Vitamin D enhances IL-1ß secretion and restricts growth of Mycobacterium tuberculosis in macrophages from TB patients.

The emergence of multidrug-resistant strains of Mycobacterium tuberculosis (MTB), the bacterium responsible for tuberculosis (TB), has rekindled the interest in the role of nutritional supplementation of micronutrients, such as vitamin D, as adjuvant treatment. Here, the growth of virulent MTB in macrophages obtained from the peripheral blood of patients with and without TB was studied. The H37Rv strain genetically modified to express Vibrio harveyi luciferase was used to determine the growth of MTB by luminometry in the human monocyte-derived macrophages (hMDMs) from study subjects. Determination of cytokine levels in culture supernatants was performed using a flow cytometry-based bead array technique. No differences in intracellular growth of MTB were observed between the different study groups. However, stimulation with 100nM 1,25-dihydroxyvitamin D significantly enhanced the capacity of hMDMs isolated from TB patients to control the infection. This effect was not observed in hMDMs from the other groups. The interleukin (IL)-1ß and IL-10 release by hMDMs was clearly increased upon stimulation with 1,25-dihydroxyvitamin D. Furthermore, the 1,25-dihydroxyvitamin D stimulation also led to elevated levels of TNF-α (tumor necrosis factor-alpha) and IL-12p40. It was concluded that vitamin D triggers an inflammatory response in human macrophages with enhanced secretion of cytokines, as well as enhancing the capacity of hMDMs from patients with active TB to restrict mycobacterial growth.

Identification of drug susceptibility pattern and mycobacterial species in sputum smear positive pulmonary tuberculosis patients with and without HIV co-infection in north west Ethiopia.

BACKGROUND: Ethiopia is among the high-burden countries of tuberculosis (TB) in the world Since mycobacterial culture and susceptibility testing are not routinely performed in Ethiopia, recent data on susceptibility patterns and the mycobacterial species cultured from sputum smear positive patients are limited. OBJECTIVES: The aim was to determine first line anti-TB drug susceptibility of Mycobacterium tuberculosis isolates obtained from consecutive newly diagnosed smear positive pulmonary TB patients in north west Ethiopia. METHODOLOGY: A retrospective cross sectional study was conducted using previously collected sputum samples (n=180) kept at the referral hospital of the University of Gondar at -20 degrees C. Sputum samples were cultured on Lowenstein Jensen (LJ) medium. Conventional Polymerase Chain Reaction (PCR) using RD4 primers to identify the M. tuberculosis complex was performed on cultured isolates. Ninety eight (84.4%) of the 116 isolates identified as M. tuberculosis were tested for their drug susceptibility pattern using the proportion method Clinical baseline data including body mass index, body temperature, clinical symptoms and erythrocyte sedimentation rate were obtained. RESULTS: The culture retrieval rate of previously frozen sputum samples was 64.4% (116/180). All the isolated mycobacterial species (n=116) were confirmed as belonging to the M. tuberculosis complex by PCR. Of 98 isolates for which the drug susceptibility test was done, 15.3% (15/98) were found to be resistant to one or more antimycobacterial drugs, and resistance to isoniazid and streptomycin was most common with 8.2% (8/98) and 6.1% (6/98) respectively. TB patients co infected with HIV had increased erythrocyte sedimentation rate, higher age and lower sputum smear grade than HIV negative TB patients. CONCLUSIONS: No mycobacteria other than M. tuberculosis were detected in sputum smear positive TB-patients. Although no multi drug resistant strain was observed, relatively high rates of INH resistance were found in this region. Culture facilities are urgently needed in regional centers to increase diagnostic sensitivity and monitor developing trends of drug resistance in Ethiopia.

Nitric oxide production in the exhaled air of patients with pulmonary tuberculosis in relation to HIV co-infection.

BACKGROUND: Nitric oxide (NO) is essential for host defense in rodents, but the role of NO during tuberculosis (TB) in man remains controversial. However, earlier observations that arginine supplementation facilitates anti-TB treatment, supports the hypothesis that NO is important in the host defense against TB. Local production of NO measured in fractional exhaled air (FeNO) in TB patients with and without HIV co-infection has not been reported previously. Thus, our aim was to investigate levels of FeNO in relation to clinical symptoms and urinary NO metabolites (uNO). METHODS: In a cross sectional study, FeNO and uNO were measured and clinical symptoms, chest x-ray, together with serum levels of arginine, tumor necrosis factor alpha (TNF-alpha) and interleukin 12 (IL-12) were evaluated in sputum smear positive TB patients (HIV+/TB, n = 36, HIV-/TB, n = 59), their household contacts (n = 17) and blood donors (n = 46) from Gondar University Hospital, Ethiopia. RESULTS: The proportion of HIV-/TB patients with an increased FeNO level (> 25 ppb) was significantly higher as compared to HIV+/TB patients, but HIV+/TB patients had significantly higher uNO than HIV-/TB patients. HIV+ and HIV-/TB patients both had lower levels of FeNO compared to blood donors and household contacts. The highest levels of both uNO and FeNO were found in household contacts. Less advanced findings on chest x-ray, as well as higher sedimentation rate were observed in HIV+/TB patients as compared to HIV-/TB patients. However, no significant correlation was found between FeNO and uNO, chest x-ray grading, clinical symptoms, TNF-alpha, IL-12, arginine levels or sedimentation rate. CONCLUSION: In both HIV negative and HIV co infected TB patients, low levels of exhaled NO compared to blood donors and household were observed. Future studies are needed to confirm whether low levels of exhaled NO could be a risk factor in acquiring TB and the relative importance of NO in human TB.