Whole Blood Stimulation Assay as a Treatment Outcome Monitoring Tool for VL Patients in Ethiopia: A Pilot Evaluation.

Visceral leishmaniasis (VL) is a lethal disease if left untreated. Current treatments produce variable rates of treatment failure and toxicity without sterile cure, rendering treatment efficacy monitoring essential. To avoid repeated invasive tissue aspirates as well as empirical treatment, there is a need for new tools that allow a less-invasive and early assessment of treatment efficacy in the field. Cross-sectional studies have suggested levels of cytokines/chemokines after whole blood stimulation as good markers of cure, but longitudinal studies are lacking. In this study, we followed 13 active VL cases in an endemic area in Ethiopia by measuring the production of IFN-γ, TNF-α, IP-10, IL-2, IL-10, MCP-1, and MIG before, during, and at the end of treatment. After 24 hours of stimulation of whole blood with soluble Leishmania antigen, we observed an early, robust, and incremental increase of IFN-γ, TNF-α, and IP-10 levels in all patients during treatment. Moreover, based on the IFN-γ levels that showed an average 13-fold increase from the time of diagnosis until the end of treatment, we could almost perfectly discriminate active from cured status. Similar concentrations and patterns were found in stimulation assays with the two main Leishmania species. The levels of IFN-γ, IP-10, or TNF-α also seemed to be inversely associated with the parasite load at baseline. Despite a 1/10 drop in concentrations, similar patterns were observed in IFN-γ and IP-10 levels when dried plasma spots were stored at 4°C for an average of 225 days. All the above evidence suggests a detectable restoration of cell-mediated immunity in VL and its association with parasite clearance. With a potential application in rural settings by means of dried plasma spots, we recommend to further explore the early diagnostic value of such assays for treatment efficacy monitoring in large cohort studies including treatment failure cases.Funding was provided by the Belgian Directorate-General for Development Cooperation under the ITM-DGDC framework agreement FA-IIII. WA is personally supported by a Research Foundation Flanders postdoctoral fellowship. In addition, this work was funded by the Instituto de Salud Carlos III via the project PI18CIII/00029 and via the Red de Enfermedades Tropicales, Subprograma RETICS del Plan Estatal de I+D+I 2013-2016, which is cofunded by FEDER “Una manera de hacer Europa” funds, via projects RD16/0027/0017 and RD16CIII/0003/0002.S

Opportunistic Cryptococcal Antigenemia in the HAART Era at HIV Epidemic Settings of Northwest Ethiopia

Background. Cryptococcus neoformans is a frequent opportunistic infection in patients with the acquired immunodeficiency syndrome. While the advent of ART reduces the occurrence of cryptococcal meningitis in HIV patients, cryptococcal disease remains a leading cause of morbidity and mortality in the developing world especially in sub-Saharan Africa which is the epicenter of HIV. This study aimed to assess the cryptococcal antigenemia, CD4+ Th cell counts, HIV RNA viral load, and clinical presentations among HIV-positive patients in Northwest Ethiopia. Method. A total of two hundred (200) HIV-positive patients were recruited for this study. Cryptococcus antigenemia prevalence in plasma samples of HIV‐positive patients was determined by using Antigen lateral flow assay (CrAg‐LFA) also, and CD4+ Th cell counts and HIV‐RNA levels were quantified from blood specimen. Patients’ demographic data, clinical manifestation, and concurrent opportunistic infection were recorded. Result. The sex distributions of study participants were 105(52.5%) male and 94(47.5%) female with an age range of 15–65 (mean 39.42 ± 9) years. All patients had a CD4+ T-cell count 10,000 copies/ml, respectively, as well; Tuberculosis, Candidiasis, and herpes zoster are the most often observed concurrent infections while cryptococcal antigenemia is significantly associated with oral candidiasis (p<0.001). Conclusion. Although the advent of ART, early diagnosis of cryptococcosis, and application of antifungal interventions, HIV-induced cryptococcal antigenemia positivity in HIV infected individuals is still the countries’ big challenge. Thus, stringent follow-up and case management should be considered

Diagnostic performance of CL Detect rapid-immunochromatographic test for cutaneous leishmaniasis: a systematic review and meta-analysis

Abstract Background Sensitive, robust, and fast point-of-care tests are needed for cutaneous leishmaniasis (CL) diagnosis. The recently developed CL Detect rapid test (InBios) for detecting Leishmania peroxidoxin antigen has been evaluated in several studies. However, diagnostic performances were controversial. Therefore, this systematic review and meta-analysis aimed to determine the pooled sensitivity and specificity of CL Detect for CL diagnosis. Methods PubMed, Scopus, EMBASE, ScienceDirect, and Google Scholar were sources of articles. We included studies reporting the diagnostic accuracy of CL Detect and CL-suspected patients in the English language. The methodological qualities of the included studies were appraised using the quality assessment of diagnostic accuracy studies-2 (QUADAS‐2). Meta-analysis was conducted using Stata 14.2 and R software. Results A total of 9 articles were included. The study sample size ranged from 11 to 274. The sensitivities of the individual studies ranged from 23 to 100%, and the specificities ranged from 78 to 100%. Pooled sensitivity and specificity were 68% (95% CI, 41–86%) and 94% (95% CI, 87–97%), respectively. AUC displayed 0.899. Pooled sensitivity was lower (47%, 95% CI, 34–61%) when PCR was used as a reference than microscopy (83%, 95% CI, 39–97%). Pooled sensitivity was lower (48%, 95% CI, 30–67%) for all lesion durations compared to ≤ 4 months (89%, 95% CI, 43–99%). Conclusions CL Detect has poor sensitivity and does not meet the minimal sensitivity of 95% of target product profiles designed for CL point-of-care tests. Currently, the CL Detect test looks unsuitable for CL diagnosis, despite its high specificity. Findings are limited by the low number of studies available. Further large-scale studies are recommended. Systematic review registration PROSPERO CRD42022323497

Seroprevalence of HIV among pregnant women in Ethiopia: a systematic review and meta-analysis

Abstract Objective This systematic review and meta-analysis aimed to determine the pooled prevalence of HIV among pregnant women in Ethiopia. Result PubMed, EMBASE, Science Direct and Google scholar databases were searched to retrieve 15 relevant articles based on the inclusion criteria. A total of 13,746 participants were included in the original studies and considered in this analysis. Among subjects, 717 were infected with HIV only, and 12 were HIV-HBV co-infected pregnant women. In this meta-analysis, the pooled prevalence of HIV among pregnant women in Ethiopia was 5.74% (95% CI 3.96–7.53%). Regional analysis showed that 9.50% (95% CI 7.76–11.23%) in Amhara, 4.80% (95% CI 3.12–6.49%) in Addis Ababa, 2.14% (95% CI − 0.54 to 4.82%) in SNNP and 4.48% (95% CI 2.56–6.41%) in Oromia region. Besides, six studies reported HIV-HBV co-infection and the pooled prevalence was 0.68% (95% CI 0.27–1.08%) among pregnant women in Ethiopia

Helminth species-specific effects on IFN-gamma producing T cells during active and latent tuberculosis

BackgroundInterferon-gamma (IFN-gamma) is a key cytokine inducing protective immune responses during tuberculosis (TB) infection. Helminth-induced immune responses may affect IFN-gamma production by T cells, although its connection with disease severity and immune recovery during treatment is unexplored. We investigated the species-specific effect of helminths on the IFN-gamma production by T cells in relation to disease severity during active and latent TB infection (LTBI). MethodsIn this study, 69 active pulmonary TB patients (PTB), 28 with LTBI and 66 healthy controls were included. Active TB was diagnosed using GenXpert MTB/RIF while QuantiFERON test (QFT) was used for the screening of healthy community controls (CCs) and for the diagnosis of LTBI. Helminth infection was identified by routine diagnosis whereas clinical disease severity was evaluated by the TB score. Intracellular IFN-gamma production of T cells in stimulated peripheral blood mononuclear cells (PBMCs) was analyzed by flow cytometry using TB antigens (PPD), the polyclonal T cell activator staphylococcal enterotoxin B (SEB), or medium as unstimulated control. ResultsHelminth infected CCs and LTBI subjects showed a significant reduction of IFN-gamma(+) CD4(+) T cells by PPD-stimulation compared to non-helminth infected control groups. The significant reduction in the frequency of IFN-gamma(+) T cells in both latent and active PTB patients following SEB stimulation was mostly attributed to Schistosoma mansoni infection, whereas Ascaris lumbricoides, Schistosoma mansoni, and hookworm infection contributed equally in CCs. Following anti-helminthic and anti-TB treatment for 2 months, the frequency of IFN-gamma(+) CD4 T cells in helminth coinfected PTB was restored to levels of helminth negative PTB before treatment. Helminth coinfected PTB patients with an intermediate and severe clinical course had reduced capacity for production of IFN-gamma(+) T cells compared to the corresponding non-helminth infected PTB. ConclusionWe found a reduction in IFN-gamma producing T cells by helminth coinfection which was restored following anti-helminthic treatment. This reduction was helminth species-dependent in an exploratory sub-analysis and correlated to increased disease severity. Author summaryProtective immunity against tuberculosis (TB) requires a Th-1 response with cytokines like TNF and IFN-gamma which plays a key role in the recruitment and activation of immune cells. Helminth infection, on the other hand, can lead to induction of regulatory T cells and a Th-2 skewed response decreasing IFN-gamma in T cells. Decreased Th-1 responses could favor reactivation of latent TB infection (LTBI), although the helminth species-specific effect on IFN-gamma(+)CD4(+) T cells and the link to TB disease severity in patients with active pulmonary TB (PTB) have not been fully investigated. Therefore, blood cells (PBMCs) from healthy controls, LTBI individuals, and PTB patients were used to evaluate the impact of different helminths on the frequency of IFN-gamma(+)CD4(+) T cells, in Gondar Ethiopia. Ascaris lumbricoides, Schistosoma mansoni, and hookworm infection in healthy controls contributed equally to decreasing the frequency of IFN-gamma(+)CD4(+) T cells, whereas in both LTBI and PTB patients S. mansoni coinfection had the greatest impact on reducing IFN-gamma producing capacity of T cells. Decreased IFN-gamma producing capacity of T cells was correlated with increased TB disease severity, only in helminth coinfected PTB patients, and anti-helminthic therapy restored the IFN-gamma producing capacity of T cells at the 2 months follow-up.Funding Agencies|Swedish Heart-Lung Foundation (Hjart-Lungfonden); Swedish Research Council (Vetenskapsradet)</p

Helminth species dependent effects on Th1 and Th17 cytokines in active tuberculosis patients and healthy community controls

Despite that the impact of different helminth species is not well explored, the current dogma states that helminths affect the Th1/Th2 balance which in turn affects the risk of tuberculosis (TB) reactivation and severity of disease. We investigated the influence of helminth species on cytokine profiles including IL-17A in TB patients and healthy community controls (CCs). In total, 104 newly diagnosed pulmonary TB patients and 70 HIV negative and Quanti-FERON negative CCs in Gondar, Ethiopia were included following helminth screening by stool microscopy. Plasma samples and ex vivo stimulation of peripheral blood mononuclear cells (PBMCs) with purified protein derivative (PPD) and Staphylococcus enterotoxin B (SEB) was used to determine cytokine profiles by cytometric bead array. In CCs, Ascaris lumbricoides or Schistosoma mansoni infections were associated with an impaired Th1-type response (IFN-gamma, IL-6 and TNF-alpha) in PBMCs mainly with SEB stimulations, whereas in TB patients only hookworm infection showed a similar pattern. Among CCs, the IL-17A response in PBMCs stimulated with SEB was higher only for S. mansoni, whereas in TB patients, the elevated systemic IL-17A plasma level was significantly suppressed in hookworm infected TB patients compared to patients without helminth coinfection. Following treatment of TB and helminth infection there was a general decrease in ex vivio IL-10 and TNF-alpha production in unstimulated, PPD or SEB stimulated PBMCs that was the most pronounced and significant in TB patients infected with S. mansoni, whereas the follow-up levels of IFN-gamma and IL-17A was significantly increased only in TB patients without helminth coinfection from PBMCs stimulated mainly with SEB. In summary, in addition to confirming helminth specific effects on the Th1/Th2 response before and after TB treatment, our novel finding is that IL-17A was impaired in helminth infected TB patients especially for hookworm, indicating a helminth species-specific immunoregulatory effect on IL-17A which needs to be further investigated.Funding Agencies|Swedish Heart-Lung Foundation; Swedish Research Council</p

Comparison of complete blood count.

<p>Comparison of complete blood count.</p

The plasma from VL patients without intestinal parasite co-infections (IP-) and in VL patients co-infected with intestinal parasites (IP+) was tested by ELISA (CRP, IL-6, IL-8, myeloperoxidase and elastase) or enzymatic activity (arginase) as described in Materials and Methods.

<p>Statistical significance was established using a Mann-Whitney test, ns = not significant (<i>p</i><0.05).</p

Intestinal parasite distribution amongst VL patients.

<p>Intestinal parasite distribution amongst VL patients.</p

Comparison of Th2 cytokine levels.

<p>Comparison of Th2 cytokine levels.</p