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Cd4+ccr9+ peripheral blood lymphocytes contain differentiated t regulatory 1 cells and subset that provides b cell help for immunoglobulin production



Cd4+ccr9+ peripheral blood lymphocytes contain differentiated t regulatory 1 cells and subset that provides b cell help for immunoglobulin production



Digestive Disease Week Abstracts & Itinerary Planner : Abstract No S1095



Background/Aims: Lymphocytes expressing the chemokine receptor CCR9 (a small bowel specific chemokine receptor) has been suggested to play an important role in Small Bowel (SB) immune responses. We have previously reported that CCR9 expression in peripheral blood lymphocytes (PBL) defines a subset of cells with mucosal T cell phenotype and a Th1 cytokine profile (Gastroenterology 2002;A31: 256). Moreover, the percentage of this cell subset is increased in the peripheral blood of patients with SB inflammation (Gastroenterology 2001; 121:246-254). The purpose of the current study was to further analyze the functional characteristics of CCR9+CD4+ PBL from normal donors. Methods: PBL from normal donors were analyzed by flow cytometry. Freshly sorted CD4+CCR9+ and CD4+CCR9- PBL were activated with PMA plus ionomycin and intracellular staining for IL-10, IL-2, and IFN-g was performed. B cell help in immunoglobulin (Ig) production was performed by co-culturing freshly sorted CD4+CD45RO+CCR9+ or CD4+CD45RO+CCR9- with autologous CD19+ B cells for 11 days. Culture supernatants were analyzed for IgM, IgA, and IgG by ELISA. Results: Compared to CD4+CCR9- subset of PBL, CD4+CCR9+ contain IL-10+ cells which are distinct from IFN-g + or IL-2+ cells suggesting that a subset of cells within CD4+CCR9+ exhibit the cytokine profile of T regulatory 1 (Tr1) cells. The percentage of CCR9+IL-10+ CD4+ cells varied from different donors and ranged between 2.5% to 13%. Moreover, co-culture of memory CCR9+, but not CCR9-CD4+ cells, with autologous B cells induces the production of IgM, IgA, and IgG without prior T cell activation or addition of exogenous cytokines. The level of IgA and IgG in culture supernatants ranged from <100 ng/ml from B cells cultured alone or with CCR9-CD4+ cells, to 500 ng/ml from B cells cultured with CCR9+CD4+ cells. Conclusions: The CD4+CCR9+ memory subset of PBL from normal donors contains differentiated Tr1 cells and a subset that supports B cells for Ig, including IgA, production. Analysis of the cytokine profile of this subset in the peripheral blood of patients with SB Crohn's disease will determine whether an imbalance of Th1/Tr1 cytokine profile is operative in the pathogenesis of this disease.

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