Ascaris lumbricoides coinfection reduces tissue damage by decreasing IL-6 levels without altering clinical evolution of pulmonary tuberculosis or Th1/Th2/Th17 cytokine profile

Abstract INTRODUCTION: Immunological control of Mycobacterium tuberculosis infection is dependent on the cellular immune response, mediated predominantly by Th1 type CD4+ T cells. Polarization of the immune response to Th2 can inhibit the host immune protection against pathogens. Patients with tuberculosis coinfected with helminths demonstrate more severe pulmonary symptoms, a deficiency in the immune response against tuberculosis, and an impaired response to anti-tuberculosis therapy. METHODS: We evaluated the cellular immune response and the impact of the presence of Ascaris lumbricoides on the immune and clinical response in pulmonary tuberculosis patients. Ninety-one individuals were included in the study: 38 tuberculosis patients, 11 tuberculosis patients coinfected with Ascaris lumbricoides and other helminths, 10 Ascaris lumbricoides patients, and 34 non-infected control individuals. Clinical evolution of pulmonary tuberculosis was studied on 0, 30, 60, and 90 days post-diagnosis of Mycobacterium tuberculosis and Ascaris lumbricoides. Furthermore, immune cells and plasma cytokine profiles were examined in mono/coinfection by Mycobacterium tuberculosis and Ascaris lumbricoides using flow cytometry. RESULTS: There were no statistical differences in any of the evaluated parameters and the results indicated that Ascaris lumbricoides infection does not lead to significant clinical repercussions in the presentation and evolution of pulmonary tuberculosis. CONCLUSIONS: The association with Ascaris lumbricoides did not influence the Th1, Th2, and Th17 type responses, or the proportions of T lymphocyte subpopulations. However, higher serum levels of IL-6 in tuberculosis patients may explain the pulmonary parenchymal damage.

Seropositivity for ascariosis and toxocariosis nd cytokine expression among the indigenous people in the Venezuelan delta region / Seropositividad para ascariosis y toxocariosis y expresión de citocinas entre la población indígena de la región del delta Venezolano

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.

El objetivo del presente estudio fue determinar la seropositividad de infección por Ascaris suum y Toxocara canis, utilizando antígenos de excreción/secreción (E/S) de Ascaris suum (AES) y Toxocara canis (TES) en una población indígena. Adicionalmente, se cuantificó la expresión de citocinas a partir de células de sangre periférica. Un total de 50 indígenas Warao se incluyeron en el estudio; 43 fueron adultos y 7 niños. Entre los adultos, 44,1% fueron seropositivos para ambos parásitos; mientras que los niños sólo mostraron seropositividad a uno u otro de los helmintos. Para ascariosis, el porcentaje de seropositividad para los antígenos AES fue alto tanto en adultos como en niños; 23,3% y 57,1%, respectivamente. Para toxocariosis, el porcentaje de seropositividad para los antígenos TES fue bajo en adultos así como en niños; 9,3% y 14,3%, respectivamente. El porcentaje de seronegatividad fue similar tanto para los antígenos AES como para TES en adultos (27,9%) y niños (28,6%). Cuando la seropositividad fue analizada a través de la técnica de Western blotting utilizando los antígenos AES; 3 bandas de 97,2, 193,6 y 200,2 kDas fueron principalmente reconocidas. Para los antígenos TES, 9 bandas fueron mayormente identificadas; 47,4, 52,2, 84,9, 98,2, 119,1, 131,3, 175,6, 184,4 y 193,6 kDas. Los análisis coproparasitológicos mostraron que los parásitos Blastocystis hominis, Hymenolepis nana y Entamoeba coli fueron los parásitos intestinales más comúnmente observados. La cuantificación de la expresión de las citocinas IFN-γ, IL-2, IL-6, TGF-β, TNF-α, IL-10 e IL-4 mostró que hubo un significante incremento de la expresión de IL-4 entre los indígenas con seropositividad para los antígenos TES (p < 0.002). La seropositividad para Ascaris y Toxocara fue prevalente entre los indígenas Warao.

Severity of atopic dermatitis and Ascaris lumbricoides infection: an evaluation of CCR4+ and CXCR3+ helper T cell frequency / Gravidade da dermatite atópica e infecção por Ascaris lumbricoides: uma avaliação da frequência de células T auxiliares CCR4+ e CXCR3+

INTRODUCTION: Ascaris lumbricoides-infected patients present lower prevalence of severe atopic dermatitis. METHODS: Peripheral blood of infected children with atopic dermatitis was assessed by flow cytometry of the frequency of Th1 and Th2 cells through the expression of CXCR3 and CCR4 chemokine receptors, respectively. RESULTS: Helminth-free patients with atopic dermatitis presented a high frequency of CCR4+Th2 cells. Parasitized patients with atopic dermatitis showed a lower frequency of CXCR3+Th1 cells compared to infected individuals only. CONCLUSIONS: Ascariasis modifies the blood traffic of Th2 cells in atopic dermatitis patients, while the allergic disease down-regulates the traffic of Th1 cells in parasitized patients.

INTRODUÇÃO: Pacientes infectados com Ascaris lumbricoides apresentam menor prevalência de dermatite atópica grave. MÉTODOS: Sangue periférico de crianças infectadas com dermatite atópica foi analisado por citometria de fluxo quanto à frequência de células Th1 e Th2 pela expressão de receptores de quimiocina CXCR3 e CCR4, respectivamente. RESULTADOS: Pacientes sem helmintos com dermatite atópica apresentaram alta frequência de células Th2CCR4+. Pacientes parasitados com dermatite atópica apresentaram menor frequência de células Th1CXCR3+ comparados aos indivíduos apenas infectados. CONCLUSÕES: Ascaridiases altera o tráfego sanguíneo de células Th2 em pacientes com dermatite atópica, enquanto a doença alérgica diminui o tráfego de células Th1 em pacientes parasitados.

Ascaris lumbricoides: Mimetismo molecular por absorción de epitopes P1 / Ascaris lumbricoides: Molecular mimicry by P1 epitope absorption

El mimetismo molecular se ha asociado con la cronicidad de las infecciones parasitarias. Este mecanismo puede deberse a absorción de moléculas del hospedador o a síntesis de moléculas similares. P1 ha sido detectado en diversas especies parásitas. Experiencias previas han demostrado epitopes P1 en ejemplares adultos de Ascaris lumbricoides. El objetivo de este trabajo fue estudiar la absorción de estos epitopes por estadios larvales del nematodo, a los fines de determinar el mecanismo de expresión de estos antígenos sobre la cutícula parasitaria. Las larvas fueron eclosionadas a partir de heces con huevos de este helminto. Fueron cultivadas en medio RPMI con eritrocitos P1. Se realizó la Técnica de Inhibición de la Aglutinación Semicuantitativa usando cuatro anticuerpos monoclonales anti-P1. El sistema revelador fue una suspensión de eritrocitos frescos P1 en medio enzimático de bromelina. Se investigaron por la misma técnica productos de excreción-secreción símil P1 usando larvas cultivadas en medio RPMI en ausencia de eritrocitos P1. Las larvas cultivadas con glóbulos rojos inhibieron la aglutinación de todos los anticuerpos monoclonales con los eritrocitos P1. No se identificaron productos de excreción- secreción símil P1. Se ha demostrado que las larvas de A. lumbricoides absorben epitopes P1 del medio de cultivo. Se concluye que el parásito puede absorber este antígeno durante su ciclo de vida para usarlo en el mimetismo molecular.

Molecular mimicry has been associated with chronicity of parasitic infections. This mechanism can be due to absorption of the host's molecules or to synthesis of similar molecules. P1 has been detected in various parasite species. Previous experiences have shown P1 epitopes in Ascaris lumbricoides's adult parasites. The aim of this study was to analyse absoption of these epitopes by nematode larvae in order to determine the mechanism by which the parasite can express these antigens on their cuticle. Larvae were hatched from the faeces with helminth eggs. Larvae were cultivated in RPMI medium with P1 red cells. The Quantitative Inhibition Agglutination Test was performed using 4 anti- P1 monoclonal antibodies. P1 fresh red cells in bromelin enzymatic medium was used as revealing system. Simil P1 excretory-secretory products were investigated using larvae cultivated in RPMI medium without P1 erythrocytes by the same technique. The larvae which were cultivated with red cells inhibited the agglutination between all anti- P1 monoclonal antibodies and P1 erythrocytes. Simil P1 excretory-secretory products were not identified. It was demonstrated that A. lumbricoides larvae absorb P1 epitopes from the culture medium. It can be concluded that the parasite can absorb this antigen during its life cycle in order to use it in molecular mimicry.

Host immune response to Toxoplasma gondii and Ascaris lumbricoides in a highly endemic area: evidence of parasite co-immunomodulation properties influencing the outcome of both infections

Toxoplasmosis and ascaridiasis evoke polar Th-1 and Th-2 host immune responses, respectively. A study to investigate the specific cytokine profile production by in vitro cultures of peripheral blood mononuclear cells from individuals living under precarious sanitary conditions in a highly endemic area for the parasites Toxoplasma gondii and Ascaris lumbricoides was conducted. High levels of both IFN-³ (Th-1) and IL-13 (Th-2) were observed in groups of co-infected individuals presenting toxoplasmic ocular lesions. Significantly lower IL-10 and TGF-² levels were produced by co-infected individuals in comparison with groups of individuals not infected with A. lumbricoides and either positive or negative for T. gondii living under good sanitary conditions (control groups). The possible influence of co-parasitism on the clinical presentation of ocular toxoplasmosis is discussed.

Antígenos de larvas pulmonares de Ascaris suum reconocidos por anticuerpos producidos en Oryctolagus cuniculus / Ascaris suum lung larvae antigens recognized by antibodies produced in Oryctolagus cuniculus

El presente trabajo tuvo como objetivo determinar mediante la técnica de Western Blot los antígenos de larvas pulmonares de Ascaris suum que son detectados por anticuerpos producidos en Oryctolagus cuniculus inmunizado experimentalmente. Las larvas pulmonares (L3 y L4) fueron obtenidas en 120 ejemplares de Mus musculus cepa BALB/c ratón infectados experimentalmente por vía oral con huevos infectivos de A. suum. Parte de estas larvas fueron cultivadas en el medio Eagle (MEM) para la obtención de antígenos de excreción/secreción y la otra fue sonificada para la obtención de antígenos somáticos, los cuales sirvieron para inmunizar dos ejemplares de O. cuniculus, utilizando Adyuvante Completo e Incompleto de Freund. A las 5 semanas de inmunización se obtuvo sangre de los conejos por punción cardiaca a fin de recuperar el suero, parte del cual fue purificado parcialmente por precipitación salina y diálisis. Mediante la técnica de electroinmuno-transferencia (Western Blot) y usando sueros de los conejos inmunizados se detectaron en los antígenos de excreción/secreción de 20 horas de cultivo reducidos con dithiothreitol, 12 bandas antigénicas de 100, 72.4, 56.2, 42.7, 39.8, 34.6, 31.6, 30.2, 19.5, 16.9, 15.5 y 14.9 KDa, siendo las más reactivas las de 100, 72.4, 16.9, 15.5 y 14.9 KDa. En los antígenos somáticos bajo condiciones de reducción, se detectaron solamente seis bandas de 42.7, 39.8, 34.6, 30.2, 28 y 25.2 KDa de poca reactividad. Estos resultados permiten afirmar que los antígenos de excreción/secreción de A. suum de 20 horas de incubación en el medio MEM inducen la producción de un mayor número de anticuerpos de tipo IgG en conejos inmunizados experimen-talmente.

Excretory/secretory antigens (E/SAg) and somatic antigens (SAg) of Ascaris suum lung larvae that induce the immunoglobulin G antibodies production in Oryctolagus cuniculus experimentally immunized was determined. For this purposes, specimens of Mus musculus BALB/c were inoculated orally with infective eggs of A. suum obtained from pigs naturally parasitized in order to obtain the lung larvae. Part of these larvae was cultured in Minimum Essential Medium (MEM) to obtain E/SAg and another part was sonicated to obtain the SAg too. Both, E/SAg and SAg mixed with complete and incomplete Freund's adjuvant were used for rabbits immunization. Five weeks after the immunization, the rabbits were bled by cardiac puncture obtaining the immunosera by centrifugation, which was purified partially by saline precipitation and dialysis. By using an Western blot technique with purified immunoserum and E/SAg obtained to 20 hours and reduced with dithiothreitol, fourteen antigens bands of 100, 72.4, 56.2, 42.7, 39.8, 34.6, 31.6, 30.2, 19.5, 16.9, 15.5 y 14.9 KD, were detected. The bands of 100, 72.4, 16.9, 15.5 and 14.9 KDa were been the most reactives. Thereby the SAg, also reduced with dithiothreitol, seven bands of 42.7, 39.8, 34.6, 30.2, 28.0 and 25.2 KDa were detected. They were a bit clear. In conclusion, the E/SAg induce the highest production of immunoglobulin G antibodies in rabbits experimentally immunized.

Antifilarial and anti PPD IgM antibodies in cord blood.

Tuberculosis and Filariasis are commonly encountered in South India, more so in the low socio-economic group. This study was undertaken to find out the fetal immune response to maternal filariasis and tuberculosis. The presence of anti-filarial and anti-PPD IgG and IgM antibodies in the mother and cord blood at the time of delivery was identified by ELISA. Six out of 29 cases showed IgM against Brugia malayi antigen in the cord blood. In both instances, 6 out of 29 for PPD and 7 out of 29 for B. malayi, the maternal blood too showed the presence of IgM to these two antigens. Comparison with anti-Ascaris lumbricoides IgM antibodies in mother and cord blood in these (6 out of 29 against PPD and 7 out of 29 against B. malayi) cases did not show a similar distribution of IgM antibodies in mother and cord blood, indicating that there is a fetal response to maternal filariasis and tuberculosis.

comparative study of immunoglobulins E,A,M,G, and eosinophilia in ascariasis and schistosomiasis

Serum Igs E, A, M, G, and blood eosinophils were determined in 31 helminth-infested subjects [ascaris 15, bilharzia 13, mixed infection 3]. A highly significant rise [4 times the control mean value] was obtained with IgE and eosinophils. The former was more elevated in ascariasis and vice versa. Correlation study suggests that the eosinophil response is not solely IgE-mediated. While IgA was significantly raised, IgM was raised in ascaris cases and the significant rise in IgG was more evident in bilharzial cases. cases with mixed infection revealed no summation effect. The wide range, in the pathological group was pointed out and the possible mechanism and meaning of the findings were discussed