Evaluation of the transcriptional immune biomarkers in peripheral blood from warao indigenous associate with the infection by mycobacterium tuberculosis

Abstract INTRODUCTION: Biomarkers are critical tools for finding new approaches for controlling the spread of tuberculosis (TB), including for predicting the development of TB therapeutics, vaccines, and diagnostic tools. METHODS: Expression of immune biomarkers was analyzed in peripheral blood cells stimulated and non-stimulated with M. tuberculosis antigens ESAT-6, CFP10 and TB7.7. in Warao indigenous individuals. These biomarkers may be able to differentiate TB states, such as active tuberculosis (ATB) cases and latent tuberculosis infection (LTBI) from non-infected controls (NIC). A real-time reverse transcription polymerase chain reaction (RT-qPCR) assay was performed on 100 blood samples under non-stimulation or direct ex vivo conditions (NS=50) and stimulation conditions (S=50). RESULTS: The findings are shown as the median and interquartile range (IQR) of relative gene expression levels of IFN-γ, CD14, MMP9, CCR5, CCL11, CXCL9/MIG, and uPAR/PLAUR immune biomarkers. MMP9 levels were significantly higher in the LTBI-NS and LTBI-S groups compared with the NIC-NS and NIC-S groups. However, CCR5 levels were significantly lower in the LTBI-S group compared with both NIC-NS and NIC-S groups. CCL11 levels were significantly lower in the LTBI-S group compared with the NIC-NS group. CONCLUSIONS: Preliminary findings showed that MMP9 immune biomarkers separated LTBI indigenous individuals from NIC indigenous individuals, while CCR5, CCL11, CD14, and IFN-γ did not differentiate TB states from NIC. MMP9 may be useful as a potential biomarker for LTBI and new infected case detection among Warao indigenous individuals at high risk of developing the disease. It may also be used to halt the epidemic, which will require further validation in larger studies.

Concordance between IFNγ gene +874 A/T polymorphism and interferon-γ expression in a TB-endemic indigenous setting

Abstract INTRODUCTION: Interferon-γ (IFN-γ) plays a crucial role in resistance to mycobacterial diseases; accordingly, variants of the gene encoding this cytokine may be associated with elevated risk of contracting pulmonary tuberculosis (TB). METHODS: Blood samples were collected from 135 Warao indigenous individuals with newly diagnosed sputum culture-positive TB. Of these, 24 were diagnosed with active tuberculosis (ATB). The study comprised 111 participants, who were grouped as follows: 1) 14 tuberculin skin test (TST)-positive Warao indigenous individuals and 4 that were QuantiFERON-TB?Gold In-Tube (QFT-IT) test-positive, collectively comprising the latent TB infection group (LTBI), n = 18), and 2) healthy controls who were QFT-IT- and TST-negative, comprising the control group (CTRL, n = 93). Detection of the IFN γ gene (IFNG) +874A/T polymorphism was performed via PCR and quantification of IFNG expression via qPCR. RESULTS: Relative to indigenous and white Americans, ATB and CTRL groups had a higher frequency of the IFNG SNP (+874A): 23 (95.8%) and 108 (97.3%), respectively. Indigenous Warao individuals homozygous for the IFNG (+874) A allele exhibited 3.59-fold increased risk of developing TB (95% confidence interval, 2.60-4.96, p =0.0001). A decreased frequency of the AT genotype was observed in individuals with pulmonary TB (4.16%) and controls (0.90%). The frequency of the TT genotype was decreased among controls (1.80%); none of the patients with TB were found to have this genotype. The differences in IFNG expression between the groups, under unstimulated and stimulated conditions, were not statistically significant. CONCLUSIONS: Preliminary results demonstrate concordance between IFNG +874 A/A genotype and low expression of IFNG.

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.

Immunologic evaluation and validation of methods using synthetic peptides derived from Mycobacterium tuberculosis for the diagnosis of tuberculosis infection

The goal of this study was to demonstrate the usefulness of an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of pulmonary tuberculosis (PTB) and extrapulmonary TB (EPTB). This assay used 20 amino acid-long, non-overlapped synthetic peptides that spanned the complete Mycobacterium tuberculosis ESAT-6 and Ag85A sequences. The validation cohort consisted of 1,102 individuals who were grouped into the following five diagnostic groups: 455 patients with PTB, 60 patients with EPTB, 40 individuals with non-EPTB, 33 individuals with leprosy and 514 healthy controls. For the PTB group, two ESAT-6 peptides (12033 and 12034) had the highest sensitivity levels of 96.9% and 96.2%, respectively, and an Ag85A-peptide (29878) was the most specific (97.4%) in the PTB groups. For the EPTB group, two Ag85A peptides (11005 and 11006) were observed to have a sensitivity of 98.3% and an Ag85A-peptide (29878) was also the most specific (96.4%). When combinations of peptides were used, such as 12033 and 12034 or 11005 and 11006, 99.5% and 100% sensitivities in the PTB and EPTB groups were observed, respectively. In conclusion, for a cohort that consists entirely of individuals from Venezuela, a multi-antigen immunoassay using highly sensitive ESAT-6 and Ag85A peptides alone and in combination could be used to more rapidly diagnose PTB and EPTB infection.

Diagnostic accuracy of immunological methods in patients with tuberculous pleural effusion from Venezuela / Precisión diagnóstica de métodos inmunológicos en pacientes con tuberculosis pleural de Venezuela

In recent years, better diagnostics for tuberculosis (TB) has received increasing attention, especially the diagnosis of tuberculous pleural effusion, which is difficult and at present the main tool in TPE diagnostic is pleural effusion smear and culture, but unfortunately, sensitivities are low, therefore better TPE diagnostic tools are needed. The aim of this study was to find a diagnostic algorithm to assess the progress in TPE diagnostic at the Hospital Vargas de Caracas, that permits identification of the majority of patients, at a satisfactory cost-benefit ratio, evaluating the levels of IFN-g and IL-12p40 in pleural effusion and serum, as well as the antibody reactivity in order to compare it with microbiological tests. A total of 60 individuals with pleural effusion were studied; 20 patients with tuberculous pleural effusion (TPE) formed the patient group and 40 patients with non-tuberculous pleural effusion (NTPE) formed the control group. The levels of IFN-g and IL-12p40 in effusion and serum and class and subclasses of IgG reactivity to Mycobacterium tuberculosis antigens were measured by ELISA. The utility of these methods for diagnosis of TPE was evaluated using receiver operating characteristic (ROC) curve analysis. The results of the 11 immunological methods evaluated showed that the anti-PPD IgG2 method was able to reach the highest specificity of 95% (CI: 88.3-101.8), positive predictive value (PPV)=75 (at 30% sensitivity); while that the overall sensitivity of methods was between 95% and 30%, of these, two methods reached higher sensitivities; increased levels of pleural IFN-g, with a sensitivity of 95% (CI: 85.5-104.5) with the highest negative predictive value (NPV)=97, (at 82.5% specificity), followed by decreased levels of serum IL-12p40 with a sensitivity of 95% (CI: 85.5-104.5), NPV=95.2 (at 50% specificity). In contrast, microbiological methods showed that smear had a sensitivity of only 20%, while smear plus ...

Recientemente existe un gran interés hacia un mejor y más rápido diagnóstico de tuberculosis (TB), especialmente de tuberculosis pleural, el cual es difícil. Al presente las principales herramientas diagnósticas son la baciloscopia y el cultivo de líquido pleural; desafortunadamente, las sensibilidades de estos métodos son bajas, por lo que el desarrollo de nuevas herramientas diagnósticas es necesario. El objetivo del presente estudio consistió en encontrar un algoritmo que permita la rápida identificación de la mayoría de los pacientes con TB pleural que ingresan en el Hospital Vargas de Caracas a un buen costo-beneficio. Para esto se evaluaron los niveles de las citocinas Interferón-gamma (IFN-g) y la Interleucina 12p40 (IL-12p40) en líquido pleural y suero, así como la reactividad de anticuerpos contra antígenos de Mycobacterium tuberculosis. Se estudiaron 60 individuos con derrame pleural; 20 individuos con líquido pleural tuberculoso (LPT) conformaron el grupo de pacientes y 40 individuos con líquido pleural no tuberculoso (LPNT) el grupo de controles. La técnica de inmunoensayo de ELISA fue utilizada para medir los niveles de IFN-g y IL-12p40; así como las reactividades de los diversos isotipos y subclases de inmunoglobulina G (IgG) frente a antígenos del bacilo. La utilidad de los métodos fue evaluada utilizando el análisis de las curvas ROC (receiver operating characteristic). Los resultados de los 11 métodos inmunológicos evaluados mostraron que el método IgG2 anti-PPD alcanzó la mayor especificidad de 95%, (CI: 88,3-101,8) con un valor predictivo positivo (VPP) de 75. La sensibilidad de los métodos estuvo entre 30% y 95%; dos métodos alcanzaron altas sensibilidades: los altos niveles de IFN-g en líquido pleural, con sensibilidad de 95% (CI: 85,5-104,5), con un valor predictivo negativo (VPN) de 97, seguido de los bajos niveles de IL-12p40 en suero, con una sensibilidad de 95% (CI: 85,5-104,5) con un VPN de 95,2. En contraste, ...

Comparison of serological responses in two different populations with pulmonary tuberculosis

Observational studies on the humoural immune responses of the Warao indigenous people from Delta Amacuro, an isolated area, were compared with urban residents of the Venezuelan capital. Mycobacterium tuberculosis-specific reactivities (IgM, IgE, sIgA, IgG and IgG subclasses) were measured by ELISA using PPD and 38-kDa M. tuberculosis antigens. A total of 294 individuals were studied, 162 Warao (indigenous people) and 132 Creole (non-indigenous people). The patient group consisted of 87 Warao patients and 58 Creole patients, while the control group consisted of 75 Warao controls and 74 Creole controls. Combinations among the isotypes studied were performed. The findings showed that for the Warao people, sensitivity to the combination including anti-PPD IgG and IgE was 92.0 percent, while for the Creole people, sensitivity to the combination including anti-PPD IgG but more so anti-PPD IgG1 and IgG2 was 90.0 percent. Simple tests were able to show higher specificities, which were population-specific; specificities were anti-PPD IgG3, 100.0 percent and anti-PPD IgM, 97.4 percent for the Warao and Creole peoples, respectively. In conclusion, while simple tests reached high specificity, the multi-isotype tests improved sensitivity; the latter shows this approach may be useful in diagnostic testing.

Respuesta inmunitaria en tuberculosis y el papel de los antígenos de secreción de Mycobacterium tuberculosis en la protección, patología y diagnóstico: revisión / Immunitary response in Tuberculosis and the role of Mycobacterium tuberculosis secretion antigens in its protection, pathology and diagnosis: review

La tuberculosis (TB) es una de las principales causas de muerte por infección con Mycobacterium tuberculosis en seres humanos en el ámbito mundial, y es preocupante que su incidencia esté aumentado en los últimos años, debido principalmente a la infección por el virus de inmunodeficiencia humano (VIH). Considerando que los métodos diagnósticos utilizados actualmente para detectar la enfermedad no son totalmente efectivos y rápidos, debido a que el cultivo no es útil como primera opción diagnóstica porque ya que requiere más de 4 semanas y el examen microscópico tiene una sensibilidad de 50 a 60% y es aún más baja en pacientes pediátricos, múltiples esfuerzos se realizan con el objetivo de caracterizar cada vez más los antígenos de M. tuberculosis asociados con protección y para desarrollar rápidos y mejores métodos de diagnóstico. Entre los antígenos secretados de M. tuberculosis y que han sido descritos como inductores de la secreción de mediadores asociados a protección contra la infección por M. tuberculosis están CPF-10, ESAT-6, 27 kDa y 38 kDa, por inducir la producción de IFN-g, TNF-a y óxido nítrico, todos estos relacionados con una respuesta inmunitaria protectora. A través del estudio de las funciones y composición de estos antígenos, se hacen esfuerzos por encontrar métodos más efectivos en el diagnóstico de la enfermedad, en sus diversas etapas de evolución. La presente revisión tiene como finalidad describir los antígenos que han sido reportados como antígenos relevantes de M. tuberculosis por participar en la respuesta inmunitaria protectora frente a la infección y su posible utilidad en el diagnóstico.

Inmunidad humoral y celular específica: y niveles de adenosindesaminasa en niños con tuberculosis: Hospital de Niños "JM de Los Ríos" Caracas-Venezuela / Specific humoral and cellular immunity: and adenosine deaminase levels in children with tuberculosis: Hospital de Niños J.M. de Los Ríos Caracas-Venezuela

La tuberculosis (TBC) constituye un problema de salud pública en Venezuela. Pese a los avances en la micobacteriología, el diagnóstico en niños continúa siendo un gran reto. Estudiar la inmunidad celular y humoral específicas y niveles de Adenosindesaminasa(ADA) en suero en niños con diagnóstico de tuberculosis y evaluar su utilidad como prueba diagnóstica. Estudio prospectivo, diseño no experimental. Se evaluaron dos grupos (Pacientes-Controles), edades entre un mes-14 años, ambos sexos. Niños hospitalizados en Hospital J.M de Los Ríos (2004–2005), con tuberculosis activa; Controles: asintomáticos, radiografía de tórax normal, sin evidencia de contacto epidemiológico. Se midió reactividad de isotipos IgG, IgG1, IgG2, IgG3 e IgA en suero y la IgA secretora (IgAs) en saliva específicos contra el antígeno PPD de M. tuberculosis, niveles de producción de IL-12 e IL-5, (técnica ELISA); niveles séricos de ADA (Prueba de Giusti modificado). Total 25 niños, 11 Pacientes y 14 Controles. Sensibilidad prueba tuberculina 45,5 por ciento. Niveles de ADA sérica sin diferencia entre los grupos. Niveles de IgG específica e IgG2 fueron mayores en los pacientes (P<0,002). Sensibilidad y especificidad para IgG2 fue de 80 por ciento y 100 por ciento, respectivamente. Medición IL-12 reportó sensibilidad 50 por ciento, especificidad 85,7 por ciento, diferencia entre los grupos significativa (P<0,05). Niveles IL-5 mayores en los pacientes (P<0,05), sensibilidad 44.4 por ciento, especificidad 100 por ciento. Resto de las pruebas serológicas sin diferencias entre los grupos. Las determinaciones inmunológicas y enzimática mostraron en general elevada especificidad con una sensibilidad variable. La determinación de IgG2 anti PPD en suero fue la prueba más sensible y específica. Niveles de ADA en suero tienen utilidad limitada debido a su baja sensibilidad.

The secretory immunoglobulin A response to Mycobacterium tuberculosis in a childhood population

We report on the measurement of saliva anti-Purified Protein Derivative sIgA and 38kDa antibodies from 127 children, of whom 31 were strong tuberculosis suspects and 96 were healthy contact children. The results concerning the percentage of children with antibody reactivity to PPD and 38kDa antigens showed that, of these 2 antigens, 38kDa induced higher reactivity in patients positive and negative for the Tuberculin Skin Test (28 percent and 16.6 percent, respectively) in comparison to controls positive and negative for the TST (11.7 percent and 7.1 percent, respectively). There was a statistically significant difference between patients positive and controls negative for the TST. In relation to the Purified Protein Derivative antigen, while 14.2 percent of patients positive for the TST showed antibody reactivity to the PPD antigen, no patients negative for the TST had reactivity to this antigen. The findings suggest that these two antigens seem be associated with a different development of the mucosal defence mechanisms mediated by sIgA against Mycobacterium tuberculosis.

Foram dosados anticorpos sIgA anti-Purified Protein Derivative e 38kDa da saliva de 127 crianças, das quais 31 eram de pacientes altamente suspeitos de tuberculose e 96 eram provenientes de crianças saudáveis, que tiveram contato com pacientes. Os resultados referentes à porcentagem de crianças, reativas ao PPD e ao antígeno 38kDa, mostraram que destes dois antígenos, o 38kDa induziu maior reatividade em pacientes positivos e negativos ao Tuberculin Skin Test (28 por cento e 16,6 por cento, respectivamente), em comparação aos controles positivos e negativos ao TST (11,7 por cento e 7,1 por cento, respectivamente). Houve diferença estatisticamente significativa entre pacientes positivos e controles negativos ao Tuberculin Skin Test. Em relação ao antígeno PPD, enquanto 14,2 por cento de pacientes positivos ao TST mostraram anticorpos reativos ao antígeno Purified Protein Derivative, nenhum paciente negativo ao TST foi reativo ao antígeno. Os achados sugerem que, aparentemente, estes dois antígenos estão associados a desenvolvimento distinto dos mecanismos de defesa da mucosa mediados por sIgA contra Mycobacterium tuberculosis.

Células epiteliales en la inmunidad del pulmón / Immunological role of lung epithelial cells

El sistema respiratorio se encuentra en contacto con patógenos, sin embargo, gracias a la respuesta inmune innata de este, solo en raras ocasiones se establece la enfermedad. Las células epiteliales del tracto respiratorio juegan un papel importante para evitar la colonización del pulmón por agentes infecciosos, identificando a los microorganismos a través de receptores especializados, como los Toll-like,. Las células epiteliales son capaces de secretar citocinas, péptidos antimicrobianos y otras moléculas proinflamatorias las cuales evitan el establecimiento de patógenos

Levels of complement C3 and C4 components in Amerindians living in an area with high prevalence of tuberculosis

The levels of complement C3 and C4 components were determined in non-indigenous (creoles) and indigenous (Warao) populations, the latter with an extremely high tuberculosis (TB) rate. Serum samples from 209 adults were studied and classified in 4 groups taking into account tuberculin skin tests (TST): (1) the group of Warao patients (58 positive for the TST, WP TST+ and 9 negative for the TST, WP TST-), (2) the group of creole patients (34 positive for the TST, CP TST+ and 9 negative for the TST, CP TST-), (3) the group of healthy Warao controls (38 positive and 14 negative for TST, WC TST+ and WC TST-, respectively), (4) the creole controls (26 positive and 21 negative for the TST, CC TST+ and CC TST-, respectively). With respect to the results concerning the measurement of both complement C3 and C4 components with the exception of the WC TST and the CC groups, the WP TST+ and WP TST- as well as WC TST+ groups showed a significant frequency of individuals with decreased levels of complement C3 component (20.6, 33.3, and 26.3 percent, respectively) and also C4 component (12.0, 11.1, and 13.3 percent, respectively) in comparison to both creole patients (CP TST+, 8.82 percent and CP TST-, 0 percent and CP TST+, 5.88 percent and CP TST-, 0 percent) for C3 and C4, respectively. The study of these parameters carried out in 15 Warao subjects with active infection, before and after anti-TB chemotherapy,statisticallyconfirmedthat the effective chemotherapy did not restore normal levels of the complement C3 and C4 components among Warao patients. Aditional tests for hepatitis B or hepatitis C infection, and the profile of the hepatic proteins were not associated to the deficiency in production of the complement components.In conclusion, the results show that within the Warao population, a high percentage of subjects exhibit decreased levels of both complement C3 and C4 components independent of latent or active infection and the status of TST.

Respuesta de inmunidad celular en la tuberculosis pulmonar: revisión / Cell immunity response in human pulmonary tuberculosis: review

La tuberculosis (TBC) pulmonar humana, un problema de salud pública en el mundo, es una enfermedad infecciosa causada por Mycobacterium tuberculosis. Se conoce que una minoría de las personas que son infectadas por M. tuberculosis son capaces de progresar a enfermedad clínica. Se puede decir, en términos generales, que el 90 por ciento de las personas tendrán controlados los bacilos en estado latente para toda la vida, por medio de su sistema inmunológico. Un 5 por ciento presentará TBC primaria progresiva y el otro 5 por ciento presentará la enfermedad en estados tardíos de la vida, lo que se denomina TBC de reactivación o post-primaria. En los individuos resistentes, el control de la infección o de los bacilos tuberculosos que se encuentran en la región alveolar requiere principalmente del desarrollo de una respuesta de inmunidad celular (RIC) del tipo Th1. Este tipo de respuesta incluye la participación de los macrófagos alveolares, los linfocitos T CD4+ y CD8+, principalmente los linfocitos T gd y la producción de citocinas como: IL-2, IFN-g, IL-12, IL-18 y TNF-a. Aunado están las quimiocinas como RANTES, MCP-1, MIP-1a e IL-8 que juegan un papel muy importante en la migración de las diferentes subpoblaciones celulares al sitio de infección para la formación del granuloma. Además, es primordial el papel de las células "natural killer" (NK), y de las células epiteliales como parte de la respuesta de inmunidad innata

Estudio de la respuesta inmunitaria en niños Warao de comunidades con alta prevalencia de tuberculosis.

Resumen. En una población de niños Warao menores de 15 años con una tasa de prevalencia de tuberculosis (TBC) de 3190/100.000 se planteó como objetivo, estudiar ciertos parámetros de la respuesta inmunitaria en una población de 107 niños, de éstos, 32 presentaban TBC (27 PPD positivos y 5 PPD negativos) y 75 eran controles sin clínica de TBC (45 PPD positivos y 30 PPD negativos). Se evaluaron la inmunidad humoral, celular y algunos aspectos de la inmunidad innata. Entre los niños pacientes y controles, no se encontraron diferencias significativas en los componentes C3 y C4 del complemento. En cuanto a la inmunidad humoral, el grupo de pacientes presentó un alto porcentaje de niños con niveles totales de IgA e IgG elevados (13,8% y 79,3%, respectivamente) en comparación con los niños controles (0% y 69,2%). En el grupo control, un alto porcentaje de niños presentaron niveles totales de IgM e IgAs elevados (69,2% y 56,16%, respectivamente) en comparación con los pacientes (48,3% y 31,25%). El 100% de los niños presentó altos niveles séricos de IgE. Los estudios de la respuesta celular retardada mostraron que los niños de mayor edad presentaron las mayores medidas de induraciones. Los niños menores de 7 años presentaron similar reactividad positiva al PPD independientemente de la presencia o no de cicatriz de BCG. Un alto porcentaje de niños sin y con cicatriz (46,8% y 27,6% respectivamente) presentaron medidas de induraciones a la tuberculina de 0 mm. La reactividad a Candida mostró un 80% de niños con estados de anergia no específica. Sin embargo, el mayor porcentaje de niños con TBC estaba vacunado y presentaba positividad a la prueba de tuberculina. En conclusión, en los niños Warao se observó un aumento en los niveles de los componentes C3 y C4 del complemento y una hipergammaglobulinemia. Los resultados de los isotipos mostraron que, la disminución de IgAs podría ser un marcador de enfermedad activa, mientras que el aumento de IgM un marcador de infección reciente. La policlonalidad fue independiente de la reactividad al PPD.

Abstract. The present study was carried out in a Warao childhood population with extremely high tuberculosis (TB) rate of 3190/100,000 in 0-15 years old children. One hundred seven serum and saliva samples were tested, 32 from patients with active TB (27 positive and 5 negative for the tuberculin skin test, TST) and 75 apparently healthy contact children (45 positive and 30 negative for the TST). The innate, immunoglobulin and cellular responses were studied. The results showed that both, patients and controls, had a high percentage of children with increased levels of complement C3 and C4 components. A high percentage of children with increased total serum IgA and IgG (13.8% and 79.3% respectively) was observed in children with TB in comparison to control children (0% and 69.2%). A high percentage of control children had increased levels of IgM and sIgA (69.2% and 56.16%, respectively) in comparison to patients (48.3% and 31.25%). Both groups showed children with increased levels of IgE. The results concerning to the cellular immune response to PPD and the BCG vaccination status showed that there was a correlation between an increase in PPD reactivity and age. The PPD reactivity in children less than 7 years old was similar and also independent of the BCG vaccination status. A significant number of children without or with scars (46.8% and 27.6%, respectively) showed induration values of 0 mm to tuberculin skin test. The Candida reactivity showed a high percentage of children (80%) with anergy status. In conclusion, an increase in the levels of complement components C3 and C4 and hypergammaglobulinemia was observed in Warao children, and these results were independent from PPD reactivity and BCG vaccination. The isotype results showed that the decrease in sIgA could be and active disease marker, while the increase in IgM levels could represent a marker of recent disease.

Hematologic values among warao indians with tuberculosis from the Orinoco Delta of Venezuela

La tuberculosis (TBC) es un problema de salud pública entre las comunidades indígenas de Venezuela. Este estudio muestra los valores de los parámetros hematológicos de dos poblaciones diferentes con TBC, los Warao y los criollos. Se estudiaron pacientes Warao (PW) y criollos (PC), controles adultos tanto Warao (CW) como criollos (CC) y niños Warao (NW). Los PW y los NW presentaron valores significativamente bajos de la hemoglobina (Hb) en comparación con los CC y CW. Todos los grupos Warao presentaron concentraciones significativamente disminuidas de los valores de la concentración de hemoglobina corpuscular media (CHCM) en comparación con los PC y CC. En cuanto a los glóbulos blancos, los grupos Warao, incluyendo los niños, presentaron un incremento significativo del porcentaje de monocitos y linfocitos en comparación con los PC y CC. Sin embargo, el porcentaje de los neutrófilos se encontró significativamente disminuido en comparación con ambos grupos criollos. Ambos grupos de pacientes así como los niños Warao presentaron incremento significativo en el número de plaquetas en comparación con los CW y CC. Los resultados del índice de masa corporal (IMC) en la población Warao, el cual fue utilizado como indicador del estado nutricional de la población mostraron que, éste alcanzó el máximo en el grupo correspondiente a la edad de los adolescentes y luego disminuyó a medida que aumentaba la edad de los individuos. En conclusión, tanto los PW como los NW presentan disminución de los valores de la hemoglobina, la cual se correlacionó con una hipocromía eritrocitaria. Al igual que los NW, los adultos Warao, aún en ausencia de enfermedad presentaron una fórmula leucocitaria caracterizada por monocitosis, aumento del porcentaje de linfocitos y disminución del porcentaje de neutrófilos

Tuberculosis en menores de 15 años en la población Warao de Venezuela / Tuberculosis in subjects under 15 years of age in the population of Warao in Venezuela

It is difficult to establish a definitive diagnosis of tuberculosis in rural areas where there is no access to a large hospital. The Warao people of the Delta Amacuro State in Venezuela, have a very high prevalence of adult TB, and we suspected that the Warao children would also have a high prevalence of the disease, almost entirely undiagnosed. We applied a simple methodology to select children suspicious for tuberculosis that is based on a rating system using clinical criteria, reactivity to tuberculin and intradomicilliary contacts. Of the 502 children under the age of 15 that were evaluated with this rating system, 27 were determined to be suspicious of TB and were further evaluated by a chest X-ray. Radiologic confirmation of TB was found in 16 (60) of the 27 suspicious children. Of these 16 patients, 13 (81) were PPD positive and 3 were PPD negative. Additionally, 7 of the 16 children with pathologic x-ray changes had one or more confirmatory findings: 3 were positive by culture or smear examination and 5 had a positive serologic B diagnostic test. In conclusion this methodology proved to be highly efficient in diagnosing childhood tuberculosis in this population, and should also be useful in other rural populations with a high prevalence of adult TB.

Characterization of the cellular immune response in chagas'disease patients / Caracterización de la respuesta inmunológica celular en pacientes con la enfermedad de chagas

The in vitro lymphocyte proliferative responses and some mechanisms possibly involved in immunoregulation, were evaluated in 57 patients with Chagas'disease. The patients were classified as being cardiomyophathic (CDM), with positive complement fixation tests (CFT) for T. cruzi, or as infected chagasic patients (INF) with a positive CFT but without any clinical evidence of cardiac involvement. The in vitro responses to T. cruzi antigens were significantly greater in both groups of patients than in the controls, the intensity of the reactions being greatest in CDM patients. Trypanosoma cruzi antigen-induced suppression of in vitro mitogen responsiveness was not detected in either group of Chagas'patients. The mitogenic responses to PHA of CDM patients were,in fact,significantly lower than those of healthly controls. Con A responses were normal in these patients,as was also the case in INF patients for both mitogens. Cell mixingexperiments, showed an augmentation in responsiveness to PHA by co-cultures of mononuclear cells from CDM patients with those from healthtly controls. This was not observed in combinations of cells from controls and INF patients. Due to thepossible modulatory influences of circulating immune complexes,these were also evaluated,higher levels being found in INF and CDM patients than in controls. Thesignificance of these results in the physiopathology of Chagas'is discussed