scholarly journals Modulation of Mycobacterium bovis-Specific Responses of Bovine Peripheral Blood Mononuclear Cells by 1,25-Dihydroxyvitamin D3

2001 ◽  
Vol 8 (6) ◽  
pp. 1204-1212 ◽  
Author(s):  
W. R. Waters ◽  
B. J. Nonnecke ◽  
T. E. Rahner ◽  
M. V. Palmer ◽  
D. L. Whipple ◽  
...  

ABSTRACT Historically, administration of vitamin D has been considered beneficial in the treatment of tuberculosis. The interaction of this vitamin {i.e., 1,25-dihdroxyvitamin D3[1,25(OH)2D3]} with the antitubercular immune response, however, is not clear. In the present study, in vitro recall responses of peripheral blood mononuclear cells (PBMC) from cattle infected with Mycobacterium bovis were used to study the immune-modulatory effects of 1,25(OH)2D3 on M. bovis-specific responses in vitro. Addition of 1 or 10 nM 1,25(OH)2D3 inhibited M. bovis-specific proliferative responses of PBMC from M. bovis-infected cattle, affecting predominately the CD4+ cell subset. In addition, 1,25(OH)2D3 inhibited M. bovis-specific gamma interferon (IFN-γ) production yet enhanced M. bovis-specific nitric oxide (NO) production. Lymphocyte apoptosis, measured by flow cytometry using annexin-V staining, was diminished by addition of 1,25(OH)2D3 to PBMC cultures. These findings support the current hypothesis that 1,25(OH)2D3enhances mycobacterial killing by increasing NO production, a potent antimicrobial mechanism of activated macrophages, and suggest that 1,25(OH)2D3 limits host damage by decreasingM. bovis-induced IFN-γ production.

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S758-S758
Author(s):  
Aviva Szigeti ◽  
Margaret Hammerschlag ◽  
Diana Weaver ◽  
Tamar Smith-Norowitz ◽  
Stephan Kohlhoff

Abstract Background Chlamydia pneumoniae (Cpn) is unique in its ability to cause chronic infections, potentially triggering asthma exacerbations as well as subsequent asthma development. Th1-mediated immunity and IFN-γ are critical for clearing chlamydial infections. Persistent or recent Cpn infection may be identified in vitro by detecting T-helper cytokine IFN-γ produced by peripheral blood mononuclear cells (PBMC) stimulated by Cpn. Inhaled corticosteroids (ICS) may have an inhibitory effect on IFN-γ. Prior studies have shown increased Th2 responses upon in vitro Cpn stimulation with increased age. Our aim was to determine whether age and inhaled corticosteroid (ICS) use affect Cpn-induced PBMC produced IFN-γ levels. Methods Pediatric and adult subjects with (n = 23) and without (n = 10) asthma were enrolled. PBMC obtained from all subjects were stimulated with Cpn (MOI = 0.1 x48h) in vitro. IFN-γ levels in culture supernatants were determined by ELISA and reported as pg/mL. Nasopharyngeal (NP) swabs were tested for Cpn using Real-Time PCR. Statistical analysis for continuous variables was performed using the Mann–Whitney U test. Results None of the subjects were positive for Cpn by PCR on NP swab. Levels of IFN-γ produced by PBMC stimulated by Cpn were similar between asthmatic vs. control subjects (41.7 vs. 68.8, respectively; P = 0.72) and between pediatric and adult subjects with asthma (IFN-γ 54 vs. 20.1 respectively, P = 0.95). Pediatric subjects with asthma who received ICS had lower IFN-γ levels than those who did not (median IFN-γ 25.5 vs. 209; P = 0.003). Conclusion Our finding of lower IFN-γ levels among asthma patients on ICS compared with those not on ICS suggests that ICS use may dampen the systemic inflammatory response. While we did not find a statistically significant difference between pediatric and adult age groups in this pilot study, there was a trend to higher Cpn-induced IFN-γ levels among younger pediatric subjects. Future prospective studies should further define predictors of diminished IFN-γ responses in patients with asthma. Disclosures All authors: No reported disclosures.


Pteridines ◽  
2013 ◽  
Vol 24 (3) ◽  
pp. 237-243
Author(s):  
Sebastian Schroecksnadel ◽  
Elena-Sophia Ledjeff ◽  
Johanna Gostner ◽  
Christiana Winkler ◽  
Katharina Kurz ◽  
...  

AbstractIn vitro, large amounts of neopterin are released from human monocyte-derived macrophages and dendritic cells primarily upon stimulation with Th1-type cytokine interferon-γ (IFN-γ). IFN-γ also induces the enzyme indoleamine 2,3-dioxygenase (IDO), which degrades tryptophan (TRP) to form kynurenine (KYN). IDO-mediated TRP catabolism is very effective in suppressing the proliferation of T lymphocytes as well as of pathogens in vitro and in vivo. In this study, we investigated whether exogenously added neopterin may influence IDO activity in resting and in stimulated peripheral blood mononuclear cells (PBMC). PBMC were isolated from healthy donors, and neopterin was added in a concentration range from 0.01 to 50 μmol/L. After 30 min, PBMC were stimulated or not with 10 μg/mL of mitogen phytohemagglutinin (PHA). After 48 h, culture supernatants were collected, KYN and TRP concentrations were measured by high-performance liquid chromatography, and the ratio of KYN vs. TRP was calculated as an estimate of IDO activity. Spontaneous as well as PHA-induced TRP breakdown was suppressed by exogenously added neopterin in a dose-dependent way; the lowest active concentration of neopterin was <100 nmol/L. As neopterin concentrations in the nanomolar range are commonly observed in patients suffering from infections, sepsis, or uremia, our results suggest that neopterin formation might also serve as a feedback mechanism to slow down TRP degradation in vivo.


2019 ◽  
Vol 67 (7) ◽  
pp. 1053-1060
Author(s):  
Elżbieta Kozłowska ◽  
Paulina Żelechowska ◽  
Adam Wysokiński ◽  
Paweł Rasmus ◽  
Anna Łucka ◽  
...  

Increasing evidence has shown that the immune system is involved in the schizophrenia development, with alterations in immune cell reactivity being one possible factor contributing to its pathogenesis. The purpose of the study was to evaluate in vitro the capability of peripheral blood mononuclear cells (PBMCs) obtained from subjects with schizophrenia and controls to engage in spontaneous and phytohemagglutinin (PHA)-stimulated cytokine production. The concentrations of various cytokines (interleukin (IL)-1β, IL-17A, tumor necrosis factor (TNF), interferon (IFN)-γ and IL-10) in supernatants from cultured PBMCs were measured using the cytometric bead array. No significant differences in the spontaneous production of IL-1β, IL-17A, IFN-γ and IL-10 by PBMCs were detected between individuals with schizophrenia and controls. TNF synthesis by PBMCs was found to be lower among those with schizophrenia. In all subjects and controls, greater cytokine generation was associated with PBMCs treated with PHA compared with those that were not. The PBMCs from people with schizophrenia displayed considerably higher sensitivity to mitogen stimulation, as the production of IL-17A, TNF and IFN-γ was at least threefold of that observed in healthy subjects, which may be driven by antipsychotics taken by patients with schizophrenia. Correlation was observed between spontaneous production of IFN-γ and Positive and Negative Syndrome Scale G subscore (which measures the general symptoms of schizophrenia) and between PHA-stimulated synthesis of IL–17A and G subscore. Our data confirm that the immune system dysregulation may underlie schizophrenia pathophysiology. There is a potential possibility that immunological tests could be used as a diagnostic, therapeutic and side-effects biomarker for schizophrenia, but further studies are needed.


Vaccines ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 109
Author(s):  
Muhammad Ehsan ◽  
Muhammad Haseeb ◽  
Ruisi Hu ◽  
Haider Ali ◽  
Muhammad Ali Memon ◽  
...  

During host-parasite interactions, binding of excretory/secretory proteins (ESPs) on the host immune cells is considered the fundamental phase for regulation of immune responses. In this study, gene encoding Haemonchus contortus tropomyosin (Hc-TpMy), was successfully cloned and expressed, and the recombinant protein after host cell surface attachment was evaluated for immune functional analysis with goat peripheral blood mononuclear cells (PBMCs) in vitro. The isopropyl-β-D-thiogalactopyranoside (IPTG)-induced recombinant protein was successfully recognized by the sera of rat experimentally infected with rHc-TpMy. The immunofluorescence assay detected attachment of rHc-TpMy on the surface of host PBMCs. Furthermore, immunoregulatory roles of rHc-TpMy on cytokines expression, PBMC proliferation, migration, nitric oxide (NO) production, apoptosis and monocytes phagocytosis were observed. The results showed that expression of IL-4 and IFN-γ cytokines, cell proliferation, NO production and PBMC migration were significantly suppressed by goat PBMCs after co-incubation with rHc-TpMy protein. However, the productions of IL-10, IL-17 and TGF-β1 cytokines, PBMCs apoptosis and monocytes phagocytosis were elevated at dose dependent manner. Our findings indicated that rHc-TpMy is an important ES binding protein exhibit distinct immuno-suppressive roles on goat PBMCs which might be a potential molecular target to control haemonchosis in future.


2009 ◽  
Vol 127 (3-4) ◽  
pp. 382-388 ◽  
Author(s):  
Cleusa Alves Theodoro Rodrigues ◽  
Luís Fábio da Silva Batista ◽  
Roberto Santos Teixeira Filho ◽  
Claire da Silva Santos ◽  
Cristiane Garboggini Pinheiro ◽  
...  

2016 ◽  
Vol 42 (04) ◽  
pp. 195-201
Author(s):  
Yafen Cheng ◽  
Chung-Hsi Chou ◽  
Hsiang-Jung Tsai

Mycobacterium bovis (M. bovis) is causative agent of tuberculosis in cattle and humans populations. To understand its effects on gene expression profiles, we conducted an in vitro time-course study to identify transcriptional changes in infected bovine peripheral blood mononuclear cells (PBMCs), using quantitative RT-PCR. We discovered a likely involvement of C-type lectin domain family 4, member E (CLEC4E) in triggering a series of negative intracellular signaling via Syk/CARD9 pathway for cytokines, as early as 24 h post-infection (hpi). This is the first report confirming induction of CLEC4E and the Syk/CARD9 pathway in PBMCs in response to M. bovis infection, and these findings support the view that M. bovis inhibits signaling pathways of antimycobacterial host defense in bovine cells. In addition, M. bovis infection in PBMCs may suppress apoptosis by interfering with TNF-[Formula: see text] signaling. This study, contributes to a better understanding of M. bovis-reduced signal transduction and microbial changes in PBMCs earlier than 24 hpi.


2018 ◽  
Vol 68 (2) ◽  
pp. 201-210
Author(s):  
Hiromichi Ohtsuka ◽  
Maki Inoue ◽  
Yosuke Maeda ◽  
Taishi Tanabe ◽  
Motoshi Tajima

Abstract The expressions of cytokines mRNA, including interleukin-4 (IL-4), interleukin- 17A (IL-17A) and interferon-gamma (IFN-γ), their master regulatory transcription factors, and signal transducers and activator of transcription (STAT) stimulated in vitro with Pasteurella (P.) multocida soluble antigen were examined in peripheral blood mononuclear cells (PBMC) from Holstein calves. The healthy Holstein calves were divided into three groups; 2 weeks old (2W Group, N=8), 6 weeks old (6W Group, N=8), and 10 weeks old (10W Group, N=8). PBMC were stimulated in vitro by soluble antigen of P. multocida. There were significantly lower expressions of IFN-γ, IL-4, and STAT-6 mRNA of PBMC stimulated with P. multocida soluble antigen in the 2W Group compared to that in the 10W Group. Expression of IL-17A and IFN-γ in PBMC stimulated with P. multocida soluble antigen were significantly higher compared with the PBMC without stimulation in the 6W groups. The results of the present study demonstrated that 2W old calves had decreased cytokine expression of PBMC when in vitro stimulated with P. multocida soluble antigen in vitro.


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