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1.
Parasite Immunol ; 46(7): e13059, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-39039790

RESUMEN

Immunosuppressed patients, particularly transplant recipients, can develop severe strongyloidiasis. This study aimed to detect anti-Strongyloides IgG antibodies in a panel of sera from liver transplant patients. Two techniques were used: ELISA as the initial screening test and Western blotting as a confirmatory test. ELISA reactivity of 10.9% (32/294) was observed. The 40-30 kDa fraction was recognised in 93.7% (30/32) of the patients, resulting in a positivity rate of 10.2%. These data highlight the importance of serological screening for Strongyloides stercoralis infection in liver transplant recipients.


Asunto(s)
Anticuerpos Antihelmínticos , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G , Trasplante de Hígado , Strongyloides stercoralis , Estrongiloidiasis , Receptores de Trasplantes , Humanos , Estrongiloidiasis/diagnóstico , Estrongiloidiasis/inmunología , Estrongiloidiasis/sangre , Anticuerpos Antihelmínticos/sangre , Animales , Strongyloides stercoralis/inmunología , Inmunoglobulina G/sangre , Western Blotting , Masculino , Tamizaje Masivo/métodos , Persona de Mediana Edad , Femenino , Adulto , Enfermedades Desatendidas/diagnóstico , Enfermedades Desatendidas/epidemiología , Enfermedades Desatendidas/inmunología , Huésped Inmunocomprometido , Anciano
2.
Korean J Gastroenterol ; 83(6): 247-252, 2024 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-38918038

RESUMEN

Toxocariasis, a zoonotic infection transmitted by Toxocara canis (from dogs) and Toxocara cati (from cats) larvae, poses rare but severe risks to humans. We present a case of hepatic visceral larva migrans (VLM) caused by Toxocara canis in a 21-year-old male with a history of close contact with a pet dog. Initial symptoms and imaging findings mimicked a pyogenic liver abscess. The initial laboratory investigations revealed neutrophilia and elevated levels of IgE. Despite broad-spectrum antibiotics, persistent fever prompted further investigation. Subsequent serological testing for Toxocara antibodies and histopathological analysis of liver tissue demonstrating eosinophil infiltrates and Charcot-Leyden crystals led to a confirmed diagnosis of a liver abscess caused by Toxocara canis. Serological testing for Toxocara antibodies and histopathological analysis of liver tissue confirmed a Toxocara canis-induced liver abscess. Albendazole treatment yielded significant clinical improvement. This case highlights the necessity of considering toxocariasis in liver abscess differentials, particularly in high-seroprevalence regions like Vietnam. Relying solely on serological tests may be insufficient, emphasizing the need for corroborative evidence, including invasive procedures like liver biopsy, for accurate hepatic toxocariasis diagnosis.


Asunto(s)
Albendazol , Larva Migrans Visceral , Tomografía Computarizada por Rayos X , Toxocara canis , Humanos , Toxocara canis/aislamiento & purificación , Larva Migrans Visceral/diagnóstico , Larva Migrans Visceral/tratamiento farmacológico , Masculino , Animales , Adulto Joven , Albendazol/uso terapéutico , Perros , Hígado/parasitología , Hígado/patología , Anticuerpos Antihelmínticos/sangre , Ultrasonografía , Absceso Hepático/diagnóstico , Absceso Hepático/parasitología , Absceso Hepático/tratamiento farmacológico , Toxocariasis/diagnóstico , Toxocariasis/tratamiento farmacológico , Inmunoglobulina E/sangre , Antihelmínticos/uso terapéutico
3.
Mol Biochem Parasitol ; 259: 111621, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38705360

RESUMEN

Neurocysticercosis is the leading cause for acquired epilepsy worldwide, and it is caused by the larval stage of the parasite Taenia solium. Several proteins of this stage have been characterized and studied to understand the parasite-host interaction, however, the proteins from the early cysticercus stages (the postoncospheral form) have not yet been characterized. The study of the postoncospheral form proteins is important to understand the host-parasite relationship in the early stages of infection. The aim of this work was to identify postoncospheral form antigenic proteins using sera from neurocysticercosis patients. T. solium activated oncospheres were cultured in HCT-8 cells to obtain the postoncospheral form. Soluble total and excretory/secretory proteins were obtained from the postoncospheral form and were incubated with both pool sera and individual serum of neurocysticercosis positive human patients. Immunoblotting showed target antigenic proteins with apparent molecular weights of 23 kDa and 46-48 kDa. The 46-48 kDa antigen bands present in soluble total and excretory/secretory postoncospheral form proteins were analyzed by LC-MS/MS; proteins identified were: nuclear elongation factor 1 alpha, enolase, unnamed protein product/antigen diagnostic GP50, calcium binding protein calreticulin precursor and annexin. The postoncospheral form expresses proteins related to interaction with the host, some of these proteins are predicted to be exosomal proteins. In conclusion, postoncospheral proteins are consistent targets of the humoral immune response in human and may serve as targets for diagnosis and vaccines.


Asunto(s)
Antígenos Helmínticos , Proteínas del Helminto , Neurocisticercosis , Taenia solium , Taenia solium/inmunología , Taenia solium/genética , Antígenos Helmínticos/inmunología , Animales , Humanos , Neurocisticercosis/inmunología , Neurocisticercosis/parasitología , Neurocisticercosis/diagnóstico , Proteínas del Helminto/inmunología , Proteínas del Helminto/genética , Proteínas del Helminto/química , Espectrometría de Masas en Tándem , Anticuerpos Antihelmínticos/sangre , Anticuerpos Antihelmínticos/inmunología , Cromatografía Liquida , Peso Molecular
4.
Microbes Infect ; 26(5-6): 105364, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38777107

RESUMEN

Cystic echinococcosis, a zoonosis caused by cestodes belonging to the Echinococcus granulosus sensu lato (s.l.) genetic complex, affects humans and diverse livestock species. Although a veterinary vaccine exhibiting high levels of antibody-mediated protection has successfully reached the market, the large genetic diversity among parasite isolates and their particular host preferences, makes still necessary the search for novel vaccine candidates. Glutathione transferases (GSTs) constitute attractive targets for immunoprophylaxis due to their outstanding relevance in helminth detoxification processes, against both exogenous and endogenous stressors. Among the six GSTs known to be expressed in E. granulosus s.l., EgGST1 (Mu-class), EgGST2 (Sigma-class), and EgGST3 (a still non-classifiable isoenzyme), show the highest proteomic expression. Therefore, their recombinant forms -rEgGST1, rEgGST2 and rEgGST3- were herein analyzed regarding their potential to induce long-term antiparasite protection in mice. Only immunization with rEgGST1 induced long-lasting protection; and accordingly, rEgGST1-specific antibodies enhanced the parasite killing through both the classical activation of the host complement system and the antibody-dependent cellular cytotoxicity by macrophages. These results support further testing of rEgGST1 as a vaccine candidate in diverse hosts due to the broad expression of EgGST1 in different parasite stages and tissues.


Asunto(s)
Anticuerpos Antihelmínticos , Equinococosis , Echinococcus granulosus , Glutatión Transferasa , Echinococcus granulosus/inmunología , Echinococcus granulosus/genética , Echinococcus granulosus/enzimología , Animales , Equinococosis/prevención & control , Equinococosis/inmunología , Equinococosis/parasitología , Glutatión Transferasa/inmunología , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Ratones , Anticuerpos Antihelmínticos/inmunología , Formación de Anticuerpos/inmunología , Femenino , Ratones Endogámicos BALB C , Inmunización , Proteínas del Helminto/inmunología , Proteínas del Helminto/genética
5.
Parasit Vectors ; 17(1): 206, 2024 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-38715089

RESUMEN

BACKGROUND: Opisthorchiasis and cholangiocarcinoma (CCA) continue to be public health concerns in many Southeast Asian countries. Although the prevalence of opisthorchiasis is declining, reported cases tend to have a light-intensity infection. Therefore, early detection by using sensitive methods is necessary. Several sensitive methods have been developed to detect opisthorchiasis. The immunological detection of antigenic proteins has been proposed as a sensitive method for examining opisthorchiasis. METHODS: The Opisthorchis viverrini antigenic proteins, including cathepsin B (OvCB), asparaginyl endopeptidase (OvAEP), and cathepsin F (OvCF), were used to construct multi-antigenic proteins. The protein sequences of OvCB, OvAEP, and OvCF, with a high probability of B cell epitopes, were selected using BepiPred 1.0 and the IEDB Analysis Resource. These protein fragments were combined to form OvCB_OvAEP_OvCF recombinant DNA, which was then used to produce a recombinant protein in Escherichia coli strain BL21(DE3). The potency of the recombinant protein as a diagnostic target for opisthorchiasis was assessed using immunoblotting and compared with that of the gold standard method, the modified formalin-ether concentration technique. RESULTS: The recombinant OvCB_OvAEP_OvCF protein showed strong reactivity with total immunoglobulin G (IgG) antibodies against light-intensity O. viverrini infections in the endemic areas. Consequently, a high sensitivity (100%) for diagnosing opisthorchiasis was reported. However, cross-reactivity with sera from other helminth and protozoan infections (including taeniasis, strongyloidiasis, giardiasis, E. coli infection, enterobiasis, and mixed infection of Echinostome spp. and Taenia spp.) and no reactivity with sera from patients with non-parasitic infections led to a reduced specificity of 78.4%. In addition, the false negative rate (FNR), false positive rate (FPR), positive predictive value (PPV), negative predictive value (NPV), and diagnostic accuracy were 0%, 21.6%, 81.4%, 100%, and 88.9%, respectively. CONCLUSIONS: The high sensitivity of the recombinant OvCB_OvAEP_OvCF protein in detecting opisthorchiasis demonstrates its potential as an opisthorchiasis screening target. Nonetheless, research on reducing cross-reactivity should be undertaken by detecting other antibodies in other sample types, such as saliva, urine, and feces.


Asunto(s)
Antígenos Helmínticos , Opistorquiasis , Opisthorchis , Opistorquiasis/diagnóstico , Opisthorchis/inmunología , Opisthorchis/genética , Animales , Antígenos Helmínticos/genética , Antígenos Helmínticos/inmunología , Humanos , Anticuerpos Antihelmínticos/sangre , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/genética , Sensibilidad y Especificidad , Proteínas del Helminto/inmunología , Proteínas del Helminto/genética , Epítopos/inmunología , Epítopos/genética , Catepsina B/genética , Catepsina B/inmunología , Escherichia coli/genética , Cisteína Endopeptidasas
6.
BMC Vet Res ; 20(1): 182, 2024 May 08.
Artículo en Inglés | MEDLINE | ID: mdl-38720329

RESUMEN

BACKGROUND: Porcine cysticercosis, a serious zoonotic parasitic disease, is caused by the larvae of Taenia solium and has been acknowledged by the World Organization for Animal Health. The current detection methods of Cysticercus cellulosae cannot meet the needs of large-scale and rapid detection in the field. We hypothesized that the immunofluorescence chromatography test strip (ICS) for detecting Cysticercus cellulosae, according to optimization of a series of reaction systems was conducted, and sensitivity, specificity, and stability testing, and was finally compared with ELISA. This method utilizes Eu3+-labeled time-resolved fluorescent microspheres (TRFM) coupled with TSOL18 antigen to detect TSOL18 antibodies in infected pig sera. RESULTS: ICS and autopsy have highly consistent diagnostic results (n = 133), as determined by Cohen's κ analysis (κ = 0.925). And the results showed that the proposed ICS are high sensitivity (0.9459) with specificity (0.9792). The ICS was unable to detect positive samples of other parasites. It can be stored for at least six months at 4℃. CONCLUSIONS: In summary, we established a TRFM-ICS method with higher sensitivity and specificity than indirect ELISA. Results obtained from serum samples can be read within 10 min, indicating a rapid, user-friendly test suitable for large-scale field detection.


Asunto(s)
Anticuerpos Antihelmínticos , Antígenos Helmínticos , Cisticercosis , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Sensibilidad y Especificidad , Enfermedades de los Porcinos , Animales , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/parasitología , Enfermedades de los Porcinos/sangre , Cisticercosis/veterinaria , Cisticercosis/diagnóstico , Anticuerpos Antihelmínticos/sangre , Antígenos Helmínticos/sangre , Antígenos Helmínticos/inmunología , Técnica del Anticuerpo Fluorescente/veterinaria , Técnica del Anticuerpo Fluorescente/métodos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Ensayo de Inmunoadsorción Enzimática/métodos , Cysticercus/inmunología , Taenia solium/inmunología
7.
Parasit Vectors ; 17(1): 172, 2024 Apr 02.
Artículo en Inglés | MEDLINE | ID: mdl-38566124

RESUMEN

BACKGROUND: Antigen detection in Taenia solium cysticercosis confirms viable infection in the intermediate host (either pig or human). The reference B158/B60 monoclonal antibody (mAb)-based Ag-enzyme-linked immunosorbent assay (ELISA) has acceptable levels of sensitivity and specificity in human neurocysticercosis with multiple brain cysts, although its sensitivity is lower in cases with single brain cysts, whereas in porcine cysticercosis the assay specificity is affected by its frequent cross-reaction with Taenia hydatigena, another common cestode found in pigs. Our group has produced 21 anti-T. solium mAbs reacting against antigens of the whole cyst, vesicular fluid, and secretory/excretory products, identifying TsW8/TsW5 as the most promising pair of mAbs for an Ag-ELISA. METHODS: We report the use of the TsW8/TsW5 Ag-ELISA to measure cysticercus antigen levels [expressed as optical density (OD) values] in two panels of sera collected from day 0 (baseline) to day 90 postinfection (PI) from pigs experimentally infected with T. solium (n = 26) and T. hydatigena (n = 12). At baseline and on days 28 and 90 PI, we used Bland-Altman (BA) analysis and Lin's concordance correlation coefficients (CCC) to determine the concordance between the TsW8/TsW5 and the B158/B60 Ag-ELISA. RESULTS: The TsW8/TsW5 Ag-ELISA was able to efficiently measure circulating antigen levels in T. solium-infected pigs, similar to that obtained with the B158/B60 Ag-ELISA. Almost all paired log-OD differences between assays were within the limits of agreement (LoA) in the BA analysis at baseline and on days 28 and 90 PI (92.3%, 100%, and 100%, respectively), and a high concordance of log-ODs between assays was also found (Lin's CCC: 0.69, 0.92, and 0.96, respectively, all P < 0.001). In pigs infected with T. hydatigena, almost all paired log-OD differences were within the LoA in the BA analysis, whereas the concordance of log-ODs between assays was low at baseline (Lin's CCC: 0.24) but increased on days 28 and 90 PI (Lins' CCC: 0.88 and 0.98, P < 0.001). CONCLUSIONS/SIGNIFICANCE: The TsW8/TsW5 Ag-ELISA recognizes antigens in pigs with T. solium cysticercosis and is highly concordant with the B158/B60 Ag-ELISA. However, its diagnostic use is hampered by cross-reactions with T. hydatigena, as in other mAb-based Ag-ELISAs.


Asunto(s)
Cisticercosis , Quistes , Enfermedades de los Porcinos , Taenia solium , Taenia , Animales , Humanos , Porcinos , Cysticercus , Anticuerpos Monoclonales , Enfermedades de los Porcinos/diagnóstico , Cisticercosis/veterinaria , Ensayo de Inmunoadsorción Enzimática/veterinaria , Antígenos , Antígenos Helmínticos , Anticuerpos Antihelmínticos
8.
Acta Trop ; 254: 107208, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38621620

RESUMEN

The study aimed to elicit protective immune responses against murine schistosomiasis mansoni at the parasite lung- and liver stage. Two peptides showing amino acid sequence similarity to gut cysteine peptidases, which induce strong memory immune effectors in the liver, were combined with a peptide based on S. mansoni thioredoxin peroxidase (TPX), a prominent lung-stage schistosomula excretory-secretory product, and alum as adjuvant. Only one of the 2 cysteine peptidases-based peptides in a multiple antigenic peptide construct (MAP-3 and MAP-4) appeared to adjuvant protective immune responses induced by the TPX peptide in a MAP form. Production of TPX MAP-specific IgG1 serum antibodies, and increase in lung interleukin-1 (IL-1), uric acid, and reactive oxygen species (ROS) content were associated with significant (P < 0.05) 50 % reduction in recovery of lung-stage larvae. Increase in lung triglycerides and cholesterol levels appeared to provide the surviving worms with nutrients necessary for a stout double lipid bilayer barrier at the parasite-host interface. Surviving worms-released products elicited memory responses to the MAP-3 immunogen, including production of specific IgG1 antibodies and increase in liver IL-33 and ROS. Reduction in challenge worm burden recorded 45 days post infection did not exceed 48 % associated with no differences in parasite egg counts in the host liver and small intestine compared to unimmunized adjuvant control mice. Alum adjuvant assisted the second peptide, MAP-4, in production of IgG1, IgG2a, IgG2b and IgA specific antibodies and increase in liver ROS, but with no protective potential, raising doubt about the necessity of adjuvant addition. Accordingly, different vaccine formulas containing TPX MAP and 1, 2 or 3 cysteine peptidases-derived peptides with or without alum were used to immunize parallel groups of mice. Compared to unimmunized control mice, significant (P < 0.05 to < 0.005) 22 to 54 % reduction in worm burden was recorded in the different groups associated with insignificant changes in parasite egg output. The results together indicated that a schistosomiasis vaccine able to entirely prevent disease and halt its transmission still remains elusive.


Asunto(s)
Adyuvantes Inmunológicos , Anticuerpos Antihelmínticos , Inmunoglobulina G , Hígado , Pulmón , Schistosoma mansoni , Esquistosomiasis mansoni , Vacunas de Subunidad , Animales , Schistosoma mansoni/inmunología , Esquistosomiasis mansoni/prevención & control , Esquistosomiasis mansoni/inmunología , Esquistosomiasis mansoni/parasitología , Pulmón/parasitología , Pulmón/inmunología , Ratones , Anticuerpos Antihelmínticos/inmunología , Anticuerpos Antihelmínticos/sangre , Hígado/parasitología , Hígado/inmunología , Inmunoglobulina G/sangre , Adyuvantes Inmunológicos/administración & dosificación , Vacunas de Subunidad/inmunología , Vacunas de Subunidad/administración & dosificación , Femenino , Antígenos Helmínticos/inmunología , Modelos Animales de Enfermedad , Compuestos de Alumbre/administración & dosificación , Ratones Endogámicos BALB C , Vacunas de Subunidades Proteicas
9.
Parasitol Res ; 123(4): 196, 2024 Apr 25.
Artículo en Inglés | MEDLINE | ID: mdl-38662084

RESUMEN

Many pathogens are related to carcinogenesis. Chronic inflammation, as a result of persistent infection, leads to DNA damage, higher expression of oncogenes, decreased apoptosis and immunosuppression, which are some of the reasons for cancer induction. Among parasites, Schistosoma, Opistorchis and Clonorchis are recognised as infectious agents which contribute to cancer. A relationship between Anisakis and cancer was hypothesised because cellular responses to Anisakis products could result in inflammation and DNA damage. Previous research has shown a decrease in CD8+ γδ T-cells and an increase in αß and γδ T-cell apoptosis in colon cancer (CC) samples. Ninety-two CC patients and 60 healthy subjects were recruited. γδ and αß T-cells were analysed, and their apoptosis was evaluated. Anti-Anisakis antibodies were tested in sera from CC patients and controls. Anti-Anisakis IgG, IgM, IgA and IgE antibodies were significantly higher in CC patients. A significant increase in anti-Anisakis IgA levels was observed in patients with angiolymphatic invasion. The number of all γδ T-cells, as well as CD3+ CD4+ αß T-cells, was significantly lower in CC patients. The apoptosis of all T-cells was significantly increased in patients with CC. We observed a significantly higher percentage of anti-Anisakis IgE positive patients having a deficit of CD3+ γδ T-cells. Our results suggest a relationship between Anisakis and CC.


Asunto(s)
Anisakis , Anticuerpos Antihelmínticos , Neoplasias del Colon , Humanos , Masculino , Persona de Mediana Edad , Anticuerpos Antihelmínticos/sangre , Anticuerpos Antihelmínticos/inmunología , Femenino , Neoplasias del Colon/inmunología , Neoplasias del Colon/parasitología , Anciano , Animales , Anisakis/inmunología , Adulto , Apoptosis , Anciano de 80 o más Años , Subgrupos de Linfocitos T/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Linfocitos T/inmunología
10.
Acta Parasitol ; 69(2): 1122-1131, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38551763

RESUMEN

PURPOSE: Cystic echinococcosis (CE) is caused by the larval form of Echinococcus granulosus. Clinical, radiologic, pathologic, and serologic findings should be evaluated together for the diagnosis of CE. The sensitivity and specificity oalf serologic tests may vary depending on the method used. In this study, we aimed to detect IgG antibodies specific to E. granulosus using indirect hemagglutination assay (IHA), enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibodies (IFA) and western blot (WB) tests. METHODS: In our study, the serum samples of 74 patients sent to our laboratory with suspicion of CE were studied using two different commercial IHA tests, ELISA, IFA and WB test. The test results were evaluated along with radiological findings and histopathological examinations, the latter being the gold standard. RESULTS: Of all the patients, 51 (69%) were female and 23 (31%) were male. There was a statistically significant difference between males and females (χ2 = 9.7, p = 0.002). Out of 74 patients, positivity rates for Siemens IHA, Fumouze IHA, ELISA, IFA and WB test were positive as 33 (44.6%), 35 (47.3%), 43 (58.1%), 42 (56.7%) and 38 (51.3%), respectively. The sensitivity and specificity of the tests were as follows: 66.67 and 2.31% for Siemens IHA; 70.83% and 96.15% for Fumouze IHA; 85.42%, and 88.46% for ELISA; 83.33% and 88.46% for IFA; 72.92% and 88.46% for WB test. CONCLUSION: There were statistically significant differences in between all five methods (p < 0,001). While the tests with the highest specificity was Fumouze IHA, the test with the highest sensitivity was the ELISA test. It was concluded that IHA and ELISA tests were more practical in practice because of their greater applicability.


Asunto(s)
Anticuerpos Antihelmínticos , Equinococosis , Echinococcus granulosus , Ensayo de Inmunoadsorción Enzimática , Pruebas de Hemaglutinación , Inmunoglobulina G , Sensibilidad y Especificidad , Pruebas Serológicas , Humanos , Equinococosis/diagnóstico , Equinococosis/sangre , Femenino , Masculino , Echinococcus granulosus/inmunología , Animales , Anticuerpos Antihelmínticos/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Pruebas Serológicas/métodos , Adulto , Persona de Mediana Edad , Inmunoglobulina G/sangre , Western Blotting , Técnica del Anticuerpo Fluorescente Indirecta , Adulto Joven , Adolescente , Anciano , Niño
11.
Acta Trop ; 254: 107199, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38552996

RESUMEN

BACKGROUND: Opisthorchis viverrini infection is a significant health problem in several countries, especially Southeast Asia. The infection causes acute gastro-hepatic symptoms and also long-term infection leading to carcinogenesis of an aggressive bile duct cancer (cholangiocarcinoma; CCA). Hence, the early diagnosis of O. viverrini infection could be the way out of this situation. Still, stool examination by microscopic-based methods, the current diagnostic procedure is restricted by low parasite egg numbers in the specimen and unprofessional laboratorians. The immunological procedure provides a better chance for diagnosis of the infection. Hence, this study aims to produce single-chain variable fragment (scFv) antibodies for use as a diagnostic tool for O. viverrini infection. METHODS: This study uses phage display technologies to develop the scFv antibodies against O. viverrini cathepsin F (OvCatF). The OvCatF-deduced amino acid sequence was analyzed and predicted for B-cell epitopes used for short peptide synthesis. The synthetic peptides were used to screen the phage library simultaneously with OvCatF recombinant protein (rOvCatF). The potentiated phages were collected, rescued, and reassembled in XL1-blue Escherichia coli (E. coli) as a propagative host. The positive clones of phagemids were isolated, and the single-chain variable (scFv) fragments were sequenced, computationally predicted, and molecular docked. The complete scFv fragments were digested from the phagemid, subcloned into the pOPE101 expression vector, and expressed in XL1-blue E. coli. Indirect ELISA and Western analysis were used to verify the detection efficiency. RESULTS: The scFv phages specific to OvCatF were successfully isolated, subcloned, and produced as a recombinant protein. The recombinant scFv antibodies were purified and refolded to make functional scFv. The evaluation of specific recognition of the particular epitopes and detection limit results by both computational and laboratory performances demonstrated that all three recombinant scFv antibodies against OvCatF could bind specifically to rOvCatF, and the lowest detection concentration in this study was only one hundred nanograms. CONCLUSION: Our produced scFv antibodies will be the potential candidates for developing a practical diagnostic procedure for O. viverrini infection in humans in the future.


Asunto(s)
Opisthorchis , Anticuerpos de Cadena Única , Anticuerpos de Cadena Única/inmunología , Anticuerpos de Cadena Única/genética , Opisthorchis/inmunología , Animales , Anticuerpos Antihelmínticos/inmunología , Opistorquiasis/inmunología , Catepsinas/inmunología , Epítopos/inmunología , Humanos , Proteínas Recombinantes/inmunología , Técnicas de Visualización de Superficie Celular , Epítopos de Linfocito B/inmunología , Ensayo de Inmunoadsorción Enzimática , Biblioteca de Péptidos
12.
Trans R Soc Trop Med Hyg ; 118(8): 514-519, 2024 Aug 05.
Artículo en Inglés | MEDLINE | ID: mdl-38401052

RESUMEN

BACKGROUND: Control interventions recommended by the World Health Organization have successfully resulted in low-intensity schistosomiasis transmission areas. To achieve elimination of transmission, new diagnostic screening tools are needed to overcome less than adequate sensitivity of the currently used Kato-Katz faecal thick smear method. Ideally, in-house serological tests should be avoided due to not having a continuous supply of kits as would be necessary for large population studies. Quality assurance provided by manufacturers and proper performance evaluations are also needed. We evaluated the accuracy of two commercially available serology tests as screening methods for detecting light schistosomiasis infections. METHODS: Serum samples were collected in 2015 from individuals living in a low-endemicity locality in northeastern Brazil and deposited in a biorepository. We evaluated immunoglobulin G (IgG) and IgM enzyme-linked immunosorbent assays (ELISAs) and an immunochromatographic test (ICT). The Helmintex method was used to define true-positive samples. RESULTS: Overall sensitivity was close to 90% for both the IgG ELISA and ICT, yet specificity was 28% and 18%, respectively. For the IgM ELISA, the values were estimated to be 55% and 43%, respectively. CONCLUSIONS: Poor specificity and positive predictive values prevent these tests from being recommended for screening populations in low-intensity schistosomiasis-endemic areas.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G , Inmunoglobulina M , Sensibilidad y Especificidad , Humanos , Brasil/epidemiología , Inmunoglobulina G/sangre , Femenino , Masculino , Inmunoglobulina M/sangre , Adulto , Persona de Mediana Edad , Adolescente , Esquistosomiasis/diagnóstico , Esquistosomiasis/epidemiología , Anticuerpos Antihelmínticos/sangre , Tamizaje Masivo/métodos , Niño , Adulto Joven , Animales , Anciano , Cromatografía de Afinidad
13.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 35(6): 633-637, 2024 Jan 30.
Artículo en Chino | MEDLINE | ID: mdl-38413025

RESUMEN

OBJECTIVE: To investigate the prevalence and epidemiological characteristics of taeniasis and cysticercosis among residents in Tibetan agricultural areas of Sichuan Province, so as to provide insights for the prevention and control of taeniasis and cysticercosis. METHODS: From 2016 to 2022, Kangding City, Daocheng County, Derong County, Ruoergai County and Muli Tibetan Autonomous County were sampled from Tibetan agricultural areas of Ganzi Tibetan Autonomous Prefecture, Aba Tibetan and Qiang Autonomous Prefecture and Liangshan Yi Autonomous Prefecture in Sichuan Province, and 1 to 6 townships were sampled from each county (district), followed by 4 to 7 villages sampled from each township. Primary school children were sampled using a cluster sampling method, and permanent residents at ages of over 16 years were randomly sampled from each village. Participants' demographics, history of tapeworm excretion during the past year and clinical symptoms and signs of cysticercosis were collected through questionnaire surveys, and participants' stool and venous blood samples were collected. Taenia eggs were detected in stool samples using the direct smear method, and deworming was performed among taeniasis patients with areca nut-squash seeds. The tapeworm species were identified using a multiplex PCR assay, and serum specific IgG antibody against cysticercus was detected using enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 5 249 respondents participated in the questionnaire survey, including 603 respondents (11.5%) with a self-reported history of proglottids secretion during the past year. A total of 3 976 residents were subjected to stool examinations, and the detection of Taenia eggs was 6.5%. Of 258 participants undergoing deworming, there were 403 cases (94.2%) with excretions of Taenia worms or proglottids. The mean prevalence of taeniasis was 10.9% (439/4 043), and there were gender-, age- and region-specific prevalence rates of taeniasis (χ2 = 36.73, 126.31 and 163.41, all P values < 0.05). Multiplex PCR assays detected 41 cases with T. solium infections (12.5%), 197 cases with T. saginata infections (59.9%) and 91 cases with T. asiatica infections (27.6%) among 329 patients undergoing deworming, and there were region-specific prevalence rates of T. solium, T. saginata and T. asiatica infections (χ2 = 45.39, P < 0.05). In addition, the sero-prevalence of anti-cysticercus IgG antibody was 7.0% (345/4 933), and there were age- and region-specific sero-prevalence rates of anti-cysticercus IgG antibody (χ2 = 13.49 and 51.76, both P values < 0.05). CONCLUSIONS: Multiple Taenia species are prevalent in Tibetan agricultural areas of Sichuan Province and the sero-prevalence of anti-cysticercus antibody is high among residents. Monitoring and control of taeniasis and cysticercosis should be strengthened.


Asunto(s)
Cisticercosis , Taenia solium , Teniasis , Niño , Animales , Humanos , Cysticercus , Tibet/epidemiología , Prevalencia , Teniasis/epidemiología , Cisticercosis/epidemiología , Cisticercosis/diagnóstico , Anticuerpos Antihelmínticos , Inmunoglobulina G
14.
Parasit Vectors ; 17(1): 9, 2024 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-38178167

RESUMEN

BACKGROUND: The excretory/secretory (ES) antigen of Trichinella spiralis muscle larvae (ML) is currently the most widely used diagnostic antigen to detect T. spiralis infection. However, this antigen has certain drawbacks, such as a complicated ES antigen preparation process and lower sensitivity during the early phase of infection. The aim of this study was to investigate the features of a novel T. spiralis trypsin (TsTryp) and evaluate its potential diagnostic value for trichinellosis. METHODS: The TsTryp gene was cloned and recombinant TsTryp (rTsTryp) expressed. Western blotting and an enzyme-linked immunosorbent assay (ELISA) were performed to confirm the antigenicity of rTsTryp. The expression pattern and distribution signature of TsTryp at various life-cycle stages of T. spiralis were analyzed by quantitative PCR, western blotting and the immunofluorescence test. An ELISA with rTsTryp and ML ES antigens was used to detect immunoglobulins G and M (IgG, IgM) in serum samples of infected mice, swine and humans. The seropositive results were further confirmed by western blot with rTsTryp and ML ES antigens. RESULTS: TsTryp expression was observed in diverse T. spiralis life-cycle phases, with particularly high expression in the early developmental phase (intestinal infectious larvae and adults), with distribution observed mainly at the nematode outer cuticle and stichosome. rTsTryp was identified by T. spiralis-infected mouse sera and anti-rTsTryp sera. Natural TsTryp protease was detected in somatic soluble and ES antigens of the nematode. In mice infected with 200 T. spiralis ML, serum-specific IgG was first detected by rTsTryp-ELISA at 8 days post-infection (dpi), reaching 100% positivity at 12 dpi, and first detected by ES-ELISA at 10 dpi, reaching 100% positivity at 14 dpi. Specific IgG was detected by rTsTryp 2 days earlier than by ES antigens. When specific IgG was determined in serum samples from trichinellosis patients, the sensitivity of rTsTryp-ELISA and ES antigens-ELISA was 98.1% (51/52 samples) and 94.2% (49/52 samples), respectively (P = 0.308), but the specificity of rTsTryp was significantly higher than that of ES antigens (98.7% vs. 95.4%; P = 0.030). Additionally, rTsTryp conferred a lower cross-reaction, with only three serum samples in total testing positive from 11 clonorchiasis, 20 cysticercosis and 24 echinococcosis patients (1 sample from each patient group). CONCLUSIONS: TsTryp was shown to be an early and highly expressed antigen at intestinal T. spiralis stages, indicating that rTsTryp represents a valuable diagnostic antigen for the serodiagnosis of early Trichinella infection.


Asunto(s)
Trichinella spiralis , Triquinelosis , Adulto , Humanos , Porcinos , Ratones , Animales , Triquinelosis/diagnóstico , Tripsina , Antígenos Helmínticos , Proteínas del Helminto , Ensayo de Inmunoadsorción Enzimática/métodos , Larva/fisiología , Estadios del Ciclo de Vida , Pruebas Serológicas , Inmunoglobulina G , Anticuerpos Antihelmínticos
15.
PLoS Negl Trop Dis ; 17(12): e0011776, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38039267

RESUMEN

BACKGROUND: CLA (conjugated linoleic acid)-mediated activation of the schistosome tegument-associated sphingomyelinase and consequent disruption of the outer membrane might allow host antibodies to access the apical membrane antigens. Here, we investigated a novel approach to enhance specific antibody delivery to concealed surface membrane antigens of Schistosoma mansoni utilising antibody-conjugated-CLA nanomicelle technology. METHODOLOGY/PRINCIPAL FINDINGS: We invented and characterised an amphiphilic CLA-loaded whey protein co-polymer (CLA-W) as an IV injectable protein nanocarrier. Rabbit anti-Schistosoma mansoni infection (anti-SmI) and anti-Schistosoma mansoni alkaline phosphatase specific IgG antibodies were purified from rabbit sera and conjugated to the surface of CLA-W co-polymer to form antibody-conjugated-CLA-W nanomicelles (Ab-CLA-W). We investigated the schistosomicidal effects of CLA-W and Ab-CLA-W in a mouse model of Schistosoma mansoni against early and late stages of infection. Results showed that conjugation of nanomicelles with antibodies, namely anti-SmI, significantly enhanced the micelles' schistosomicidal and anti-pathology activities at both the schistosomula and adult worm stages of the infection resulting in 64.6%-89.9% reductions in worm number; 72.5-94% and 66.4-85.2% reductions in hepatic eggs and granulomas, respectively. Treatment induced overall improvement in liver histopathology, reducing granuloma size and fibrosis and significantly affecting egg viability. Indirect immunofluorescence confirmed CLA-W-mediated antigen exposure on the worm surface. Electron microscopy revealed extensive ultrastructural damage in worm tegument induced by anti-SmI-CLA-W. CONCLUSION/SIGNIFICANCE: The novel antibody-targeted nano-sized CLA delivery system offers great promise for treatment of Schistosoma mansoni infection and control of its transmission. Our in vivo observations confirm an immune-mediated enhanced effect of the schistosomicidal action of CLA and hints at the prospect of nanotechnology-based immunotherapy, not only for schistosomiasis, but also for other parasitic infections in which chemotherapy has been shown to be immune-dependent. The results propose that the immunodominant reactivity of the anti-SmI serum, Schistosoma mansoni fructose biphosphate aldolase, SmFBPA, merits serious attention as a therapeutic and vaccine candidate.


Asunto(s)
Esquistosomiasis mansoni , Esquistosomiasis , Esquistosomicidas , Ratones , Animales , Conejos , Esquistosomiasis mansoni/parasitología , Schistosoma mansoni , Esquistosomiasis/tratamiento farmacológico , Anticuerpos Antihelmínticos , Esquistosomicidas/farmacología , Polímeros/farmacología , Polímeros/uso terapéutico , Antígenos Helmínticos
16.
Front Cell Infect Microbiol ; 13: 1306567, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38145042

RESUMEN

Human trichinellosis is a parasitic infection caused by roundworms belonging to the genus Trichinella, especially Trichinella spiralis. Early and accurate clinical diagnoses of trichinellosis are required for efficacious prognosis and treatment. Current drug therapies are limited by antiparasitic resistance, poor absorption, and an inability to kill the encapsulating muscle-stage larvae. Therefore, reliable biomarkers and drug targets for novel diagnostic approaches and anthelmintic drugs are required. In this study, metabolite profiles of T. spiralis adult worms and muscle larvae were obtained using mass spectrometry-based metabolomics. In addition, metabolite-based biomarkers of T. spiralis excretory-secretory products and their related metabolic pathways were characterized. The metabolic profiling identified major, related metabolic pathways involving adenosine monophosphate (AMP)-dependent synthetase/ligase and glycolysis/gluconeogenesis in T. spiralis adult worms and muscle larvae, respectively. These pathways are potential drug targets for the treatment of the intestinal and muscular phases of infection. The metabolome of larva excretory-secretory products was characterized, with amino acid permease and carbohydrate kinase being identified as key metabolic pathways. Among six metabolites, decanoyl-l-carnitine and 2,3-dinor-6-keto prostaglandin F1α-d9 were identified as potential metabolite-based biomarkers that might be related to the host inflammatory processes. In summary, this study compared the relationships between the metabolic profiles of two T. spiralis growth stages. Importantly, the main metabolites and metabolic pathways identified may aid the development of novel clinical diagnostics and therapeutics for human trichinellosis and other related helminthic infections.


Asunto(s)
Trichinella spiralis , Triquinelosis , Animales , Humanos , Triquinelosis/diagnóstico , Antígenos Helmínticos , Proteínas del Helminto/metabolismo , Larva/fisiología , Ensayo de Inmunoadsorción Enzimática , Anticuerpos Antihelmínticos , Músculos , Biomarcadores
17.
J Clin Microbiol ; 61(12): e0076023, 2023 12 19.
Artículo en Inglés | MEDLINE | ID: mdl-37966225

RESUMEN

Neurocysticercosis (NCC) is the most common helminthic infection of the human central nervous system. The antibody detection assay of choice is the enzyme-linked immunoelectrotransfer blot assay using lentil-lectin purified parasite antigens (LLGP-EITB, Western blot), an immunoassay with exceptional performance in clinical samples. However, its use is mainly restricted to a few research laboratories because the assay is labor-intensive and requires sophisticated equipment, expertise, and large amounts of parasite material for preparation of reagents. We report a new immunoprint assay (MAPIA) that overcomes most of these barriers. We initially compared the performance of five different antigen combinations in a subset of defined samples in the MAPIA format. After selecting the best-performing assay format (a combination of rGP50 + rT24H + sTs14 antigens), 148 archived serum samples were tested, including 40 from individuals with parenchymal NCC, 40 with subarachnoid NCC, and 68 healthy controls with no evidence of neurologic disease. MAPIA using three antigens (rGP50 + rT24H + sTs14) was highly sensitive and specific for detecting antibodies in NCC. It detected 39 out of 40 (97.5%) parenchymal NCC cases and 40/40 (100%) subarachnoid cases and was negative in 67 out of 68 (98.53%) negative samples. MAPIA using three recombinant and synthetic antigens is a simple and economical tool with a performance equivalent to the LLGP-EITB assay for the detection of specific antibodies to NCC. The MAPIA overcomes existing barriers to adoption of the EITG LLGP and is a candidate for worldwide use.


Asunto(s)
Neurocisticercosis , Taenia solium , Animales , Humanos , Neurocisticercosis/diagnóstico , Neurocisticercosis/parasitología , Perú , Antígenos Helmínticos , Sensibilidad y Especificidad , Inmunoensayo , Ensayo de Inmunoadsorción Enzimática , Anticuerpos Antihelmínticos
18.
Emerg Infect Dis ; 29(7): 1340-1348, 2023 07.
Artículo en Inglés | MEDLINE | ID: mdl-37347506

RESUMEN

Neurocysticercosis prevalence estimates often are based on serosurveys. However, assessments of Taenia solium seropositivity durability in patients with various neurocysticercosis types are lacking. We optimized a triplex serologic ELISA by using synthetic GP50, T24H, and Ts18var3 antigens for T. solium. We used that assay to test sequential serologic responses over several years after neurocysticercosis cure in 46 patients, 9 each with parenchymal or ventricular neurocysticercosis and 28 with subarachnoid disease. Triplex results were concordant with 98% of positive and 100% of negative enzyme-linked immunoelectrotransfer blots. Eight years after neurocysticercosis cure, 11.1% of patients with parenchymal, 47.3% with subarachnoid, and 41.7% with ventricular disease were still seropositive. Median time to seroreversion after cure in this cohort in a T. solium nonendemic area was 2 years for parenchymal disease, 4 years for ventricular disease, and 8 years for subarachnoid disease. Our findings can inform epidemiologic models that rely on serosurveys to estimate disease burden.


Asunto(s)
Neurocisticercosis , Taenia solium , Taenia , Animales , Humanos , Neurocisticercosis/diagnóstico , Neurocisticercosis/epidemiología , Antígenos Helmínticos , Anticuerpos Antihelmínticos , Ensayo de Inmunoadsorción Enzimática/métodos
19.
Microb Pathog ; 179: 106110, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-37060967

RESUMEN

Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by the metacestode larva of Echinococcus granulosus. In this study, two-dimensional gel electrophoresis (2-DE) coupled with immunoblot analysis revealed that E. granulosus severin and 14-3-3zeta proteins (named EgSeverin and Eg14-3-3zeta, respectively) might be two potential biomarkers for serological diagnosis of echinococcosis. The recombinant EgSeverin (rEgSeverin, 45 kDa) and Eg14-3-3zeta (rEg14-3-3zeta, 35 kDa) were administered subcutaneously to BALB/c mice to obtain polyclonal antibodies for immunofluorescence analyses (IFAs). And IFAs showed that both proteins were located on the surface of protoscoleces (PSCs). Western blotting showed that both proteins could react with sera from E. granulosus-infected sheep, dog, and mice. Indirect ELISAs (rEgSeverin- and rEg14-3-3zeta-iELISA) were developed, respectively, with sensitivities and specificities ranging from 83.33% to 100% and a coefficient of variation (CV %) of less than 10%. The rEgSeverin-iELISA showed cross-reaction with both E. granulosus and E. multilocularis, while the rEg14-3-3zeta-iELISA showed no cross-reaction with other sera except for the E. granulosus-infected ones. The field sheep sera from Xinjiang and Qinghai were analyzed using rEgSeverin-iELISA, rEg14-3-3zeta-iELISA, and a commercial kit respectively, and no significant differences were found among the three methods (p > 0.05). However, the CE positive rates in sheep sera from Qinghai were significantly higher than those from Xinjiang (p < 0.01). Overall, the results suggest that EgSeverin and Eg14-3-3zeta could be promising diagnostic antigens for E. granulosus infection.


Asunto(s)
Equinococosis , Echinococcus granulosus , Perros , Animales , Ovinos , Ratones , Echinococcus granulosus/genética , Proteínas 14-3-3/metabolismo , Equinococosis/diagnóstico , Equinococosis/veterinaria , Western Blotting , Ensayo de Inmunoadsorción Enzimática/métodos , Zoonosis , Anticuerpos Antihelmínticos
20.
Appl Microbiol Biotechnol ; 107(5-6): 1609-1619, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-36773062

RESUMEN

In many parts of the tropics, schistosomiasis is a major parasitic disease second only to malaria as a cause of morbidity and mortality. Diagnostic approaches include microscopic sampling of excreta such as the Kato-Katz method, radiography, and serology. Due to their vital role in many stages of the parasitic life cycle, proteases have been under investigation as targets of immunological or chemotherapeutic anti-Schistosoma agents. Five major classes of protease have been identified on the basis of the peptide hydrolysis mechanism: serine, cysteine, aspartic, threonine, and metalloproteases. Proteases of all five catalytic classes have been identified from S. mansoni through proteomic or genetic analysis. The study aimed to produce polyclonal antibodies (pAbs) against schistosomal cysteine proteases (CP) to be used in the diagnosis of schistosomiasis. This study was conducted on S. mansoni-infected patients from highly endemic areas and from outpatients' clinic and hospitals and other patients infected with other parasites (Fasciola, hookworm, hydatid, and trichostrongyloids). In this study, the produced polyclonal antibodies against S. mansoni cysteine protease antigens were labeled with horseradish peroxidase (HRP) conjugate and used to detect CP antigens in stool and serum samples of S. mansoni-infected patients by sandwich ELISA. The study involved 200 S. mansoni-infected patients (diagnosed by finding characteristic eggs in the collected stool samples), 100 patients infected with other parasites (Fasciola, hookworm, hydatid, and trichostrongyloids), and 100 individuals who served as parasite-free healthy negative control. The prepared pAb succeeded in detecting CP antigens in stool and serum samples of S. mansoni-infected patients by sandwich ELISA with a sensitivity of 98.5% and 98.0% respectively. A positive correlation was observed between S. mansoni egg counts and both stool and serum antigen concentrations. Purified 27.5 kDa CP could be introduced as a suitable candidate antigen for early immunodiagnosis using sandwich ELISA for antigen detection. KEY POINTS: • Detection of cysteine protease antigens can replace parasitological examination. • Sandwich ELISA has a higher sensitivity than microscopic examination of eggs. • Identification of antigens is important for the goal of obtaining diagnostic tools.


Asunto(s)
Proteasas de Cisteína , Esquistosomiasis mansoni , Animales , Esquistosomiasis mansoni/epidemiología , Esquistosomiasis mansoni/parasitología , Proteómica , Antígenos Helmínticos , Sensibilidad y Especificidad , Anticuerpos Antihelmínticos , Schistosoma mansoni , Heces , Endopeptidasas , Péptido Hidrolasas , Diagnóstico Precoz
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