ABSTRACT
Mesenchymal stem cells (MSCs) can inhibit T cell proliferation; however, the underlying mechanisms are not clear. In this study, we investigated the mechanisms of the immunoregulatory activity of MSCs on T cells. Irradiated MSCs co-cultured with either naive or pre-activated T cells in a mixed lymphocyte reaction (MLR) significantly suppressed T cell proliferation in a dose-dependent manner, irrespective of allogeneic disparity between responders and MSCs. Transwell assays revealed that the suppressive effect was primarily mediated by soluble factors that induced apoptosis. Splenocytes stimulated with alloantigen in the presence of the MSC culture supernatant (CS) produced a significant amount of IL-10, which was attributed to an increase in the number of IL-10 secreting cells, confirmed by an ELISPOT assay. The blockade of IL-10 and IL-10 receptor interaction by anti-IL-10 or anti-IL-10-receptor antibodies abrogated the suppressive capacity of MSC CS, indicating that IL-10 plays a major role in the suppression of T cell proliferation. The addition of 1-methyl-DL-tryptophan (1-MT), an indoleamine 2,3-dioxygenase (IDO) inhibitor, also restored the proliferative capacity of T cells. In conclusion, we demonstrated that soluble mediators from culture supernatant of MSCs could suppress the proliferation of both naive and pre-activated T cells in which IL-10 and IDO play important roles.
Subject(s)
Animals , Mice , Cell Proliferation , Cells, Cultured , Indoleamine-Pyrrole 2,3,-Dioxygenase/antagonists & inhibitors , Interleukin-10/biosynthesis , Lymphocyte Activation , Lymphokines/pharmacology , Mesenchymal Stem Cells/cytology , Mice, Inbred BALB C , Mice, Inbred C57BL , Receptors, Interleukin-10/metabolism , T-Lymphocytes/cytology , Tryptophan/analogs & derivativesABSTRACT
Mesenchymal stem cells (MSCs) can inhibit T cell proliferation; however, the underlying mechanisms are not clear. In this study, we investigated the mechanisms of the immunoregulatory activity of MSCs on T cells. Irradiated MSCs co-cultured with either naive or pre-activated T cells in a mixed lymphocyte reaction (MLR) significantly suppressed T cell proliferation in a dose-dependent manner, irrespective of allogeneic disparity between responders and MSCs. Transwell assays revealed that the suppressive effect was primarily mediated by soluble factors that induced apoptosis. Splenocytes stimulated with alloantigen in the presence of the MSC culture supernatant (CS) produced a significant amount of IL-10, which was attributed to an increase in the number of IL-10 secreting cells, confirmed by an ELISPOT assay. The blockade of IL-10 and IL-10 receptor interaction by anti-IL-10 or anti-IL-10-receptor antibodies abrogated the suppressive capacity of MSC CS, indicating that IL-10 plays a major role in the suppression of T cell proliferation. The addition of 1-methyl-DL-tryptophan (1-MT), an indoleamine 2,3-dioxygenase (IDO) inhibitor, also restored the proliferative capacity of T cells. In conclusion, we demonstrated that soluble mediators from culture supernatant of MSCs could suppress the proliferation of both naive and pre-activated T cells in which IL-10 and IDO play important roles.
Subject(s)
Animals , Mice , Cell Proliferation , Cells, Cultured , Indoleamine-Pyrrole 2,3,-Dioxygenase/antagonists & inhibitors , Interleukin-10/biosynthesis , Lymphocyte Activation , Lymphokines/pharmacology , Mesenchymal Stem Cells/cytology , Mice, Inbred BALB C , Mice, Inbred C57BL , Receptors, Interleukin-10/metabolism , T-Lymphocytes/cytology , Tryptophan/analogs & derivativesSubject(s)
Animals , Endothelial Growth Factors/pharmacology , Fibroblast Growth Factors/pharmacology , Humans , Intercellular Signaling Peptides and Proteins/pharmacology , Lymphokines/pharmacology , Neovascularization, Physiologic/drug effects , Platelet-Derived Growth Factor/pharmacology , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Angiostatin is a potent angiogenesis inhibitor that is composed of the first four kringles of plasminogen fragment. Angiostatin with one less kringle molecule (kringle 1 to 3) was recently demonstrated to be an effective angiogenic inhibitor. To determine whether recombinant plasminogen kringle 1-3 (rPK1-3) can inhibit the corneal neovascularization induced by potent angiogenic factors; angiogenin, bFGF, or VEGF, hydron polymer discs each containing 2.0 microg of angiogenin, 500 ng of bFGF, or 500 ng of VEGF respectively were implanted into the corneal stroma of 138 rabbit eyes, and then discs each containing 10 microg, 12.5 microg, 20 microg or 30 microg of rPK1-3 were implanted randomly. Discs containing phosphate buffered saline were also implanted as a control. The angiogenesis score on number and length of newly formed vessels on the each of the rabbit's cornea were recorded daily by two observers (blinded). The treated corneas were also examined histologically. Recombinant PK1-3 treated corneas showed less neovascularization induced by all angiogenic factors (p < 0.05). and the extent of inhibition of neovascularization was proportional to the concentration of rPK1-3 (p < 0.05). Histologic examination showed leukocyte infiltration into the corneal stroma on the PBS treated eyes whereas rPK1-3 treated eyes showed only traces of leukocytes. These results of the effective rPK1-3 inhibition of corneal neovascularization induced by angiogenin, bFGF, or VEGF suggest that this angiostatin related fragment, rPK1-3, may be useful in the treatment of various neovascular diseases. Copyright 2000 Academic Press.
Subject(s)
Chick Embryo , Rabbits , Angiogenesis Inhibitors/pharmacology , Angiogenesis Inhibitors/genetics , Animals , Chorion/drug effects , Chorion/blood supply , Cornea/pathology , Cornea/drug effects , Cornea/blood supply , Endothelial Growth Factors/pharmacology , Fibroblast Growth Factor 2/pharmacology , Kringles/genetics , Lymphokines/pharmacology , Microscopy/methods , Neovascularization, Pathologic/drug therapy , Plasminogen/pharmacology , Plasminogen/genetics , Recombinant Proteins/pharmacology , Recombinant Proteins/genetics , Ribonuclease, Pancreatic/pharmacologyABSTRACT
Intravenous injections of lipopolysaccharide (LPS, 20 microng/Kg) and of a factor originating from LPS-stimulated macrophage monolayers (Neutrophil Recruitment Inhibitory Factor, NRIF) inhibited neutrophil migration into peritoneal cavities induced by carrageenin in rats for up to 24 h. Mononuclear cell migration induced by thioglycollate was also inhibited by the same treatment with LPS but was not affected by NRIF. We conclude that NRIF specifically blocks neutrophil migration and we suggest that NRIF released into the circulation may constitute an important determinant of septicemia