Treatment of gastric cancer (GC) includes gastrectomy and chemoradiotherapy. The tumor immune microenvironment (TIM) has implications for potential immunotherapy. We analyzed the PD-L1 status of GC and compared it with the number of CD8+ T-lymphocytes, the density of macrophage infiltration and the number of CTLA-4 T-cells and other clinicalpathological parameters. The research included 97 patients who underwent gastrectomy/lymphadenectomy due to GC from 2014 to 2018 at University Hospital Split. Clinical data were collected from hospital records. Paraffin blocks of tumor tissue and draining lymph nodes were collected from the archives of the Department of Pathology. Immunohistochemistry (IHC) with TP53 and MLH1 antibodies, double IHC with CD8 and PD-L1 antibodies, double immunofluorescence with CTLA-4 IFN-γ antibodies and in situ hybridization with EBER probe were performed. Statistical significance was set at p < 0.05. Positive PD-L1 status was associated with intestinal type (p = 0.046), density of macrophages and CD8 + T cells (p < 0.001, both). The median number of CD8+ T cells was higher in PD-L1-positive than in negative tumors. A cutoff of 28.5 CD8 + T cells in one highmagnification field predicted PD-L1-positive tumors (AuROC 0.797, sensitivity 74.2%, specificity 77.3%). IFN-γ expression in tumor cells was found in 37 GCs and was positively associated with CTLA4+ T lymphocytes in the lymph node (p = 0.027), as well as CTLA4+/IFN-γ+ T lymphocytes in the tumor and lymph node (all p < 0.001). Median overall survival (OS) was 17 months. In those who died, IFN-γ expression in nodal metastases was associated with shorter OS (p = 0.008). PD-L1 expression in tumor cells is associated with CD8 + T cells and macrophages in the TME, and IFN-γ in tumor cells with CTLA4+ /IFN-γ + T cells in the tumor and draining lymph node.