Effect of Sertraline in a murine model of allogenic skin transplantation
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1
Hospital Universitario San Ignacio, Departamento de Cirugía Plástica, Colombia
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2
Pontificia Universidad Javeriana, Instituto de Genética Humana, Colombia
Sertraline (SRT) is a selective Serotonin reuptake inhibitor (SSRI) and increases the concentration of serotonin at neuronal synapsis. It is used for treatment of symptoms related to depression, obsessive-compulsive disease, panic disorders and anxiety, among others. SSRIs, like SRT, have been described to have anti-inflammatory properties in vitro and in a few in vivo models. Moreover several SSRI have been shown to have an inhibitory effect on Toll like receptors (TLR- 3, 4, 7 and 8). A relation has been described between TLR and the activation of the innate immune response in acute transplant rejection and in particular in skin and composite tissue allotransplantation(CTA). In CTA, patients require immunosuppressive therapy to prevent and control rejection with diverse medications, with variable non desirable side effects. There is a constant quest for alternative methods to minimize the negative impact of immunosuppression. Frequently patients who need a CTA, because of their initial lesions, also show symptoms of anxiety and depression, often requiring pharmacological treatment and psychological support. Given that SRT is a first line drug in treatment of depression with anti-inflammatory effects, our goal was to evaluate the immunomodulatory effect of this drug in a series of in vitro and in an in vivo murine skin allotransplantation model.
Objectives
General
To evaluate the immunomodulatory effect of SRT in vitro and in an in vivo animal model.
Specific
- To compare the production of cytokines produced by human peripheral blood mononuclear cells (PBMC) stimulated with LPS in the presence or abcence of SRT.
-To compare the percentages of cytokine secreting T cells in PBMC stimulated with SEB in the presence or abcence of SRT.
- To evaluate differences in acute rejection of allogenic skin in mice treated or not with SRT.
- To compare the frequency of cytokine secreting T cells in splenocytes of mice with allogenic skin transplants treated or not with SRT.
Experimental design
Human PBMC cultures were stimulated with LPS in the presence or not of SRT and the production of cytokines (IL-1β, IL-6, IL-10, TNF-α and IL-12p70) measured by CBA. PBMC cultures were stimulated with SEB and the percentages of CD4+ and CD8+ cytokine (IL-10, IFN-γ, IL-2) producing cells was evaluated by an intracellular cytokine assay. For in vivo experiments, murine skin from the ear of a donor BALBc mouse was transplanted to the back of a C57BL/6 mouse. Three groups were considered: The control group and two groups treated intraperitoneally with SRT at different doses (10mg/Kg and 30mg/Kg), administered every 3 days. Daily monitoring of skin grafts looking for acute rejection signs was performed. Splenocytes were obtained at the end of the study, and stimulated with PMA/Ionomycin and the frequencies of cytokine secreting T cells (IL-2, IL-10, TNF-α, IFN-γ) were evaluated by Flow cytometry. We performed 2 experiments: In the first 10 mice were studied (4 controls and 3 mice for each treatment dose of SRT). The second experiment consisted of 13 mice, (4 controls, 6 individuals for SRT at 10mg/kg dose and 3 for 30mg/kg dose). In the second experiment treatment was assigned in an aleatory fashion and clinical outcome was evaluated in blind. The experiments of human PBMC were analyzed with a paired Wilcoxon test. For evaluation of the clinical results a Fisher Yates test was performed to establish statistical differences. In the splenocyte experiments the Mann Whitney test was applied. P <0,05 was considered significant.
Results
In the experiments of human PBMC stimulated with LPS and treated with SRT, we noted a significant decreased production of IL-12p70, TNF-α and IL-1β in comparison with those treated with the control.
The PBMC stimulated with SEB in the presence of sertraline showed a significant decrease in the percentage of CD4+ T cells producing IL-2, IL-10 and IFN-γ and decrease of CD8+ producing IL-2, IFN-γ.
In the first but not the second in vivo experiments with the murine allotransplantation model mice treated with SRT at 30mg/kg dose (p=0,029, Fisher Yates test) but not with the 10mg/kg, showed a reduction of graft rejection compared to the control group (p=0,14). When both experiments are considered together a similar conclusion is obtained, (p= 0,021, for Sertraline at 30mg/kg and p=0,051, for SRT at 10mg/kg) .
In the splenocyte culture experiments a significant decrease in the percentage of CD4+ T cells secreting IL-2 was observed, but only in the first of the two experiments done.
Conclusion
Our results support reports in the literature that propose that the antinflammatory effect of SRT is due to its capacity to modulate the production of cytokines by cells of the immune system. Moreover we show for the first time that this effect can be translated in to a clinical effect in the murine skin allotransplantation model. However the dose of SRT used to obtain the clinical effect seems to be relatively high. It remains to be determined if SRT can have an immunosupresive effect in humans treated with the doses used clinically.
Acknowledgements
Juan C. Zambrano and Sindy M. Muñoz contributed equally to this work. This work was supported by grants from HUSI 001 and PUJ (ID 4969). Sindy M. Muñoz was funded by scholarship from COLCIENCIAS.
Keywords:
Sertraline,
Allogenic,
Skin Transplantation,
Mice,
Immunomodulation
Conference:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología, Medellin, Colombia, 13 Oct - 16 Oct, 2015.
Presentation Type:
Poster Presentation
Topic:
Transplantation immunology
Citation:
Zambrano
JC,
Muñoz
SM,
Franco
MA and
Rodriguez
L
(2015). Effect of Sertraline in a murine model of allogenic skin transplantation.
Front. Immunol.
Conference Abstract:
IMMUNOCOLOMBIA2015 - 11th Congress of the Latin American Association of Immunology - 10o. Congreso de la Asociación Colombiana de Alergia, Asma e Inmunología.
doi: 10.3389/conf.fimmu.2015.05.00138
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Received:
18 Apr 2015;
Published Online:
14 Sep 2015.
*
Correspondence:
Dr. Luz-Stella Rodriguez, Pontificia Universidad Javeriana, Instituto de Genética Humana, Bogotá, Colombia, luz-rodriguez@javeriana.edu.co