Anti-mycobacterial recall responses differentiate female patients with genital tuberculosis from patients with other gynecological problems

Background: Female Genital Tuberculosis (FGTB) is one form of extra pulmonary tuberculosis affecting the female reproductive organs, most commonly the fallopian tubes and the endometrium. It affects young women aged between 20 and 40 years of age and is an important cause of infertility. It often occurs as a secondary complication following pulmonary tuberculosis. Diagnosis depends mainly on clinical suspicion in countries where facilities for mycobacterial culture and histopathology are unavailable. Even in places where these facilities exist, diagnosis still remains difficult because of the lower sensitivity and specificity of the methods as well as the invasive procedure of acquiring biopsy specimens. Objective: To explore the immunological profiles of female genital tuberculosis (FGTB) patients in response to mycobacterial antigens. Methods: Twenty-five clinically suspected cases of FGTB and 12 control subjects who came to the Black Lion hospital for unrelated gynecological problems were included in the study. Peripheral blood samples were collected from each subject. Plasma was separated by centrifugation and PBMC were isolated over ficoll-hypaque and stimulated in vitro with mycobacterial antigens to examine their proliferative response as incorporation of tritiated thymidine using a β-counter. HIV status and total IgG-, IgA- and IgM- antibody levels were determined by ELISA tests.Results: In vitro recall responses to M. tuberculosis antigens (PPD and BCG sonicate) as well as plasma levels of IgGIgA- and IgM-antibodies to MPT59 showed statistically significant differences between the patients and the controls (

Ex vivo cytokine mRNA levels correlate with changing clinical status of ethiopian TB patients and their contacts over time.

There is an increasing body of evidence which suggests that IL-4 plays a role in the pathogenesis of TB, but a general consensus on its role remains elusive. We have previously published data from a cohort of Ethiopian TB patients, their contacts, and community controls suggesting that enhanced IL-4 production is associated with infection with M. tuberculosis, rather than overt disease and that long-term protection in infected community controls is associated with co-production of the IL-4 antagonist IL-4d2, alongside elevated IL-4. Here, for the first time, we compare data on expression of IFN-gamma, IL-4 and IL-4delta2 over time in TB patients and their household contacts. During the follow-up period, the TB patients completed therapy and ceased to display TB-like symptoms. This correlated with a decrease in the relative amount of IL-4 expressed. Over the same period, the clinical status of some of their contacts also changed, with a number developing TB-like symptoms or clinically apparent TB. IL-4 expression was disproportionately increased in this group. The findings support the hypothesis that elevated IL-4 production is generally associated with infection, but that TB disease is associated with a relatively increased expression of IL-4 compared to IFN-gamma and IL-4delta2. However, the data also suggest that there are no clear-cut differences between groups: the immune response over time appears to include changes in the expression of IFN-gamma, IL-4 and IL-4delta2, and it is the relative, not absolute levels of cytokine expression that are characteristic of clinical status

Polymorphism in killer cell immunoglobulin-like receptors and human leukocyte antigen-c and predisposition to preeclampsia in Ethiopian pregnant women population.

INTRODUCTION: Preeclampsia (PE) is a human specific pregnancy-related syndrome of unknown etiology that affects 2-8 % of pregnancies. Polymorphism in maternal Killer Cell Immunoglobulin-like Receptors (KIRs) and the ligand fetal Human Leukocyte Antigen-C (HLA-C) may predispose pregnant mothers for PE due to defective trophoblast invasion into the maternal decidua. Our study aimed to investigate the association between maternal KIR and fetal HLA-C polymorphism and PE in Ethiopian pregnant women. METHODS: We included a total of 288 (157 controls and 131 PE cases) in a case-controls study at Adama Regional Referral Hospital, Ethiopia. The KIR and HLA-C genotyping was done using traditional polymerase chain reaction on genomic DNA extracted form maternal venous and cord blood followed by 2% agarose gel electrophoresis. RESULTS: The statistical associations between variables were evaluated using Pearson's Chi-square test. P < 0.05, with 95 % confidence interval was considered statistically significant. A significant association was observed between the KIR2DS1 and PE, with a higher frequency (60.5 %) of the gene in the control group. Similarly, a significant association was observed between KIR AA genotype and PE, with a higher frequency (38.2 %) of this genotype in the PE group. Ethiopians share the same risk genotype for PE as seen in previous African and European studies, namely homozygosity of a maternal KIR AA genotype. However, Ethiopians differ from other East African populations by sharing the same protective KIR2DS1 gene as Europeans

Breast cancer in Ethiopia: evidence for geographic difference in the distribution of molecular subtypes in Africa

Abstract Background Breast cancer is a heterogeneous disease with several morphological and molecular subtypes. Widely accepted molecular classification system uses assessment of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and proliferation marker Ki67. Few studies have been conducted on the incidence and molecular types of breast cancer in Sub-Saharan Africa. Previous studies mainly from Western and Central Africa, showed breast cancer to occur at younger ages and to present with aggressive features, such as high-grade, advanced stage and triple-negative phenotype (negative for ER, PR and HER2). Limited data from East Africa including Ethiopia however shows hormone receptor negative tumors to account for a lower proportion of all breast cancers than has been reported from elsewhere in Africa. Methods In this study from Tikur Anbessa Specialized Hospital, 114 breast cancer patients diagnosed between 2012 and 2015 were enrolled. ER, PR, Ki67 and HER2 receptor status were assessed using immunohistochemistry from tissue microarrays. FISH was used for assessment of gene amplification in all equivocal tumor samples and for confirmation in HER2-enriched cases. Results The distribution of molecular subtypes was: Luminal A: 40%; Luminal B: 26%; HER2-enriched: 10%; TNBC: 23%. ER were positive in 65% of all tumors and 43% the cases were positive for PR. There was statistically significant difference in median age at diagnosis between the molecular subtypes (P < 0.05). There was a bimodal distribution of molecular subtypes in different age ranges with Luminal B subtype being more common at younger ages (median = 36) and Luminal A subtype more prevalent at older ages (median = 42). There were no statistically significant differences in tumor grade, histology, and stage between the molecular subtypes of breast cancer. Conclusion The present study detected Luminal A breast cancer to be the most common subtype and reveals a relatively low rate of hormone receptor negative and TNBC. Our findings and results from other East African studies suggest geographic variability in the distribution of the molecular subtypes of breast cancer in Africa and hence have important clinical and policy implications for breast cancer control and treatment in Ethiopia.https://deepblue.lib.umich.edu/bitstream/2027.42/142374/1/12905_2018_Article_531.pd

The Effects of Prednisolone Treatment on Cytokine Expression in Patients with Erythema Nodosum Leprosum Reactions.

Erythema nodosum leprosum (ENL) is a systemic inflammatory complication occurring mainly in patients with lepromatous leprosy (LL) and borderline lepromatous leprosy. Prednisolone is widely used for treatment of ENL reactions but clinical improvement varies. However, there is little good in vivo data as to the effect of prednisolone treatment on the pro-inflammatory cytokines in patients with ENL reactions. As a result, treatment and management of reactional and post-reactional episodes of ENL often pose a therapeutic challenge. We investigated the effect of prednisolone treatment on the inflammatory cytokines TNF, IFN-γ, IL-1β, IL-6, and IL-17 and the regulatory cytokines IL-10 and TGF-β in the skin lesion and blood of patients with ENL and compared with non-reactional LL patient controls. A case-control study was employed to recruit 30 patients with ENL and 30 non-reactional LL patient controls at ALERT Hospital, Ethiopia. Blood and skin biopsy samples were obtained from each patient before and after prednisolone treatment. Peripheral blood mononuclear cells from patients with ENL cases and LL controls were cultured with M. leprae whole-cell sonicates (MLWCS), phytohemagglutinin or no stimulation for 6 days. The supernatants were assessed with the enzyme-linked immunosorbent assay for inflammatory and regulatory cytokines. For cytokine gene expression, mRNA was isolated from whole blood and skin lesions and then reverse transcribed into cDNA. The mRNA gene expression was quantified on a Light Cycler using real-time PCR assays specific to TNF, IFN-γ, IL-β, TGF-β, IL-17A, IL-6, IL-8, and IL-10. The ex vivo production of the cytokines: TNF, IFN-γ, IL-1β, and IL-17A was significantly increased in untreated patients with ENL. However, IL-10 production was significantly lower in untreated patients with ENL and significantly increased after treatment. The ex vivo production of IL-6 and IL-8 in patients with ENL did not show statistically significant differences before and after prednisolone treatment. The mRNA expression in blood and skin lesion for TNF, IFN-γ, IL-1β, IL-6, and IL-17A significantly reduced in patients with ENL after treatment, while mRNA expression for IL-10 and TGF-β was significantly increased both in blood and skin lesion after treatment. This is the first study examining the effect of prednisolone on the kinetics of inflammatory and regulatory cytokines in patients with ENL reactions before and after prednisolone treatment. Our findings suggest that prednisolone modulates the pro-inflammatory cytokines studied here either directly or through suppression of the immune cells producing these inflammatory cytokines

Population-based Screening for Pulmonary Tuberculosis utilizing Community Health Workers in Ethiopia

Objective To evaluate the utility of a volunteer health development army in conducting population screening for active TB in a rural community in Southern Ethiopia. Methods A population-based cross-sectional survey was conducted in sixKebeles (the lowest administrative units).Volunteer women community workers led a symptom screening program to identify adults ≥15 years with TB in the community. Individuals with cough ≥ 2 weeks had spot and morning sputum samples and were examined using AFB smear microscopy, culture and Xpert MTB/RIF. Results All 24,517 adults in the study area had a symptom screen performed; 544 (2.2%) had cough ≥ 2 weeks. Among a positive symptom screen, 13 (2.4%) had a positive sputum AFB smear microscopy, 13 (2.4%) a positive culture and 32 (5.8%) a positive Xpert MTB/RIF test. Overall, 34 TB cases (6%) were identified by culture and/or Xpert which corresponds to a prevalence of 139 per 100,000 persons. Conclusion We demonstrate the capability of community health workers (volunteer and paid) to rapidly conduct a large-scale population TB screening evaluation and highlight the high yield of such a program to detect previously undiagnosed cases when combined with Xpert MTB/RIF testing. This could be a model to implement in other similar settings

Robust barcoding and identification of Mycobacterium tuberculosis lineages for epidemiological and clinical studies.

BACKGROUND: Tuberculosis, caused by bacteria in the Mycobacterium tuberculosis complex (MTBC), is a major global public health burden. Strain-specific genomic diversity in the known lineages of MTBC is an important factor in pathogenesis that may affect virulence, transmissibility, host response and emergence of drug resistance. Fast and accurate tracking of MTBC strains is therefore crucial for infection control, and our previous work developed a 62-single nucleotide polymorphism (SNP) barcode to inform on the phylogenetic identity of 7 human lineages and 64 sub-lineages. METHODS: To update this barcode, we analysed whole genome sequencing data from 35,298 MTBC isolates (~ 1 million SNPs) covering 9 main lineages and 3 similar animal-related species (M. tuberculosis var. bovis, M. tuberculosis var. caprae and M. tuberculosis var. orygis). The data was partitioned into training (N = 17,903, 50.7%) and test (N = 17,395, 49.3%) sets and were analysed using an integrated phylogenetic tree and population differentiation (FST) statistical approach. RESULTS: By constructing a phylogenetic tree on the training MTBC isolates, we characterised 90 lineages or sub-lineages or species, of which 30 are new, and identified 421 robust barcoding mutations, of which a minimal set of 90 was selected that included 20 markers from the 62-SNP barcode. The barcoding SNPs (90 and 421) discriminated perfectly the 86 MTBC isolate (sub-)lineages in the test set and could accurately reconstruct the clades across the combined 35k samples. CONCLUSIONS: The validated 90 SNPs can be used for the rapid diagnosis and tracking of MTBC strains to assist public health surveillance and control. To facilitate this, the SNP markers have now been incorporated into the TB-Profiler informatics platform ( https://github.com/jodyphelan/TBProfiler )

Prevalence of pulmonary TB and spoligotype pattern of Mycobacterium tuberculosis among TB suspects in a rural community in Southwest Ethiopia

<p>Abstract</p> <p>Background</p> <p>In Ethiopia where there is no strong surveillance system and state of the art diagnostic facilities are limited, the real burden of tuberculosis (TB) is not well known. We conducted a community based survey to estimate the prevalence of pulmonary TB and spoligotype pattern of the <it>Mycobacterium tuberculosis </it>isolates in Southwest Ethiopia.</p> <p>Methods</p> <p>A total of 30040 adults in 10882 households were screened for pulmonary TB in Gilgel Gibe field research centre in Southwest Ethiopia. A total of 482 TB suspects were identified and smear microscopy and culture was done for 428 TB suspects. Counseling and testing for HIV/AIDS was done for all TB suspects. Spoligotyping was done to characterize the <it>Mycobacterium tuberculosis </it>isolates.</p> <p>Results</p> <p>Majority of the TB suspects were females (60.7%) and non-literates (83.6%). Using smear microscopy, a total of 5 new and 4 old cases of pulmonary TB cases were identified making the prevalence of TB 30 per 100,000. However, using the culture method, we identified 17 new cases with a prevalence of 76.1 per 100,000. There were 4.3 undiagnosed pulmonary TB cases for every TB case who was diagnosed through the passive case detection mechanism in the health facility. Eleven isolates (64.7%) belonged to the six previously known spoligotypes: T, Haarlem and Central-Asian (CAS). Six new spoligotype patterns of <it>Mycobacterium tuberculosis</it>, not present in the international database (SpolDB4) were identified. None of the rural residents was HIV infected and only 5 (5.5%) of the urban TB suspects were positive for HIV.</p> <p>Conclusion</p> <p>The prevalence of TB in the rural community of Southwest Ethiopia is low. There are large numbers of undiagnosed TB cases in the community. However, the number of sputum smear-positive cases was very low and therefore the risk of transmitting the infection to others may be limited. Active case finding through health extension workers in the community can improve the low case detection rate in Ethiopia. A large scale study on the genotyping of <it>Mycobacterium tuberculosis </it>in Ethiopia is crucial to understand transmission dynamics, identification of drug resistant strains and design preventive strategies.</p

Two-Thirds of Smear-Positive Tuberculosis Cases in the Community Were Undiagnosed in Northwest Ethiopia: Population Based Cross-Sectional Study

Background: Tuberculosis (TB) case detection rate remains low in Ethiopia. One of the underlying reasons is the emphasis on passive case finding strategy which may seriously underestimate the burden of the disease. Estimating the prevalence of smear-positive pulmonary TB through active case finding at population level can help assessing the degree to which passive case detection is successful. Methods and findings: This is population based cross-sectional study. The study population was all individuals aged $14 years. Interviews using a uniform questionnaire were done initially to identify individuals with chronic cough ($15 days) and the two sputum (spot and morning) samples were gathered for standard smear microscopy. A total of 23,590 individuals aged $14 years were interviewed and 984 had a chronic cough for$15 days. Of 831 individuals who provided two sputum samples for acid fast bacilli (AFB), 41 had positive smears. A total of 22 smear-positive TB cases detected through passive case finding were on anti-TB treatment. The prevalence of new smear-positive TB was 174 per 100,000 in persons aged $14 years (95$45 years [AOR: 2.26, 95 % CI (1.12–4.59). Conclusions: The study revealed that about two-thirds of patients with active TB remain undiagnosed and thus untreated. This may indicate the need for strengthening case detection at the community level. Furthermore, the high burden of T