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In mice, experimental infection with Trypanosoma brucei causes decreased bone marrow B-cell development, abolished splenic B-cell maturation and loss of antibody mediated protection including vaccine induced memory responses. Nothing is known about this phenomenon in human African trypanosomiasis (HAT), but if occurring, it would imply the need of revaccination of HAT patients after therapy and abolish hope for a HAT vaccine. The effect of gambiense HAT on peripheral blood memory T- and B-cells and on innate and vaccine induced antibody levels was examined. The percentage of memory B- and T-cells was quantified in peripheral blood, prospectively collected in DR Congo from 117 Trypanosoma brucei gambiense infected HAT patients before and six months after treatment and 117 controls at the same time points. Antibodies against carbohydrate antigens on red blood cells and against measles were quantified. Before treatment, significantly higher percentages of memory B-cells, mainly T-independent memory B-cells, were observed in HAT patients compared to controls (CD20+CD27+IgM+, 13.0% versus 2.0%, p<0.001). The percentage of memory T-cells, mainly early effector/memory T-cells, was higher in HAT (CD3+CD45RO+CD27+, 19.4% versus 16.7%, pâ=â0.003). After treatment, the percentage of memory T-cells normalized, the percentage of memory B-cells did not. The median anti-red blood cell carbohydrate IgM level was one titer lower in HAT patients than in controls (p<0.004), and partially normalized after treatment. Anti-measles antibody concentrations were lower in HAT patients than in controls (medians of 1500 versus 2250 mIU/ml, pâ=â0.02), and remained so after treatment, but were above the cut-off level assumed to provide protection in 94.8% of HAT patients, before and after treatment (versus 98.3% of controls, pâ=â0.3). Although functionality of the B-cells was not verified, the results suggest that immunity was conserved in T.b. gambiense infected HAT patients and that B-cell dysfunction might not be that severe as in mouse models.
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Anticorpos Antiprotozoários/sangue , Linfócitos B/imunologia , Imunidade Humoral/imunologia , Memória Imunológica/imunologia , Tripanossomíase Africana/imunologia , Adulto , Anticorpos Antiprotozoários/imunologia , Feminino , Citometria de Fluxo , Humanos , Masculino , Fenótipo , Trypanosoma brucei gambiense/imunologia , Tripanossomíase Africana/sangueRESUMO
BACKGROUND: Aedes albopictus has been reported in several Central African countries, including the Democratic Republic of the Congo (DRC). The establishment of this mosquito species poses a serious threat as a vector of various infectious diseases. Although Ae. albopictus has been reported in the western region of the DRC, information about its distribution is still scarce in the country. The aim of this study was to investigate the current nationwide distribution of the invasive Ae. albopictus, as well as other native Aedes mosquitoes, in the DRC and to identify suitable areas for its future expansion. METHODS: Two entomological surveys were conducted in 2017-2019 and 2022. Based on the occurrence sites of Ae. albopictus, important environmental variables were identified. Then, geographical areas suitable for Ae. albopictus establishment were determined using the maximum entropy model. The distribution and abundance of Ae. albopictus were also compared with those of the major native Aedes species. RESULTS: Aedes albopictus was found in the western, northern, central, and eastern regions of the DRC, but it was not found in the southeastern region. The maximum entropy model predicted that most parts of the DRC are suitable for the establishment of this mosquito. The unsuitable areas encompassed the eastern highlands, known for their low temperatures, and the southeastern highlands, which experience both low temperatures and a long dry season. The native Aedes species found were Aedes aegypti, Aedes simpsoni, Aedes africanus, and Aedes vittatus. Aedes albopictus dominated in the western and northern regions, while Ae. aegypti was more prevalent in other regions. CONCLUSIONS: Aedes albopictus has been well established in the western and northern regions of the DRC. This mosquito is expanding its distribution while replacing the native Aedes species. Most of the country is suitable for the establishment of this mosquito species, except the highlands of the eastern and the southeastern regions.
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Aedes , Animais , República Democrática do Congo , Mosquitos Vetores , Meio AmbienteRESUMO
OBJECTIVES: Partial artemisinin resistance, mediated by Plasmodium falciparum K13 (PfK13) mutations, has been confirmed in certain areas of East Africa that are historically associated with high-level antimalarial resistance. The Democratic Republic of Congo (DRC) borders these areas in the East. This study aimed to determine the prevalence of resistance markers in six National Malaria Control Program surveillance sites; Boende, Kabondo, Kapolowe, Kimpese, Mikalayi, and Rutshuru. METHODS: The single nucleotide polymorphisms (SNPs) in P. falciparum genes PfK13, Pfdhfr, Pfdhps, Pfmdr1, and Pfcrt were assessed using targeted next-generation sequencing of isolates collected at enrollment in therapeutic efficacy studies. RESULTS: PfK13 SNPs were detected in two samples: in Kabondo (R561H) and in Rutshuru (P441L), both areas near Uganda and Rwanda. The Pfdhps ISGEGA haplotype, associated with reduced sulfadoxine-pyrimethamine chemoprevention efficacy, ranged from 0.8% in Mikalayi (central DRC) to 42.2% in Rutshuru (East DRC). CONCLUSIONS: R561H and P441L observed in eastern DRC are a concern, as they are associated with delayed artemisinin-based combination therapies-clearance and candidate marker of resistance, respectively. This is consistent with previous observations of shared drug resistance profiles in parasites of that region with bordering areas of Rwanda and Uganda. The likely circulation of parasites has important implications for the ongoing surveillance of partial artemisinin-resistant P. falciparum and for future efforts to mitigate its dispersal.
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Antimaláricos , Artemisininas , Malária Falciparum , Humanos , Plasmodium falciparum/genética , República Democrática do Congo/epidemiologia , Antimaláricos/farmacologia , Antimaláricos/uso terapêutico , Malária Falciparum/tratamento farmacológico , Malária Falciparum/epidemiologia , Malária Falciparum/parasitologia , Artemisininas/farmacologia , Artemisininas/uso terapêutico , Mutação , Uganda , Proteínas de Protozoários/genéticaRESUMO
OBJECTIVES: Diagnosis of the neurological stage of human African trypanosomiasis is performed by examination of cerebrospinal fluid (CSF) for the presence of trypanosomes and numbers of white blood cells (WBC). Both CSF parameters are also used to assess treatment outcome during follow-up. In view of the importance of CSF examination, and the practical problems encountered with it, we compared the sensitivity of two trypanosome concentration techniques and the repeatability of two cell counting methods, as well as occurrence of systematic differences between them. METHODS: Patients were recruited at Dipumba hospital, in Mbuji-Mayi in the Democratic Republic of the Congo. In 94 CSF samples, trypanosome detection was performed with modified single centrifugation (MSC) and double centrifugation (DC). On 189 CSF samples with ≤30 cells/µl, cell counting was performed in duplicate in a Fuchs-Rosenthal counting chamber and in a disposable Uriglass counting chamber. RESULTS: Modified single centrifugation detected trypanosomes in significantly (P < 0.0001) more patients (85) than DC (46). Cell counts did not differ systematically in the two methods. Variability in the differences between duplicate cell counts was significantly higher (P = 0.002) in Uriglass (SD of differences 2.03) than in Fuchs-Rosenthal (SD of differences 1.62). CONCLUSIONS: For analysis of CSF in the context of sleeping sickness stage determination and follow-up after treatment, we strongly recommend the MSC for parasite detection and the application of disposable counting chambers. When the first cell count is ≤20 cells/µl, we recommend repeating the counting procedure on the same CSF specimen and taking the average of both countings.
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Líquido Cefalorraquidiano/parasitologia , Leucócitos/parasitologia , Trypanosoma brucei gambiense/isolamento & purificação , Tripanossomíase Africana/líquido cefalorraquidiano , Tripanossomíase Africana/classificação , República Democrática do Congo , Humanos , Contagem de Leucócitos , Padrões de ReferênciaRESUMO
OBJECTIVE: To identify the risk factors associated with ocular toxoplasmosis (OT) in a cohort of Congolese patients with uveitis. METHODS AND ANALYSIS: A cross-sectional study was conducted between March 2020 and July 2021 in two ophthalmic clinics in Kinshasa. Patients with a diagnosis of uveitis were enrolled in the study. Each patient underwent an interview, an ophthalmological examination and serology testing. Logistic regression was performed to identify risk factors for OT. RESULTS: 212 patients were included in the study with a mean age at presentation of 42.1±15.9 years (limits: 8-74 years) and a sex ratio of 1.1:1. OT concerned 96 patients (45.3%). The age of the patients below 60 years (p=0.001, OR=9.75 CI 95% 2.51 to37.80)), the consumption of cat meat (p=0.01, OR=2.65 CI 95% 1.18 to 5.96)) and undercooked meat (p=0.044, OR=2.30 CI 95% 1.02 to 5.21)) and living in rural area (p=0.021, OR=11.4 (CI 95% 1.45 to 89.84])) were identified as risk factors for OT. CONCLUSION: OT affects more young people. It is associated with dietary habits. Informing and educating the population is necessary to avoid infection.
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Toxoplasmose Ocular , Uveíte , Humanos , Toxoplasmose Ocular/epidemiologia , Estudos Transversais , República Democrática do Congo/epidemiologia , Uveíte/epidemiologia , Fatores de RiscoRESUMO
PURPOSE: To describe demographic data, clinical features, and serological profiles in a cohort of Congolese patients with ocular toxoplasmosis (OT). METHOD: Cross-sectional study, carried out between March 2020 and July 2021 in two ophthalmic clinics in Kinshasa. RESULTS: The study comprised 95 participants with OT. Fifty-three patients were male (55.8%). The mean age at presentation was 35.6 ± 14.1 years (range 8-69 years); 71 had active OT (74.7%), among them, 33 had primary OT (46.5%), and 38 had recurrences (53.5%). At presentation, 51 patients (53.7%) had visual impairment (VA < 6/18). Retinochoroidal lesions were located in the central retina in 60 patients (63.1%). Patients with primary OT tend to have higher IgG levels than those with recurrent OT (P = .01). CONCLUSION: We report the largest cohort of patients with OT in sub-Saharan Africa. In our setting, most patients had recurrent OT with multiple, extensive, and central retinochoroidal lesions.
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Toxoplasmose Ocular , Baixa Visão , Humanos , Masculino , Criança , Adolescente , Adulto Jovem , Adulto , Pessoa de Meia-Idade , Idoso , Feminino , Estudos Transversais , República Democrática do CongoRESUMO
Gambiense human African trypanosomiasis (gHAT), also known as gambiense sleeping sickness, is a parasitic infection caused by Trypanosoma brucei gambiense. During the last decades, gHAT incidence has been brought to an all-time low. Newly developed serological tools and drugs for its diagnosis and treatment put the WHO goal of interruption of transmission by 2030 within reach. However, further research is needed to efficiently adapt these new advances to new control strategies. We assessed the serological evolution of cured gHAT patients over a two-year period using four different tests: the rapid diagnostic test (RDT) HAT Sero K-SeT, ELISA/T.b. gambiense, Trypanosoma brucei gambiense inhibition ELISA (iELISA), and the immune trypanolysis test. High seropositive rates were observed in all the tests, although sero-reversion rates were different in each test: ELISA/T.b. gambiense was the test most likely to become negative two years after treatment, whereas RDT HAT Sero-K-SeT was the least likely. iELISA and trypanolysis showed intermediate and comparable probabilities to become negative. Stage 1 patients were also noted to be more likely to become negative than Stage 2 patients in all four serological tests. Our results confirm previous findings that trypanosome-specific antibody concentrations in blood may persist for up to two years, implying that HAT control programs should continue to take the history of past HAT episodes into consideration.
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PURPOSE: To present a narrative review about ocular toxoplasmosis epidemiology, disease burden and prevalent African parasitic strains. METHODS: An initial search for MeSH terms was conducted with a posterior advanced search in two electronic databases. Full text reading was performed. RESULTS: Animal African studies have identified Toxoplasma gondii type II, type III, Africa 1, and Africa 3 strains. Seroprevalence varies from 6.4% to 74.5%. Nevertheless, there is a scarcity of epidemiology and serotyping information about ocular toxoplasmosis. African studies have demonstrated that uveitis patients present high frequencies of ocular toxoplasmosis. There is a lack of studies describing specific clinical characteristics, which can be related, to environmental and socioeconomic factors, parasite serotype and genotype, and genetic susceptibility of the host. CONCLUSION: As Toxoplasma gondii has more virulent strains in the Southern hemisphere, it is relevant to determine African strain types and the correlation between the infecting strains and the clinical manifestations.
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Toxoplasma , Toxoplasmose Ocular , África/epidemiologia , Animais , Genótipo , Humanos , Estudos Soroepidemiológicos , Toxoplasma/genética , Toxoplasmose Ocular/diagnóstico , Toxoplasmose Ocular/epidemiologia , Toxoplasmose Ocular/parasitologiaRESUMO
BACKGROUND: Detection of spliced leader (SL)-RNA allows sensitive diagnosis of gambiense human African trypanosomiasis (HAT). We investigated its diagnostic performance for treatment outcome assessment. METHODS: Blood and cerebrospinal fluid (CSF) from a consecutive series of 97 HAT patients, originating from the Democratic Republic of the Congo, were prospectively collected before treatment with acoziborole, and during 18 months of longitudinal follow-up after treatment. For treatment outcome assessment, SL-RNA detection was compared with microscopic trypanosome detection and CSF white blood cell count. The trial was registered under NCT03112655 in clinicaltrials.gov. FINDINGS: Before treatment, respectively 94.9% (92/97; CI 88.5-97.8%) and 67.7% (65/96; CI 57.8-76.2%) HAT patients were SL-RNA positive in blood or CSF. During follow-up, one patient relapsed with trypanosomes observed at 18 months, and was SL-RNA positive in blood and CSF at 12 months, and CSF positive at 18 months. Among cured patients, one individual tested SL-RNA positive in blood at month 12 (Specificity 98.9%; 90/91; CI 94.0-99.8%) and 18 (Specificity 98.9%; 88/89; CI 93.9-99.8%). INTERPRETATION: SL-RNA detection for HAT treatment outcome assessment shows ≥98.9% specificity in blood and 100% in CSF, and may detect relapses without lumbar puncture. FUNDING: The DiTECT-HAT project is part of the EDCTP2 programme, supported by Horizon 2020, the European Union Funding for Research and Innovation (grant number DRIA-2014-306-DiTECT-HAT).
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Antiprotozoários , Trypanosoma , Tripanossomíase Africana , Animais , Humanos , Antiprotozoários/uso terapêutico , Seguimentos , RNA Líder para Processamento , Resultado do Tratamento , Trypanosoma brucei gambiense/genética , Tripanossomíase Africana/diagnóstico , Tripanossomíase Africana/tratamento farmacológicoRESUMO
BACKGROUND. Clinical management of human African trypanosomiasis requires patient follow-up of 2 years' duration. At each follow-up visit, cerebrospinal fluid (CSF) is examined for trypanosomes and white blood cells (WBCs). Shortening follow-up would improve patient comfort and facilitate control of human African trypanosomiasis. METHODS. A prospective study of 360 patients was performed in the Democratic Republic of the Congo. The primary outcomes of the study were cure, relapse, and death. The WBC count, immunoglobulin M level, and specific antibody levels in CSF samples were evaluated to detect treatment failure. The sensitivity and specificity of shortened follow-up algorithms were calculated. RESULTS. The treatment failure rate was 37%. Trypanosomes, a WBC count of > or = 100 cells/microL, and a LATEX/immunoglobulin M titer of 1:16 in CSF before treatment were risk factors for treatment failure, whereas human immunodeficiency virus infection status was not a risk factor. The following algorithm, which had 97.8% specificity and 94.4% sensitivity, is proposed for shortening the duration of follow-up: at 6 months, patients with trypanosomes or a WBC count of > or = 50 cells/microL in CSF are considered to have treatment failure, whereas patients with a CSF WBC count of > or = 5 cells/microL are considered to be cured and can discontinue follow-up. At 12 months, the remaining patients (those with a WBC count of > or = 6-49 cells/microL) need a test of cure, based on trypanosome presence and WBC count, applying a cutoff value of > or = 20 cells/microL. CONCLUSION. Combining criteria for failure and cure allows follow-up of patients with second-stage human African trypanosomiasis to be shortened to a maximum duration of 12 months.
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Antiprotozoários/uso terapêutico , Trypanosoma brucei gambiense/isolamento & purificação , Tripanossomíase Africana/tratamento farmacológico , Adulto , Algoritmos , Animais , Líquido Cefalorraquidiano/citologia , República Democrática do Congo , Feminino , Humanos , Contagem de Leucócitos , Masculino , Fatores de Risco , Sensibilidade e Especificidade , Fatores de Tempo , Tripanossomíase Africana/líquido cefalorraquidiano , Tripanossomíase Africana/epidemiologia , Adulto JovemRESUMO
Cassava cyanide-related neurocognitive impairment may persist for years in Central African children who rely on cassava as a dietary staple. In the Democratic Republic of the Congo, a cassava processing method, the 'wetting method', reduced cyanide in cassava, prevented konzo, and proved a cost-effective intervention to improve children's cognitive development. Scaling up use of the wetting method may help prevent neurocognitive impairment in millions of at-risk children in sub-Saharan Africa.
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BACKGROUND: Spliced Leader (SL) trypanosome RNA is detectable only in the presence of live trypanosomes, is abundant and the Trypanozoon subgenus has a unique sequence. As previously shown in blood from Guinean human African trypanosomiasis (HAT) patients, SL-RNA is an accurate target for diagnosis. Detection of SL-RNA in the cerebrospinal fluid (CSF) has never been attempted. In a large group of Congolese gambiense HAT patients, the present study aims i) to confirm the sensitivity of SL-RNA detection in the blood and; ii) to assess the diagnostic performance of SL-RNA compared to trypanosome detection in CSF. METHODOLOGY/PRINCIPAL FINDINGS: Blood and CSF from 97 confirmed gambiense HAT patients from the Democratic Republic of Congo were collected using PAXgene blood RNA Tubes. Before RNA extraction, specimens were supplemented with internal extraction control RNA to monitor the extraction, which was performed with a PAXgene Blood RNA Kit. SL-RNA qPCR was carried out with and without reverse transcriptase to monitor DNA contamination. In blood, 92/97 (94.8%) HAT patients tested SL-RNA positive, which was significantly more than combined trypanosome detection in lymph and blood (78/97 positive, 80.4%, p = 0.001). Of 96 CSF RNA specimens, 65 (67.7%) were SL-RNA positive, but there was no significant difference between sensitivity of SL-RNA and trypanosome detection in CSF. The contribution of DNA to the Cq values was negligible. In CSF with normal cell counts, a fraction of SL-RNA might have been lost during extraction as indicated by higher internal extraction control Cq values. CONCLUSIONS/SIGNIFICANCE: Detection of SL-RNA in blood and CSF allows sensitive demonstration of active gambiense HAT infection, even if trypanosomes remain undetectable in blood or lymph. As this condition often occurs in treatment failures, SL-RNA detection in blood and CSF for early detection of relapses after treatment deserves further investigation. TRIAL REGISTRATION: This study was an integral part of the diagnostic trial "New Diagnostic Tools for Elimination of Sleeping Sickness and Clinical Trials: Early tests of Cure" (DiTECT-HAT-WP4, ClinicalTrials.gov Identifier: NCT03112655).
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RNA de Protozoário/genética , RNA de Protozoário/isolamento & purificação , Trypanosoma brucei gambiense , Tripanossomíase Africana/parasitologia , República Democrática do Congo/epidemiologia , Humanos , RNA de Protozoário/sangue , RNA de Protozoário/líquido cefalorraquidiano , Tripanossomíase Africana/sangue , Tripanossomíase Africana/líquido cefalorraquidiano , Tripanossomíase Africana/epidemiologiaRESUMO
Beside diagnostic uncertainties due to the lack of a perfect gold standard test for Helicobacter pylori infection, the diagnosis and the prevalence estimation for this infection encounter particular challenges in Africa including limited diagnostic tools and specific genetic background. We developed and evaluated the accuracy of an enzyme-linked immunosorbent assay (ELISA) system tailored for H. pylori genetics in Africa (HpAfr-ELISA). Strains belonging to main genetic populations infecting Africans were exploited as sources for whole-cell antigens to establish in-house the ELISA system. A phase II unmatched case-control study explored the diagnostic accuracy of the HpAfr-ELISA using a training set of samples collected from dyspeptic patients from Kinshasa, the Democratic Republic of Congo (DRC) who had been tested with invasive standard tests (i.e., histology, culture, and rapid urease test) in 2017. Then the assay was cross-validated through a community-based survey assessing the prevalence of H. pylori and associated factors in 425 adults from Mbujimayi, DRC in 2018. Bayesian inferences were used to deal with statistical uncertainties of estimates (true prevalence, sensitivity, and specificity) in the study population. At its optimal cut-off-value 20.2 U/mL, the assay achieved an estimated sensitivity of 97.6% (95% credible interval [95%CrI]: 89.2; 99.9%) and specificity of 90.5% (95%CrI: 78.6; 98.5). Consistent outcomes obtained at repeated tests attested the robustness of the assay (negative and positive agreements always > 70%). The true prevalence of H. pylori was estimated 53.8% [95%CrI: 42.8; 62.7%]. Increasing age (adjusted odds ratio [aOR] > 1.0 [95% confidence interval (CI): > 1.0; 1.1]; p<0.001), overcrowding households (aOR = 3.2 [95%CI: 2.0; 5.1]; p<0.001), and non-optimal hand hygiene (aOR = 4.5 [95%CI: 2.0; 11.4]; p = 0.001) were independently associated with the H. pylori-seropositivity. The novel ELISA system has demonstrated good diagnostic accuracy and potential usefulness for management and mitigation strategies for H. pylori infection in African settings.
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Ensaio de Imunoadsorção Enzimática/métodos , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/microbiologia , Helicobacter pylori/isolamento & purificação , Adolescente , Adulto , África/epidemiologia , Idoso , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
Konzo, a distinct upper motor neuron disease associated with a cyanogenic diet and chronic malnutrition, predominately affects children and women of childbearing age in sub-Saharan Africa. While the exact biological mechanisms that cause this disease have largely remained elusive, host-genetics and environmental components such as the gut microbiome have been implicated. Using a large study population of 180 individuals from the Democratic Republic of the Congo, where konzo is most frequent, we investigate how the structure of the gut microbiome varied across geographical contexts, as well as provide the first insight into the gut flora of children affected with this debilitating disease using shotgun metagenomic sequencing. Our findings indicate that the gut microbiome structure is highly variable depending on region of sampling, but most interestingly, we identify unique enrichments of bacterial species and functional pathways that potentially modulate the susceptibility of konzo in prone regions of the Congo.
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Suscetibilidade a Doenças/microbiologia , Comportamento Alimentar , Microbioma Gastrointestinal/fisiologia , Manihot/efeitos adversos , Doença dos Neurônios Motores/microbiologia , Criança , República Democrática do Congo/epidemiologia , Fezes/microbiologia , Feminino , Humanos , Manihot/química , Metagenômica , Doença dos Neurônios Motores/epidemiologia , Nitrilas/efeitos adversosRESUMO
Background: Human African trypanosomiasis (HAT) is a protozoal disease transmitted by tsetse flies. Infection with trypanosomes can lead directly to active HAT or latent infection with no detectable parasites, which may progress to active HAT or to spontaneous self-cure. Genetic variation could explain these differences in the outcome of infection. To test this hypothesis, polymorphisms in 17 candidate genes were tested ( APOL1 [ G1 and G2], CFH, HLA-A, HPR, HP, IL1B, IL12B, IL12RB1, IL10, IL4R, MIF, TNFA , IL6, IL4, IL8, IFNG, and HLA-G). Methods: Samples were collected in Democratic Republic of the Congo. 233 samples were genotyped: 100 active HAT cases, 33 from subjects with latent infections and 100 negative controls. Commercial service providers genotyped polymorphisms at 96 single nucleotide polymorphisms (SNPs) on 17 genes. Data were analyzed using Plink V1.9 software and R. Loci, with suggestive associations (uncorrected p < 0.05) validated using an additional 594 individuals, including 164 cases and 430 controls. Results: After quality control, 87 SNPs remained in the analysis. Two SNPs in IL4 and two in IFNG were suggestively associated (uncorrected p<0.05) with a differential risk of developing a Trypanosoma brucei gambiense infection in the Congolese population. The IFNG minor allele (rs2430561, rs2069718) SNPs were protective in comparison between latent infections and controls. Carriers of the rs2243258_T and rs2243279_A alleles of IL4 and the rs2069728_T allele of IFNG had a reduced risk of developing illness or latent infection, respectively. None of these associations were significant after Bonferroni correction for multiple testing. A validation study using more samples was run to determine if the absence of significant association was due to lack of power. Conclusions: This study showed no evidence of an association of HAT with IL4 and IFNG SNPs or with APOL1 G1 and G2 alleles, which have been found to be protective in other studies.
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Human African trypanosomiasis (HAT) caused by the extracellular protozoon Trypanosoma brucei, is a neglected tropical disease affecting the poorest communities in sub-Saharan Africa. HAT progresses from a hemolymphatic first stage (S1) to a meningo-encephalitic late stage (S2) when parasites reach the central nervous system (CNS), although the existence of an intermediate stage (Int.) has also been proposed. The pathophysiological mechanisms associated with the development of S2 encephalopathy are yet to be fully elucidated. Here we hypothesized that HAT progression toward S2 might be accompanied by an increased release of microvesicles (MVs), sub-micron elements (0.1-1 µm) involved in inflammatory processes and in the determination of the outcome of infections. We studied the morphology of MVs isolated from HAT cerebrospinal fluid (CSF) by transmission electron microscopy (TEM) and used flow cytometry to show that total-MVs and leukocyte derived-CD45+ MVs are significantly increased in concentration in S2 patients' CSF compared to S1 and Int. samples (n = 12 per group). To assess potential biological properties of these MVs, immortalized human astrocytes were exposed, in vitro, to MVs enriched from S1, Int. or S2 CSF. Data-independent acquisition mass spectrometry analyses showed that S2 MVs induced, compared to Int. or S1 MVs, a strong proteome modulation in astrocytes that resembled the one produced by IFN-γ, a key molecule in HAT pathogenesis. Our results indicate that HAT S2 CSF harbors MVs potentially involved in the mechanisms of pathology associated with HAT late stage. Such vesicles might thus represent a new player to consider in future functional studies.
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Astrócitos/metabolismo , Vesículas Extracelulares/metabolismo , Trypanosoma brucei gambiense , Tripanossomíase Africana/metabolismo , Biomarcadores , Congo , Vesículas Extracelulares/ultraestrutura , Feminino , Citometria de Fluxo , Interações Hospedeiro-Parasita , Humanos , Masculino , Doenças Negligenciadas , Proteoma , Proteômica , Tripanossomíase Africana/líquido cefalorraquidiano , Tripanossomíase Africana/diagnóstico , Tripanossomíase Africana/parasitologiaRESUMO
The impact of concurrent exposure to neurotoxic metals is a significant threat to brain function, mostly in contexts of multiple exposures as seen in the developing world. Ninety-five children (46 boys and 49 girls, 6 to 11-year old) from Congo-Kinshasa were assessed for cognition using the Kaufman Assessment Battery for Children (2nd edition) and exposure to Cr, Cu, Zn, Co, Mn, As, Cd, Se, Hg, Fe, and Pb by inductively coupled plasma mass spectrometry (ICPMS) in serum and urine collections. Concentrations of elements were all above normal ranges except for Cd, Se and Hg. General linear mixed effects models were used to predict neurocognitive outcomes with variable selection methods including backward elimination, elastic net, or subsets identified based on subject matter expertise. After adjusting for sex, age, and SES, urinary Coâ¯>â¯5⯵g/l was associated with poor simultaneous processing (memory) (pâ¯=â¯0.0237). Higher excretion but normal concentration of Cd in serum was associated with better memory (pâ¯=â¯0.03), planning (pâ¯=â¯0.05), and overall performance scores (pâ¯<â¯0.01); thus appeared to be neuroprotective. However, higher excretion of Zn had negative influence on the overall performance scores (pâ¯=â¯0.02). Predictive neurotoxicology is a challenging task in contexts of multiple and concurrent exposures. Urinary Coâ¯>â¯5⯵g/l is a risk factor for poor neurodevelopmental outcomes in such contexts. The impact of heavy metals on cognition is dependent on concentrations of and interactions between toxic and essential elements.
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Intoxicação por Metais Pesados/epidemiologia , Metais Pesados/efeitos adversos , Metais Pesados/toxicidade , Criança , Pré-Escolar , Cognição/efeitos dos fármacos , Cognição/fisiologia , Estudos Transversais , República Democrática do Congo/epidemiologia , Feminino , Intoxicação por Metais Pesados/fisiopatologia , Humanos , MasculinoRESUMO
BACKGROUND: To date, no biological marker for treatment outcome in human African trypanosomiasis (HAT) has been described. The accuracy of biological markers for prediction of treatment outcome of HAT caused by Trypanosoma brucei gambiense was assessed. METHODS: Cerebrospinal fluid (CSF) white blood cell (WBC) count and immunoglobulin M (IgM), trypanosome-specific antibody, total protein, and interleukin-10 levels were determined before and up to 24 months after treatment of late-stage HAT. RESULTS: Treatment failure was experienced by 48 of 260 patients. Pretreatment CSF WBC counts > or = 102 cells/microL, IL-10 concentrations > or = 37 pg/mL, LATEX/IgM end titers > or = 1:32, LATEX/T. b. gambiense end titers > or = 1:2, and protein concentrations > or = 674 mg/L were associated with treatment failure. Six months after treatment, patients with CSF WBC counts < or = 5 cells/microL were at low risk of HAT recurrence (negative predictive value, >0.93). After 12 months, the combination of CSF WBC count > or = 8 cells/microL and LATEX/IgM end titer > or = 1:4 predicted treatment failure with 97% specificity and 79% sensitivity. Eighteen months after treatment, each marker accurately predicted treatment outcome. The combination of CSF WBC count > or = 8 cells/microL and LATEX/IgM end titer > or = 1:4 was 100% specific for treatment failure after 18 and 24 months. CONCLUSIONS: HAT-affected patients with elevated pretreatment CSF levels of WBC, interleukin-10, IgM, trypanosome-specific antibody, and total protein are at risk of treatment failure. Six months after treatment, patients with CSF WBC counts < or = 5 cells/microL can be considered to be cured. The assessment of a combination of CSF WBC count and LATEX/IgM level allowed accurate prediction of outcome beginning at 12 months after treatment, as did each individual marker at 18 months after treatment.
Assuntos
Trypanosoma brucei gambiense/isolamento & purificação , Tripanossomíase Africana/tratamento farmacológico , Tripanossomíase Africana/parasitologia , Animais , Anticorpos Antiprotozoários/líquido cefalorraquidiano , Biomarcadores , Líquido Cefalorraquidiano/química , Líquido Cefalorraquidiano/imunologia , Humanos , Imunoglobulina M/líquido cefalorraquidiano , Interleucina-10/líquido cefalorraquidiano , Contagem de Leucócitos , Melarsoprol/uso terapêutico , Valor Preditivo dos Testes , Proteínas/análise , Resultado do Tratamento , Tripanossomicidas/uso terapêuticoRESUMO
Trypanosoma brucei (T. b.) gambiense is the parasite subspecies responsible for most reported cases of human African trypanosomiasis (HAT) or sleeping sickness. This severe infection leads to characteristic disruption of the sleep-wake cycle, recalling attention on the circadian timing system. Most animal models of the disease have been hitherto based on infection of laboratory rodents with the T. b. brucei subspecies, which is not infectious to humans. In these animal models, functional, rather than structural, alterations of the master circadian pacemaker, the hypothalamic suprachiasmatic nucleus (SCN), have been reported. Information on the SCN after infection with the human pathogenic T. b. gambiense is instead lacking. The present study was aimed at the examination of the SCN after T. b. gambiense infection of a susceptible rodent, the multimammate mouse, Mastomys natalensis, compared with T. b. brucei infection of the same host species. The animals were examined at 4 and 8 weeks post-infection, when parasites (T. b. gambiense or T. b. brucei) were detected in the brain parenchyma, indicating that the disease was in the encephalitic stage. Neuron and astrocyte changes were examined with Nissl staining, immunophenotyping and quantitative analyses. Interestingly, significant neuronal loss (about 30% reduction) was documented in the SCN during the progression of T. b. gambiense infection. No significant neuronal density changes were found in the SCN of T. b. brucei-infected animals. Neuronal cell counts in the hippocampal dentate gyrus of T. b. gambiense-infected M. natalensis did not point out significant changes, indicating that no widespread neuron loss had occurred in the brain. Marked activation of astrocytes was detected in the SCN after both T. b. gambiense and T. b. brucei infections. Altogether the findings reveal that neurons of the biological clock are highly susceptible to the infection caused by human pathogenic African trypanosomes, which have the capacity to cause permanent partial damage of this structure.
RESUMO
Neuron populations of the lateral hypothalamus which synthesize the orexin (OX)/hypocretin or melanin-concentrating hormone (MCH) peptides play crucial, reciprocal roles in regulating wake stability and sleep. The disease human African trypanosomiasis (HAT), also called sleeping sickness, caused by extracellular Trypanosoma brucei (T. b.) parasites, leads to characteristic sleep-wake cycle disruption and narcoleptic-like alterations of the sleep structure. Previous studies have revealed damage of OX and MCH neurons during systemic infection of laboratory rodents with the non-human pathogenic T. b. brucei subspecies. No information is available, however, on these peptidergic neurons after systemic infection with T. b. gambiense, the etiological agent of 97% of HAT cases. The present study was aimed at the investigation of immunohistochemically characterized OX and MCH neurons after T. b. gambiense or T. b. brucei infection of a susceptible rodent, the multimammate mouse, Mastomysnatalensis. Cell counts and evaluation of OX fiber density were performed at 4 and 8 weeks post-infection, when parasites had entered the brain parenchyma from the periphery. A significant decrease of OX neurons (about 44% reduction) and MCH neurons (about 54% reduction) was found in the lateral hypothalamus and perifornical area at 8 weeks in T. b. gambiense-infected M. natalensis. A moderate decrease (21% and 24% reduction, respectively), which did not reach statistical significance, was found after T. b. brucei infection. In two key targets of diencephalic orexinergic innervation, the peri-suprachiasmatic nucleus (SCN) region and the thalamic paraventricular nucleus (PVT), densitometric analyses showed a significant progressive decrease in the density of orexinergic fibers in both infection paradigms, and especially during T. b. gambiense infection. Altogether the findings provide novel information showing that OX and MCH neurons are highly vulnerable to chronic neuroinflammatory signaling caused by the infection of human-pathogenic African trypanosomes.