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Seropositivity for ascariosis and toxocariosis and cytokine expression among the indigenous people in the Venezuelan Delta region.

Araujo Z, Brandes S, Pinelli E, Bochichio MA, Palacios A, Wide A, Rivas-Santiago B, Jiménez JC - Rev. Inst. Med. Trop. Sao Paulo (2015 Jan-Feb)

Bottom Line: Stool examinations showed that Blastocystis hominis, Hymenolepis nana and Entamoeba coli were the most commonly observed intestinal parasites.Quantification of cytokines IFN-γ, IL-2, IL-6, TGF-β, TNF-α, IL-10 and IL-4 expressions showed that there was only a significant increased expression of IL-4 in indigenous with TES seropositivity (p < 0.002).Ascaris and Toxocara seropositivity was prevalent among Warao indigenous.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Inmunología de Enfermedades Infecciosas, Instituto de Biomedicina, Universidad Central de Venezuela, Caracas, Venezuela.

ABSTRACT
The present study aimed at measuring seropositivities for infection by Ascaris suum and Toxocara canis using the excretory/secretory (E/S) antigens from Ascaris suum (AES) and Toxocara canis (TES) within an indigenous population. In addition, quantification of cytokine expressions in peripheral blood cells was determined. A total of 50 Warao indigenous were included; of which 43 were adults and seven children. In adults, 44.1% were seropositive for both parasites; whereas children had only seropositivity to one or the other helminth. For ascariosis, the percentage of AES seropositivity in adults and children was high; 23.3% and 57.1%, respectively. While that for toxocariosis, the percentage of TES seropositivity in adults and children was low; 9.3% and 14.3%, respectively. The percentage of seronegativity was comparable for AES and TES antigens in adults (27.9%) and children (28.6%). When positive sera were analyzed by Western blotting technique using AES antigens; three bands of 97.2, 193.6 and 200.2 kDas were mostly recognized. When the TES antigens were used, nine major bands were mostly identified; 47.4, 52.2, 84.9, 98.2, 119.1, 131.3, 175.6, 184.4 and 193.6 kDas. Stool examinations showed that Blastocystis hominis, Hymenolepis nana and Entamoeba coli were the most commonly observed intestinal parasites. Quantification of cytokines IFN-γ, IL-2, IL-6, TGF-β, TNF-α, IL-10 and IL-4 expressions showed that there was only a significant increased expression of IL-4 in indigenous with TES seropositivity (p < 0.002). Ascaris and Toxocara seropositivity was prevalent among Warao indigenous.

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Related in: MedlinePlus

Primers sequences. The following primer pairs were used: IL-2, IL-4, IL-6,IL-10, IL-12p35, IFN-γ, TGF-β, TNF-α and β-actin as internal control.
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f01: Primers sequences. The following primer pairs were used: IL-2, IL-4, IL-6,IL-10, IL-12p35, IFN-γ, TGF-β, TNF-α and β-actin as internal control.

Mentions: Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) was performed, 25nanograms of cDNA generate as above was amplified and made up to 10.5 µL with sterileand nuclease-free water. Twelve and a half µL of the master-mix containing the PCRbuffer (50 mMTris-HCl, pH 9; 50 mMNaCl; 5 mM MgCl2; 200 µM of eachdeoxynucleoside trisphosphate, dATP, dGTP, dCTP and dTTP) and the Taq DNA polymerase (50U/µL) and one µL of each primer (five mM) was added in a final volume of 25 µL.Sequences of the used primer pair are shown in Figure1. Mixtures with cDNA were placed in a MJ mini Personal Thermal Cycler (BioRadLaboratories, CA, US) preheated to 95 °C for 10 min. Cycling parameters were 40 cyclesof denaturation at 95 °C for 15 sec, annealing at 60 °C for one min, extension at 70°Cfor one min and a final extension for seven min at 72 °C. For IL-2 and β-actin theannealing was carried out at 58 °C and 55 °C respectively. Amplified products wereseparated by 2% agarose gel electrophoresis, stained with SYBR Green I (Sigma-AldrichCo, St. Louis MO, US) and visualized by a Benchtop UV transilluminator, MultiDoc-ItDigital Imaging System camera combination.


Seropositivity for ascariosis and toxocariosis and cytokine expression among the indigenous people in the Venezuelan Delta region.

Araujo Z, Brandes S, Pinelli E, Bochichio MA, Palacios A, Wide A, Rivas-Santiago B, Jiménez JC - Rev. Inst. Med. Trop. Sao Paulo (2015 Jan-Feb)

Primers sequences. The following primer pairs were used: IL-2, IL-4, IL-6,IL-10, IL-12p35, IFN-γ, TGF-β, TNF-α and β-actin as internal control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4325523&req=5

f01: Primers sequences. The following primer pairs were used: IL-2, IL-4, IL-6,IL-10, IL-12p35, IFN-γ, TGF-β, TNF-α and β-actin as internal control.
Mentions: Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) was performed, 25nanograms of cDNA generate as above was amplified and made up to 10.5 µL with sterileand nuclease-free water. Twelve and a half µL of the master-mix containing the PCRbuffer (50 mMTris-HCl, pH 9; 50 mMNaCl; 5 mM MgCl2; 200 µM of eachdeoxynucleoside trisphosphate, dATP, dGTP, dCTP and dTTP) and the Taq DNA polymerase (50U/µL) and one µL of each primer (five mM) was added in a final volume of 25 µL.Sequences of the used primer pair are shown in Figure1. Mixtures with cDNA were placed in a MJ mini Personal Thermal Cycler (BioRadLaboratories, CA, US) preheated to 95 °C for 10 min. Cycling parameters were 40 cyclesof denaturation at 95 °C for 15 sec, annealing at 60 °C for one min, extension at 70°Cfor one min and a final extension for seven min at 72 °C. For IL-2 and β-actin theannealing was carried out at 58 °C and 55 °C respectively. Amplified products wereseparated by 2% agarose gel electrophoresis, stained with SYBR Green I (Sigma-AldrichCo, St. Louis MO, US) and visualized by a Benchtop UV transilluminator, MultiDoc-ItDigital Imaging System camera combination.

Bottom Line: Stool examinations showed that Blastocystis hominis, Hymenolepis nana and Entamoeba coli were the most commonly observed intestinal parasites.Quantification of cytokines IFN-γ, IL-2, IL-6, TGF-β, TNF-α, IL-10 and IL-4 expressions showed that there was only a significant increased expression of IL-4 in indigenous with TES seropositivity (p < 0.002).Ascaris and Toxocara seropositivity was prevalent among Warao indigenous.

View Article: PubMed Central - PubMed

Affiliation: Laboratorio de Inmunología de Enfermedades Infecciosas, Instituto de Biomedicina, Universidad Central de Venezuela, Caracas, Venezuela.

ABSTRACT
The present study aimed at measuring seropositivities for infection by Ascaris suum and Toxocara canis using the excretory/secretory (E/S) antigens from Ascaris suum (AES) and Toxocara canis (TES) within an indigenous population. In addition, quantification of cytokine expressions in peripheral blood cells was determined. A total of 50 Warao indigenous were included; of which 43 were adults and seven children. In adults, 44.1% were seropositive for both parasites; whereas children had only seropositivity to one or the other helminth. For ascariosis, the percentage of AES seropositivity in adults and children was high; 23.3% and 57.1%, respectively. While that for toxocariosis, the percentage of TES seropositivity in adults and children was low; 9.3% and 14.3%, respectively. The percentage of seronegativity was comparable for AES and TES antigens in adults (27.9%) and children (28.6%). When positive sera were analyzed by Western blotting technique using AES antigens; three bands of 97.2, 193.6 and 200.2 kDas were mostly recognized. When the TES antigens were used, nine major bands were mostly identified; 47.4, 52.2, 84.9, 98.2, 119.1, 131.3, 175.6, 184.4 and 193.6 kDas. Stool examinations showed that Blastocystis hominis, Hymenolepis nana and Entamoeba coli were the most commonly observed intestinal parasites. Quantification of cytokines IFN-γ, IL-2, IL-6, TGF-β, TNF-α, IL-10 and IL-4 expressions showed that there was only a significant increased expression of IL-4 in indigenous with TES seropositivity (p < 0.002). Ascaris and Toxocara seropositivity was prevalent among Warao indigenous.

Show MeSH
Related in: MedlinePlus