞是机体各种组织和肿瘤微环境中必不可少的细胞成分。巨噬细胞在肿瘤微环境中的高度浸润决定了用重组细胞毒性t淋巴细胞相关蛋白4 (rCTLA-4)、程序性死亡配体1 (rPD-L1)和程序性细胞死亡蛋白1 (rPD-1)蛋白对个体化巨噬细胞进行体外治疗以阻断免疫检查点的重要性。方法:通过将巨噬细胞引入小鼠体内,研究其对CTLA-4、PD-L1和PD-1受体的体液免疫的发展。BALB/c小鼠腹腔巨噬细胞在含有重组人CTLA-4、PD-L1和PD-1蛋白的培养基中培养。利用CTLA-4、PD-L1和PD-1抗体免疫荧光染色分析巨噬细胞处理重组蛋白。将处理后的巨噬细胞腹腔注射小鼠,诱导抗ctla -4、抗pd - l1和抗pd -1抗体。通过酶联免疫吸附法测定接种小鼠的抗体滴度,然后对结果进行统计分析。抗体在MCF7细胞中通过免疫荧光染色测定特异性。结果:用rCTLA-4、rPD-L1和rPD-1体外处理巨噬细胞可诱导免疫小鼠产生特异性抗体。不同浓度的rPD-L1和rPD-1治疗巨噬细胞对特异性抗体滴度无显著影响,而抗rctla -4滴度依赖于培养基中的蛋白浓度。免疫荧光分析显示抗ctla -4和PD-L1抗体与MCF7细胞发生反应。结论:rCTLA-4、rPD-L1和rPD-1体外治疗巨噬细胞有助于诱导体液免疫,为肿瘤免疫治疗开辟新的途径。
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来源期刊
Reports of Biochemistry and Molecular Biology
Reports of Biochemistry and Molecular Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-
CiteScore
2.80
自引率
23.50%
发文量
60
审稿时长
10 weeks
期刊介绍: The Reports of Biochemistry & Molecular Biology (RBMB) is the official journal of the Varastegan Institute for Medical Sciences and is dedicated to furthering international exchange of medical and biomedical science experience and opinion and a platform for worldwide dissemination. The RBMB is a medical journal that gives special emphasis to biochemical research and molecular biology studies. The Journal invites original and review articles, short communications, reports on experiments and clinical cases, and case reports containing new insights into any aspect of biochemistry and molecular biology that are not published or being considered for publication elsewhere. Publications are accepted in the form of reports of original research, brief communications, case reports, structured reviews, editorials, commentaries, views and perspectives, letters to authors, book reviews, resources, news, and event agenda.
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