Fig. 5: IL-35 drives the conversion of human NK cells into an irreversible ILC1-like phenotype.

a Two-dimensional T-sne plots showing NK cell clustering based on residency surface marker profiles (CD103, CD9, CD49a) after 2, 4, and 8 days in culture (D2, D4, D8, respectively) with IL-2 + /− IL-35. Representative T-sne (upper left and lower) and quantification (%) (upper right) of ILC1-like cells as defined by the combined expression of CD103, CD9 and CD49a. Mean values ± S.D are shown (n = 3 individual donors). b, c Representative flow cytometry plots (left) and quantification of EOMES and T-BET MFI expression in NK cells cultured for 8 days in the presence of IL-2 with or without IL-35. Mean values ± S.D are shown (n = 6 and 5 individual donors, respectively). d Representative flow cytometry plots (left) and quantification (right) of % and MFI CD9, CD103, and CD49a expression in ILC1-like cells after 8 days of culture in the presence of IL-2 with IL-35 according to their proliferation status (cycling vs non-cycling) based on CTV fluorescence intensity. Mean values ± S.D are shown (n = 3 individual donors). e Quantification of IFN-γ expression in NK cells at day 7 following 48 h (2 d) in indicated culture conditions. Results are expressed as relative MFI compared to control condition IL-2 (5 d) -> IL-2 (2 d). Mean values ± S.D are shown (n = 5 individual donors). f Representative flow cytometry plots (left) and quantification (%) of CD9⁺ CD103⁺ NK (right) after 5 days (5 d) in culture with IL-2 with or without IL-35 (D5) and at day 7 (D7) following 48 h (2 d) in indicated culture conditions. Mean values ± S.D are shown (n = 5 individual donors). Statistical significance was determined using paired T test. Source data are provided as a Source Data file.