MHCI类(MHC-I)损失在非小细胞肺癌(NSCLC)中是常见的,使得肿瘤细胞对T细胞裂解具有抗性。NK细胞杀死MHC-I缺陷的肿瘤细胞,尽管以前的工作表明它们存在于NSCLC边缘,他们功能受损。内,我们评估了NK细胞和CD8T细胞的浸润和活化是否随MHC-I表达而变化。
■我们使用单染色免疫组织化学(IHC)和Kaplan-Meier分析来测试NK细胞和CD8T细胞浸润对总体和无病存活的影响。为了描述MHC-I不同肺癌的免疫协变量,我们使用多重免疫荧光(mIF)成像,然后进行多变量统计建模.为了确定IFNγ激活和非激活淋巴细胞之间的浸润和细胞间通讯的差异,我们开发了一个计算管道,从mIF图像中枚举单细胞邻域,然后进行多元判别分析。
■肿瘤细胞MHC-I表达的空间定量揭示了肿瘤内和肿瘤间的异质性,这与局部淋巴细胞景观有关。IHC分析显示,患者肿瘤中的高CD56+细胞数量与无病生存率(DFS)(HR=0.58,p=0.064)和总体生存率(OS)(HR=0.496,p=0.041)呈正相关。OS相关性随着CD56+和CD8+细胞的高计数而增强(HR=0.199,p<1×10-3)。mIF成像和多变量判别分析显示在MHC-I携带肿瘤中CD3+CD8+T细胞和CD3-CD56+NK细胞富集(p<0.05)。为了推断功能细胞状态和本地细胞间通信的关联,我们分析了空间单细胞邻域谱来描绘IFNγ+/-NK细胞和T细胞的细胞环境。我们发现,与IFNγ-NK细胞和CD8T细胞相比,IFNγNK和CD8T细胞与其他IFNγ淋巴细胞的相关性更高(p<1×10-30)。此外,IFNγ+淋巴细胞最常聚集在MHC-I+肿瘤细胞附近。
■肿瘤浸润NK细胞和CD8T细胞共同影响NSCLC肿瘤进展的控制。NK和CD8T细胞的联合在携带MHC-I的肿瘤中最为明显,尤其是在IFNγ的存在下。在近邻分析中IFNγ+NK细胞与其他IFNγ+淋巴细胞的频繁共定位表明NSCLC淋巴细胞活化是协同调节的。
■MHC-I丢失在NSCLC中经常发生,并且与肿瘤微环境(TME)中免疫力下降相对应。NK细胞识别“自身缺失”靶标,并可用于靶向MHC-I缺失的NSCLC肿瘤。虽然NK细胞在肿瘤边缘的存在已被证明在NSCLC中,他们被证明在这种环境中失去了功能。
我们开发了空间分析管道,利用TME在单细胞分辨率下的局部异质性来测试NK细胞和T细胞是否共同促进NSCLC的抗肿瘤免疫。我们发现高密度的肿瘤浸润NK细胞与DFS相对应,这种相关性在高符合CD8T细胞的患者中增加,尤其是中央肿瘤。有趣的是,在带有MHC-I的肿瘤中发现两种细胞类型聚集在一起,特别是当两者都表达IFNγ时,提示协调的淋巴细胞活性可能增强NSCLC的免疫控制。
■这项研究为开发同时增加NK和T细胞抗肿瘤免疫的新型免疫疗法提供了理论基础。NK细胞与NSCLC患者生存和免疫效应活性增加的关联,甚至在MHC-I缺陷的肿瘤中,进一步强调需要设计和部署NK细胞激活策略,这些策略可能在CD8T细胞难治性NSCLC中非常有效.
UNASSIGNED: MHC class I (MHC-I) loss is frequent in non-small cell lung cancer (NSCLC) rendering tumor cells resistant to T cell lysis. NK cells kill MHC-I-deficient tumor cells, and although previous work indicated their presence at NSCLC margins, they were functionally impaired. Within, we evaluated whether NK cell and CD8 T cell infiltration and activation vary with MHC-I expression.
UNASSIGNED: We used single-stain immunohistochemistry (IHC) and Kaplan-Meier analysis to test the effect of NK cell and CD8 T cell infiltration on overall and disease-free survival. To delineate immune covariates of MHC-I-disparate lung cancers, we used multiplexed immunofluorescence (mIF) imaging followed by multivariate statistical modeling. To identify differences in infiltration and intercellular communication between IFNγ-activated and non-activated lymphocytes, we developed a computational pipeline to enumerate single cell neighborhoods from mIF images followed by multivariate discriminant analysis.
UNASSIGNED: Spatial quantitation of tumor cell MHC-I expression revealed intra- and inter-tumoral heterogeneity, which was associated with the local lymphocyte landscape. IHC analysis revealed that high CD56+ cell numbers in patient tumors were positively associated with disease-free survival (DFS) (HR=0.58, p=0.064) and overall survival (OS) (HR=0.496, p=0.041). The OS association strengthened with high counts of both CD56+ and CD8+ cells (HR=0.199, p<1×10-3). mIF imaging and multivariate discriminant analysis revealed enrichment of both CD3+CD8+ T cells and CD3-CD56+ NK cells in MHC-I-bearing tumors (p<0.05). To infer associations of functional cell states and local cell-cell communication, we analyzed spatial single cell neighborhood profiles to delineate the cellular environments of IFNγ+/- NK cells and T cells. We discovered that both IFNγ+ NK and CD8 T cells were more frequently associated with other IFNγ+ lymphocytes in comparison to IFNγ- NK cells and CD8 T cells (p<1×10-30). Moreover, IFNγ+ lymphocytes were most often found clustered near MHC-I+ tumor cells.
UNASSIGNED: Tumor-infiltrating NK cells and CD8 T cells jointly affected control of NSCLC tumor progression. Co-association of NK and CD8 T cells was most evident in MHC-I-bearing tumors, especially in the presence of IFNγ. Frequent co-localization of IFNγ+ NK cells with other IFNγ+ lymphocytes in near-neighbor analysis suggests NSCLC lymphocyte activation is coordinately regulated.