Table 3 Characteristics of hot, cold and mixed clusters (N = 514 for T-cell priming marker expression; N = 388 for other variables including histologies, MSI, TMB and PD-L1 status) (see Supplemental Table 1 for T-cell priming marker functions)^a.

T-cell priming markers	Hot cluster (N = 89)^b high expression N (%)	Cold cluster (N = 249)^c high expression N (%)	Mixed cluster (N = 176)^d high expression N (%)
CD27	61 (68.5%)	8 (3.2%)	29 (16.5%)
CD28	58 (65.2%)	4 (1.6%)	40 (22.7%)
CD40	36 (40.4%)	21 (8.4%)	57 (32.4%)
CD40LG	53 (60.0%)	7 (2.8%)	44 (25.0%)
CD80	54 (60.7%)	10 (4.0%)	51 (29.0%)
CD86	47 (52.8%)	1 (0.4%)	36 (20.5%)
CD137	48 (53.9%)	5 (2.0%)	24 (13.6%)
GITR	46 (51.7%)	25 (10.0%)	28 (15.9%)
GZMB	45 (50.6%)	20 (8.0%)	20 (11.4%)
ICOS	50 (56.2%)	3 (1.2%)	17 (9.7%)
ICOSLG	44 (49.4%)	75 (30.1%)	73 (41.5%)
IFNG	34 (38.2%)	3 (1.2%)	14 (8.0%)
OX40	51 (57.3%)	20 (8.0%)	51 (29.0%)
OX40LG	32 (36.0%)	33 (13.3%)	54 (30.7%)
TBX21	52 (58.4%)	9 (3.6%)	26 (14.8%)

Other variables	Hot cluster (N = 61) N (%)	Cold cluster (N = 203) N (%)	Mixed cluster (N = 124) N (%)	P value
MSI Unstable	4 (6.6%)	3 (1.5%)	7 (5.6%)	0.059
TMB ≥ 10 mutations/mb	6 (9.8%)	15 (7.4%)	10 (8.1%)	0.83
PD-L1 ≥ 1%	27 (44.3%)	44 (21.7%)	50 (40.3%)	<0.001^e
Colorectal Cancer	15 (24.6%)	62 (30.5%)	34 (27.4%)	0.94
Pancreatic Cancer	8 (13.1%)	14 (6.9%)	15 (12.1%)	0.17
Breast Cancer	9 (14.8%)	20 (9.9%)	11 (8.9%)	0.44

For the genes, the numbers and percent of patients in each cluster with high expression are demonstrated. For other variables, the number and percentages of patients in each cluster meeting each criterion are demonstrated. For instance, 48 patients (53.9%) in Hot cluster have high expression of CD137; high expression is defined as rank value ≥75th percentile. 4 patients (6.6%) in Hot cluster had MSI unstable status.
CD Cluster of differentiation, GITR Glucocorticoid-Induced TNFR-Related, GZMB Granzyme B, ICOS Inducible T Cell Costimulator, IFNG Interferon-gamma, LG ligand, MSI microsatellite instability, PD-L1 programmed death ligand 1, TBX T-Box Transcription Factor, TMB tumor mutational burden.
^a514 patients were evaluated for T-cell priming marker expression; 388/514 had all data of T-cell priming marker expression, MSI status, TMB, and PD-L1 status.
^bHot cluster is one of the three clusters identified by Ward’s hierarchical clustering, and has characteristics of generally high expression of T-cell priming markers.
^cCold cluster has characteristics of generally low expressions of T-cell priming markers.
^dMixed cluster has characteristics of mixed expression levels of T-cell priming markers.
^eSignificant p value (chi square test) was defined ≤0.008 (0.05/6 variables) for other variables.

Search