Direct correlation between Th1 and Th17 responses in immunity to Brucella infection

Th1 cells play a central role in immunity to brucellosis, while the exact role of Th17 cells has remained unknown. This study aimed to evaluate the peripheral distributions of Th1 and Th17 cells and serum levels of IFN-γ, IL-17A and IL-22 cytokines in brucellosis patients. One hundred patients (36 acute, 41 under-treatment and 23 relapsed) and 30 age- and sex-matched healthy controls were included. The frequencies of Th1 and Th17 cells were determined by flow cytometric analysis. Serum levels of IFN-γ, IL-17A and IL-22 were measured by multi-analyte flow assay. Increased frequencies of Th1 and Th17 cells were observed in acute and relapsed brucellosis versus under-treatment patients and healthy controls (P < 0.05). The mean serum levels of IFN-γ were significantly elevated in acute and relapsed groups compared to under-treatment patients (P = 0.002 and P = 0.01 respectively). Acute patients showed higher levels of IL-22 than under-treatment (P = 0.008). Direct correlations were found between increased frequencies of Th1 and Th17 cells in acute and relapsed patients (P = 0.007 and P = 0.001 respectively) and between IL-17A and IL-22 in both groups of patients. Our findings indicate a cooperative role for Th1 and Th17 cells in immunity to brucellosis which is more evident during acute and relapse phases of brucellosis.     

T mu and T gamma-lymphocyte counts and their ratio in patients with brucellosis

The levels of circulating T mu- and T gamma-lymphocytes in brucellosis patients have been studied in relation to the clinical form of the disease, the severity of the process and the allergic transformation of the body. A decrease in the content of lymphocytes, affecting mainly T mu-cells and less commonly T gamma-cells, as well as a change in their ratio has been revealed. The level of this decrease depended on the clinical form of brucellosis, the severity of the course of acute and subacute brucellosis, the phase of chronic brucellosis and the allergic transformation of the body. The disproportion of immunoregulating cells in brucellosis, revealed in this study, is supposed to be of pathogenetic importance.     

Ciprofloxacin in the treatment of patients with brucellosis]

With the aim to estimate the clinical and immunological efficiency of the ciprofloxacin (cifloxinal) 105 patients with acute (51), subacute (19) and chronic (35) brucellosis were studied. Control group (17 patients with acute and 30 patients with chronic brucellosis) have been treated with combination of two antibiotics: doxycycline and rifampicin. Ciprofloxacin in a dose 500 mg bid within 14 days in acute stage and 20 days in chronic stage of disease essentially reduced duration of local inflammatory processes of brucellosis with simultaneous treatment of the chronic infection focus, provides good proximate and distant outcomes of treatment. Ciprofloxacin can be considered as an alternative drug for the treatment of brucellosis, more effective (clinically and immunologically) than a combination of two antibiotics: doxycycline and rifampicin.     

Diagnostic value of serum concentrations of high‐mobility group‐box protein 1 and soluble hemoglobin scavenger receptor in brucellosis

Both cluster of differentiation (CD)4⁺ and CD8⁺ T lymphocytes play key roles in immunity to Brucella, in part because they secrete interferon (IFN)‐γ and activate bactericidal functions in macrophages. Therefore, use of markers of macrophage activation may have diagnostic and prognostic significance. High‐mobility group‐box 1 protein (HMGB1), a late‐onset pro‐inflammatory cytokine, is secreted by activated macrophages. Soluble hemoglobin scavenger receptor (sCD163) is a specific marker of anti‐inflammatory macrophages. The aim of this study was to investigate the diagnostic value of HMGB1 and sCD163 concentrations in brucellosis and its various clinical forms. Serum HMGB1 and sCD163 concentrations in 49 brucellosis patients were compared with those in 52 healthy control subjects. Both serum HMGB1 and sCD163 concentrations were significantly higher in brucellosis patients than in healthy controls (P < 0.001). There were no statistically significant differences in serum concentrations of HMGB1 and sCD163 between cases of acute, subacute and chronic brucellosis. Additionally, serum HMGB1 concentrations were positively correlated with sCD163 concentrations, whereas neither HMGB1 nor sCD163 concentrations were correlated with C‐reactive protein concentrations, white cell counts or erythrocyte sedimentation rates. Therefore, serum concentrations of HMGB1 and sCD163 may be diagnostic markers for brucellosis, but neither can be used to differentiate the three different forms of this disease (acute, subacute and chronic).     

Indices of phagocytosis of S and R forms of Brucella in patients with acute and chronic brucellosis caused by Brucella abortus

The characteristics of phagocytosis (phagocytic activity and phagocytic index) in patients with acute and chronic brucellosis have been studied with B. abortus S- and R-forms used as its objects. The study has revealed that higher characteristics of phagocytosis with respect to B. abortus S-forms are observed in patients with acute brucellosis, whereas in patients with chronic brucellosis taking a benign course phagocytosis is more intensive in respect of R-forms. In patients with frequent exacerbations of chronic brucellosis differences in the characteristics of phagocytosis with respect to B. abortus S- and R-forms are not statistically valid.     

Serum cytokine levels in patients with acute brucellosis and their relation to the traditional inflammatory markers

The number of clinical studies on gamma interferon (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin-4 (IL-4) in human brucellosis is limited. The present study was focused on IFN-gamma, TNF-alpha and IL-4 levels in acute brucellosis cases, in the acute phase and at the end of the treatment. The relation of these cytokines to traditional inflammation markers was also investigated. The study included 27 cases of acute brucellosis and 20 healthy volunteers who had no complaints. It was found that mean IFN-gamma and TNF-alpha levels, CRP (C-reactive protein) levels and ESR (erythrocyte sedimentation rate) values were significantly higher in acute brucellosis cases as compared to post-treatment values and values measured in the control group. In addition, IFN-gamma and TNF-alpha levels measured in the acute phase correlated with the increase in CRP levels and ESR values. Our results confirmed that IFN-gamma and TNF-alpha are involved in the pathophysiology of brucellosis and are closely related to the inflammatory activation of the disease. In view of the present findings, it is suggested that IFN-gamma and TNFalpha may be used for monitoring brucellosis.     

Dynamics of lymphocyte subpopulations, immunoglobulins and specific antibodies in acute brucellosis

The main lymphocyte subpopulations (T-, B-, T mu-, T gamma, and L-lymphocytes), serum immunoglobulins (A, M, G), and specific antibodies have been analyzed at different periods of the development of acute brucellosis. The heterogeneous pattern of changes in the main characteristics of both humoral and cell-mediated immunity has been revealed. A definite interrelation of cell-mediated and humoral reactions in the immunological process in brucellosis has been established.     

Investigation of CTLA‐4 and CD86 gene polymorphisms in Iranian patients with brucellosis infection

The protective immune response against Brucella involves T‐cell‐mediated immunity. T‐lymphocyte receptors, CD28 and cytotoxic T‐lymphocyte‐associated protein‐4 (CTLA‐4), bind the same ligands, CD80 (B7‐1) and CD86 (B7‐2) on antigen‐presenting cells and regulate T cell activation. CD28 delivers stimulatory signals whereas CTLA‐4 provides inhibitory signals for T cell activation. Here, we investigated the association of four polymorphisms in CTLA4 (+49A/G [rs231775] and ?318 C/T [rs5742909]) and its ligand CD86 (+1057 G/A [rs1129055] and +2379G/C [rs17281995]) with brucellosis infection. The study included 153 Iranian patients with active brucellosis and 128 healthy individuals as the control group. Genotyping of the CTLA4 and CD86 variants was performed using tetra amplification refractory mutation system‐polymerase chain reaction (T‐ARMS‐PCR) and PCR–restriction fragment length polymorphism analysis, respectively. It was found that the CTLA4 ?318 CT genotype and T allele were present more frequently in cases than in controls and are therefore associated with an increased risk for brucellosis (?318 TT genotype; OR = 2.544, P = 0.002). Likewise, the CD86 +1057 GA and AA genotypes and A allele were associated with an increased risk of brucellosis (+1057 AA genotype; OR = 3.81, P = 0.001). However, no statistically significant difference between brucellosis patients and controls in the allele and genotype distributions of CTLA4, +49A/G (P = 0.859) and CD86, +2379G/C (P = 0.476) was found. In conclusion, CTLA4 ?318 CT genotype and T allele and the CD86 +057 GA and AA genotypes and A allele play roles as risk factors for developing brucellosis infection in Iran.

     

A simple enzyme immunoassay for detecting brucellosis antibodies

Abstract A simple enzyme immunoassay was developed and evaluated for serological diagnosis of brucellosis in 25 patients with various forms of brucellosis and 292 control patients with other conditions and disorders. All brucellosis patients gave a positive test with the initial sample. In 3 acute, febrile brucellosis patients with follow-up sera taken during therapy a sharp drop in specific antibody was noted. There was a less pronounced antibody reduction in 1 chronic and 2 relapse patients and an antibody increase in 1 chronic and 1 relapse case. All control samples gave negative results. In addition, the assay was evaluated as a screening test with 315 sera from 'healthy' individuals living in a brucellosis focus and representing 15% of that population. 11.5% (34/293) of subjects with no reported history of brucellosis and 45% (10/22) of cases treated in the past gave a positive test result. The agreement in those samples between the assay and the serum agglutination test was 95.5%.     

Detection of IgM Antibrucella Antibody in the Absence of IgGs: A Challenge for the Clinical Interpretation of Brucella Serology

The use of enzyme-linked immunosorbent assay (ELISA) for the detection of IgG and IgM antibodies antibrucella has become widespread in the diagnosis of human brucellosis. IgM anti-Brucella antibodies are indicative of acute infection. Between 2009–2013, 5307 patients were evaluated for serologic diagnosis at the Microbiology Laboratory of the Albacete General Hospital. A ELISA IgM-positive, IgG-negative anti-Brucella antibody serology pattern was detected in 17 of those patients. Epidemiology data, symptoms, laboratory data, treatment and outcome from these patients were reviewed. Sixteen patients presented with musculoskeletal pain, fatigue and/or fever and 1 was asymptomatic. Five patients received treatment with doxycycline combined with rifampin, gentamycin or streptomycin during 6–12 weeks, with no improvement. None of the 17 patients were finally diagnosed with brucellosis. Our results indicate that anti-Brucella IgM positive serology, per se, is not enough to diagnose acute brucellosis and other methods should be used for confirmation. Brucella serology data should be interpreted taking into account the patient''s clinical history and epidemiological context.     

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

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.     

Circulating L-lymphocytes in brucellosis patients

The content of L-lymphocytes in patients with different clinical forms of brucellosis has been studied. The degree of a decrease in the level of these cells has been found to depend on the activity of the infectious process and its clinical manifestations in the form of polyarthritis. The severity of the course of acute and subacute brucellosis has been found to produce no essential influence on the level of L-cells. These cells are supposed to participate in the processes of pathogenesis and the elimination of the pathogen in brucellosis.     

Interleukin-18 single nucleotide polymorphisms contribute to the susceptibility to brucellosis in Iranian patients

It seems that IL-18 has a crucial role in immunity against Brucella infection. Since the expression of IL-18 can be affected by polymorphisms in its gene, we decided to investigate any probable relationship between the six different IL-18 gene polymorphisms and brucellosis. A total of 193 patients with brucellosis and 83 healthy farmers who consumed contaminated raw milk and dairy products from the animals with brucellosis, were included in this study. All the individuals were genotyped for six IL-18 polymorphisms at positions -656, -607, -137, +113, +127 and codon 35/3, using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The distributions of alleles for IL-18 polymorphisms at positions -137G/+113T/+127C/codon 35/3A (correlated with higher production of IL-18) were significantly higher in healthy controls than in patients (P=0.012, 0.012, 0.012 and 0.0018, respectively). It could be suggested that individuals who inherited the aforementioned genotypes/alleles are able to produce higher levels of IL-18 at the onset of infection, and it leads to more IFN-gamma production and control Brucella infection before the emerging brucellosis.     

Impact of massive dose of vitamin A given to preschool children with acute diarrhoea on subsequent respiratory and diarrhoeal morbidity.

OBJECTIVE--To assess the impact of vitamin A supplementation on morbidity from acute respiratory tract infections and diarrhoea. DESIGN--Double blind randomised placebo controlled field trial. SETTING--An urban slum area in New Delhi, India. SUBJECTS--900 children aged 12-60 months attending a local health facility for acute diarrhoea of less than seven days'' duration randomly allocated to receive vitamin A 200,000 IU or placebo. MAIN OUTCOME MEASURES--Incidence and prevalence of acute lower respiratory tract infections and diarrhoea during the 90 days after termination of the enrolment diarrhoeal episode measured by twice weekly household surveillance. RESULTS--The incidence (relative risk 1.07; 95% confidence interval 0.92 to 1.26) and average number of days spent with acute lower respiratory tract infections were similar in the vitamin A supplementation and placebo groups. Among children aged 23 months or less there was a significant reduction in the incidence of measles (relative risk 0.06; 95% confidence interval 0.01 to 0.48). The incidence of diarrhoea was also similar (relative risk 0.95; 0.86 to 1.05) in the two groups. There was a 36% reduction in the mean daily prevalence of diarrhoea associated with fever in the vitamin A supplemented children older than 23 months. CONCLUSIONS--Results were consistent with a lack of impact on acute lower respiratory tract related mortality after vitamin A supplementation noted in other trials and a possible reduction in the severity of diarrhoea.     

Assessment of evidence for a protective role of vitamin D in multiple sclerosis

Evidence for a role of vitamin D insufficiency in determining risk in Multiple Sclerosis (MS) is supported by studies in both pediatric- and adult-onset patients. The potential role of vitamin D in modulating MS disease activity is an area of active clinical trials research, and the possibility of primary disease prevention with vitamin D supplementation in early life is an emerging concept. With Sir Austin Bradford Hill's criteria as a framework, the present review assesses the evidence for a causal relationship between vitamin D insufficiency and the pathobiology of MS, and discusses rationale for future clinical trials with vitamin D.     

Prevalence and relevant factors of positive RF in brucellosis patients with arthralgia

BackgroundBrucellosis is a critical zoonotic disease in the world, it is the non-specific arthralgia that make brucellosis patients easily misdiagnosed as rheumatoid arthritis (RA) in endemic regions. Elevated rheumatoid factor (RF) is an essential indicator of RA, and the RF in brucellosis patients is significantly higher than healthy people. Therefore, this study further explored the distribution of RF and the relevant factors of the RF positivity in brucellosis patients with arthralgia, in order to strengthen the recognition of physicians for brucellosis patients with RF positivity, especially in brucellosis-endemic areas, so as to avoid misdiagnosis and untimely treatment that may lead to malignant outcomes.Methodology and principal findingsThe medical records of all 572 brucellosis inpatients were collected in the Sixth People’s Hospital of Shenyang, China from 2015 to 2016. After excluding 106 patients without arthralgia, 5 patients who unwilling to perform RF testing and 16 patients with diseases that may affect RF, 445 brucellosis inpatients with arthralgia were involved in this retrospective cross-sectional study. 143 (32.1%) patients with RF >10 IU/ml were classified into the RF positive group, with an average level of 16.5[12.2, 34.7] IU/ml, of which 45 (10.1%) patients were high-positive with RF >30 IU/ml. Multivariate logistic regression model was used to further analyze the relevant factors of the RF positivity and found that age, wrist joint pain and elevated C-reactive protein (CRP) were positively associated with RF positivity, with OR of 1.02 (P = 0.024), 8.94 (P = 0.008) and 1.79 (P = 0.019), respectively.ConclusionThe prevalence of positive RF in brucellosis patients with arthralgia was critical, nearly one-third of patients had RF positive. Elderly men brucellosis patients with arthralgia, wrist joint pain and elevated CRP were at high risk of positive RF. It is reminded that physicians should focus on differential diagnosis during clinical diagnosis and treatment, especially in brucellosis-endemic regions.     

Free and bound Brucella antigen and the level of circulating immune complexes in brucellosis patients

The aggregate hemagglutination test has been shown to be a highly sensitive and specific method for the detection of infectious antigenemia in different forms of brucellosis, permitting the determination of free Brucella antigen in 34% of patients with the acute form of the disease and in 53% of patients with the chronic course of brucellosis. The results of the determination of the quantitative level of circulating immune complexes (CIC) indicate that in the acute form of the disease their level exceeds the CIC level observed in the chronic course of brucellosis. The preliminary dissociation of CIC in the patients' blood sera has been found to increase the overall release of Brucella antigen to 80%.     

Interferon-mediated immunopathological events are associated with atypical innate and adaptive immune responses in patients with severe acute respiratory syndrome

It is not understood how immune inflammation influences the pathogenesis of severe acute respiratory syndrome (SARS). One area of strong controversy is the role of interferon (IFN) responses in the natural history of SARS. The fact that the majority of SARS patients recover after relatively moderate illness suggests that the prevailing notion of deficient type I IFN-mediated immunity, with hypercytokinemia driving a poor clinical course, is oversimplified. We used proteomic and genomic technology to systematically analyze host innate and adaptive immune responses of 40 clinically well-described patients with SARS during discrete phases of illness from the onset of symptoms to discharge or a fatal outcome. A novel signature of high IFN-alpha, IFN-gamma, and IFN-stimulated chemokine levels, plus robust antiviral IFN-stimulated gene (ISG) expression, accompanied early SARS sequelae. As acute illness progressed, SARS patients entered a crisis phase linked to oxygen saturation profiles. The majority of SARS patients resolved IFN responses at crisis and expressed adaptive immune genes. In contrast, patients with poor outcomes showed deviated ISG and immunoglobulin gene expression levels, persistent chemokine levels, and deficient anti-SARS spike antibody production. We contend that unregulated IFN responses during acute-phase SARS may culminate in a malfunction of the switch from innate immunity to adaptive immunity. The potential for the use of the gene signatures we describe in this study to better assess the immunopathology and clinical management of severe viral infections, such as SARS and avian influenza (H5N1), is therefore worth careful examination.     

In vivo studies of altered expression patterns of p53 and proliferative control genes in chronic vitamin A deficiency and hypervitaminosis.

Several clinical trials have revealed that individuals who were given beta-carotene and vitamin A did not have a reduced risk of cancer compared to those given placebo; rather, vitamin A could actually have caused an adverse effect in the lungs of smokers [Omenn, G.S., Goodman, G.E., Thornquist, M.D., Balmes, J., Cullen, M.R., Glass, A., Keogh, J.P., Meyskens, F.L., Valanis, B., Williams, J.H., Barnhart, S. & Hammar, S. N. Engl. J. Med (1996) 334, 1150-1155; Hennekens, C.H., Buring, J.E., Manson, J.E., Stampfer, M., Rosner, B., Cook, N.R., Belanger, C., LaMotte, F., Gaziano, J.M., Ridker, P.M., Willet, W. & Peto, R. (1996) N. Engl. J. Med. 334, 1145-1149]. Using differential display techniques, an initial survey using rats showed that liver RNA expression of c-H-Ras was decreased and p53 increased in rats with chronic vitamin A deficiency. These findings prompted us to evaluate the expression of c-Jun, p53 and p21WAF1/CIF1 (by RT-PCR) in liver and lung of rats. This study showed that c-Jun levels were lower and that p53 and p21WAF1/CIF1 levels were higher in chronic vitamin A deficiency. Vitamin A supplementation increased expression of c-Jun, while decreasing the expression of p53 and p21WAF1/CIF1. Western-blot analysis demonstrated that c-Jun and p53 showed a similar pattern to that found in the RT-PCR analyses. Binding of retinoic acid receptors (RAR) to the c-Jun promoter was decreased in chronic vitamin A deficiency when compared to control hepatocytes, but contrasting results were found with acute vitamin A supplementated cells. DNA fragmentation and cytochrome c release from mitochondria were analyzed and no changes were found. In lung, an increase in the expression of c-Jun produced a significant increase in cyclin D1 expression. These results may explain, at least in part, the conflicting results found in patients supplemented with vitamin A and illustrate that the changes are not restricted to lung. Furthermore, these results suggest that pharmacological vitamin A supplementation may increase the risk of adverse effects including the risk of oncogenesis.     

Effects of the multi-kinase inhibitor midostaurin in combination with chemotherapy in models of acute myeloid leukaemia

Recently, several targeted agents have been developed for specific subsets of patients with acute myeloid leukaemia (AML), including midostaurin, the first FDA-approved FLT3 inhibitor for newly diagnosed patients with FLT3 mutations. However, in the initial Phase I/II clinical trials, some patients without FLT3 mutations had transient responses to midostaurin, suggesting that this multi-targeted kinase inhibitor might benefit AML patients more broadly. Here, we demonstrate submicromolar efficacy of midostaurin in vitro and efficacy in vivo against wild-type (wt) FLT3-expressing AML cell lines and primary cells, and we compare its effectiveness with that of other FLT3 inhibitors currently in clinical trials. Midostaurin was found to synergize with standard chemotherapeutic drugs and some targeted agents against AML cells without mutations in FLT3. The mechanism may involve, in part, the unique kinase profile of midostaurin that includes proteins implicated in AML transformation, such as SYK or KIT, or inhibition of ERK pathway or proviability signalling. Our findings support further investigation of midostaurin as a chemosensitizing agent in AML patients without FLT3 mutations.