Abstract 6468: Dissecting the immune landscape in EBV+ classical Hodgkin lymphoma following IL-23 inhibition using single-cell hyperplex immunofluorescence analysis: A case report

Abstract Background: The immune tumor microenvironment (TME) in Epstein-Barr Virus (EBV)+ classic Hodgkin lymphoma (cHL) displays a T helper (Th) 1 profile typical of effective antitumor response with increased infiltration of CD8+ T-cells. cHL cases showing evidence of interleukin (IL)-23 /IL-17 axis activation are usually EBV-. The TME of EBV+ cHL developed after IL-23 repressive therapy has not been reported. We report the TME characteristics of EBV+ cHL in a psoriasis patient on IL-23 inhibition therapy. Case Presentation: A 40-year-old male with chronic plaque psoriasis underwent Guselkumab (IL-23 inhibitor) treatment for four months before developing bulky cervical lymphadenopathy. Excisional biopsy revealed Hodgkin and Reed-Sternberg (HRS) cells, predominantly in the interfollicular area, with weak to moderate expression of B-cell markers (PAX-5, CD20, CD79a) along with MUM1, CD30 and CD200. Importantly, HRS cells were strongly positive for EBER by in situ hybridization, showing a latency pattern IIa (LMP-1+, EBNA-2-) with a viral load of 75 IU/ml. A diagnosis of EBV+ cHL, possibly iatrogenic/related to IL23 inhibitor therapy was proposed. Hypothesis: IL-23/IL-17 axis promoted the reprogramming of the TME, fostering the emergence of EBV-driven HRS cells. Methods: Excised lymph node sections were analyzed using an automated sequential immunofluorescence (seqIF™) protocol (Lunaphore COMET™), tailored for the TME of cHL. This enabled the spatial mapping of 9 markers (CD20, T-bet, CD4, CD163, CCR4, CCR6, HLA-DR, Ki67, CTLA4). Utilizing the HORIZON™ image analysis software, which integrates single-cell segmentation and rule-based classification algorithms, we identified 8 distinct cell populations, including HRS cells, B cell subsets, Th1, Th2, Th17, Tregs, and macrophage subsets. Additionally, this unbiased approach allowed for the precise measurement of immune exhaustion and proliferation markers expression. Findings: The seqIF™ analysis revealed an immunosuppressive TME showing predominance of Th2 cells (CCR4+, CCR6-), numerous CTLA-4+ immunosuppressive CD4+ T cells, a very low Th1/Th2 ratio and a significant reduction of Th17 cells. Our study suggests reprogramming of Th17 to Th2 response following IL-23 inhibitory therapy, promoting the development of EBV+ cHL. This contrasts with the Th1 profile reported for the TME of EBV+ cHL. Conclusions: This single case study highlights the utility of spatial proteomics mapping immune interactions in the TME of EBV+ cHL. A larger study of EBV+ and EBV- cHL cases is planned to further characterize these TMEs, including fibroblastic populations, and identify potential immuno-oncology targets. These findings could guide personalized treatments and identify EBV+ cases unsuitable for IL-23 inhibitors to mitigate cHL risk. Citation Format: Janusz Franco-Barraza, Bhaumik Shah, Mariia Dmitrieva, Joanna Kowal, Erica Geneletti, Mariusz Wasik, Reza Nejati, Edna Cukierman. Dissecting the immune landscape in EBV+ classical Hodgkin lymphoma following IL-23 inhibition using single-cell hyperplex immunofluorescence analysis: A case report [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6468.