Immune modulation and increased neurotrophic factor production in multiple sclerosis patients treated with testosterone

Stefan M Gold, Sara Chalifoux, Barbara S Giesser, Rhonda R Voskuhl, Stefan M Gold, Sara Chalifoux, Barbara S Giesser, Rhonda R Voskuhl

Abstract

Background: Multiple sclerosis is a chronic inflammatory disease of the central nervous system with a pronounced neurodegenerative component. It has been suggested that novel treatment options are needed that target both aspects of the disease. Evidence from basic and clinical studies suggests that testosterone has an immunomodulatory as well as a potential neuroprotective effect that could be beneficial in MS.

Methods: Ten male MS patients were treated with 10 g of gel containing 100 mg of testosterone in a cross-over design (6 month observation period followed by 12 months of treatment). Blood samples were obtained at three-month intervals during the observation and the treatment period. Isolated blood peripheral mononuclear cells (PBMCs) were used to examine lymphocyte subpopulation composition by flow cytometry and ex vivo protein production of cytokines (IL-2, IFNgamma, TNFalpha, IL-17, IL-10, IL-12p40, TGFbeta1) and growth factors (brain-derived neurotrophic factor BDNF, platelet-derived growth factor PDGF-BB, nerve growth factor NGF, and ciliary neurotrophic factor CNTF). Delayed type hypersensitivity (DTH) skin recall tests were obtained before and during treatment as an in vivo functional immune measure.

Results: Testosterone treatment significantly reduced DTH recall responses and induced a shift in peripheral lymphocyte composition by decreasing CD4+ T cell percentage and increasing NK cells. In addition, PBMC production of IL-2 was significantly decreased while TGFbeta1 production was increased. Furthermore, PBMCs obtained during the treatment period produced significantly more BDNF and PDGF-BB.

Conclusion: These results are consistent with an immunomodulatory effect of testosterone treatment in MS. In addition, increased production of BDNF and PDGF-BB suggests a potential neuroprotective effect.

Trial registration: NCT00405353 http://www.clinicaltrials.gov.

Figures

Figure 1
Figure 1
Immunomodulation and growth factor induction by testosterone treatment in 10 male MS patients. A, Testosterone treatment significantly decreased CD4+ T cell and increased CD16/56+ NK cell percentages. B, Treatment also significantly decreased delayed type hypersensitivity recall responses. C-F, In addition, treatment significantly decreased IL-2 and increased TGFβ1, BDNF and PDGF-BB levels produced by PHA stimulated peripheral blood mononuclear cells (PBMCs) during testosterone treatment (months 3–12) compared to baseline (base). Protein levels are expressed as the mean percent change compared with the mean from two pretreatment baseline time points. Mean month 12 concentrations were 670.5 ± 223.4 pg/ml for IL-2, 1552.0+273.3 pg/ml for TGFβ1, 246.1 ± 40.37 pg/ml for BDNF and 42.6 ± 15.6 pg/ml for PDGF-BB, respectively.
Figure 2
Figure 2
Growth factors reduce neurotoxicity in vitro. Neuroprotection against glutamate toxicity (1 mM for 2 h) by growth factors at concentrations similar to those at treatment month 12 in the clinical trial (2000 pg/ml TGFβ1, 500 pg/ml BDNF, 100 pg/ml PDGF-BB). A-C, Brightfield microscopy images indicate that addition of growth factors during glutamate exposure induced partial protection of axonal integrity in those cultures (black arrows) and decreased the number of TUNEL positive cells (white triangles), representative images shown from 8 experiments. D, a protective effect was confirmed quantitatively by a significant reduction in LDH release. LDH data are shown from 8 independent experiments with wells run in duplicate for each condition.

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