Whole blood assay to access T cell-immune responses to Mycobacterium tuberculosis antigens in healthy Brazilian individuals

Antas, Paulo Renato Zuquim; Cardoso, Fernando L. L.; Oliveira, E. B. de; Gomes, Patrícia K. C.; Cunha, K S; Sarno, Euzenir Nunes; Sampaio, Elizabeth P.

doi:10.1590/s0074-02762004000100009

Cited by 8 publications

(4 citation statements)

References 9 publications

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“…It is noteworthy to mention that the cytokine responses to mycobacterial stimulation observed in the whole blood assay might differ from those detected in other culture systems. While the levels of some cytokines such as IL-2, IL-5, IL-6, IL-7, IL-13, TNF-α, MCP-1, MIP-1β (Silva, et al 2013) and IFN-γ (Antas, et al 2004);Deenadayalan, et al 2013;Silva, et al 2013) were comparable between whole blood and PBMC cultures stimulated with specific M. tuberculosis antigens, levels of other cytokines like IL-1β, IL-4, IL-10, IL-12 (p70), IL-17, G-CSF and GM-CSF were significantly different between the two culture models (Silva, et al 2013).…”

Section: Discussionmentioning

confidence: 99%

“…However, other studies employing longer stimulation periods of 5 and 6 days were capable of detecting IFN-γ in whole blood cultures stimulated with specific M. tuberculosis antigens (Antas, et al 2004;Deenadayalan, et al 2013;Silva, et al 2013). Thus, future kinetic studies employing additional and extended time points might be useful in addressing the effect of mycobacterial stimulation on the expression of other surface receptors and on the release of cytokines.…”

Section: Discussionmentioning

confidence: 99%

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Analysis of the immunomodulatory properties of two heat-killed mycobacterial preparations in a human whole blood model

et al. 2015

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The significant role played by mycobacteria in modulating immune responses through enhancing the crosstalk between innate and adaptive immunity has been highlighted in several studies. Owing to their unique antigenic profile, heat killed (HK) preparations of rapid-growing mycobacteria, currently undergoing clinical development, have been assessed as adjuvant therapy in various diseases. The purpose of this study is to investigate the regulation of leukocyte surface receptors, in whole blood from healthy donors, following in vitro stimulation with HK Mycobacterium vaccae (M. vaccae) or M. obuense. We have demonstrated the ability of both mycobacterial preparations to target monocytes and neutrophils and to regulate the surface expression of selected adhesion receptors, antigen-presenting and costimulatory receptors, pattern recognition receptors, complement and Fc receptors, as well as cytokine/chemokine receptors. Toll-like receptors (TLRs) 1 and 2 were also shown to be involved in mediating the M. obuense-induced upregulation of selected surface receptors on monocytes. Whole blood stimulation with M. vaccae or M. obuense resulted in a significant increase in the secretion of a specific set of cytokines and chemokines. Both mycobacterial preparations induced strong antigen-specific proliferative responses in peripheral blood mononuclear cells. Collectively, our data shows that M. vaccae and M. obuense have the potential to act as potent immunomodulators. Future research based on these findings may reveal novel immune pathways induced by these preparations with potential implication for their use in diverse immunotherapeutic approaches.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Analysis of the immunomodulatory properties of two heat-killed mycobacterial preparations in a human whole blood model

et al. 2015

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“…However, because there were no children aged < 13 years and HIV-positive patients enrolled in this study, we could not determine whether they were risk factors for false negative IGRA results. Third, the growing research on comparative studies of cytokine levels in cultures of human whole blood and PBMCs demonstrated that whole blood assays correlated well with PBMC cultures ( 27 – 29 ); inversely, poor correlation was noted in the IFN-γ levels in the two culture systems by Silberer and colleagues ( 30 ). In this study, we only detected the cytokine levels using PBMC cultures rather than whole blood, which may limit tis application.…”

Section: Discussionmentioning

confidence: 99%

Significant difference in Th1/Th2 paradigm induced by tuberculosis-specific antigens between IGRA-positive and IGRA-negative patients

Ren²,

Yuan³

et al. 2022

Front. Immunol.

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False negative interferon-γ release assay (IGRA) results constitute the major dilemma for the diagnosis of tuberculosis (TB) infections. Herein, we conducted a cohort study to compare the host immunological response to TB-specific antigens between active TB patients with positive and negative IGRA results and control groups. A total of 274 laboratory-confirmed TB patients were included in our analysis, consisting of 221 were IGRA positive and 53 were IGRA negative. Patients with the elderly were identified as an independent risk factor for negative IGRA results. In addition, the elevated level of IL-4 and the decreased levels of IFN-γ, IL-2, IL-6, IL-1β, and IL-12 in IGRA negative TB relative to IGRA positive TB group, demonstrating a significant difference in Th1/Th2 paradigm between two groups. The IFN-γ&IL-2 based assay could correctly identify 247 out of 307 MTB-infected individuals [271 TB patients and 36 individuals with latent TB infection (LTBI)], demonstrating a sensitivity of 80.5%. Then the IFN-γ and IL-4 were applied to distinguish healthy control and IGRA-negative group. When using the stepwise algorithm, the sensitivity for detecting Mycobacterium tuberculosis (MTB) infections was significantly increased from 80.5% to 89.6%. Additionally, patients with negative IGRA results had a conversion to culture-negative status longer than those with positive IGRA results. In conclusion, a stepwise algorithm outperforms IGRA assays to accurately identify MTB infections by the combination IFN-γ, IL-2, and IL-4. Further study is needed to evaluate the accuracy of our diagnostic algorithm in the LTBI population.

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“…Based on the alteration of EBV-specific T-cell function in MS patients, the aim of the current study was to establish a comparison in immune cell response, after their exposure to EBV peptides. To provide a culture that is physiologically similar to in vivo condition, we used whole blood (WB), a suitable substitute for peripheral blood mononuclear cells (PBMCs), because purification of PBMC using Ficoll results in selective loss of gamma delta T lymphocytes and other important components impacting cellular response and IFN-γ levels [12,13]. Furthermore, WB assay is an easy and fast method which sustains the integrity of cellular components of blood and allows different elements to have essential interactions equivalent to in vivo situations [14,15].…”

Section: Introductionmentioning

confidence: 99%

Investigation of IL-2 and IFN-γ to EBV Peptides in Stimulated Whole Blood among Multiple Sclerosis Patients and Healthy Individuals

Rafiee¹,

Ravanshad²,

Asadi³

et al. 2021

Intervirology

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Introduction: Epstein-Barr virus (EBV), a double-stranded DNA virus, has 2 phases of lytic and latent infection in host cells. After infecting B lymphocytes, EBV becomes persistent in these cells. In healthy individuals, T lymphocytes play a key role in killing EBV-infected B cells. Statistical studies have shown that symptomatic EBV infection increases the risk of MS. Methods: This study intended to measure the immune system’s response against the different components of EBV, focusing particularly on T lymphocytes’ reaction. Consequently, the mRNA level of IL-2 and IFN-γ, liable for impressing autoimmune diseases and as indicators of T-cell function, was compared in EBNA1- and BRLF1-treated whole blood (WB) cultures of 10 healthy individuals and 10 MS patients using real-time RT-PCR. Results: The analysis of the results demonstrated a significant increased level of IL-2 in MS patients than healthy subjects after exposure to both peptides. Also, the mRNA level of IFN-γ increased in MS patients in EBNA1-treated WB culture. Conclusion: According to the study’s results, EBV peptides can reactivate immune cells, especially T lymphocytes, and may indirectly induce inflammation and develop MS; however, it seems that long-time exposure to these peptides has reducing effect on T-cell function and faces the control of infected B lymphocytes with difficulties.

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Whole blood assay to access T cell-immune responses to Mycobacterium tuberculosis antigens in healthy Brazilian individuals

Abstract: The production of interferon γ (IFNγ) guarantees effective T cell-mediated immunity against

Cited by 8 publications

References 9 publications

Analysis of the immunomodulatory properties of two heat-killed mycobacterial preparations in a human whole blood model

Analysis of the immunomodulatory properties of two heat-killed mycobacterial preparations in a human whole blood model

Significant difference in Th1/Th2 paradigm induced by tuberculosis-specific antigens between IGRA-positive and IGRA-negative patients

Investigation of IL-2 and IFN-γ to EBV Peptides in Stimulated Whole Blood among Multiple Sclerosis Patients and Healthy Individuals

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