Human thymic hormones increase in vitro IL-4 production in atopic patients.

BACKGROUND: There is little information about the relationship between thymic hormones and atopy. METHODS: Human thymostimulin was obtained from thymus of children who died in car crashes. These polypeptides were purified by a Sephadex G-50 column fractionation and incubated in vitro with human lymphocytes obtained from atopic and non-atopic subjects of different ages. The SDS-PAGE revealed at least the presence of three broad bands of proteins with 20, 30 and 60 kDa of molecular weight approximately. Levels of IL-4 from lymphocytic cultures were measured by ELISA and correlated with atopic and non-atopic status and with age. The non-atopic controls showed 5.20 UI/ml +/- 1.14 UI/ml of IL-4 meanwhile the non-atopic cells stimulated showed 8.15 UI/ml +/- 2.438 UI/ml. On the other hand, the atopic cells revealed a spontaneous release of 12 +/- 1.812 UI/ml meanwhile those stimulated by the thymostimulin showed 18.53 UI/ml +/- 1.40 UI/ml. RESULTS: Thymic polypeptides were able to increase the levels of IL-4 in both groups although the atopic subjects showed the greater increase (p > 0.001) independently of their age. CONCLUSIONS: As it has been suggested that these hormones could be used therapeutically in atopic subjects, our results warn about the adverse effects that could be produced with them.

Proteinase and gelatinolytic properties of a Triatoma infestans extract.

An extract of Triatoma infestans has previously been demonstrated to produce specific IgG and IgE both in animals and in atopic humans with rhinitis/asthma as well as hypersensitivity pneumonitis in guinea pigs aerosolized with T. infestans. We attempted to determine whether the antigen or antigens responsible belonged to the protease group, as occurs with other allergens such as house dust mites and cockroaches. To do this, T. infestans was studied by SDS-PAGE, Western blots and gelatinolysis with and without the use of specific protease inhibitors such as E-64, TLCK, TPCK, PMSF, leupeptin, o-phenanthrolene and pepstatin-A. These assays revealed serine-like proteolytic and gelatinolytic activities. The presence of 10 to 12 bands of between 14 and 100 kDa was detected. The proteolytic activity pattern of T. infestans was greatest at pH 8.5 and gelatinolytic activity was highly sensitive to PMSF, suggesting that this enzyme could be characterized as a serine protease. Western blots revealed that two bands of 17 and 58 kDA reacted with the sera of atopic humans with respiratory diseases and anti-IgE. However, whether these bands correlated with allergenicity is unclear since the presence of several proteins in each of these bands does not rule out the possibility that this correlation could exist, especially because cross-reactions with antigens from the cockroach Periplaneta americana and its specific antiserum in animals and atopic humans have been demonstrated. The role of proteases in the etiopathogenesis of perennial rhinitis and bronchial asthma in inhabitants of the area of Argentina infested by T. infestans requires further investigation.

Proteinase and gelatinolytic properties of a bat feces extract.

It was previously demonstrated that a bat feces extract (BAT) was able to produce a specific IgG in animals, a specific IgE in respiratory atopic humans and a hypersensitivity pneumonitis in guinea pigs. As numerous allergens (such as house-dust mite, cockroaches and pollens) revealed a enzymatic activity measured by different assays we decided to study the proteinase and the gelatinolytic activities of the BAT. Several protease inhibitors such as E-64, TLCK, TPCK, PMSF, leupeptin, o-phenantroline and pepstatin-A were applied to establish the chemical properties of the enzymatic activity. These assays revealed a serine-trypsin-like proteolytic and gelatinolytic activities specially at pH 8,5. On the other hand, two bands of 21 and 40 kDa reacted with the human atopic sera suggesting a possible correlation between allergenicity and proteinase activity. Their role in the etiology of perennial rhinitis and asthma requires further investigations.

[Hypersensitivity pneumonitis induced in guinea pigs by antigens to bat feces]. / Neumonitis por hipersensibilidad inducida en cobayos por los antígenos de las heces del murciélago.

A bat feces extract was able to induce the synthesis of specific IgG in rabbits and IgE in atopics. These findings were corroborated by skin tests, RAST and RAST-inhibition. Nowadays, data concerning the experimental induction of hypersensitivity pneumonitis using a glycoprotein of bat feces are presented. This antigen was aerosolized for inhalation by adult guinea pigs for 12 weeks to detect specific serum IgM, IgG and IgE antibodies as well as sensitized T-CD4 cells. Histopathological studies of the lungs showed interstitial infiltrates of macrophages and lymphocytes, cellular bronchiolitis and single non-necrotizing granulomas from the seventh to the ninth weeks. From the tenth week to the end of the experiment the lesions of the lungs progressively worsened. The results from this animal model suggest that the chronic contact with the bat feces'antigen can induce other inflammatory lung reactions than those IgE dependant.

[Hypersensitiviy to Trichophyton rubrum antigens in atopic and non-atopic podiatrists]. / Hipersensibilidad a los antígenos del Trichophyton rubrum en podólogos atópicos y no atópicos.

BACKGROUND: Podiatrists as well as manicurists are exposed to inhale nail dust contaminated with mycotic particles when filing and burring the onychomycotic nails of their patients. As some of them with atopic background suffered worsening of their symptoms we decided to study the immune response to the fungus Trichophyton rubrum (Tr) that was isolated from the nail dust obtained from the podiatrists office. METHODOLOGY AND RESULTS: A detailed clinical record, intracutaneous skin tests with a extract of Tr and the serum levels of total and specific IgE were performed in rhinitis-asthma as well as control patients. As the asthmatic group refused to perform the challenge bronchial test with Tr we developed a guinea pig experimental model with daily aerosolization of Tr during 12 weeks studying the levels of specific IgE and IgG as well as the lung's histopathology. Atopic patients showed positive immediate skin tests with Tr and both groups revealed delayed hypersensitivity to the antigen.RAST-IgE-anti-Tr and RAST-inhibition confirmed the specificity of the antibodies. Guinea pigs also synthetized IgG and IgE anti-Tr and suffered different degrees of lung lesions similar to those of hypersensitivity pneumonitis. CONCLUSION: Atopic podiatrists are exposed to fungal allergens that may participate or aggravate their previous respiratory conditions.

Immunological changes during cockroach immunotherapy.

Data obtained in a 3-year survey of specific immunotherapy (SIT) with a Periplaneta americana antigen (Pa-1) are presented. Parameters such as serum IgE-paperadioimmunosorbent test, specific IgE and IgG, interleukin (IL)-2, IL-4 and IL-4R levels were recorded before and after SIT. While serum IgE levels and IgE-RAST-anti-Pa-1 did not change throughout SIT (p = not significant), IgG-RAST-anti-Pa-1 showed a marked increase from the first year (p < 0.002 to p < 0.001). Only after 3 years of SIT did the serum levels of IL-2, IL-4 and IL-4R show lower values than before this period (p = 0.05, p = 0.05, p = 0.01, respectively). The comparative statistical analysis of the cytokine data between the nonatopic subjects and the atopic treated patients revealed no significant differences (p = 0.02). The symptomatic scores showed significant results at the third year of SIT in sneezing attacks, nose blowing and nasal obstruction (p = 0.001, p < 0.001, p = 0.001, respectively).

Proteinase and gelatinolytic activities of house dust mite and cockroach extracts.

The presence of gelatinolytic activity in dust mite and Periplaneta americana allergenic crude extracts were studied. The former presented major activity in a broad band between 45 and 66 kDa and minor activity at 32 kDa, while the latter showed a more complex pattern with gelatinolytic activity at 90, 78, 65, 34, 32 and 24 kDa. When the proteolytic activity patterns of dust mites and cockroach crude extracts were analyzed at three different pH levels, the proteases in both cases were optimally active at pH 6, showed no activity at pH 3.5 and little activity at pH 8.5. The susceptibility of both extracts to a set of well-known protease inhibitors suggested that they are composed of cysteine and serine proteinases, the latter probably being a trypsin-like type. When immunochemical properties were studied, dust mite bands of about 200, 110, 65, 60 and 43 kDa showed immunoreactivity against a polyclonal human anti-dust mite serum, with the band of approximately 200 kDa presenting the highest antigenicity. A similar analysis was applied to the cockroach extract, which exhibited immunoreactive bands at 90, 78, 65 and 34 kDa when incubated with a polyclonal rabbit anti-Blatta serum. Only those of 90, 78 and 65 kDa reacted against a polyclonal human anti-Blatta serum. These results suggested a correlation between some proteases with gelatinolytic activity and the allergenicity of both extracts.

Hypersensitivity pneumonitis induced by Penicillium notatum antigens in guinea pigs.

Data concerning the experimental induction of hypersensitivity pneumonitis in guinea pigs using a glycoprotein derived from Penicillium notatum are presented. This antigen was obtained from the mycelial and metabolic products of the cultures passed through a Sephadex G-50 column. It was aerosolized for inhalation by adult guinea pigs for 12 weeks to detect specific serum IgM, IgG and IgE antibodies as well as sensitized leukotriene CD4 cells. Histopathological studies of the lungs showed interstitial infiltrates of macrophages and leukotriene cells, cellular bronchiolitis and single non-necrotizing granulomas. The results from this animal model suggest that this hypersensitivity pneumonitis is a typical delayed-type reaction due to chronic contact with the heterologous glycoprotein of Penicillium.

Interstitial pneumonitis induced in guinea pigs by the antigens of Rhizopus nigricans.

Rhizopus nigricans (Rn) is one of the most common members of the Mucorales that produces opportunistic infections and hypersensitivity states. Data concerning experimental induction in guinea pigs of hypersensitivity pneumonitis with a glycoprotein antigen are presented. This antigen was obtained from the mycelial and metabolic products of the cultures and was aerosolized during 12 weeks. The presence of specific antibodies (IgG and/or IgE) was detected by serological techniques; histopathological studies of the lungs showed interstitial infiltrates of macrophages and LTCD8+ cells, as revealed by the MoAb used. Single non-necrotizing granulomas were characteristic from the tenth week to the end of the experiment. The results from this animal model suggest that hypersensitivity pneumonitis is a typical delayed-type reaction due to chronic contact with the heterologous glycoprotein of Rn. The relation of Rn antigen and the development of occupational diseases of the lung such as malt-worker's lung and wood-trimmer's disease is proposed and discussed.

Cross reactivity between the antigens of Periplaneta americana and Triatoma infestans.

The cockroach Periplaneta americana (Pa) and the reduviid Triatoma infestans (Ti) are two very common insects in Argentina that pollute indoor and outdoor environments. They are also strongly immunogenic in animals and in atopic men. Several immunological methods such as serological absorption procedures, diffusion gel methods and capillary electrophoresis were applied in order to establish the existence of common glycoprotein epitopes in the cockroach and in the reduviid. The presence of specific antibodies (IgG & IgE) were detected in rabbits as well as in human sera. The RAST and RAST-inhibition confirmed the cross-reactivity between these two antigens. Capillary electrophoresis evoked similar patterns among Pa, Ti and the moults of the nymphs of Ti. All these findings have clinical and epidemiological importance.

Interstitial pneumonitis induced in guinea-pigs by the antigens of Periplaneta americana.

Data concerning the experimental induction in guinea-pigs of hypersensitivity pneumonitis with a cockroach antigen are presented. A glycoprotein obtained from the chitinous structures of Periplaneta americana was aerosolized daily to guinea-pigs during 12 weeks. The presence of specific antibodies (IgG) was detected by serological techniques; histopathological studies of the lungs showed interstitial infiltrates of macrophages and LT-CD8+ cells, as revealed by the MoAb used. Single non-necrotizing granulomas were characteristic from the 10th week to the end of the experiment. The results from this animal model suggest that this hypersensitivity pneumonitis is a typical delayed-type reaction due to chronic contact with the heterologous glycoproteins of P. americana.

Papel de la Blattaria sudamericana en la rinitis y asma alérgicos / Role of Blattaria sudamericana in rhinitis and allergic asthma

Entre los factores que producen la sensibilización del organismo humano, un lugar destacado lo ocupan los componentes del polvillo habitacional. Este incluye un sistema ecológico complejo cuya composición integran productos orgánicos de origen animal y vegetal así como elementos onorgánicos. La presencia de un ácaro del género Dermatophagoides pteronnysinus en dicho polvillo constituyó un hallazgo trascedental que impulsó la investigación hacia la búsqueda de productos provenientes de insectos que indujeros sensibilización por vía inhalatoria. Pareció de interés estudiar las propriedades inmunoquímicas de la cucaracha Blattaria sudamericana, insecto muy común en la Argentina, y valorar su papel en las alegrias respiratórias. De tal manera, un extracto de dicho insecto con 42 mg/ml de proteínas fue pasado por columnas de Sephadex G-200 y de DEAE celulosa cuantificándose sus fracciones proteicas y azucaradas a 280 nm y a 470 nm respectivamente por espectrofotometría. A su vez, dicho extracto fue inyectado a conejos juntamente con el adyuvante de Freund durante 13 semanas lográndose la producción de anticuerpos circulantes del tipo IgG, precipitantes y hemaglutinantes revelados por Outchterlony, inmunoelectroforesis y hemaglutinación pasiva. Los cotenidos proteicos medidos por la técnica del Lowry fueron elevados por las diferentes fracciones obtenidas al igual que los de hexosas lo que movió a sospechar la composición glucoproteica de los antígenos mayores. El peso molecular del extracto de Blattaria fue establecido en 180.000 daltons por comparación con marcadores proteicos estandardizados...

Papel de la Blattaria sudamericana en la rinitis y asma alérgicos / Role of Blattaria sudamericana in rhinitis and allergic asthma

Entre los factores que producen la sensibilización del organismo humano, un lugar destacado lo ocupan los componentes del polvillo habitacional. Este incluye un sistema ecológico complejo cuya composición integran productos orgánicos de origen animal y vegetal así como elementos onorgánicos. La presencia de un ácaro del género Dermatophagoides pteronnysinus en dicho polvillo constituyó un hallazgo trascedental que impulsó la investigación hacia la búsqueda de productos provenientes de insectos que indujeros sensibilización por vía inhalatoria. Pareció de interés estudiar las propriedades inmunoquímicas de la cucaracha Blattaria sudamericana, insecto muy común en la Argentina, y valorar su papel en las alegrias respiratórias. De tal manera, un extracto de dicho insecto con 42 mg/ml de proteínas fue pasado por columnas de Sephadex G-200 y de DEAE celulosa cuantificándose sus fracciones proteicas y azucaradas a 280 nm y a 470 nm respectivamente por espectrofotometría. A su vez, dicho extracto fue inyectado a conejos juntamente con el adyuvante de Freund durante 13 semanas lográndose la producción de anticuerpos circulantes del tipo IgG, precipitantes y hemaglutinantes revelados por Outchterlony, inmunoelectroforesis y hemaglutinación pasiva. Los cotenidos proteicos medidos por la técnica del Lowry fueron elevados por las diferentes fracciones obtenidas al igual que los de hexosas lo que movió a sospechar la composición glucoproteica de los antígenos mayores. El peso molecular del extracto de Blattaria fue establecido en 180.000 daltons por comparación con marcadores proteicos estandardizados...(AU)

Immunochemical properties of the antigens of Blattaria sudamericana.

An extract of the cockroach Blattaria sudamericana (BS), a very common insect in Argentina, was passed through Sephadex G-200 and DEAE cellulose ion exchange columns. This method yielded 2 proteins and 6 hexoses whose quantities were determined by absorbance in a LKB Uvicord spectrophotometer at 280 nm and 470 nm respectively. Protein contents were also recorded by the Lowry method and the micro-Kjehldal technique. Adult albino rabbits were immunized with a mixture of BS and complete Freund's adjuvant over a period of thirteen weeks. The antisera obtained were studied by the Ouchterlony method, the immunoelectrophoresis and the passive hemagglutination technique against BS and the eluated fractions with the highest protein content. Molecular weights were studied with the usual marker proteins subjected to gel filtration by a Sephadex G-200 column. Each protein concentration was 13.5 mg/ml in a volume of 1.5 ml meanwhile BS was 42 mg/ml. This antigen showed a molecular weight of 180,000 daltons. Fifty untreated atopic patients of both sexes with ages between 19 and 74 years suffering perennial rhinitis and bronchial asthma were chosen according to the American Thoracic Society criteria. Their IgE levels ranged between 110 and 3000 KU/L with an average of 620 KU/L. IgE anti-BS-RAST showed positive results in half of the patients while type I positive skin tests with BS 1:100 appeared in 62% of the atopic group.