IFN-γ stimulation of dental follicle mesenchymal stem cells modulates immune response of CD4+ T lymphocytes in Der p1+ asthmatic patients in vitro (original) (raw)
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Clinical & Experimental Allergy, 2018
SummaryBackgroundAsthma is a chronic inflammatory disease in which inflammatory responses have the polarisation of CD4+ T cells to Th2 cells. Dental follicle mesenchymal stem cells (DFSCs) have strong anti‐inflammatory properties comparable to other mesenchymal stem cells.ObjectiveWe investigated the immunomodulatory effects of DFSCs on CD4+ T helper cell responses of asthmatic patients and compared the results with those obtained with asthmatic subjects on immunotherapy and with healthy individuals.MethodPeripheral blood mononuclear cells (PBMC) were isolated from immunotherapy naïve asthmatics, asthmatics on subcutaneous Der p1 immunotherapy and from healthy individuals. PBMC were pre‐conditioned with anti‐CD3/anti‐CD28 mAbs, Der p1 or IFN‐γ in the presence and absence of DFSCs and analysed for T cell viability and proliferation, CD4+CD25+FOXP3+ regulatory T cell frequencies, cytokine expression, and GATA3, T bet and FoxP3 expressions. Neutralisation of TGF‐β and blockade of IDO a...
Biomedicine & Pharmacotherapy, 2017
Background: The major feature of asthma is governed by chronic airway inflammation. This investigation was proposed to achieve the suitable candidate for ameliorating long-term chronic asthmatic changes of respiratory tract. Methods: 36 rats were classified into healthy (C) and ovalbumin (OVA)-sensitized animals (S). To sensitize, the rats were exposed to OVA over a course of 32 AE 1 days. One day after sensitization, equal six different groups were subjected to experimental procedure (n = 6); Rats only received intratracheally 50 ml PBS (CPT and SPT groups), 50 ml conditioned medium (CM) (CST and SST groups) and 50 ml PBS containing 2 Â 10 6 rat bone marrow-derived mesenchymal stem cells (rBMMSCs) (CCT and SCT groups). Two weeks after treatment, tracheal responsiveness, immunologic responses and recruitment of rBMMSCs into the lung as well as pathological changes were evaluated. Results: A high degree of tracheal responsiveness, total white blood cell and percentages of eosinophil and neutrophil was significantly recorded in all sensitized groups rather than of controls (p < 0.001 to p < 0.05). Of interest, all above-mentioned parameters decreased significantly in SST and notably SCT groups as compared to S group (p < 0.001 to p < 0.05). The results revealed decrease number of blood CD3 + CD4 + and concurrent increase in CD3 + CD8 + in all sensitized rats as compared to control (p < 0.001 to p < 0.05). Noticeably, no significant modulatory effects of either cell or CM administration were achieved on the CD3 + CD4 + and CD3 + CD8 + populations in non-asthmatic rats. Moreover, the number of CD3 + CD4 + in SST and SCT groups tended to increase, which coincided with a decreased manner of CD3 + CD8 + populations as compared with S group (p < 0.001 to p < 0.05). However, the CD3 + CD4 + cells in SCT rats were significantly higher than the group SST (p < 0.01) whereas CD3 + CD8 + cells diminished simultaneously (p < 0.001). Real-time PCR analysis further showed that both CM and particularly MSCs changed the expression of interleukin (IL)-4 and IL-10 in the asthmatic groups to the near level of control rats (p<0.001 to p < 0.05). Histopathological analysis revealed a profound reduction of lungs injuries in asthmatic rats when received CM and peculiarly mesenchymal stem cells (p < 0.01 to p < 0.05). Conclusion: Our study shed light on the superior effects of rBMMSCs, rather than CM, in attenuating of chronic asthmatic changes in the rat model.
Clinical <html_ent glyph="@amp;" ascii="&"/> Experimental Allergy, 1998
Background The underlying mechanisms of immunotherapy (IT) are still unknown but may be related to modifications of cytokine production of T lymphocytes. Objective In this study we determined the in vitro allergen-induced production of IL-2, IL-4, IL-5, IL-12 and IFNg of peripheral blood mononuclear cells (PBMC) of eight young asthmatics, aged 15 Ϯ 2 years, receiving IT (IT group) and of eight comparable asthmatics, aged 13 Ϯ 3.5 years, who never received IT (non-IT group). Methods All patients suffered from perennial asthma and were allergic to house dust mite (HDM). They were selected if they showed a positive stimulation index (SI) of PBMC after in vitro incubation with HDM (i.e. SI > 2). Cells were incubated with and without HDM (10 mg/mL) during 24 h, 48 h and 7 days. Cytokines were determined in the supernatant at the three time points and are expressed as median values in pg/mL. Results In the IT group the secretion of IL-2 was lower compared with the non-IT group after 7 days incubation of PBMC with HDM (0 vs 33.2, P ¼ 0.008). In both groups maximal secretion of IL-2 was observed after 48 h. In the non-IT group a high value of IL-2 persisted after 7 days, whereas in the IT group a significant decline of IL-2 occurred after 7 days. Although IL-4 secretion was low in all subjects, more patients of the non-IT group showed detectable IL-4 in the HDM cultures after 24 h and 48 h, although the difference was not statistically significant (P ¼ 0.08 and P ¼ 0.28, respectively). Furthermore, IL-4 secretion was lower in the HDM cultures after 24 h in the IT group (1.75 vs 4.1, P ¼ 0.011) and 48 h (2.2 vs 4.1, P ¼ 0.035). IL-5 secretion was lower in the HDM cultures after 24 h (12.4 vs 47.6, P ¼ 0.035) and 48 h (26.8 vs 135, P ¼ 0.046) in the IT group than in the non-IT group. After 7 days of incubation with HDM there was no difference between the groups. There was no difference between both groups in secretion of IFNg and IL-12. Conclusions These results show a difference in vitro cytokine secretion of PBMC of asthmatics receiving IT compared with asthmatics who never received IT. PBMC of patients receiving IT secrete less IL-2 and IL-5 after in vitro incubation with HDM and show a tendency to secrete less IL-4. The efficacy of IT may be attributed to a modified cytokine secretion of PBMC.
Medical Archives
Background: Allergic Rhinitis (AR) is the most common immunological disease that has been associated with inflammatory responses and is characterized by sneezing. Previous studies found that AR's allergen exposure significantly induces plasma cells and reduces regulatory T (Treg) cells, a population that contributes to control AR. Therefore, upregulating Treg expression can regulate plasma cells leading to inhibit sneezing in AR. Mesenchymal stem cells (MSCs) are multipotent stem cells that have the immunoregulatory and antiinflammation ability by secreting various cytokines including IL-10 and TGF-β which potent as a promising therapeutic modality for allergic airway diseases, including AR. Objective: To investigate the role of MSCs in generating CD4+, CD25+, and Foxp3+ Regulatory T cells associated with suppressing plasma cell in AR model. Methods: In this study, fifteen male Wistar rats (6 to 8 weeks old) were randomly divided into three groups (control group, sham group, and MSCs treatment group). OVA nasal challenge was conducted daily from day 15 to 21, and MSCs (1x10 6) were administrated intraperitoneally to OVA-sensitized rats on day 21. Sneezing was observed from day 22 to 28. The rats were sacrificed on day 22 and day 28. The expression of CD4+ CD25+ Foxp3+ in Treg and plasma cells was analyzed by flow cytometry assay. Results: This study showed that the percentage of plasma cell and sneezing times significantly decreased in MSCs treatment. This finding was aligned with the significant increase of CD4+CD25+Foxp3+ Treg level. Conclusion: MSCs administration suppress plasma cells population and sneezing times by up regulating Treg to control AR.
Clinical and Experimental Health Sciences, 2019
Objective: The purpose of our study is to investigate the immunomodulatory effects of Dental Follicle Mesenchymal Stem Cells (DF-MSCs) on lymphocytes isolated from peripheral blood of Atopic Dermatitis (AD) patients, a Th2 disease and psoriasis, a Th1 / Th17 disease and compare them with healthy individuals in vitro. Methods: Patients with the AD (n = 9) and psoriasis (n = 6) who are followed up in Marmara University Pediatric Allergy and Immunology and Dermatology outpatient clinics and healthy subjects (n = 6) were included. Peripheral Blood Mononuclear Cells (PBMCs) were isolated from 20 ml of venous blood of all participants. Cells were cultured for 72 hours in the absence and presence of DF-MSCs with anti-CD3/anti-CD28 stimulation or without stimulation. At the end of this period, CD4+ and CD8+ T lymphocyte proliferation and cytokine levels from the culture supernatants were analyzed by flow cytometry. Results: In the presence of DF-MSCs, proliferation ratio was suppressed in both CD4+ and CD8+ cells in AD and psoriasis patients (p<0,05). IFN-γ levels significantly increased in AD patients in the presence of DF-MSCs (p<0,05) whereas decreased significantly in psoriasis patients in the presence of DF-MSCs (p<0,05). IL-4 levels significantly decreased in AD patients in the presence of DF-MSCs (p<0,05) but remained unchanged in psoriasis patients (p>0,05). IL-10 increased significantly in both groups in the presence of DF-MSCs (p<0,05). Conclusion: Our results support immunoregulatory effects of DF-MSCs on both AD and psoriasis which are Th2 and Th1 / Th17 dominant diseases respectively. Our evidence-based results demonstrated that DF-MSCs could have a beneficial therapeutic implication for inflammatory skin diseases.
American Journal of Respiratory and Critical Care Medicine, 2005
Increased Th2 cytokine production in asthma is widely accepted, but excess production by asthmatic human airway CD4 ϩ T cells has not been demonstrated, nor has a relationship with disease severity. The importance of airway CD8 ϩ T cell type 1 and type 2 cytokine production in asthma is unknown. We investigated frequencies of IFN-␥, interleukin (IL)-4 and IL-5 producing CD4 ϩ and CD8 ϩ blood and sputum T cells from normal subjects and subjects with asthma and compared between cell subsets, subject groups, and body compartments with and without in vitro stimulation and investigated relationships between cytokine production and asthma severity. Production of IL-4, IL-5, and IFN-␥ by unstimulated sputum CD4 ϩ and CD8 ϩ T cells was increased in subjects with asthma and related to disease severity, more for CD8 ϩ than for CD4 ϩ T cells. Frequencies of sputum CD8 ϩ T cells producing type 1 and type 2 cytokines were similar to those of CD4 ϩ T cells. In vitro stimulation polarized peripheral blood cytokine production toward IFN-␥ production, significantly more in subjects with asthma than in normal subjects. These data demonstrate increased type 1 and 2 cytokine production in CD4 ϩ and CD8 ϩ T cells in sputum and relate production to disease severity. Findings in blood did not reflect those in airways.
IL4, IL13, and dexamethasone augment fibroblast proliferation in asthma
Journal of Allergy and Clinical Immunology, 2001
Background: IL-4 and IL-13 have been shown to be critical for expression of the asthma phenotype in a murine model and may modulate human fibroblast function. Objective: We hypothesized that IL-4 and IL-13 would increase airway fibroblast proliferation and reduce the ability of dexamethasone to decrease this proliferation. Methods: Six subjects with severe asthma, 5 subjects with mild asthma, and 5 healthy subjects underwent bronchoscopy with endobronchial biopsy. Biopsy specimens were placed in Dulbecco modified Eagle medium and cultured, and only fibroblasts from the first and second passages were evaluated. Cells were incubated with IL-4 (50 ng/mL), IL-13 (10 ng/mL), and the combination for 48 hours in the presence and absence of dexamethasone, 10 -7 mol/L, and 10 -8 mol/L. Fibroblasts were also incubated with IFN-γ at 50 ng/mL to assess the response of a T H 1 cytokine on proliferation.
Mesenchymal stem cell (MSC) immunosuppressive functions make them attractive candidates for anti-inflammatory therapy in allergic asthma. However, the mechanisms by which they ensure therapeutic effects remain to be elucidated. In an acute mouse model of house dust mite (Der f)-induced asthma, one i.v. MSC injection was sufficient to normalize and stabilize lung function in Der f-sensitized mice as compared to control mice. MSC injection decreased in vivo airway responsiveness and decreased ex vivo carbachol-induced bronchial contraction, maintaining bronchial expression of the inhibitory type 2 muscarinic receptor. To evaluate in vivo MSC survival, MSCs were labeled with PKH26 fluorescent marker prior to i.v. injection, and 1 to 10 days later total lungs were digested to obtain single-cell suspensions. 91.5 6 2.3% and 86.6 6 6.3% of the recovered PKH26 1 lung cells expressed specific macrophage markers in control and Der f mice, respectively, suggesting that macrophages had phagocyted in vivo the injected MSCs. Interestingly, only PKH26 1 macrophages expressed M2 phenotype, while the innate PKH26 2 macrophages expressed M1 phenotype. Finally, the remaining 0.5% PKH26 1 MSCs expressed 10-to 100-fold more COX-2 than before injection, suggesting in vivo MSC phenotype modification. Together, the results of this study indicate that MSCs attenuate asthma by being phagocyted by lung macrophages, which in turn acquire a M2 suppressive phenotype.
Bone marrow-derived mononuclear cells vs. mesenchymal stromal cells in experimental allergic asthma
Respiratory Physiology & Neurobiology, 2013
We compared the effects of bone marrow-derived mononuclear cells (BMMCs) and mesenchymal stromal cells (MSCs) on airway inflammation and remodeling and lung mechanics in experimental allergic asthma. C57BL/6 mice were sensitized and challenged with ovalbumin (OVA group). A control group received saline using the same protocol. Twenty-four hours after the last challenge, groups were further randomized into subgroups to receive saline, BMMCs (2 × 10 6) or MSCs (1 × 10 5) intratracheally. BMMC and MSC administration decreased cell infiltration, bronchoconstriction index, alveolar collapse, collagen fiber content in the alveolar septa, and interleukin (IL)-4, IL-13, transforming growth factor (TGF)- and vascular endothelial growth factor (VEGF) levels compared to OVA-SAL. Lung function, alveolar collapse, collagen fiber deposition in alveolar septa, and levels of TGF- and VEGF improved more after BMMC than MSC therapy. In conclusion, intratracheal BMMC and MSC administration effectively modulated inflammation and fibrogenesis in an experimental model of asthma, but BMMCs was associated with greater benefit in terms of reducing levels of fibrogenesis-related growth factors.