Abstract
To understand how corticosteroids act; a characterization of their effects on lymphocytes is necessary. The effect of in vivo corticosteroids on lymphocyte subpopulations, their surface molecules and externalization of phosphatidylserine (apoptosis) is examined. In a crossover study, a single, intravenous dose of 2 mg/kg prednisolone or saline was given to six male adult human volunteers. Blood samples were withdrawn before and 30 min, 2, 5, 23 and 29 h thereafter. Lymphocyte subsets were determined by FACS analysis. Externalization of phosphatidylserine was measured by Annexin-V; cell fragments were excluded by propidium iodide staining. Lymphocyte number decreased from 2,007 ± 473 to 634 ± 119 μl after 5 h and rose to 3,112 ± 436 μl after 23 h. CD4, CD8 and B cell counts declined significantly after 5 h (P ≤ 0.01). The expression of CD28 or CD95 on T cells and the natural killer cells were unaffected. There was a significant rebound of lymphocyte numbers above baseline 23 h after prednisolone. At baseline 9.9 ± 3.8% of cells in the lymphocyte gate did not stain for CD3, CD20 or CD56 (referred to as “null cells”). 5 h after application of prednisolone, there was a significant increase of “null cells” (28 ± 12%, P = 0.018). The percentage of phosphatidylserine positive CD4 cells rose from 8.1 ± 3.3 to 19.8 ± 8% after intravenous prednisolone, while the percentage of phosphatidylserine positive CD8, B and NK cells remained largely unchanged. Prednisolone induces a most significant depletion of CD4 cells, which to some degree is associated with apoptosis. The net increase of lymphocyte numbers 23 h after prednisolone application may be a beneficial late effect of a single i.v. prednisolone shot.
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Abbreviations
- CD:
-
Cluster of differentiation
- DNA:
-
Deoxy ribonucleotide acid
- FACS:
-
Fluorescence activating cell sorting
- FITC:
-
Fluorescein isothiocyanate
- GCR:
-
Glucocorticoid receptor
- NK-cells:
-
Natural killer cells
- PE:
-
Phycoerythrin
- PS:
-
Phosphatidylserine
- SD:
-
Standard deviation
References
Almawi WY (2001) Molecular mechanisms of glucocorticoid effects. Mod Asp Immunobiol 2:78–82
Benito JM, Lopez M, Martin JC, Lozano S, Martinez P, Gonzalez-Lahoz J, Soriano V (2002) Differences in cellular activation and apoptosis in HIV-infected patients receiving protease inhibitors or nonnucleoside reverse transcriptase inhibitors. AIDS Res Hum Retroviruses 18:1379–1388
Boss B, Neeck G, Engelhardt B, Riedel W (1999) Influence of corticosteroids on neutrophils, lymphocytes, their subsets, and T-cell activity markers in patients with active rheumatoid arthritis, compared to healthy controls. Ann NY Acad Sci 876:198–200
Butcher EC, Picker LJ (1996) Lymphocyte homing and homeostasis. Science 272(5258):60–66
Buttgereit F, Wehling M, Burmester GR (1998) A new hypothesis of modular glucocorticoid actions. Arthritis Rheum 41:761–767
Distelhorst CW (2002) Recent insights into the mechanism of glucocorticoid induced apoptosis. Cell Death Differ 9:6–19
Fauci AS, Dale DC (1975) The effect of hydrocortisone on the kinetics of normal human lymphocytes. Blood 46(2):235–243
Haynes BF, Fauci AS (1979) Mechanism of corticosteroid action on lymphocyte subpopulations. IV. Effects of in vitro hydrocortisone on naturally occurring and mitogen-induced suppressor cells in man. Cell Immunol 44:157–168
Knipp S, Feyen O, Ndagijimana J, Niehues T (2003) Ex vivo apoptosis, CD95 and CD28 expression in T cells of children with juvenile idiopathic arthritis. Rheumatol Int 23:112–115
Lanza L, Scudeletti M, Puppo F, Bosco O, Peirano L, Filaci G, Fecarotta E, Vidali G, Indiveri F (1996) Prednisone increases apoptosis in in vitro activated human peripheral blood T lymphocytes. Clin Exp Immunol 103(3):482–9019
Niehues T, McCloskey TW, Ndagijimana J et al (2001) Apoptosis in T-Lymphocyte subsets in human immunodeficieny virus-infected children measured immediately ex vivo and following in vitro activation. Clin Diagn Lab Immunol 8:74–78
Pitzalis C, Pipitone N, Perretti M (2002) Regulation of leukocyte–endothelial interactions by glucocorticoids. Ann NY Acad Sci 966:108–118
Scudeletti M, Lanza L, Monaco E, Monetti M, Puppo F, Filaci G, Indiveri F (1999) Immune regulatory properties of corticosteroids: prednisone induces apoptosis of human T lymphocytes following the CD3 down-regulation. Ann NY Acad Sci 876:164–179
Underwood JL, Murphy CG, Chen J, Franse-Carman I, Wood I, Epstein DL, Alvarado JA (1999) Glucocorticoids regulate transendothelial fluid flow resistance and formation of intercellular junctions. Am J Physiol 272:C330–C342
van Engeland M, Nieland LJ, Ramaekers FC et al (1998) Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry 31:1–9
Wasmuth JC, Hackbarth F, Rockstroh JK, Sauerbruch T, Spengler U (2003) Changes of lymphocyte apoptosis associated with sequential introduction of highly active antiretroviral therapy. HIV Med 4(2):111–119
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The authors thank Annette Seibt and Nico Vente for technical assistance with FACS analysis and data interpretation.
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Financial support used included only departmental funds of the University Hospital Duesseldorf.
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Jetzek-Zader, M., Gudowius, S., Feyen, O. et al. A single intravenous dose of prednisolone induces phosphatidylserine externalization, loss of surface marker expression and a 24-h net increase in human peripheral blood lymphocytes ex vivo. Rheumatol Int 27, 667–673 (2007). https://doi.org/10.1007/s00296-007-0319-4
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DOI: https://doi.org/10.1007/s00296-007-0319-4