外周血t淋巴细胞对模型系统肾细胞癌细胞作用的研究

A. Kuzevanova, O. Khalmurzaev, A. A. Borunova, N. Apanovich, T. Zabotina, A. A. Alimov, V. Matveev, A. Karpukhin
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摘要

背景。基于免疫检查点抑制剂的免疫疗法的引入显著提高了肾癌治疗的有效性。然而,并非所有患者都对这种治疗有反应,也没有可靠的预测指标。因此,开发一种模型系统来评估细胞对肿瘤的免疫反应似乎是一项紧迫的任务。开发一种评估t细胞免疫反应的模型是本研究的重点。材料和方法。在标准无菌条件下获得原代肿瘤细胞培养和外周血t细胞部分。通过抗cd3和抗cd28抗体激活t细胞。采用RTCA xCELLigence生物传感器技术(ACEA Biosciences, USA)评估细胞指数。将同一患者的肿瘤细胞和t细胞一起培养,以评估肿瘤细胞群的生长速度。每隔30分钟测量一次。观察时间为24小时。研究表明,非活化t细胞不影响培养癌细胞的增殖特性。相反,活化的t细胞抑制了癌细胞的增殖特性,这与Т-cells携带HLA-DR受体(CD3+ HLA-DR +)的比例因活化而增加有关。肿瘤特异性t细胞活性可导致三种后果:缺乏作用、增殖特性的部分抑制和肿瘤细胞的完全死亡。后一种情况下,用流式细胞术检测这些细胞的缺失。开发的方法使得有可能评估T细胞的细胞毒性特性与肿瘤细胞在一个特定的病人。这种方法的优点是可以在免疫检查点抑制剂存在的情况下进行测量。所提出的方法可用于评估个性化治疗框架内的治疗方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Study of the action of peripheral blood T-lymphocytes on renal cell carcinoma cells in model systems
Background. The introduction of immunotherapy based on immune checkpoint inhibitors has significantly improve the effectiveness of kidney cancer treatment. Nevertheless, not all patients respond to such treatment and there are no reliable predictive markers. Therefore, the development of a model system for assessing the cellular immune response to a tumor seems to be an urgent task.Aim. Development of a model to assess the T-cell immune response was the focus of this study.Materials and methods. Primary tumor cell culture and peripheral blood T-cell fraction were obtained under standard sterile conditions. T-cell activation were perform via anti-CD3 and anti-CD28 antibodies. The cell index was assessed using the RTCA xCELLigence biosensor technology (ACEA Biosciences, USA).Results. Tumor and T-cells from the same patient were cultured together to assess the growth rate of the tumor cell population. Measurements were taken at 30-minute intervals. The duration of observation was 24 hours. It has been shown that non-activated T-cells do not affect the proliferative properties of cultured cancer cells. On the contrary, activated T-cells suppressed the proliferative properties of cancer cells, which was associated with an increase in the proportion of Т-cells carrying the HLA-DR receptor (CD3+HLA–DR+) because of activation. Tumor-specific T-cell activity can lead to three consequences: lack of effect, partial suppression of proliferative properties, and complete death of tumor cells. In the latter case, the absence of such cells was determined by flow cytometry.Conclusion. The developed approach makes it possible to evaluate the cytotoxic properties of T cells in relation to tumor cells in a particular patient. The advantage of this method is that the measurement can be carried out in the presence of immune checkpoint inhibitors. The proposed method may be useful for evaluating the treatment regimen within the framework of personalized therapy.
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