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Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
Published on: November 23, 2010
[Immune response and anergy. Study in tuberculosis patients from the Universitary Hospital, Maracaibo, Venezuela ]
Milagros Montiel1, Sergio Rivera, Rosaura Hernández
1Universidad del Zulia.
Abstract:
With the aim of evaluate the anergy in Mycobacterium tuberculosis individuals infected we study 57 patients and 15 healthy controls, both sex and with different age, which to practice Sputum Bacteriology culture (BK) and the la reaction intradermic with PDD. The test of tuberculine on skin was achieve inoculate 0.1 ml de PPD intradérmic vía, the results were obtained at 72 h. 28 patient total result BK+ and 29 BK-. The Group BK+, 15 result PPD+ and 13 PPD-, the lastet were classify as anergic patients (BK+ PPD-). From the 29 patient residual (BK-), 20 was PPD+ and 9 PPD-. All the controls resulted BK- PPD-. Only the anergy in patient BK was evaluated + (28 individuals) patient. Was classified from the point of clinical view in three series: N series: 27 patient TBC+ BK-, P Series: 28 patient TBC+ BK+ and EP Series: 2 patient with extra pulmonary TBC. According to the grade of pulmonary complication, the patient separated in two groups, according to the extension and type of lesion found in the evaluation with X. rays The "group 1" with 9 patient (32.14%) it introduced lesions on one side with or without caverns and the "group 2" with 19 patient (67.85%) with bilateral lesions with or without caverns. Was carried out the Lymphoblastic Transformation Test (TTL) stimulating with PHA and PPD, for which patterns of outlying blood with heparina were gathered. The concentrations of IL-4, IL-6 and INF-gamma was gotten inmunoenzimatic in supernatants of cultivations for rehearsals and in plasm, hind to the stimulation with the PPD. The statistical analysis was carried out utilizing the Test T of Student in pair, Varianza Analysis and the square Chi. Only the controls and the patient BK-, they showed normal response to the PHA, in contrast, the patient BK+ PPD+ they introduced a suppression of the response opposite the PHA of aprox. 90%. The patient PPD+ they showed a superior response in cultivation that the patient PPD-. The concentration of IL-4 in the plasm of the anergic patient (PPD-) showed a significant difference (P < 0.05) with concerning the PPD group+. The concentration of INF-gamma was lower in the anergic patient significantly different to the of the patient PPD+. Similar result was gotten in the mensuration of IL-4 and INF-gamma in supernatants of cultivation. One could observe the presence of patient with IL-4+ INF-gamma, and INF-gamma or IL-4 only. In the patient classified clinical like series "P", the concentration of IL-4 was old in the "group 2" with concerning the "group 1." the opposite occurred with the concentrations of the INF-gamma which were old in the "group 1" (both differences were not significant statistical). The increase of IL-4 next to low concentrations of INF-gamma in anergic patient and committed clinical, it suggest a prevalence of the pattern Th2 (T2) in the immune response.
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