Purpose: Immunotherapies have led to a paradigm shift in the treatment of hepatocellular carcinoma (HCC). Studies have revealed the single-cell catalogues of tumor-infiltrating immune cells and the trajectories of their differentiation. Nevertheless, the spatial distribution of these immune cells with distinct phenotypes in tumor microenvironment and their clinicopathological significance in resectable and unresectable HCCs are still largely unclear. Experimental design: We analyzed the spatial dynamics of intratumoral CD4 and CD8 T cells and their association with B and plasma cells using 283 surgically resected HCCs, 58 unresectable HCC samples before combined immunotherapy (atezolizumab plus bevacizumab [Atezo+Bev]), and autopsy specimens from 50 cases of advanced-stage HCCs through multiplex immunohistochemistry combined with transcriptomic and driver gene mutation analysis. Classification based on the spatial dynamics of T and B cell responses (refined immunosubtype) was developed and its clinicopathological significance was analyzed. Results: We found that stem-like CD4 and CD8 T cells were mainly observed in T cell aggregates. Differentiation of T follicular helper cells were associated with the development of tertiary lymphoid structures, whereas differentiation of CD4 TCXCL13 cells with phenotype resembling T peripheral helper cells were associated with the development of lymphoplasmacytic microenvironment. The refined immunosubtype could predict clinical outcomes of resectable HCC after surgery and unresectable HCC after Atezo+Bev therapy. The immune microenvironment of metastatic lesions tended to reflect those of primary lesions. Conclusions: We revealed the spatial dynamics of T and B cell response in HCC, which is closely associated with the clinical outcome after surgical resection or Atezo+Bev therapy.

中文翻译：

---

T 细胞和 B 细胞反应的空间动力学预测可切除和不可切除肝细胞癌的临床结局


目的：免疫疗法导致肝细胞癌 （HCC） 治疗的范式转变。研究揭示了肿瘤浸润免疫细胞的单细胞目录及其分化轨迹。然而，这些在肿瘤微环境中具有不同表型的免疫细胞的空间分布及其在可切除和不可切除的 HCC 中的临床病理意义在很大程度上仍不清楚。实验设计： 我们使用 283 例手术切除的 HCC、58 例联合免疫治疗前不可切除的 HCC 样本 （atezolizumab 加贝伐珠单抗 [Atezo + Bev]）和 50 例晚期 HCC 的尸检标本，通过多重免疫组化结合转录组学和驱动基因突变分析，分析了瘤内 CD4 和 CD8 T 细胞的空间动力学及其与 B 细胞和浆细胞的关联。开发了基于 T 和 B 细胞反应空间动力学 （精细免疫亚型） 的分类，并分析了其临床病理意义。结果： 我们发现干细胞样 CD4 和 CD8 T 细胞主要在 T 细胞聚集体中观察到。滤泡辅助性 T 细胞的分化与三级淋巴结构的发育有关，而具有类似于 T 外周辅助细胞表型的 CD4 TCXCL13细胞的分化与淋巴浆细胞微环境的发育有关。精细免疫亚型可以预测术后可切除 HCC 和 Atezo+Bev 治疗后不可切除 HCC 的临床结局。转移病灶的免疫微环境往往反映原发病灶的免疫微环境。 结论： 我们揭示了 HCC 中 T 细胞和 B 细胞反应的空间动力学，这与手术切除或 Atezo+Bev 治疗后的临床结局密切相关。