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Immunosuppressive drugs affect high-mannose/hybrid N-glycans on human allostimulated leukocytes.

Pocheć E, Bocian K, Ząbczyńska M, Korczak-Kowalska G, Lityńska A - Anal Cell Pathol (Amst) (2015)

Bottom Line: The N-glycosylation process is very sensitive to different environmental agents, among them the pharmacological environment of immunosuppressive drugs.Some results show that high-mannose oligosaccharides have the ability to suppress different stages of the immune response.This is the first study indicating that β1 and β3 integrins bearing high-mannose/hybrid structures are affected by Rapa and CsA.

View Article: PubMed Central - PubMed

Affiliation: Department of Glycoconjugate Biochemistry, Institute of Zoology, Faculty of Biology and Earth Science, Jagiellonian University, Gronostajowa 9, 30-387 Krakow, Poland.

ABSTRACT
N-glycosylation plays an important role in the majority of physiological and pathological processes occurring in the immune system. Alteration of the type and abundance of glycans is an element of lymphocyte differentiation; it is also common in the development of immune-mediated inflammatory diseases. The N-glycosylation process is very sensitive to different environmental agents, among them the pharmacological environment of immunosuppressive drugs. Some results show that high-mannose oligosaccharides have the ability to suppress different stages of the immune response. We evaluated the effects of cyclosporin A (CsA) and rapamycin (Rapa) on high-mannose/hybrid-type glycosylation in human leukocytes activated in a two-way mixed leukocyte reaction (MLR). CsA significantly reduced the number of leukocytes covered by high-mannose/hybrid N-glycans, and the synergistic action of CsA and Rapa led to an increase of these structures on the remaining leukocytes. This is the first study indicating that β1 and β3 integrins bearing high-mannose/hybrid structures are affected by Rapa and CsA. Rapa taken separately and together with CsA changed the expression of β1 and β3 integrins and, by regulating the protein amount, increased the oligomannose/hybrid-type N-glycosylation on the leukocyte surface. We suggest that the changes in the glycosylation profile of leukocytes may promote the development of tolerance in transplantation.

No MeSH data available.


Related in: MedlinePlus

Immunosuppressive drugs alter the surface expression of high-mannose/hybrid N-glycans. Peripheral blood mononuclear cells were cultured for 6 days in the presence of CsA (200 ng per 1 mL medium), Rapa (20 ng per 1 mL medium), and the combination of CsA (150 ng per 1 mL medium) and Rapa (12 ng per 1 mL medium) in a two-way mixed leukocyte reaction (MLR). Immunosuppressive drug-treated and control cells were stained with biotinylated GNA (1 : 100), followed by incubation with FITC-conjugated ExtrAvidin. M2 region corresponds to GNA-positive leukocytes after allostimulation. Fluorescence was measured using a FACSCalibur flow cytometer (BD Biosciences). C: untreated cells, CsA: cyclosporin A, and Rapa: rapamycin.
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Related In: Results  -  Collection


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fig1: Immunosuppressive drugs alter the surface expression of high-mannose/hybrid N-glycans. Peripheral blood mononuclear cells were cultured for 6 days in the presence of CsA (200 ng per 1 mL medium), Rapa (20 ng per 1 mL medium), and the combination of CsA (150 ng per 1 mL medium) and Rapa (12 ng per 1 mL medium) in a two-way mixed leukocyte reaction (MLR). Immunosuppressive drug-treated and control cells were stained with biotinylated GNA (1 : 100), followed by incubation with FITC-conjugated ExtrAvidin. M2 region corresponds to GNA-positive leukocytes after allostimulation. Fluorescence was measured using a FACSCalibur flow cytometer (BD Biosciences). C: untreated cells, CsA: cyclosporin A, and Rapa: rapamycin.

Mentions: Glycosylation of alloantigen-activated PBMCs from two individuals mixed together in two-way MLR cell culture was analyzed using FITC-labeled Galanthus nivalis agglutinin (GNA) in flow cytometry. We found that the number of leukocytes covered by oligomannose/hybrid-type N-glycans decreased dramatically in the presence of CsA applied alone or with Rapa (Figure 1). Mean fluorescence intensity (MFI) was reduced slightly by the drugs given separately but the combination of both drugs significantly raised the MFI value. CsA was responsible for removal of high-mannose/hybrid N-glycans from the surface of over 60% of the leukocytes, but the synergistic effect of both immunosuppressive drugs markedly increased the amount of oligomannose/hybrid-type N-glycans on the remaining cells.


Immunosuppressive drugs affect high-mannose/hybrid N-glycans on human allostimulated leukocytes.

Pocheć E, Bocian K, Ząbczyńska M, Korczak-Kowalska G, Lityńska A - Anal Cell Pathol (Amst) (2015)

Immunosuppressive drugs alter the surface expression of high-mannose/hybrid N-glycans. Peripheral blood mononuclear cells were cultured for 6 days in the presence of CsA (200 ng per 1 mL medium), Rapa (20 ng per 1 mL medium), and the combination of CsA (150 ng per 1 mL medium) and Rapa (12 ng per 1 mL medium) in a two-way mixed leukocyte reaction (MLR). Immunosuppressive drug-treated and control cells were stained with biotinylated GNA (1 : 100), followed by incubation with FITC-conjugated ExtrAvidin. M2 region corresponds to GNA-positive leukocytes after allostimulation. Fluorescence was measured using a FACSCalibur flow cytometer (BD Biosciences). C: untreated cells, CsA: cyclosporin A, and Rapa: rapamycin.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4538311&req=5

fig1: Immunosuppressive drugs alter the surface expression of high-mannose/hybrid N-glycans. Peripheral blood mononuclear cells were cultured for 6 days in the presence of CsA (200 ng per 1 mL medium), Rapa (20 ng per 1 mL medium), and the combination of CsA (150 ng per 1 mL medium) and Rapa (12 ng per 1 mL medium) in a two-way mixed leukocyte reaction (MLR). Immunosuppressive drug-treated and control cells were stained with biotinylated GNA (1 : 100), followed by incubation with FITC-conjugated ExtrAvidin. M2 region corresponds to GNA-positive leukocytes after allostimulation. Fluorescence was measured using a FACSCalibur flow cytometer (BD Biosciences). C: untreated cells, CsA: cyclosporin A, and Rapa: rapamycin.
Mentions: Glycosylation of alloantigen-activated PBMCs from two individuals mixed together in two-way MLR cell culture was analyzed using FITC-labeled Galanthus nivalis agglutinin (GNA) in flow cytometry. We found that the number of leukocytes covered by oligomannose/hybrid-type N-glycans decreased dramatically in the presence of CsA applied alone or with Rapa (Figure 1). Mean fluorescence intensity (MFI) was reduced slightly by the drugs given separately but the combination of both drugs significantly raised the MFI value. CsA was responsible for removal of high-mannose/hybrid N-glycans from the surface of over 60% of the leukocytes, but the synergistic effect of both immunosuppressive drugs markedly increased the amount of oligomannose/hybrid-type N-glycans on the remaining cells.

Bottom Line: The N-glycosylation process is very sensitive to different environmental agents, among them the pharmacological environment of immunosuppressive drugs.Some results show that high-mannose oligosaccharides have the ability to suppress different stages of the immune response.This is the first study indicating that β1 and β3 integrins bearing high-mannose/hybrid structures are affected by Rapa and CsA.

View Article: PubMed Central - PubMed

Affiliation: Department of Glycoconjugate Biochemistry, Institute of Zoology, Faculty of Biology and Earth Science, Jagiellonian University, Gronostajowa 9, 30-387 Krakow, Poland.

ABSTRACT
N-glycosylation plays an important role in the majority of physiological and pathological processes occurring in the immune system. Alteration of the type and abundance of glycans is an element of lymphocyte differentiation; it is also common in the development of immune-mediated inflammatory diseases. The N-glycosylation process is very sensitive to different environmental agents, among them the pharmacological environment of immunosuppressive drugs. Some results show that high-mannose oligosaccharides have the ability to suppress different stages of the immune response. We evaluated the effects of cyclosporin A (CsA) and rapamycin (Rapa) on high-mannose/hybrid-type glycosylation in human leukocytes activated in a two-way mixed leukocyte reaction (MLR). CsA significantly reduced the number of leukocytes covered by high-mannose/hybrid N-glycans, and the synergistic action of CsA and Rapa led to an increase of these structures on the remaining leukocytes. This is the first study indicating that β1 and β3 integrins bearing high-mannose/hybrid structures are affected by Rapa and CsA. Rapa taken separately and together with CsA changed the expression of β1 and β3 integrins and, by regulating the protein amount, increased the oligomannose/hybrid-type N-glycosylation on the leukocyte surface. We suggest that the changes in the glycosylation profile of leukocytes may promote the development of tolerance in transplantation.

No MeSH data available.


Related in: MedlinePlus