RESUMO
The low prevalence levels associated with lymphatic filariasis elimination pose a challenge for effective disease surveillance. As more countries achieve the World Health Organization criteria for halting mass treatment and move on to surveillance, there is increasing reliance on the utility of transmission assessment surveys (TAS) to measure success. However, the long-term disease outcomes after passing TAS are largely untested. Using 3 well-established mathematical models, we show that low-level prevalence can be maintained for a long period after halting mass treatment and that true elimination (0% prevalence) is usually slow to achieve. The risk of resurgence after achieving current targets is low and is hard to predict using just current prevalence. Although resurgence is often quick (<5 years), it can still occur outside of the currently recommended postintervention surveillance period of 4-6 years. Our results highlight the need for ongoing and enhanced postintervention monitoring, beyond the scope of TAS, to ensure sustained success.
Assuntos
Filariose Linfática/sangue , Filariose Linfática/parasitologia , Microfilárias/isolamento & purificação , Modelos Biológicos , Animais , Simulação por Computador , Erradicação de Doenças , Filariose Linfática/epidemiologia , HumanosRESUMO
According to the World Health Organization, lymphatic filariasis (LF), a mosquito-borne neglected tropical disease (NTD), should be eliminated as a public health concern by the end of 2020. To this end, the goals of the Global Programme to Eliminate Lymphatic Filariasis (GPELF) include interrupting transmission through mass drug administration (MDA). After two decades, several countries have implemented MDA and are now ready to confirm whether transmission has been interrupted. The method for detecting the parasites in mosquito vectors known as xenomonitoring is a non-invasive tool for assessing the current transmission status of the filarial nematode Wuchereria bancrofti (which is responsible for 90% of cases) by their vectors. There are several methods available for detection of the worm in mosquito samples, such as dissection or polymerase chain reaction (PCR). However, most of these techniques still produce a considerable number of false-negative results. The present study describes a new duplex PCR protocol, which is an improvement on the traditional PCR methodology, enhanced by introducing the actin gene as an endogenous control gene. After adjusting the mosquito pool size, DNA extraction, and WbCx PCR duplex design, we achieved a reliable and sensitive molecular xenomonitoring protocol. This assay was able to eliminate 5% of false negative samples and detected less than one Wb larvae. This high sensitivity is particularly valuable after MDA, when prevalence declines. This new method could reduce the number of false-negative samples, which will enable us to improve our ability to generate accurate results and aid the monitoring strategies used by LF elimination programmes.
Assuntos
Culex/parasitologia , Filariose Linfática/transmissão , Mosquitos Vetores/parasitologia , Reação em Cadeia da Polimerase Multiplex/métodos , Wuchereria bancrofti/fisiologia , Actinas/genética , Animais , Sequência de Bases , Eletroforese em Gel de Ágar , Filariose Linfática/sangue , Filariose Linfática/tratamento farmacológico , Filariose Linfática/prevenção & controle , Feminino , Humanos , Doenças Negligenciadas/parasitologia , Sensibilidade e Especificidade , Wuchereria bancrofti/genéticaRESUMO
OBJECTIVE: The underlying problem in lymphatic filariasis is irreversible swelling of the limbs (lymphoedema), which is a unique feature of lymphatic insufficiency. It is still unclear whether the natural ability of lymphatics to form functional lymphatic vasculature is achieved or attenuated in the lymphoedemal pathology. Clinical studies have clearly shown that circulating lymphatic progenitors (CLPs), a subset of bone marrow-derived mononuclear cells (PBMCs), contribute to post-natal lymph vasculogenesis. CLP-based revascularisation could be a promising strategy to bypass the endothelial disruption and damage incurred by the filarial parasites. Thus our aim was to compare and characterise the functional prowess of PBMCs in physiological and lymphoedemal pathology. METHODS: PBMCs were isolated from venous blood sample from drug-naive endemic normals (EN) and drug-deprived filarial lymphoedema (FL) individuals using density gradient centrifugation. Adhesion, transwell migration and in vitro matrigel assays were employed to characterise the lymphvasculogenic potential of PBMCs. CLPs were phenotypically characterised using flow cytometry; expression levels of lymphatic markers and inflammatory cytokines were quantified using qRT-PCR and ELISA, respectively. RESULTS: PBMCs from FL group display poor adherence to fibronectin (P = 0.040), reduced migration towards SDF-1α (P = 0.035), impaired tubular network (P = 0.004) and branching point (P = 0.048) formation. The PBMC mRNA expression of VEGFR3 (P = 0.039) and podoplanin (P = 0.050) was elevated, whereas integrin α9 (P = 0.046) was inhibited in FL individuals; additionally, the surface expression of CD34 (P = 0.048) was significantly reduced in the FL group compared to the EN group. CONCLUSION: PBMCs from filarial lymphoedema show defective and dysregulated lymphvasculogenic function compared to endemic normals.
Assuntos
Filariose Linfática/patologia , Leucócitos Mononucleares/fisiologia , Linfedema , Adulto , Idoso , Antígenos CD34/sangue , Movimento Celular , Quimiocina CXCL12/sangue , Filariose Linfática/sangue , Doenças Endêmicas , Feminino , Fibronectinas/sangue , Humanos , Índia , Cadeias alfa de Integrinas/sangue , Linfedema/sangue , Linfedema/etiologia , Masculino , Glicoproteínas de Membrana , Pessoa de Meia-Idade , RNA Mensageiro/metabolismo , Valores de Referência , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/sangueRESUMO
OBJECTIVE: Mass drug administration (MDA) for the control of lymphatic filariasis (LF), in Ghana, started in the year 2000. While this had great success in many implementation units, there remain areas with persistent transmission, after more than 10 years of treatment. A closer examination of the parasite populations could help understand the reasons for persistent infections and formulate appropriate strategies to control LF in these areas of persistent transmission. MATERIALS AND METHODS: In a longitudinal study, we assessed the prevalence of microfilaraemia (mf) in two communities with 12 years of MDA in Ghana. In baseline surveys 6 months after the National MDA in 2014, 370 consenting individuals were tested for antigenaemia using immunochromatographic test (ICT) cards and had their mf count determined through night blood surveys. 48 ICT positives, of whom, 17 were positive for mf, were treated with 400 µg/kg ivermectin + 400 mg albendazole and subsequently followed for parasitological assessment at 3-month intervals for 1 year. This overlapped with the National MDA in 2015. RESULTS: There was a 68% parasite clearance 3 months after treatment. The pre-treatment mf count differed significantly from the post-treatment mf counts at 3 months (P = 0.0023), 6 months (P = 0.0051), 9 months (P = 0.0113) and 12 months (P = 0.0008). CONCLUSION: In these settings with persistent LF transmission, twice-yearly treatment may help accelerate LF elimination. Further large-scale evaluations are required to ascertain these findings.
Assuntos
Albendazol/uso terapêutico , Filariose Linfática/parasitologia , Filaricidas/uso terapêutico , Filarioidea/crescimento & desenvolvimento , Ivermectina/uso terapêutico , Adolescente , Adulto , Idoso , Albendazol/farmacologia , Animais , Antígenos de Helmintos/sangue , Criança , Filariose Linfática/sangue , Filariose Linfática/epidemiologia , Filariose Linfática/prevenção & controle , Feminino , Filaricidas/farmacologia , Filarioidea/efeitos dos fármacos , Gana/epidemiologia , Programas Governamentais , Humanos , Ivermectina/farmacologia , Estudos Longitudinais , Masculino , Microfilárias/efeitos dos fármacos , Microfilárias/crescimento & desenvolvimento , Pessoa de Meia-Idade , Prevalência , Adulto JovemRESUMO
Lymphatic filariasis (LF) is a serious vector-borne health problem, and Wuchereria bancrofti (W.b) is the major cause of LF worldwide and is focally endemic in Egypt. Identification of filarial infection using traditional morphologic and immunological criteria can be difficult and lead to misdiagnosis. The aim of the present study was molecular detection of W.b in residents in endemic areas in Egypt, sequence variance analysis, and phylogenetic analysis of W.b DNA. Collected blood samples from residents in filariasis endemic areas in five governorates were subjected to semi-nested PCR targeting repeated DNA sequence, for detection of W.b DNA. PCR products were sequenced; subsequently, a phylogenetic analysis of the obtained sequences was performed. Out of 300 blood samples, W.b DNA was identified in 48 (16%). Sequencing analysis confirmed PCR results identifying only W.b species. Sequence alignment and phylogenetic analysis indicated genetically distinct clusters of W.b among the study population. Study results demonstrated that the semi-nested PCR proved to be an effective diagnostic tool for accurate and rapid detection of W.b infections in nano-epidemics and is applicable for samples collected in the daytime as well as the night time. PCR products sequencing and phylogenitic analysis revealed three different nucleotide sequences variants. Further genetic studies of W.b in Egypt and other endemic areas are needed to distinguish related strains and the various ecological as well as drug effects exerted on them to support W.b elimination.
Assuntos
Sangue/parasitologia , Filariose Linfática/parasitologia , Filogenia , Wuchereria bancrofti/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Sequência de Bases , Criança , Pré-Escolar , Culex/genética , Egito/epidemiologia , Filariose Linfática/sangue , Filariose Linfática/epidemiologia , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Wuchereria bancrofti/classificação , Wuchereria bancrofti/isolamento & purificação , Adulto JovemRESUMO
Global programmes to eliminate lymphatic filariasis (GPELF) require mapping, monitoring and evaluation using filarial antigen diagnostic kits. To meet this objective, a functional single-chain fragment variable (ScFv) specific for filarial Wuchereria bancrofti SXP-1 (Wb-SXP-1) antigen was constructed for the diagnosis of active filarial infection, an alternative to the production of complete antibodies using hybridomas. The variable heavy chain (VH) and the variable light chain (kappa) (Vκ) genes were amplified from the mouse hybridoma cell line and were linked together with a flexible linker by overlap extension polymerase chain reaction (PCR). The ScFv construct (Vκ-Linker-VH) was expressed as a fusion protein with N-terminal His tag in Escherichia coli and purified using immobilized metal affinity chromatography (IMAC) without the addition of reducing agents. Immunoblotting and sandwich enzyme-linked immunosorbent assay (ELISA) were used to analyse the antigen binding affinity of purified ScFv. The purified ScFv was found to recognize recombinant and native Wb-SXP-1 antigen in microfilariae (Mf)-positive patient sera. The affinity of ScFv was comparable with that of the monoclonal antibody. The development of recombinant ScFv to replace monoclonal antibody for detection of filarial antigen was achieved. The recombinant ScFv was purified, on-column refolded and its detection ability validated using field samples.
Assuntos
Antígenos de Helmintos/sangue , Filariose Linfática/diagnóstico , Anticorpos de Cadeia Única/análise , Wuchereria bancrofti/imunologia , Animais , Antígenos de Helmintos/genética , Antígenos de Helmintos/imunologia , Filariose Linfática/sangue , Filariose Linfática/parasitologia , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Humanos , Camundongos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Anticorpos de Cadeia Única/genética , Anticorpos de Cadeia Única/imunologia , Wuchereria bancrofti/genética , Wuchereria bancrofti/isolamento & purificaçãoRESUMO
The aim of this study was to develop a low-cost antifilarial immunoglobulin (Ig) G4 detection kit for the diagnosis of lymphatic filariasis. The kit was designed to be used by minimally trained personnel without the constraints of expensive laboratory equipment. We provide a description of the development and validation of a single-serum-dilution based enzyme-linked immunosorbent assay (ELISA) kit with ready-to-use reagents for measuring antifilarial IgG4 antibodies. The kit was tested on residents in Brugia malayi-endemic areas in southern Thailand. Detection was performed by naked-eye observation of the resultant colour of the immunological reactivity. The coefficient of variation (CV) was used to assess the reproducibility of the results. Long-term stability was measured over a 6-month period. Sensitivity of the test kit was 97% when compared with microfilariae detection in thick blood smears. Specificity was 98.7% based on the sera of 57 patients living outside the endemic areas who were infected with other parasites and 100 parasite-free subjects. All positive CVs were < 10%. The test kit was remarkably stable over 6 months. Field validation was performed by the detection of antifilarial IgG4 in 4365 serum samples collected from residents of brugian filariasis-endemic areas and compared with outcome colours of the test samples by the naked eye. Subsequent ELISA evaluation of these results using an ELISA reader indicated high agreement by the kappa statistic. These results demonstrate that the test kit is efficient and useful for public health laboratories as an alternative tool for the diagnosis of lymphatic filarial infection.
Assuntos
Filariose Linfática/diagnóstico , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Anticorpos Anti-Helmínticos/sangue , Brugia Malayi/imunologia , Brugia Malayi/isolamento & purificação , Criança , Pré-Escolar , Filariose Linfática/sangue , Filariose Linfática/parasitologia , Feminino , Humanos , Imunoglobulina G/sangue , Masculino , Sensibilidade e EspecificidadeRESUMO
Lymphatic filariasis can be associated with development of serious pathology in the form of lymphedema, hydrocele, and elephantiasis in a subset of infected patients. Dysregulated host inflammatory responses leading to systemic immune activation are thought to play a central role in filarial disease pathogenesis. We measured the plasma levels of microbial translocation markers, acute phase proteins, and inflammatory cytokines in individuals with chronic filarial pathology with (CP Ag+) or without (CP Ag-) active infection; with clinically asymptomatic infections (INF); and in those without infection (endemic normal [EN]). Comparisons between the two actively infected groups (CP Ag+ compared to INF) and those without active infection (CP Ag- compared to EN) were used preliminarily to identify markers of pathogenesis. Thereafter, we tested for group effects among all the four groups using linear models on the log transformed responses of the markers. Our data suggest that circulating levels of microbial translocation products (lipopolysaccharide and LPS-binding protein), acute phase proteins (haptoglobin and serum amyloid protein-A), and inflammatory cytokines (IL-1ß, IL-12, and TNF-α) are associated with pathogenesis of disease in lymphatic filarial infection and implicate an important role for circulating microbial products and acute phase proteins.
Assuntos
Proteínas de Fase Aguda/análise , Biomarcadores/sangue , Proteínas de Transporte/sangue , Citocinas/sangue , Filariose Linfática/sangue , Lipopolissacarídeos/sangue , Glicoproteínas de Membrana/sangue , Adulto , Filariose Linfática/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.
Assuntos
DNA de Helmintos/isolamento & purificação , Filariose Linfática/diagnóstico , Microfilárias/isolamento & purificação , Wuchereria bancrofti/isolamento & purificação , Adolescente , Adulto , Animais , Antígenos de Superfície/sangue , Antígenos de Superfície/urina , Filariose Linfática/sangue , Filariose Linfática/urina , Feminino , Humanos , Limite de Detecção , Masculino , Microfilárias/genética , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/normas , Sensibilidade e Especificidade , Wuchereria bancrofti/genética , Adulto JovemRESUMO
BACKGROUND: To monitor the progress of lymphatic filariasis (LF) elimination programmes, field surveys to assess filarial antigen (Ag) prevalence require access to reliable, user-friendly rapid diagnostic tests. We aimed to evaluate the performance of the new Q Filariasis Antigen Test (QFAT) with the currently recommended Filariasis Test Strip (FTS) for detecting the Ag of Wuchereria bancrofti, the causative agent of LF, under field laboratory conditions. METHODOLOGY/PRINCIPAL FINDINGS: During an LF survey in Samoa, 344 finger-prick blood samples were tested using FTS and QFAT. Microfilariae (Mf) status was determined from blood slides prepared from any sample that reported Ag-positive by either Ag-test. Each test was re-read at 1 hour and the next day to determine the stability of results over time. Overall Ag-positivity by FTS was 29.0% and 30.2% by QFAT. Concordance between the two tests was 93.6% (kappa = 0.85). Of the 101 Mf slides available, 39.6% were Mf-positive, and all were Ag-positive by both tests. Darker test line intensities from Ag-positive FTS were found to predict Mf-positivity (compared to same/lighter line intensities). QFAT had significantly higher reported test result changes than FTS, mostly reported the next day, but fewer changes were reported between 10 minutes to 1hour. The field laboratory team preferred QFAT over FTS due to the smaller blood volume required, better usability, and easier readability. CONCLUSION/SIGNIFICANCE: QFAT could be a suitable and user-friendly diagnostic alternative for use in the monitoring and surveillance of LF in field surveys based on its similar performance to FTS under field laboratory conditions.
Assuntos
Antígenos de Helmintos , Filariose Linfática , Wuchereria bancrofti , Humanos , Filariose Linfática/diagnóstico , Filariose Linfática/sangue , Filariose Linfática/epidemiologia , Antígenos de Helmintos/sangue , Wuchereria bancrofti/imunologia , Animais , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Adolescente , Samoa , Adulto Jovem , Criança , Sensibilidade e Especificidade , Idoso , Testes Diagnósticos de Rotina/métodos , Fitas ReagentesRESUMO
Sudan is endemic for multiple neglected tropical diseases, including trachoma, onchocerciasis (OV), lymphatic filariasis (LF), and schistosomiasis (SCH). In 2019, dried blood spot samples were collected for a baseline trachoma serosurvey in three localities (El Seraif, Kotom, and Saraf Omrah) in North Darfur State. None were classified previously as OV- or LF-endemic, although low levels of SCH had been identified in all three. Approximately 30 households from 25 communities in each locality were selected by multistage cluster random sampling. Collections of DBSs were analyzed by multiplex bead assay for antibodies to multiple pathogens. This paper presents data on OV (Ov16), LF (Wb123, Bm14, Bm33), and SCH (soluble egg antigen [SEA], Sm25) antibodies among 8,322 individuals from 2,119 households. The survey-adjusted seroprevalence estimates for Ov16 were <0.3% in all localities. Lymphatic filariasis-antigen seroprevalences were discordant. Seroprevalence estimates ranged from 4.6-6.0% (Wb123), 0.99-1.4% (Bm14), and 29.2-33.3% (Bm33). Schistosomiasis seroprevalence estimates among school-aged children ranged from 2.7-8.0% (SEA) and 10.9-15.6% (Sm25). Ov16 seropositivity was low and supported the localities' classification as nonendemic. The results suggested LF exposure, but discordance between antigens, challenges defining seropositivity thresholds, and the absence of programmatic guidance based on antibody serology alone for Wuchereria bancrofti indicate a need for remapping surveys to confirm transmission. Schistosomiasis antibody levels were high enough to warrant further mapping to guide treatment decisions. The lack of gold standards limited interpretation of results, particularly for LF, but in resource-challenged areas, integrated serological surveillance offers the possibility of efficient monitoring of exposure to multiple diseases.
Assuntos
Filariose Linfática , Oncocercose , Esquistossomose , Humanos , Sudão/epidemiologia , Filariose Linfática/epidemiologia , Filariose Linfática/imunologia , Filariose Linfática/sangue , Oncocercose/epidemiologia , Oncocercose/sangue , Oncocercose/imunologia , Criança , Adolescente , Masculino , Estudos Soroepidemiológicos , Feminino , Adulto , Pré-Escolar , Adulto Jovem , Esquistossomose/epidemiologia , Esquistossomose/sangue , Pessoa de Meia-Idade , Anticorpos Anti-Helmínticos/sangue , Lactente , Animais , IdosoRESUMO
OBJECTIVES: Circulating filarial antigen (Ag) is used by elimination programs to monitor lymphatic filariasis (LF) transmission; however, antifilarial antibodies (Ab) may be more sensitive than Ag for detecting LF. Our objectives were to describe Ab seroprevalence, identify risk factors for Ab seropositivity, investigate age-specific associations between Ag and Ab, and evaluate geographic clustering of seropositivity. METHODS: Community-based serosurveys of participants aged ≥5 years were conducted in 35 primary sampling units (PSUs). Ag-positivity was detected using Alere™ Filariasis Test Strips and Ab-seropositivity using multiplex bead assays. Seroprevalence was adjusted for study design. RESULTS: Of 3795 participants (range:5-90 years), adjusted prevalence for Ag, Bm14 Ab, Wb123 Ab, and Bm33 Ab were 3.7% (n=117), 20.3% (n=583), 32.2% (n=987), and 51.0% (n=1659), respectively. Male sex, older age, and residents of suspected hotspots had higher odds of seropositivity to all seromarkers. Seroprevalence was lower in 5-9-year-olds vs ≥10-year-olds (P<0.001). Clustering was significantly higher in households (intra-cluster correlation for Ag:0.45; Bm14 Ab:0.32; Bm33 Ab:0.31; Wb123 Ab:0.29) compared to PSUs or region. CONCLUSIONS: Abs enabled identification of risk factors for seropositivity and geographical clustering to inform targeted interventions for LF programmes. Further research is needed to define Ab thresholds for active versus past infection and elimination targets.
Assuntos
Anticorpos Anti-Helmínticos , Antígenos de Helmintos , Filariose Linfática , Humanos , Filariose Linfática/epidemiologia , Filariose Linfática/transmissão , Filariose Linfática/imunologia , Filariose Linfática/sangue , Masculino , Feminino , Adolescente , Criança , Estudos Soroepidemiológicos , Adulto , Idoso , Pré-Escolar , Pessoa de Meia-Idade , Fatores de Risco , Adulto Jovem , Idoso de 80 Anos ou mais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/sangue , Antígenos de Helmintos/imunologia , Animais , Prevalência , Wuchereria bancrofti/imunologia , Hotspot de DoençaRESUMO
This study describes the properties of colloidal gold nanoparticles (AuNPs) with sizes of 20, 30 and 40 nm, which were synthesized using citrate reduction or seeding-growth methods. Likewise, the conjugation of these AuNPs to mouse anti-human IgG(4) (MαHIgG(4)) was evaluated for an immunochromatographic (ICG) strip test to detect brugian filariasis. The morphology of the AuNPs was studied based on the degree of ellipticity (G) of the transmission electron microscopy images. The AuNPs produced using the seeding-growth method showed lower ellipticity (G ≤ 1.11) as compared with the AuNPs synthesized using the citrate reduction method (G ≤ 1.18). Zetasizer analysis showed that the AuNPs that were synthesized using the seeding-growth method were almost monodispersed with a lower polydispersity index (PDI; PDI≤0.079), as compared with the AuNPs synthesized using the citrate reduction method (PDI≤0.177). UV-visible spectroscopic analysis showed a red-shift of the absorbance spectra after the reaction with MαHIgG(4), which indicated that the AuNPs were successfully conjugated. The optimum concentration of the BmR1 recombinant antigen that was immobilized on the surface of the ICG strip on the test line was 1.0 mg ml(-1). When used with the ICG test strip assay and brugian filariasis serum samples, the conjugated AuNPs-MαHIgG(4) synthesized using the seeding-growth method had faster detection times, as compared with the AuNPs synthesized using the citrate reduction method. The 30 nm AuNPs-MαHIgG(4), with an optical density of 4 from the seeding-growth method, demonstrated the best performance for labelling ICG strips because it displayed the best sensitivity and the highest specificity when tested with serum samples from brugian filariasis patients and controls.
Assuntos
Técnicas Biossensoriais/instrumentação , Brugia/isolamento & purificação , Cromatografia de Afinidade/instrumentação , Filariose Linfática/sangue , Ouro/química , Nanopartículas Metálicas/química , Fitas Reagentes , Animais , Desenho de Equipamento , Análise de Falha de Equipamento , HumanosRESUMO
Immune responses to recombinant Brugia malayi pepsin inhibitor homolog (rBm-33) were investigated in patients with human lymphatic filariasis (microfilaremics (MF) and chronic pathology (CP)) along with endemic normals (EN). Flow cytometric analysis (24 h) revealed CD4(+) T cell activation in patients (MF and CP) compared to normals (EN), with increased expression of CD69 and diminished levels of CD62L and CD127. This was associated with an elevated expression of CD154 but not CD28 and CTLA4 in CP patients. However, Bm-33-induced cytokine expression profile (IL-1ß, IL-12, IL-8, IFN-γ, IL-10 and TGF-ß) did not exhibit any significant difference between normals and patients at the same time point. Although CD4(+) T cell activation was observed initially in filarial patients (24 h), lymphoproliferation studies (96 h) suggested diminished proliferation compared to normals, indicating functional inactivation in the former upon prolonged antigen exposure. This indicates that rBm-33 induces an early T cell activation in MF and CP patients followed by a decreased lymphoproliferation that might contribute to immune suppression in these individuals.
Assuntos
Antígenos de Helmintos/uso terapêutico , Brugia Malayi/imunologia , Filariose Linfática/tratamento farmacológico , Proteínas de Helminto/imunologia , Imunidade Celular/efeitos dos fármacos , Fatores Imunológicos/uso terapêutico , Linfócitos T/efeitos dos fármacos , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/imunologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/genética , Citocinas/metabolismo , DNA Complementar/biossíntese , DNA Complementar/genética , DNA Complementar/metabolismo , Filariose Linfática/sangue , Filariose Linfática/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Helminto/uso terapêutico , Humanos , Imunidade Celular/imunologia , Fatores Imunológicos/imunologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Proteínas Recombinantes/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Linfócitos T/imunologiaRESUMO
The endemicity status of lymphatic filariasis in Oman is uncertain, with only sporadic cases reported, mostly imported. Immunochromatographic card test surveys were carried out to assess the presence of circulating Wuchereria bancrofti antigenaemia as a marker for active infection in children from suspected high-risk areas of Oman (South Batinah and Dhofar). Lot quality assurance sampling surveys were carried out on a minimum of 250 secondary-school children aged 17-18 years in each of 8 districts from February 2004 to March 2004. All tested students were negative for circulating W. bancrofti antigen. Based on these findings as well as previous data, Oman may possibly be classified as a nonendemic country, with no evidence of indigenous lymphatic filariasis transmission.
Assuntos
Filariose Linfática/epidemiologia , Doenças Endêmicas/estatística & dados numéricos , Wuchereria bancrofti/imunologia , Adolescente , Animais , Antígenos de Helmintos/sangue , Biomarcadores/sangue , Certificação , Filariose Linfática/sangue , Filariose Linfática/diagnóstico , Filariose Linfática/imunologia , Feminino , Humanos , Amostragem para Garantia da Qualidade de Lotes , Masculino , Omã/epidemiologia , Vigilância da População , Kit de Reagentes para Diagnóstico , Características de Residência , Instituições Acadêmicas , Estudos Soroepidemiológicos , Estudantes/estatística & dados numéricosRESUMO
BACKGROUND: Lymphatic filariasis (LF) infection is generally diagnosed through parasitological identification of microfilariae (mf) in the blood. Although historically the most commonly used technique for counting mf is the thick blood smear based on 20 µl blood (TBS20), various other techniques and blood volumes have been applied. It is therefore a challenge to compare mf prevalence estimates from different LF-survey data. Our objective was to standardise microfilaraemia (mf) prevalence estimates to TBS20 as the reference diagnostic technique. METHODS: We first performed a systematic review to identify studies reporting on comparative mf prevalence data as measured by more than one diagnostic test, including TBS20, on the same study population. Associations between mf prevalences based on different diagnostic techniques were quantified in terms of odds ratios (OR, with TBS20 blood as reference), using a meta-regression model. RESULTS: We identified 606 articles matching our search strategy and included 14 in our analyses. The OR of the mf prevalences as measured by the more sensitive counting chamber technique (≥ 50 µl blood) was 2.90 (95% confidence interval (CI): 1.60-5.28). For membrane filtration (1 ml blood) the OR was 2.39 (95% CI: 1.62-3.53), Knott's technique it was 1.54 (95% CI: 0.72-3.29), and for TBS in ≥ 40 µl blood it was 1.37 (95% CI: 0.81-2.30). CONCLUSIONS: We provided transformation factors to standardise mf prevalence estimates as detected by different diagnostic techniques to mf prevalence estimates as measured by TBS20. This will facilitate the use and comparison of more datasets in meta-analyses and geographic mapping initiatives across countries and over time.
Assuntos
Sangue/parasitologia , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Filariose Linfática/sangue , Filariose Linfática/diagnóstico , Microfilárias/isolamento & purificação , Animais , Humanos , Razão de Chances , PrevalênciaRESUMO
BACKGROUND: Lymphatic Filariasis (LF) is one of the incapacitating and mosquito-borne sicknesses that on progression may prompt a few recognizable types of clutters like extreme lymphedema, hydrocele, and elephantiasis. METHODS: Antigenic preparations of B. malayi adult (BmA), S. cervi adult parasites and microfilariae (mf) total parasite extract were used to analyze the serological reactivity profile with human infectious sera collected from endemic areas of Bancroftian filariasis by performing Western blot and ELISA analysis. Sera from healthy human subjects were also included in the study to determine the variation incurred in the reactivity due to the filariasis infection. Gelelectrophoresis analysis of the crude-extract of BmA revealed seven protein bands while more than ten bands were recognized in S. cervi. RESULTS: our results represent a clear variation in protein patterns among the crude-antigens. ELISA results showed highest prevalence of IgG, IgM and IgG4 antibodies against all antigen preparations when recorded among microfilaraemic chronic infected patients. In both the antigenic preparations, the positive reactions were in the order of microfilaraemic>endemic normal>chronic>acute>nonendemic normal subjects. All sera of Mf+ patients were uniformly positive, while sera of both chronic and endemic normal subjects showed less reactivity. CONCLUSION: In the present study, we endeavoured to establish the extent of cross-reactivity of antigens derived from animal filarial parasites such as B. malayi and S. cervi with W. bancrofti filariasis sera of human patients. Besides, we further analyzed antibody-isotype profile of IgG, IgG4 and IgM in various human infection sera of bancroftian filarial subjects reactive to heterologous parasite antigens derived from adult worms of S. cervi from bovine and B. malayi from bovine and jirds.
Assuntos
Anticorpos Anti-Helmínticos , Filariose Linfática , Imunoglobulina G , Imunoglobulina M , Wuchereria bancrofti , Animais , Anticorpos Anti-Helmínticos/sangue , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/química , Antígenos de Helmintos/imunologia , Reações Cruzadas , Filariose Linfática/sangue , Filariose Linfática/imunologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina M/sangue , Imunoglobulina M/imunologia , Masculino , Wuchereria bancrofti/imunologia , Wuchereria bancrofti/metabolismoRESUMO
OBJECTIVES: To determine the current status of lymphatic filariasis (LF) in Korea. METHODS: Epidemiological surveys between 2002 and 2006 in areas where LF was previously endemic: remote and coastal areas Jeollanam-do, Gyeongsangnam-do, and Jeju-do, and inland Gyeongsangbuk-do. We took night blood smears from 9426 people for microfilaria testing and assayed samples from 3049 children (10- to 13-year-olds) and 1526 adults for Brugia malayi antibodies. RESULTS: We found two cases (0.01%) with low microfilaria density in their peripheral blood (1-2/20 mul) on the remote island of Jeollanam-do in the southern part of the Korean peninsula. These patients, males over 60-years old, were treated with diethylcarbamazine (DEC). None of the 4575 people surveyed tested positive for specific B. malayi antibodies. CONCLUSION: Lymphatic filariasis appears to have been eliminated in Korea.
Assuntos
Brugia Malayi/isolamento & purificação , Filariose Linfática/epidemiologia , Adolescente , Adulto , Animais , Anticorpos Anti-Helmínticos/sangue , Criança , Filariose Linfática/sangue , Filariose Linfática/prevenção & controle , Humanos , Coreia (Geográfico)/epidemiologia , Masculino , Microfilárias/isolamento & purificação , PrevalênciaRESUMO
OBJECTIVE: To identify possible associations between selected micronutrient status indicators (serum ferritin, retinol, beta-carotene, alpha-tocopherol, and the acute phase reactant alpha-1 antichymotrypsin) and infection with human immunodeficiency virus (HIV) or Wuchereria bancrofti, and to assess the effect of the antifilarial drug diethylcarbamazine (DEC) on the micronutrient status indicators in individuals positive for one or both of the two infections. METHODS: Serum concentrations of ferritin, retinol, beta-carotene, alpha-tocopherol and the acute phase reactant alpha-1 antichymotrypsin were examined in 59 individuals with HIV, W. bancrofti infection, or both, in Tanga Region, Tanzania, before and 12 weeks after treatment with DEC. RESULTS: HIV infection, but not W. bancrofti infection, was associated with higher serum ferritin concentrations and lower beta-carotene and alpha-tocopherol. Neither HIV infection nor W. bancrofti infection was associated with serum retinol. The four micronutrient status indicators and alpha-1 antichymotrypsin were generally lower at 12 weeks after treatment both in the DEC and the placebo groups. CONCLUSIONS: The negative association between HIV infection and the antioxidant vitamins beta-carotene and alpha-tocopherol may be due to infection-induced oxidative stress, whereas W. bancrofti infection seemed not to be associated with oxidative stress. The drop in antioxidant vitamin concentrations after treatment may be due to oxidative stress induced by HIV progression (HIV infected) and inflammation around dead adult worms and microfilariae (W. bancrofti infected) rather than to an effect of DEC.