title: Sitagliptin inhibit human lymphocytes proliferation and Th1/Th17 differentiation in vitro.
abstract: Dipeptidyl peptidase-4 (DPP-4) inhibitors are a new class of anti-diabetic agents that are widely used in clinical practice to improve glycemic control in patients with type 2 diabetes. DPP-4 is also known as lymphocyte cell surface protein, CD26, and plays an important role in T-cell immunity. Recent studies suggest that DPP-4 inhibitors improve beta-cell function and attenuate autoimmunity in type 1 diabetic mouse models. To investigate the direct effect of DPP4 in immune response, human peripheral blood mononuclear cells (PBMC) from healthy volunteers were obtained by Ficoll gradient and cultivated in the absence (control) or presence of phytohemagglutinin (PHA), or stimulated with PHA and treated with sitagliptin. The immune modulation mechanisms analyzed were: cell proliferation, by MTT assay; cytokine quantification by ELISA or cytometric bead array (CBA), Th1/Th2/Th17 phenotyping by flow cytometric analysis and CD26 gene expression by real time PCR. The results showed that sitagliptin treatment inhibited the proliferation of PBMC-PHA stimulated cells in a dose dependent manner and decreased CD26 expression by these cells, suggesting that sitagliptin may interfere in CD26 expression, dimerization and cell signaling. Sitagliptin treatment not only inhibited IL-10 (p<0.05) and IFN-gamma (p=0.07) cytokines, but also completely abolish IL-6 expression by PBMCs (p<0.001). On the other hand, IL-4 were secreted in culture supernatants from sitagliptin treated cells. A statistically significant increase (p<0.05) in the ratio of TGF-beta/proliferation index after sitagliptin treatment (2627.97±1351.65), when comparing to untreated cells (646.28±376.94), was also demonstrated, indicating higher TGF-beta1 production by viable cells in cultures. Sitagliptin treatment induced a significantly (p<0.05) decrease in IL-17 and IFN-gamma intracellular expression compared with PHA alone. Also, the percentage of T CD4+IL-17+, T CD4+IFNgamma+and T CD4+IL-4+cells were significantly reduced (p<0.05) by sitagliptin. Our data demonstrated an immunosuppressive effect of sitagliptin on Th1, Th17 and Th2 lymphocytes differentiation that leads to the generation of regulatory TGF-beta1 secreting cells with low CD26 gene expression that may influence the state of pancreatic beta-cells and controlling DM1 patients.
authors.affiliation: Programa de Pós-graduação em Farmácia - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil.
authors.forename: Marcelo Maia
authors.initials: MM
authors.lastname: Pinheiro
authors.affiliation: Programa de Pós-graduação em Farmácia - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil.
authors.forename: Caroline Lais
authors.initials: CL
authors.lastname: Stoppa
authors.affiliation: Programa de Pós-graduação em Biotecnologia e Inovação em Saúde - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil.
authors.forename: Claudete Justina
authors.initials: CJ
authors.lastname: Valduga
authors.affiliation: Programa de Pós-graduação em Farmácia - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil.
authors.forename: Cristina Eunice
authors.initials: CE
authors.lastname: Okuyama
authors.affiliation: Programa de Pós-graduação Interdisciplinar em Ciências da Saúde - Universidade Cruzeiro do Sul (UNICSUL), São Paulo, Brazil.
authors.forename: Renata
authors.initials: R
authors.lastname: Gorjão
authors.affiliation: Programa de Pós-graduação em Farmácia - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil.
authors.forename: Regina Mara Silva
authors.initials: RM
authors.lastname: Pereira
authors.affiliation: Programa de Pós-graduação em Farmácia - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil; Programa de Pós-graduação em Biotecnologia e Inovação em Saúde - Universidade Anhanguera São Paulo (UNIAN), São Paulo, Brazil. Electronic address: dinizsusana@gmail.com.
authors.forename: Susana Nogueira
authors.initials: SN
authors.lastname: Diniz
mesh_terms: D000293:Adolescent; D000328:Adult; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics; D054873:Dipeptidyl-Peptidase IV Inhibitors / Q000494:pharmacology*; D005260:Female; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D000068900:Sitagliptin Phosphate / Q000494:pharmacology*; D055815:Young Adult
chemical_list: D016207:Cytokines; D054873:Dipeptidyl-Peptidase IV Inhibitors; D012333:RNA, Messenger; C042807:DPP4 protein, human; D018819:Dipeptidyl Peptidase 4; D000068900:Sitagliptin Phosphate
keywords: Cytokine; DPP-4/CD26; Immune regulation; Lymphocyte; Sitagliptin; Type 1 diabetes mellitus