Department of Mathematics and Statistics, University of Massachusetts Amherst, Amherst, MA, USA.
Sci Rep. 2021 Feb 22;11(1):4338. doi: 10.1038/s41598-021-83767-z.
Since the outcome of treatments, particularly immunotherapeutic interventions, depends on the tumor immune micro-environment (TIM), several experimental and computational tools such as flow cytometry, immunohistochemistry, and digital cytometry have been developed and utilized to classify TIM variations. In this project, we identify immune pattern of clear cell renal cell carcinomas (ccRCC) by estimating the percentage of each immune cell type in 526 renal tumors using the new powerful technique of digital cytometry. The results, which are in agreement with the results of a large-scale mass cytometry analysis, show that the most frequent immune cell types in ccRCC tumors are CD8+ T-cells, macrophages, and CD4+ T-cells. Saliently, unsupervised clustering of ccRCC primary tumors based on their relative number of immune cells indicates the existence of four distinct groups of ccRCC tumors. Tumors in the first group consist of approximately the same numbers of macrophages and CD8+ T-cells and and a slightly smaller number of CD4+ T cells than CD8+ T cells, while tumors in the second group have a significantly high number of macrophages compared to any other immune cell type (P-value [Formula: see text]). The third group of ccRCC tumors have a significantly higher number of CD8+ T-cells than any other immune cell type (P-value [Formula: see text]), while tumors in the group 4 have approximately the same numbers of macrophages and CD4+ T-cells and a significantly smaller number of CD8+ T-cells than CD4+ T-cells (P-value [Formula: see text]). Moreover, there is a high positive correlation between the expression levels of IFNG and PDCD1 and the percentage of CD8+ T-cells, and higher stage and grade of tumors have a substantially higher percentage of CD8+ T-cells. Furthermore, the primary tumors of patients, who are tumor free at the last time of follow up, have a significantly higher percentage of mast cells (P-value [Formula: see text]) compared to the patients with tumors for all groups of tumors except group 3.
由于治疗效果（尤其是免疫治疗干预）取决于肿瘤免疫微环境（TIM），因此已经开发并利用了几种实验和计算工具，例如流式细胞术、免疫组织化学和数字细胞术，以对 TIM 变化进行分类。在本项目中，我们通过使用数字细胞术估计 526 个肾肿瘤中每种免疫细胞类型的百分比，来鉴定透明细胞肾细胞癌（ccRCC）的免疫模式。这些结果与大规模质谱细胞分析的结果一致，表明 ccRCC 肿瘤中最常见的免疫细胞类型是 CD8+T 细胞、巨噬细胞和 CD4+T 细胞。突出的是，基于其相对免疫细胞数量对 ccRCC 原发性肿瘤进行无监督聚类表明存在四种不同类型的 ccRCC 肿瘤。第一组肿瘤中巨噬细胞和 CD8+T 细胞的数量大致相同，而 CD4+T 细胞的数量略低于 CD8+T 细胞，而第二组肿瘤中巨噬细胞的数量明显高于任何其他免疫细胞类型（P 值 [公式：见正文]）。第三组 ccRCC 肿瘤中 CD8+T 细胞的数量明显高于任何其他免疫细胞类型（P 值 [公式：见正文]），而第四组肿瘤中巨噬细胞和 CD4+T 细胞的数量大致相同，而 CD8+T 细胞的数量明显低于 CD4+T 细胞（P 值 [公式：见正文]）。此外，IFNG 和 PDCD1 的表达水平与 CD8+T 细胞的百分比之间存在高度正相关，且肿瘤分期和分级越高，CD8+T 细胞的百分比越高。此外，在最后一次随访时无肿瘤的患者的原发性肿瘤中，肥大细胞的百分比明显高于所有肿瘤组（除第 3 组外）的患者（P 值 [公式：见正文]）。
