Background: Gastric cancer (GC) is one of the most common and fatal cancers worldwide. Tumor mutational burden (TMB) is described as a novel powerful signature to predict response to immunotherapy in gastric cancer. Nevertheless, tumor-infiltrating immune cells (TIICs) in different TMB GC remains unknown.Results: (1) The most common missense were single nucleotide polymorphisms and C>T. Furthermore, titin (TTN), tumor protein p53 (TP53), and mucin 16 (MUC16) had a higher rate of mutations in STAD. (2) TMB-high and -low groups were not associated with survival, T stage, M stage, and grade of patients with stomach adenocarcinoma (STAD) (Psurvival=0.086, PT=0.331, PM=0.804, Pgrade=0.695). However, TMB groupings were related with N stage and sex (PN=0.003, Psex=0.003). (3) There were intricate differences in immune infiltration cells (eg, T cells, monocytes, and mast cells) between the TMB-high and TMB-low groups. (4) A total of 458 genes were identified as DEGs; Dynein cytoplasmic 1 intermediate chain 1(DYNC1I1), matrix metalloproteinase 13 (MMP13) and zinc finger and BTB domain containing 16 (ZBTB16) were correlated with overall survival (OS). (5) DEGs were mainly involved in the regulation of the extracellular matrix-receptor interaction, and the Wnt and PI3K-AKT signaling pathways.Conclusion: The present study provides a comprehensive and systematic analysis of TMB and its clinical significance, and reveals the complicated differences of 22 TIICs between TMB-high and TMB-low groups in STAD.