Elisa Vintiñi; Marcela Medina
Volume 14, Issue 4 , December 2017, , Pages 325-339
Abstract
Background: Polyinosinic:polycytidylic acid (Poly-IC) has been used as a viral stimulus to mimic in vivo and in vitro infection induced by some viruses. Objective: To determine whether non-viable Lactobacillus casei CRL431 (LcM) can modulate the immune response induced by Poly I:C in co-culture ...
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Background: Polyinosinic:polycytidylic acid (Poly-IC) has been used as a viral stimulus to mimic in vivo and in vitro infection induced by some viruses. Objective: To determine whether non-viable Lactobacillus casei CRL431 (LcM) can modulate the immune response induced by Poly I:C in co-culture models of peripheral blood mononuclear cells (PBMC) and A549 cells. Methods: T and NK cell activation was evaluated by flow cytometry and levels of TNF-α, IFN-γ, IL-10, IL-29, and IL-17 by ELISA. Cells in direct contact with A549 (PBMC-A549) and cells with no contact with it (PBMC//A549) were used for this purpose. PBMCs alone and both co-culture systems were stimulated for 24 h with the following stimuli: LPS (10 µg/ml), LcM (106 UFC/ml), Poly I:C (2 µg/ml), Poly I:C+LcM, and LcM (3 h)+Poly I:C. Moreover, unstimulated cells were used as a control. Results: Poly I:C and LcM (3 h)+Poly I:C in PBMC-A549 showed a significant increase in the percentage of CD8+ expression (p<0.05). All stimuli induced significant activation from T CD4+, CD8+ cells compared with unstimulated PBMCs in both co-culture cells system. However, activation percentages were higher in direct co-culture. Poly I:C induced a higher level of pro-inflammatory TNF-α and IFN-γ cytokines as well as IL-17 and IL-29 with lower IL-10 levels in both co-culture systems while LcM induced a beneficial pattern of cytokines that would regulate Poly I:C effect. Conclusion: This in vitro model allowed us to highlight the potential of LcM as a modulator of anti-viral immune response and suggest its potential use in formulations against RNA respiratory viruses.
Mahsa Rahmani; Hamid Reza Khorasani; Monireh Golpour; Ali Shabestani Monfared; Hosein Nattaj; Saeeid Abedian; Amrollah Mostafazadeh
Volume 13, Issue 1 , March 2016, , Pages 54-63
Abstract
Background: The human leukocyte antigen (HLA) matching between organ donor and recipient is an acceptable strategy in clinical transplantation since 1964. However, in bone marrow transplantation, finding matched donors is often problematic. Thus new method for down regulation of HLA can be an alternative ...
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Background: The human leukocyte antigen (HLA) matching between organ donor and recipient is an acceptable strategy in clinical transplantation since 1964. However, in bone marrow transplantation, finding matched donors is often problematic. Thus new method for down regulation of HLA can be an alternative strategy to solve this problem. Objective: To examine the effect of serum starvation on HLA class I expression in human peripheral blood mononuclear cells (PBMCs). Methods: PBMCs were cultured in RPMI-1640 supplemented with 10% FBS (non-starved cells) as well as in medium only (starved cells) for 16, 24, 48, 72, 96h under standard cell culture conditions. The pattern of cell death and HLA class I expression was determined by flowcytometry. Antigenicity of the starved PBMCs was evaluated in a one-way mixed lymphocyte culture by MTT assay. Results: Mean fluorescence intensity (MFI) of different indicated starved PBMCs gradually decreased and this reduction was stable after 96h of re-feeding with medium containing FBS. Under serum starvation condition, PBMCs showed apoptotic cell death pattern. There was a linear correlation between percentages of cells, which exhibited the late apoptosis death pattern and serum starvation period (r=0.88, p<0.01). Surprisingly, the starved PBMCs lost their stimulatory property in mixed culture with allo-reactive lymphocyte. Conclusions: Membrane HLA class I expression could be stably reduced in 96h starved human PBMCs culture condition, decreasing their allo-reactivity while their viability rate is enough for possible clinical application.
Seyed Javad Hasheminia; Sayyed Hamid Zarkesh-Esfahani; Sepideh Tolouei; Vahid Shaygannejad; Hedaiatallah Shirzad; Morteza Hashemzadeh Chaleshtory
Volume 11, Issue 2 , June 2014, , Pages 74-83
Abstract
Background: Multiple sclerosis (MS) is a T cell mediated autoimmune disease with unknown etiology. Appropriate MS therapeutic strategies need thorough understanding of both disease etiology and pathogenesis mechanisms. Ligation of TLR-2 and TLR-4 stimulates the production of several cytokines leading ...
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Background: Multiple sclerosis (MS) is a T cell mediated autoimmune disease with unknown etiology. Appropriate MS therapeutic strategies need thorough understanding of both disease etiology and pathogenesis mechanisms. Ligation of TLR-2 and TLR-4 stimulates the production of several cytokines leading to CNS autoimmunity and neurodegenerative diseases. Objective: To find a relationship between MS disability and TLR-2 and TLR-4 expression on mononuclear cells in the blood of MS patients. Methods: Forty-five new case (NC) MS patients (33 females and 12 males) and 45 age and gender-matched healthy controls (HC) were recruited to the study. PBMCs were prepared and the expressions of TLR-2 and TLR-4 were assessed by flowcytometry technique using appropriate monoclonal antibodies. Results: Our results showed that the expression of TLR-2 and TLR-4 proteins in the patients group was significantly higher than that of healthy controls. TLR-2 but not TLR-4 was correlated with expanded disability status scale (EDSS) scores. Conclusion: High expressions of TLR-2 and TLR-4 may represent a state of innate immune activation in patients with MS.
Mohammad Ali Rezaee; Yousef Motaharinia; Werya Hosseini; Ali Jalili; Ahmad Rashidi; Bita Mosavi; Ghasem Zamini; Mohammad Reza Rahmani
Volume 9, Issue 2 , June 2012, , Pages 109-118
Abstract
Background: Malassezia furfur is alipophilic yeast that causes skin disease. Objective: To evaluate the level of IL-10, IFN-γ and IL-12P70 in co-incubation of M. furfur grown on different forms of natural oils with PBMCs of healthy individuals. Methods: PBMCs were obtained from blood samples of ...
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Background: Malassezia furfur is alipophilic yeast that causes skin disease. Objective: To evaluate the level of IL-10, IFN-γ and IL-12P70 in co-incubation of M. furfur grown on different forms of natural oils with PBMCs of healthy individuals. Methods: PBMCs were obtained from blood samples of normal volunteers. M. furfur was cultured in different culture media containing almond oil, fish oil, walnut oil, full-fat milk, and a fat-free medium; and the yeasts grown were harvested and used for co-incubation with PBMCs in vitro. The IFN-γ, IL-10, and IL-12P70 levels were measured at different time intervals using ELISA methods. Results: Generally, IFN-γ and IL-10 levels in the coincubation of yeasts with walnut oil group (WOG) and fish oil group (FOG) were higher than those in the almond oil group (AOG) and full-fat milk group (FFMG). Although the IL-12P70 was higher in groups such as AOG, FOG, and WOG; the increase was not statistically significant. Conclusion: The results demonstrated that the type of fat used by M. furfur in the culture media can influence the immune response and increases IFN-γ and IL-10 levels in an early time point of the culture system.
