In vitro synthesis of interferon-gamma, interleukin-4, transforming growth factor-beta and interleukin-1 beta by peripheral blood mononuclear cells from tuberculosis patients: relationship with the severity of pulmonary involvement.

Given the role of cell-mediated immune responses in resistance to mycobacteria, we sought to analyse whether there was a relationship between the severity of pulmonary tuberculosis (TB) and lymphocyte proliferation as well as in vitro cytokine production. To achieve this, 25 untreated TB patients showing mild (n = 5), moderate (n = 9) or advanced (n = 11) pulmonary disease, and 12 age-matched healthy controls (mean+/-SD, 37+/-14.5 years) were studied. Peripheral blood mononuclear cells were cultured for 5 days with 10 microg/ml whole, sonicated Mycobacterium tuberculosis (WSA) or 2.5 microg/ml Concanavalin A (Con A). Supernatants were collected on day 4, from cultures grown with or without WSA, for measurement of interferon-gamma (IFN-gamma), interleukin (IL)-4, IL-1beta and transforming growth factor-beta (TGF-beta). Antigen-specific proliferation was found to be reduced among patients and more profound in those with advanced disease who also displayed a depressed response to Con A. Patients with mild TB showed a preferential production of IFN-gamma over IL-4, gave the highest level of IFN-gamma synthesis upon specific antigen stimulation and showed increased levels of IL-1beta production. Findings in patients with moderate TB appeared compatible with a mixed production of IFN-gamma and IL-4 coexisting with a higher synthesis of TGF-beta, by comparison to patients with mild TB. Advanced disease showed the highest IL-4 and TGF-beta production, with IFN-gamma synthesis readily noticeable, yet decreased in comparison with the other patient groups. Differences in cytokine response according to the amount of lung involvement suggest a role for such mediators in the immunopathogenesis underlying the distinct clinical forms of pulmonary TB, that is a predominant T helper Th)1-like or Th2-like activity in mild or in progressive TB, respectively.

Circulating profile of Th1 and Th2 cytokines in tuberculosis patients with different degrees of pulmonary involvement.

To investigate whether differences in the degree of pulmonary tuberculosis lesions could be accompanied by changes in the pattern of circulating cytokines, 29 untreated tuberculosis patients showing mild (n = 10), moderate (n = 5) or advanced (n = 14) pulmonary disease, and 12 age-matched healthy controls (mean +/- S.D., 36 +/- 15 years) were studied. ELISA methods for the evaluation of interferon-gamma, interleukin-2, interleukin-4, and interleukin-10 indicated that all patients had increased serum levels of the four cytokines in relation to controls. Mean titers of interferon-gamma and interleukin-2 in mild and moderate patients appeared higher than in those with advanced disease, whereas moderate and advanced patients showed the higher levels of IL-4 in comparison to mild cases. Raised levels of interleukin-10 were more prevalent in advanced disease, and statistically different from those in mild patients. This cytokine pattern may help to explain findings wherein mild tuberculosis is characterized by preserved cellular immune responses while advanced disease is accompanied by an impairment of such parameters.

Levels of interleukin-8 in tuberculous pleurisy and the profile of immunocompetent cells in pleural and peripheral compartments.

Our study investigated the presence of IL-8 in pleural exudates from tuberculosis patients (TBP) (n = 13), and evaluated whether it was related with the profile of major immunocompetent cells present in their pleural and peripheral compartments. To allow comparisons, an additional group of patients with parapneumonic pleural effusions (PNE) (n = 7) was included. Blood peripheral immunophenotypic studies were also carried out in 12 age-matched healthy controls (Co), and 39 tuberculosis patients classified, according to the extent of pulmonary involvement, into mild (n = 9), and advanced (n = 30) cases. Patients were recruited before starting therapy, had HIV negative serology, and showed no age differences among groups (mean +/- SD., 40.7 +/- 14.7 years). IL-8 concentrations were measured by an ELISA method while immunophenotypic analysis was performed by using FITC-conjugated monoclonal antibodies reacting against the following cell surface molecules: CD3, CD4, CD8, CD25 (IL-2R+ cells), CD19, and CD68. IL-8 was detected in all pleural exudates though levels in the TB patients, 384 +/- 110 pg/ml, appeared significantly higher than the PNE group, 185 +/- 110 pg/mg, (P < 0.015, mean +/- S.D.). In turn, the former group presented values of pleural CD3+, CD4+, and CD25, which were found increased in comparison with PNE patients (P < 0.01). Unlike the pleural compartment, patients with TBP showed a marked and significant decrease in their circulating levels of cells bearing the CD3, CD4, CD19, CD25, and CD68 phenotypes not only when comparing with Co but also with PNE and mild patients. Differences between the levels of pleural and peripheral T-cells from TBP patients may be the reflection of an important influx of T-lymphocytes from the circulatory system to the pleural cavity, probably linked to the presence of chemotactic factors within the pleural fluid like IL-8.

Adjvant effect of Staphylococcus aureus in rats infected with Salmonella Typhimurium

Estudou-se a açäo imunomoduladora do Staphylococcus aureus em ratos infectados com Salmonella typhimurium. O trabalho foi desenvolvido em duas etapas. 1- a multiplicaçäo de S. typhimurium foi observada em vários órgäos de animais pré-tratados com S. aureus. Ratos adultos endocriados receberam inoculaçäo subcutânea de 8x10(6) S.aureus e, sete dias após, receberam uma dose i.p. subletal (1,5 X 10.7) de Salmonella Typhimurium (Grupo I). Animais de um segundo grupo foram infectados somente com Salmonella typhimurium (mesma dose) e usados como controles infectados (grupo II). Os ratos foram sacrificados nos dias 1, 3, 6 e 9 após a infecçäo. Amostras de fígado e baço foram pesadas, homogeneizadas e, após diluiçöes adequadas, cultivadas em agar MacConkey. Após 24 h, contou-se o número de colônias no sangue, baço e fígado. O grupo I apresentou contagens significantemente mais baixas que o grupo II durante o período de experimentaçäo. 2- Os números absolutos e as proporçöes de macrófagos (M) e linfócitos (L) foram determinados em vários órgäos dos animais pré-tratados (grupo I) e nos controles infectados (grupo II). Após a infecçäo, os ratos receberam uma injeçäo intravenosa de 0,1 ml de Ferro Dextran (Fe 100 mgml) para coloraçäo diferencial dos macrófagos. 24 h após os animais foram sacrificados removendo-se o fígado, baço, intestino e gânglios linfáticos mesentéricos para análise histológica. Controles näo infectados foram também incluidos para comparaçäo (grupo III). Para avaliaçäo, os cortes histológicos foram corados com H-E e tratados segundo a técnica histoquímica de Bugelsky. Os números de Ms e de Ls foram mais altos no grupo I quando comparados com os valores obtidos nos grupos II e III. Os resultados das contagens, em ordem decrescente, foram: gânglios linfáticos mensentéricos (zona folicular > zona interfolicular), baço (polpa branca > polpa vermelha), intestino e fígado, sendo que no intestino (Placas de Peyer) houve predominância de Ms e relaçäo a Ls. Estes resultados confirmam o efeito adjuvante de S.aureus

Low occurrence of arthritic manifestations in patients with pulmonary tuberculosis. T cell subsets and humoral studies.

Given the suspected role of mycobacteria in the establishment of disorders with an autoimmune background and joint damage, a study was conducted to analyze whether rheumatic symptoms were likely to be present in tuberculosis (TB) patients. To this end, 330 patients with a bacteriologic confirmation of tuberculosis were investigated for the presence of arthritic complaints. The latter were recorded in five of them with rheumatic symptoms mostly involving interphalangeal and metacarpophalangeal joints, and preceding the clinical manifestations of the TB illness. Three out of these five patients remained arthritic by the time of the bacteriologic conversion and fulfilled the criteria for the diagnosis of rheumatoid arthritis. In the two remaining patients sputum negativization was accompanied by a disappearance of rheumatic manifestations. These patients were also assessed for their peripheral levels of major T cell subsets as well as for the presence of autoantibodies. Comparisons with a series of non-arthritic TB cases, rheumatoid arthritis patients, and controls revealed that presence of rheumatic manifestations was associated with a different profile of autoantibody formation and T cell subset changes. Evidence recorded in the present study indicates that joint affectation in TB is a rare event, being rather the exception than the rule.

[Inhibition of total and active E-rosettes mediated by serum from tubercular patients]. / Inhibición en la formación de rosetas E totales y activas mediada por el suero de pacientes tuberculosos.

AIM: To study the effect of tuberculous patients' serum upon normal lymphocytes in order to determine possible inhibition factors of immune response. PATIENTS AND METHODS: We studied total E-rosette formation (4 degrees C, RET) and active rosette formation (37 degrees C, REA) in tuberculous patients in different stages: mild (n:36), moderate (n:28), severe (n:24) and a control group of 32 patients. The same determinations were studied in normal circulating lymphocytes (LN) incubated before with serum from tuberculous patients (SP), serum of normal individuals (SN) and in Tc-199 medium. RESULTS: The number of RET and REA in tuberculous patients of different stages were significantly lower to the ones of the control group (p less than 0.01, p less than 0.05). This differences were also found when we compare the number of RET and REA after the incubation of LN with SP and SN respectively (p less than 0.01, p less than 0.05). SUMMARY: Tuberculous patients showed an impaired ability to form RET and REA. The incubation of normal lymphocytes with serum from tuberculous patients reduced significantly its capacity to form rosettes.

Evaluacion de las subpoblaciones de linfocitos T en pacientes tuberculosos empleando la modulacion con teofilina / T-lymphocyte subsets evaluation in patients with tuberculosis using modulation with theophylline

Se evaluaron las poblaciones y subpoblaciones linfocitarias en pacientes con tuberculosis pulmonar antes y durante la terapia relacionando estos valores con la incidencia y evolucion de la enfermedad. Pacientes en sus diversas manifestaciones clinicas, virgenes de tratamiento, se estudiaron por baciloscopia (BAAR), radiologia, i.d.r. Mantoux y analisis complementarios. Se cuantificaron mediante la prueba de Rosetas espontaneas (RE) linfocitos T totales y activos (RE a 4 graus Celsius y 37 graus Celsius), T colaboradores (RE Teofilina Resistentes: RETR) y supressores (RE Teofilina Sensibles: RETS). Los examenes se repitieron en los mismos sujetos iniciado el tratamiento y en testigos sanos (TS). Se demostro en los pacientes en todas sus formas clinicas un descenso significativo en los valores relativos y absolutos de celulas T y en la relacion RETR/RETS (menor de 1). Existe asociacion entre la forma clinica y el numero de linfocitos T colaboradores. Los pacientes en tratamiento con evolucion favorable, evidenciaron un incremento significativo en los linfocitos T totales, activos, colaboradores y en la relacion RETR/RETS. Los enfermos con baciloscopia altemente positiva presentaron i.d.r. Mantoux baja o negativa y marcado descenso de celulas inmunocompetentes...

[T-lymphocyte subsets evaluation in patients with tuberculosis using modulation with theophylline]. / Evaluación de las subpoblaciones de linfocitos T en pacientes tuberculosos empleando la modulación con teofilina.

T cells and T cells subsets in peripheral blood of patients with different forms of pulmonary tuberculosis were evaluated to explain some aspects of the immunocompromised state of these subjects. Diagnosis was made by baciloscopy (BAAR), chest roentgenography i.d.r. Mantoux, and other clinical analysis. Spontaneous E-Rosette test (RE) was used to quantify Total (RET 4 degrees C) and Active T cells (REA 37 degrees C) and the same test after incubation with Theophylline (The) for helper cells (The-resistant cells: RETR) and suppressor cells (The-sensitive cells: RETS). Patients were followed for at least 4 months after therapy. The data demonstrate a significant decrease of relative and absolute numbers of Total T-cells and a diminished T helper/T suppressor subset ratio (RETR/RETS) which dropped to less than 1 in untreated patients. Treated patients with a favourable evolution showed a restoration of Total active and helper T cells. RETR/RETS ratio was also significantly increased. In patients with highly positive BAAR, low on negative i.d.r. Mantoux, a decreased level of immunocompetent cells was observed. The 3 aspects were associated. Nutritional condition of the patients was also associated with the predisposition to acquire this disease.

[Inhibitory effect of Staphylococcus aureus on the growth of rat sarcoma]. / Efecto inhibidor de Staphylococcus aureus sobre el crecimiento de un sarcoma en ratas.

We have studied the effect upon the growth of a transplantable rat sarcoma, of an intraperitoneal inoculation of peritoneal exudate cell (PEC), spleen cells (SC) and non-adherent spleen cells (N-ASC) obtained from S. aureus previously inoculated rats and the same cells from normal rats (NPEC, NSC, NN-ASC). Inbred, adult rats were inoculated with 800 x 10(6) bacteria, killed by tyndalization. After 7 days, treated and normal animals were sacrificed and PEC, SC and N-ASC were obtained. Different groups of animals were inoculated with these cell populations. Simultaneously the rats received a s.c. inoculum of a transplantable sarcoma (S-E 100). Tumor size was measured on days 4, 7, 14, 21 and 28 after tumor challenge. A significant tumor growth inhibition was found with all three cell populations, but no effect was observed with normal cells. Tumor size on different days and tumor growth curves clearly demonstrate this effect. We conclude that Staphylococcus aureus inoculum as well as the cells from animals previously challenged with the bacteria induce tumor growth inhibition. The possibility that protein A of S. aureus may be involved in this phenomenon, or that the mechanism of tumor growth inhibition is mediated by an activation of different cell populations is discussed.

Efecto inhibidor de Staphylococcus aureus sobre el crecimiento de un sarcoma en ratas / Inhibitory effect of Staphylococcus aureus on the growth of rat sarcoma

Se investigó el efecto modulador de la respuesta antitumoral de S. aureus y diversas poblaciones celulares en ratas portadoras de un sarcoma transplantable (S-E 100). Se comprobó en estas experiencias la capacidad inhibitoria del crecimiento del S-E 100 en ratas endocriadas, a través de la inoculación i.p. de células de exudado peritoneal (CEP), células de bazo (C.B.) y células de bazo no-adherenrtes (CBN-A), provenientes de ratas previamente desafiadas con S. aureus. También se inocularon i.p. células de animales normales (CEPN, CBN, CBN-AN). Se midió el tamaño del S-E 100 en los días 4, 7, 14, 21 y 28 después de iniciado el tratamiento. Se determinó que S. aureus, así como las poblaciones celulares provenientes de animales previamente inoculados con el germen (CE, CB, CBN-A), inhibe significativamente el crecimiento del tumor. Las células de animales normales no inciden en tal efecto. Se discute si este fenómeno es producido por la proteína A de S. aureus o a través de la activación de las poblaciones celulares (linfocitos, macrófagos, NK) inoculadas

Efecto inhibidor de Staphylococcus aureus sobre el crecimiento de un sarcoma en ratas. / [Inhibitory effect of Staphylococcus aureus on the growth of rat sarcoma]

We have studied the effect upon the growth of a transplantable rat sarcoma, of an intraperitoneal inoculation of peritoneal exudate cell (PEC), spleen cells (SC) and non-adherent spleen cells (N-ASC) obtained from S. aureus previously inoculated rats and the same cells from normal rats (NPEC, NSC, NN-ASC). Inbred, adult rats were inoculated with 800 x 10(6) bacteria, killed by tyndalization. After 7 days, treated and normal animals were sacrificed and PEC, SC and N-ASC were obtained. Different groups of animals were inoculated with these cell populations. Simultaneously the rats received a s.c. inoculum of a transplantable sarcoma (S-E 100). Tumor size was measured on days 4, 7, 14, 21 and 28 after tumor challenge. A significant tumor growth inhibition was found with all three cell populations, but no effect was observed with normal cells. Tumor size on different days and tumor growth curves clearly demonstrate this effect. We conclude that Staphylococcus aureus inoculum as well as the cells from animals previously challenged with the bacteria induce tumor growth inhibition. The possibility that protein A of S. aureus may be involved in this phenomenon, or that the mechanism of tumor growth inhibition is mediated by an activation of different cell populations is discussed.

Efecto inhibidor de Staphylococcus aureus sobre el crecimiento de un sarcoma en ratas / Inhibitory effect of Staphylococcus aureus on the growth of rat sarcoma

Se investigó el efecto modulador de la respuesta antitumoral de S. aureus y diversas poblaciones celulares en ratas portadoras de un sarcoma transplantable (S-E 100). Se comprobó en estas experiencias la capacidad inhibitoria del crecimiento del S-E 100 en ratas endocriadas, a través de la inoculación i.p. de células de exudado peritoneal (CEP), células de bazo (C.B.) y células de bazo no-adherenrtes (CBN-A), provenientes de ratas previamente desafiadas con S. aureus. También se inocularon i.p. células de animales normales (CEPN, CBN, CBN-AN). Se midió el tamaño del S-E 100 en los días 4, 7, 14, 21 y 28 después de iniciado el tratamiento. Se determinó que S. aureus, así como las poblaciones celulares provenientes de animales previamente inoculados con el germen (CE, CB, CBN-A), inhibe significativamente el crecimiento del tumor. Las células de animales normales no inciden en tal efecto. Se discute si este fenómeno es producido por la proteína A de S. aureus o a través de la activación de las poblaciones celulares (linfocitos, macrófagos, NK) inoculadas (AU)

Estudio bacteriologico de la cuenca hidrica de la ciudad de Rosario: cuenca del arroyo Ludueña / Bacteriological study of the basin of Rosario city: Ludueña stram basim

El presente trabajo estudió la contaminación bacteriológica de la cuenca hídrica del arroyo Ludueña, que atraviesa la ciudad de Rosario, provincia de Santa Fe, República Argentina. Para realizar dicho estudio se dividió el curso de agua, en seis zonas, desde el ingreso en el límite oeste del municipio, hasta su desembocadura en el río Paraná (límite este). Las zonas fueron elegidas teniendo en cuenta los lugares en los que se vierten los desagües pluviales, emisarios aliviadores, fábricas, zonas de parquización, actividad náutica y recreación y finalmente la desembocadura en el río Paraná. Los resultados obtenidos dl NMP de coliformes totales, NMP de coliformes fecales y NMP de Estreptococus grupo D, no muestran valores diferentes entre si, entre las dos primeras zonas 1 y 2. En la zona 3 se produce un aumento brusco en la cantidad de bacterias en forma significativa diferentes a las otras dos zonas H = 9,84, p < 0,008 atribuibles a los emisarios aliviadores, que si bien fueron construidos para ser desagües pluviales, se le han conectado clandestinamente desagües cloacales. Los valores de las zonas (4, 5 y 6) son prácticamente los mismos que los correspondientes a la zona 3 (H = 0,44). Además en todas las muestras fueron aisladas Pseudomonas seruginosa y en una de ellas, bacterias del género Salmonellas. Estos datos nos revelan una contaminación masiva de origen fecal reciente y remota, a partir de la zona 3. Dado que este curso de agua recorre la ciudad en sus dos terceras partes, de oeste a este, y que parte de la misma, se utiliza como zona de recreación, actividades náuticas, pesca y parquización, alertamos sobre la posibilidad que se produzcan epidemias

Estudio bacteriologico de la cuenca hidrica de la ciudad de Rosario: cuenca del arroyo Ludueña / Bacteriological study of the basin of Rosario city: Ludueña stram basim

El presente trabajo estudió la contaminación bacteriológica de la cuenca hídrica del arroyo Ludueña, que atraviesa la ciudad de Rosario, provincia de Santa Fe, República Argentina. Para realizar dicho estudio se dividió el curso de agua, en seis zonas, desde el ingreso en el límite oeste del municipio, hasta su desembocadura en el río Paraná (límite este). Las zonas fueron elegidas teniendo en cuenta los lugares en los que se vierten los desag³es pluviales, emisarios aliviadores, fábricas, zonas de parquización, actividad náutica y recreación y finalmente la desembocadura en el río Paraná. Los resultados obtenidos dl NMP de coliformes totales, NMP de coliformes fecales y NMP de Estreptococus grupo D, no muestran valores diferentes entre si, entre las dos primeras zonas 1 y 2. En la zona 3 se produce un aumento brusco en la cantidad de bacterias en forma significativa diferentes a las otras dos zonas H = 9,84, p < 0,008 atribuibles a los emisarios aliviadores, que si bien fueron construidos para ser desag³es pluviales, se le han conectado clandestinamente desag³es cloacales. Los valores de las zonas (4, 5 y 6) son prácticamente los mismos que los correspondientes a la zona 3 (H = 0,44). Además en todas las muestras fueron aisladas Pseudomonas seruginosa y en una de ellas, bacterias del género Salmonellas. Estos datos nos revelan una contaminación masiva de origen fecal reciente y remota, a partir de la zona 3. Dado que este curso de agua recorre la ciudad en sus dos terceras partes, de oeste a este, y que parte de la misma, se utiliza como zona de recreación, actividades náuticas, pesca y parquización, alertamos sobre la posibilidad que se produzcan epidemias (AU)

Effect of RNA on the immunological response of rats, induced by Salmonella typhimurium.

Rats treated with RNA from normal rat spleen were challenged 5 days later with Salmonella typhimurium. The primary response was delayed and antibody titers significantly lower than controls were observed. Forty eight hours after the secondary challenge the antibody titers were also lower; this difference disappeared five days later. This study confirms the immunological depression produced by RNA. Moreover, it is demonstrated that the RNA action is transitory and that the recovery of secondary response occurs seven days after the second challenge. This action is probably explained at macrophage level.

Effect of RNA on the immunological response of rats, induced by Salmonella typhimurium.

Se trataron intraperitonealmente ratas hembras con RNA obtenido de bazo de rata normal de la misma linea, las que se inocularon 5 dias mas tarde con Salmonella typhimurium. En la respuesta primaria se observo una demora significativa en la aparicion de los anticuerpos y un titulo significativamente mas bajo que los testigos 48 horas despues de la segunda inoculacion. Esta diferencia desaparecio 5 dias mas tarde. De esta manera demostro que existe una respuesta inmunologica menor debida al acido ribonucleico. Ademas se observo que esta accion es transitoria, recuperandose la respuesta secundaria 7 dias despues de la segunda inoculacion. Esa accion probablemente se debe a bloqueo de los macrofagos