Human CD26 has dipeptidyl peptidase‐4 (DPP IV) enzyme activity and binds to adenosine deaminase (ADA). CD26 is costimulatory for lymphocytes and has a circulating soluble form (sCD26). DPP IV enzyme inhibition is a new successful type 2 diabetes therapy. We examined whether the ADA binding and catalytic functions of sCD26 contribute to its effects on T‐cell proliferation. Wildtype soluble recombinant human CD26 (srhCD26), an enzyme inactive mutant (srhCD26E‐) and an ADA non‐binding mutant (srhCD26A‐) were co‐incubated in in vitro T‐cell proliferation assays with peripheral blood mononuclear cells (PBMC) stimulated with phytohaemagglutinin (PHA), muromonab‐CD3 or Herpes simplex virus antigen (HSV Ag). Both srhCD26 and srhCD26E‐ enhanced PHA‐induced T‐cell proliferation dose‐dependently in all six subjects tested. srhCD26 and srhCD26A‐ had no overall effect on anti‐CD3‐stimulated PBMC proliferation in four of five subjects. srhCD26, srhCD26E‐ and srhCD26A‐ enhanced HSV Ag induced PBMC proliferation in low responders to HSV Ag, but had no effect or inhibited proliferation in HSV‐high responders. Thus, effects of soluble human CD26 on human T‐cell proliferation are mechanistically independent of both the enzyme activity and the ADA‐binding capability of sCD26.
Abstract:Hydrogen peroxide (H 2 O 2 ) can act as an intracellular messenger by oxidizing sulfhydryl groups in cysteines that can be oxidized at neutral pH. The oxidizing agents H 2 O 2 and pyrroloquinoline quinone and the large thiol reagents N-ethylmaleimide and 4-(hydroxymercuri) benzoate each inhibited dipeptidyl peptidase (DP) activity in the intracellular DPIV-related proteins DP8 and DP9 at pH 7.5. In contrast, these treatments did not alter activity in DPIV and fibroblast activation protein. Peptidase inhibition was completely reversed by 2-mercaptoethanol or reduced glutathione. Alkylation of DP8 by the small thiol reagent iodoacetamide prevented inhibition by H 2 O 2, N-ethylmaleimide or pyrroloquinoline quinone. Two cysteines were reactive per peptidase monomer. We exploited these properties to highly purify DP8 by thiol affinity chromatography. Homology modelling of DP8 and DP9 was consistent with the proposal that the mechanism involves decreased protein flexibility caused by intramolecular disulfide bonding. These novel data show that DP8 and DP9 are reversibly inactivated by oxidants at neutral pH and suggest that DP8 and DP9 are H 2 O 2 sensing proteins.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.