Immune infiltration is typically quantified using cellular density, not accounting for cellular clustering. Tumor-associated macrophages (TAM) activate oncogenic signaling through paracrine interactions with tumor cells, which may be better reflected by local cellular clustering than global density metrics. Using multiplex immunohistochemistry and digital pathologic analysis we quantified cellular density and cellular clustering for myeloid cell markers in 129 regions of interest from 55 samples from 35 patients with metastatic ccRCC. CD68+ cells were found to be clustered with tumor cells and dispersed from stromal cells, while CD163+ and CD206+ cells were found to be clustered with stromal cells and dispersed from tumor cells. CD68+ density was not associated with OS, while high tumor/CD68+ cell clustering was associated with significantly worse OS. These novel findings would not have been identified if immune infiltrate was assessed using cellular density alone, highlighting the importance of including spatial analysis in studies of immune cell infiltration of tumors.

中文翻译：

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CD68 +肿瘤相关巨噬细胞与肿瘤细胞的空间聚集与转移性透明细胞肾细胞癌的总体生存率下降有关。

免疫浸润通常使用细胞密度进行定量，而不考虑细胞聚集。肿瘤相关巨噬细胞（TAM）通过旁分泌与肿瘤细胞的相互作用激活致癌信号，与全局密度指标相比，局部细胞簇可能更好地反映了这一现象。使用多重免疫组织化学和数字病理分析，我们从35例转移性ccRCC患者的55个样本中，对129个目标区域中的髓样细胞标记物进行了细胞密度和细胞聚类定量分析。发现CD68 +细胞与肿瘤细胞成簇并从基质细胞分散，而发现CD163 +和CD206 +细胞与基质细胞成簇并从肿瘤细胞分散。CD68 +密度与操作系统无关，而高肿瘤/ CD68 +细胞聚集与OS明显恶化有关。如果仅用细胞密度评估免疫浸润度，就不会鉴定出这些新颖的发现，突显了在肿瘤免疫细胞浸润研究中纳入空间分析的重要性。