Extended Data Fig. 6: Intratumoral Hobit⁺ cells are dispensable for tumor control and checkpoint inhibitor response.

(a-d) AT3-OVA tumor-bearing Hobit^TomCreRosa26^DTA mice and Hobit^TomCre control (Ctrl) mice were analyzed near tumor endpoint. (a) Numbers per mg of tumor of Hobit⁺ CD69⁺ cells and Hobit⁺ CD103⁺ cells within CD44⁺ CD8 + T cells in the tumor (Hobit^TomCreRosa26^DTA n = 14 or Ctrl=17). (b) Numbers of indicated CD8⁺ T cell subsets within the tdLN (Hobit^TomCreRosa26^DTA n = 5 or Ctrl n = 4). (c) Numbers of indicated immune subsets in tumors of Hobit^TomCreRosa26^DTA (n = 5) mice and Hobit^TomCre control (Ctrl) (n = 4) mice. (d) Numbers of indicated immune subsets in tdLNs of Hobit^TomCreRosa26^DTA (n = 5) mice and Hobit^TomCre control (Ctrl) (n = 4) mice. (e,f) AT3-OVA tumor-bearing Hobit^TomCreRosa26^DTA mice and Hobit^TomCre control (Ctrl) mice were treated with ICB. (e) Individual tumor volume curves of Ctrl (upper) and Hobit^TomCreRosa26^DTA mice (lower) tumor-bearing mice treated with ICB (right) or left untreated (left) (untreated Hobit^TomCreRosa26^DTA n = 16, ICB-treated Hobit^TomCreRosa26^DTA n = 15, untreated Ctrl=16, ICB-treated Ctrl=12). (f) Contour plots of TIM-3, PD-1 (left) and GzmB (right) expression by CD44⁺ CD8⁺ T cells within the tumor. (g-i) Hobit^TomCreRosa26^DTA mice and Hobit^TomCre control (Ctrl) mice were orthotopically inoculated with AT3-OVA into the fourth MFP and were treated with ICB on days 11, 13, 16 and 19. CD44⁺ PD-1⁻ CD8⁺ cells at day 50 post-inoculation were analyzed. (g) Contour plots of Hobit versus CD69 expression within the local MFP. (h) Numbers per gram of MFP of CD69, CD49a and CD103 expressing CD44⁺ CD8⁺ T cells in Hobit^TomCreRosa26^DTA (n = 7) and Ctrl (n = 7) mice. (i) Average AT3-OVA tumor volumes till day 50 post-inoculation (left) of treated (n = 9-11) and untreated (n = 4) mice. Individual tumor volume curves of Ctrl (upper) and Hobit^TomCreRosa26^DTA mice (lower) tumor-bearing mice treated with ICB (right) or left untreated (left). (j,k) MC38-OVA tumor-bearing Hobit^TomCreRosa26^DTA mice and Hobit^TomCre control (Ctrl) mice were analyzed near tumor endpoint. (j) Contour plots (left) of Hobit versus CD69 (top, left) or CD103 (bottom left) and quantifications per mg of tumor (right) of CD69⁺ (top, right) and CD103⁺ (bottom, right) CD8⁺ CD44⁺ T cells within the tumor of Hobit^TomCreRosa26^DTA (n = 4) and Ctrl (n = 4) mice. (k) Average MC38-OVA tumor volumes till day 24 post-inoculation in ICB-treated and untreated mice and individual tumor volume curves of Ctrl (upper) and Hobit^TomCreRosa26^DTA mice (lower) tumor-bearing mice treated with ICB (right) or left untreated (left) (untreated Hobit^TomCreRosa26^DTA n = 9, ICB-treated Hobit^TomCreRosa26^DTA n = 10, untreated Ctrl=10, ICB-treated Ctrl=10). Flow cytometry plots are representative. Dots in graphs represent individual mice; horizontal lines and error bars of bar graphs indicate means and ±SEM. Data are pooled from two independent experiments or representative of two independent experiment (b-d,j). P values are from two-tailed unpaired t-tests (a-d,h,j) or 2-way ANOVA with Turkey’s comparison test (i,k). P > 0.05, not significant (n.s.).