Lymphocyte-rich classic Hodgkin lymphoma (LR-CHL) is a rare subtype of Hodgkin lymphoma. Recent technical advances have allowed for the characterization of specific cross-talk mechanisms between malignant Hodgkin Reed-Sternberg (HRS) cells and different normal immune cells in the tumor microenvironment (TME) of CHL. However, the TME of LR-CHL has not yet been characterized at single-cell resolution. Here, using single-cell RNA sequencing (scRNA-seq), we examined the immune cell profile of 8 cell suspension samples of LR-CHL in comparison to 20 samples of the mixed cellularity (MC, 9 cases) and nodular sclerosis (NS, 11 cases) subtypes of CHL, as well as 5 reactive lymph node controls. We also performed multicolor immunofluorescence (MC-IF) on tissue microarrays from the same patients and an independent validation cohort of 31 pretreatment LR-CHL samples. ScRNA-seq analysis identified a unique CD4⁺ helper T cell subset in LR-CHL characterized by high expression of Chemokine C-X-C motif ligand 13 (CXCL13) and PD-1. PD-1⁺CXCL13⁺ T cells were significantly enriched in LR-CHL compared to other CHL subtypes, and spatial analyses revealed that in 46% of the LR-CHL cases these cells formed rosettes surrounding HRS cells. MC-IF analysis revealed CXCR5⁺ normal B cells in close proximity to CXCL13⁺ T cells at significantly higher levels in LR-CHL. Moreover, the abundance of PD-1⁺CXCL13⁺ T cells in the TME was significantly associated with shorter progression-free survival in LR-CHL (P = 0.032). Taken together, our findings strongly suggest the pathogenic importance of the CXCL13/CXCR5 axis and PD-1⁺CXCL13⁺ T cells as a treatment target in LR-CHL.

富含淋巴细胞的经典型霍奇金淋巴瘤 (LR-CHL) 是一种罕见的霍奇金淋巴瘤亚型。最近的技术进步使得能够表征 CHL 肿瘤微环境 (TME) 中恶性霍奇金里德-斯腾伯格 (HRS) 细胞与不同正常免疫细胞之间的特定串扰机制。然而，LR-CHL 的 TME 尚未在单细胞分辨率下表征。在这里，我们使用单细胞 RNA 测序（scRNA-seq）检查了 8 个 LR-CHL 细胞悬液样本的免疫细胞谱，与 20 个混合细胞（MC，9 例）和结节性硬化（NS， 11 例）CHL 亚型，以及 5 例反应性淋巴结对照。我们还对来自相同患者的组织微阵列和 31 个预处理 LR-CHL 样本的独立验证队列进行了多色免疫荧光 (MC-IF)。ScRNA-seq 分析确定了一个独特的 CD4⁺ LR-CHL 中的辅助 T 细胞亚群，其特征在于趋化因子 CXC 基序配体 13 (CXCL13) 和 PD-1 的高表达。与其他 CHL 亚型相比，PD-1 ⁺ CXCL13 ⁺ T 细胞在 LR-CHL 中显着富集，空间分析显示，在 46% 的 LR-CHL 病例中，这些细胞在 HRS 细胞周围形成玫瑰花结。MC-IF 分析显示 CXCR5 ⁺正常 B 细胞与 CXCL13 ⁺ T 细胞非常接近，在 LR-CHL 中的水平显着更高。此外，TME中 PD-1 ⁺ CXCL13 ⁺ T 细胞的丰度与 LR-CHL 中较短的无进展生存期显着相关（P= 0.032）。总之，我们的研究结果强烈表明 CXCL13/CXCR5 轴和 PD-1 ⁺ CXCL13 ⁺ T 细胞作为 LR-CHL 治疗靶点的致病重要性。