Pancreatic ductal adenocarcinoma (PDAC) is characterized by a highly suppressive immune tumor microenvironment (TME), impairing the efficacy of anti-cancer treatments and resulting in poor clinical outcomes. Here, we performed comprehensive immune cellular-molecular analyses of 171 biopsies from PDAC patients, relying on multiplex immunohistochemistry (mIHC) analysis of tissue microarrays (TMA). Comparing PDAC and adjacent tissues from the same patients, we found that cancer tissues have an increased frequency of immunosuppressive FOXP3+ regulatory T cells (Tregs) and PD-L1+ cells, in support of the immunosuppressive TME in PDAC. Interestingly, a subgroup of PDAC patients with poor survival outcomes showed high frequencies of FOXP3+ Tregs and low CD8/FOXP3 and CD8/PD-L1 ratios in their cancer tissues. Transcriptomic analysis of tumor biopsies from this subgroup revealed a distinct gene expression signature associated with an overexpression of chemokines including CXCL9, CXCL10, CCL18 and CCL22. In a novel humanized PDAC xenograft murine model, we showed that CCL22 played a pivotal role in recruiting Tregs to the TME and that the administration of anti-CCL22 antibodies significantly reduced tumor progression. Collectively, this study demonstrates the prognostic value of FOXP3 as a PDAC biomarker and suggests that targeting CCL22, Tregs and the immune checkpoint axis may represent novel therapeutic avenues to treat PDAC.