Intratumoral tertiary lymphoid structures (TLSs) have been associated with improved outcome in various cohorts of patients with cancer, reflecting their contribution to the development of tumor-targeting immunity. Here, we demonstrate that high-grade serous ovarian carcinoma (HGSOC) contains distinct immune aggregates with varying degrees of organization and maturation. Specifically, mature TLSs (mTLS) as forming only in 16% of HGSOCs with relatively elevated tumor mutational burden (TMB) are associated with an increased intratumoral density of CD8+ effector T (TEFF) cells and TIM3+PD1+, hence poorly immune checkpoint inhibitor (ICI)-sensitive, CD8+ T cells. Conversely, CD8+ T cells from immunologically hot tumors like non-small cell lung carcinoma (NSCLC) are enriched in ICI-responsive TCF1+ PD1+ T cells. Spatial B-cell profiling identifies patterns of in situ maturation and differentiation associated with mTLSs. Moreover, B-cell depletion promotes signs of a dysfunctional CD8+ T cell compartment among tumor-infiltrating lymphocytes from freshly isolated HGSOC and NSCLC biopsies. Taken together, our data demonstrate that - at odds with NSCLC - HGSOC is associated with a low density of follicular helper T cells and thus develops a limited number of mTLS that might be insufficient to preserve a ICI-sensitive TCF1+PD1+ CD8+ T cell phenotype. These findings point to key quantitative and qualitative differences between mTLSs in ICI-responsive vs ICI-irresponsive neoplasms that may guide the development of alternative immunotherapies for patients with HGSOC.

3rd Department of Surgery 1st Faculty of Medicine Charles University and University Hospital Motol Prague Czech Republic

Caryl and Israel Englander Institute for Precision Medicine New York NY USA

Czech Center for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences Vestec Czech Republic

Department of Gynaecology Obstetrics and Neonatology General University Hospital Prague 1st Faculty of Medicine Charles University Prague Czech Republic

Department of Gynecology and Obstetrics 1st Faculty of Medicine Charles University University Hospital Bulovka Prague Czech Republic

Department of Gynecology and Obstetrics Charles University 3rd Faculty of Medicine and University Hospital Kralovske Vinohrady Prague Czech Republic

Department of Gynecology and Obstetrics Charles University Faculty of Medicine and University Hospital Hradec Kralove Hradec Kralove Czech Republic

Department of Immunology Charles University 2nd Faculty of Medicine and University Hospital Motol Prague Czech Republic

Department of Obstetrics and Gynecology David Geffen School of Medicine University of California Los Angeles Los Angeles CA USA

Department of Pathology 1st Faculty of Medicine Charles University and General University Hospital Prague Czech Republic

Department of Pathology 3rd Faculty of Medicine and University Hospital Kralovske Vinohrady Prague Czech Republic

Department of Pathology and Molecular Medicine 2nd Faculty of Medicine Charles University and University Hospital Motol Prague Czech Republic

Department of Pneumology University Hospital Hradec Kralove Hradec Kralove Czech Republic

Department of Radiation Oncology Weill Cornell Medical College New York NY USA

Department of Thoracic Surgery Charles University 3rd Faculty of Medicine and Thomayer University Hospital Prague Czech Republic

Departments of Surgery and Biomedical Sciences Samuel Oschin Comprehensive Cancer Institute Cedars Sinai Medical Center West Hollywood CA USA

Laboratory of Tumor Immunology Institute of Microbiology of the Czech Academy of Sciences Prague Czech Republic

Oncology and Pneumology Department 2nd Faculty of Medicine Charles University and University Hospital Motol Prague Czech Republic

Ovarian Cancer Action Research Centre Department of Surgery and Cancer Imperial College London London UK

Ovarian Cancer Research Department of Biomedicine University Hospital Basel and University of Basel Basel Switzerland

Sandra and Edward Meyer Cancer Center New York NY USA

Sotio Biotech a s Prague Czech Republic

The Fingerland Department of Pathology Charles University Faculty of Medicine and University Hospital Hradec Kralove Hradec Kralove Czech Republic

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Chemotherapy Drives Tertiary Lymphoid Structures That Correlate with ICI-Responsive TCF1+CD8+ T Cells in Metastatic Ovarian Cancer