Cystic Fibrosis(CF) patients are chronically infected with Pseudomonas aeruginosa (Pa) that is currently considered as a major cause of death. To persist in the host, bacteria will adapt to the pulmonary environment and especially to the immune system by secreting or expressing a multitude of pro / anti-inflammatory molecules. Among them, some metabolites (immuno-metabolites /metabokines) have been described as altering the activation and the polarization of subsequent immune responses. We investigate the modulation of the immune system by Pa and the evolution over time on a longitudinal clinical strains biobank of Pa. This study will allow to better understand the host/pathogen crosstalk by highlighting new metabolites and new metabolic pathways having immunomodulatory properties. 32 patients chronically infected by Pa were included in the study at the CHU Grenoble- Alpes, and for each patient two clones of the same strain were isolated 5 years apart. Clinical strains have been cultivated in SCFM (Sputum of Cystic Fibrosis Medium) then bacterial supernatants (SN) were collected during exponential and stationary growth phases. We have investigated differential immune properties of isolates by flow cytometry-based immunoassay allowing to analyze the activation and the maturation of Dendritic Cell (DC) upon TLR stimulation, in presence of bacteria SN. In parallel, SN metabolic profiles were characterized by LC-MS/MS metabolomic analysis. Then, to identify metabolites responsible of immunomodulations, we performed data analysis and correlate both results. Interestingly, we have highlighted different immunomodulatory profiles depending on strains, and on growth phases: two “immunosuppressive clusters” (one in which all DC markers expression decreases and one in which PDL2 expression increases) and one “immunostimulatory cluster”. Of note, infection of patients by “immunosuppressive strains” has been correlated with the decline of respiratory functions. In line with DC phenotypes, untargeted metabolomic analyses of bacterial SN showed 3 distinct metabolic fingerprints, highlighting the importance of Pa strains metabolism in the modulation of immunity. We identified metabokines responsible of immunomodulatory properties observed on activated DC, and notably some metabolites leading to the increase of PDL2, such as succinate or virulence factors, like siderophores. This study identifies potential immunometabolites as biomarkers for predicting CF patients’ clinical evolution. We showed that Pa isolates display differential immune properties depending on secreted metabolites and we highlighted 3 Immuno- Clusters. Characterization of the excreted metabolites by spectrometry analysis enabled us to highlight the importance of the strain's overall metabolism in its modulation of immunity. Some of the identified metabolites lead to the increase of the expression of PDL2, that constitute potential therapeutic targets for restoring host immunity in chronic infectious diseases.