Determination of intracellular cytokines produced by Th1 and Th2 cells using flow cytometry in patients with brucellosis

Department of Immunology, Faculty of Medicine, Firat Medical Center, Firat University, 23119 Elazig, Turkey.
FEMS Immunology & Medical Microbiology (Impact Factor: 3.08). 09/2005; 45(2):253-8. DOI: 10.1016/j.femsim.2005.04.001
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ABSTRACT The aim of the study was to evaluate intracellular interferon-gamma (IFN-gamma), and interleukin-4 (IL-4) levels in pre- and post-treatment periods of brucellosis patients and to determine the relationship between these parameters and patients' clinical findings. Twenty-five patients diagnosed as brucellosis and 11 aged-matched healthy volunteers were included in the study. CD3+CD4+ T lymphocytes levels were significantly lower in patients with brucellosis as compared to the control group. CD3+CD8+ T lymphocytes and CD3+IFN-gamma+ levels were increased in brucellosis patients compared with the control group. CD4+IFN-gamma+ and CD4+IL-4+ levels were no different between patients and healthy individuals. CD3+IL-4+ levels decreased in patients compared with healthy controls. Pre-treatment CD3+IFN-gamma+ levels dramatically increased in patients responsive to management compared with the unresponsive ones. In responsive cases, CD3+IFN-gamma+ levels decreased statistically after the treatment while in unresponsive cases no meaningful change was observed with respect to treatment. Adding IFN-gamma to the treatment for improving the depleted levels of IFN-gamma can be beneficial in patients with brucellosis who shows a tendency to chronicity or patients who do not respond to the treatment.

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Available from: Vedat Bulut, Jul 06, 2015
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    • "These observations suggest that once acute brucellosis has resolved, that both the number of CD4+ cells and CD4+ functional response is reduced. Whether antigen-specific memory CD4+ cells are produced during acute or chronic brucellosis remains to be determined (Moreno-Lafont et al., 2002; Akbulut et al., 2005; Kinikli et al., 2005). Recently Elfaki and Al-Hokail observed that mice deficient in β2-microglobulin produced an impaired CD8+ response associated with increased Brucella bacterial load and decreased clearance (Moreno-Lafont et al., 2002; Elfaki and Al-Hokail, 2009). "
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    ABSTRACT: Brucella spp. are facultative intracellular Gram negative bacteria with specific tropism for monocytes/macrophages. Clinical manifestations of brucellosis are primarily immune-mediated and not thought to be due to bacterial virulence factors. Acquired immunity to brucellosis has been studied through observations of naturally infected hosts (cattle, goats), laboratory mouse models, and human infection. Cell-mediated immunity drives the clinical manifestations of human disease after exposure to Brucella species but high antibody responses are not associated with protective immunity. The precise mechanisms by which cell-mediated immune responses confer protection or lead to disease manifestations remain poorly understood. Descriptive studies of immune responses in human brucellosis show that TH1 (interferon-gamma) are associated with dominant immune responses, findings consistent with animal studies. Whether these T cell responses are protective, or determine the different clinical responses associated with brucellosis is unknown, especially with regard to undulant fever manifestations, relapsing disease, or are associated with responses to distinct sets of Brucella spp. antigens are unknown. Few data regarding T cell responses in terms of specific recognition of Brucella spp. protein antigens and peptidic epitopes, either by CD4+ or CD8+ T cells, have been identified in human brucellosis patients. Additionally because current attenuated Brucella vaccines used in animals cause human disease, there is a true need for a recombinant protein subunit vaccine for human brucellosis, as well as for improved diagnostics in terms of prognosis and identification of unusual forms of brucellosis. This review will focus on current understandings of antigen-specific immune responses induced by Brucella protein antigens that has promise for yielding new insights into vaccine and diagnostics development, and for understanding pathogenetic mechanisms of human brucellosis.
    Frontiers in Cellular and Infection Microbiology 02/2012; 2:1. DOI:10.3389/fcimb.2012.00001 · 3.72 Impact Factor
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    ABSTRACT: Aim: To investigate whether cytokines are eff ective in predicting relapses among patients with acute brucellosis. Materials and methods: Th is trial was conducted in 42 patients who were being followed-up with diagnosis of acute brucellosis. Serum samples were obtained on days 0 and 45. In patients whose clinical symptoms recurred within a year of treatment and exhibited infectious parameters in compliance with brucellosis, a Rivanol standard tube agglutination (STA) test was performed and the diagnosis of relapse was based on brucella immunoglobulin M (IgM). Serum samples were evaluated for various parameters, namely tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), interleukin 2 (IL-2), IL-4, IL-6, IL-8, IL-10, and soluble IL-2 receptor (sIL-2R). Results: Relapse was seen in 7 patients. No diff erence was found between relapsing patients (RPs) and fully recovered patients (FRPs) in terms of age, sex, leukocyte levels, or C-reactive protein (CRP) values. Comparison of TNFα, IFNγ, IL-2, IL-4, IL-6, IL-8, and IL-10 values on day 0 (day of enrollment) revealed 2-fold higher IL-8 values among RPs compared to FRPs. IL-8 was suggested as signifi cant in terms of predicting relapse. Conclusion: Diagnosis and treatment of relapsing cases in acute brucellosis have not yet been clarifi ed. Predicting relapse by certain laboratory evaluations may be benefi cial in preventing clinical relapses by rearranging treatment and monitoring strategies of patients.
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    ABSTRACT: In animal models, a defective Th1 response appears to be critical in the pathogenesis of brucellosis, but the Th1 response in human brucellosis patients remains partially undefined. Peripheral blood from 24 brucellosis patients was studied before and 45 days after antibiotherapy. Twenty-four sex- and age-matched healthy donors were analyzed in parallel. Significantly increased levels of interleukin 1beta (IL-1beta), IL-2, IL-4, IL-6, IL-12p40, gamma interferon (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha), but not of IL-10, in serum and/or significantly increased percentages of samples with detectable levels of these cytokines, measured by enzyme-linked immunosorbent assays (ELISA), were found for untreated brucellosis patients, but these levels were reduced and/or normalized after treatment. Flow cytometry studies showed that the intracytoplasmic expression of IFN-gamma, IL-2, and TNF-alpha, but not that of IL-4, by phorbol myristate-activated CD4(+) CD3(+) and CD8(+) CD3(+) T lymphocytes was significantly increased in untreated brucellosis patients and was also partially normalized after antibiotherapy. The percentage of phagocytic cells, the mean phagocytic activity per cell, and the phagocytic indices for monocytes at baseline were defective and had only partially reverted at follow-up. T lymphocytes from untreated brucellosis patients are activated in vivo and show Th1 cytokine production polarization, with strikingly high serum IFN-gamma levels. In spite of this Th1 environment, we found deficient effector phagocytic activity in peripheral blood monocytes.
    Infection and immunity 07/2010; 78(7):3272-9. DOI:10.1128/IAI.01385-09 · 3.73 Impact Factor
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