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.

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 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.

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.

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).

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.

Antigenicity of Penicillium notatum in animals and in atopic patients.

A mycelial and metabolic extract of Penicillium notatum (PN) was passed through Sephadex G-50 and DEAE-cellulose columns in order to separate soluble fractions that revealed a complex composition. Their proteins and hexoses were recorded with a LKB Uvicord spectrophotometer and quantified by the Lowry and the indol techniques. On the other hand an animal model was developed in rabbits and an IgG precipitin and hemagglutinating antibodies were obtained. The PN extract as well as those fractions with the higher protein content appeared positive when they were checked against the antiserum by means of Ouchterlony, Boyden and immunoelectrophoresis. The molecular weight of PN was established in 52,000 daltons approximately in comparison with well known marker proteins. Adult human beings suffering perennial rhinitis and bronchial asthma showed positive type I skin tests with PN and its most conspicuous fractions (glycoproteins). A RAST-IgE-anti-PN was prepared following Ceska's procedure and challenged against all human sera. Only 40% of the patients revealed a positive RAST IgE-anti-PN which correlated significatively with the protein fractions (35%) and with the skin tests (43% and 39%, respectively). These results reinforced the idea that PN is a potent antigenic mold both in animals and in humans in whom we detected an IgE specific antibody presumably related to their atopic clinical condition.

[Chemical and immunologic study of pollen from the gramineous plant Lolium perenne]. / Estudio químico e inmunológico del polen de la gramínea Lolium perenne.

In order to study the chemical composition and immunologic reactivity of an extract of rye grass pollen (RGP), aliquots of this antigen were passed through Sephadex G-50 and DEAE-cellulose ion-exchange columns. This method yielded 7 proteins and 10 sugars, whose quantities were determined by absorbance in a LKB Uvicrd 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 RGP and complete Freund's adjuvant over a period of eight weeks. The antisera obtained were studied by the Ouchterlony method and the passive hemagglutination technique against RGP and all the eluated fractions. Skin tests with RGP and its fourteen most conspicuous eluated fractions were performed in 29 untreated atopic patients with seasonal rhinitis and bronchial asthma, high reactivity to a whole grass mixture and RGP and high levels of serum IgE. Results were recorded after 15 minutes for the immediate reaction when the histamine positive control had reached near maximal size and in the morning. Molecular weights were studied with the usual marker proteins subjected to gel filtration by a Sephadex G-50 column. Each protein concentration was 13,5 mg/ml in a volume of 1,5 ml, meanwhile RGP was 12 mg/ml. PRIST and RAST were carried out with human sera to determine IgE levels and specific IgE and IgG anti-RGP antibodies. Ouchterlony showed precipitin lines among the rabbit RGP-antiserum, whole grass mixture, RGP and the fractions from tubes 38 and 359, meanwhile Boyden's technique appeared positive only between the anti-serum and whole grass mixture and RGP (1:256). The other fractions gave negative results as all of the human sera of both groups. Skin tests in the atopics showed type I reactions with whole grass mixture, RGP, proteins no 33 (Sephadex), 25 and 38 (DEAE-cellulose) and sugars no 56, 192 and 360, but negative results in the controls. Molecular weights obtained were for fraction no 13 68.000, for fraction no 28 25.500 and for fraction no 33 18.400 daltons respectively. RAST revealed in 28 sera a IgG anti-RGP before hyposensitization therapy and an IgE anti-RGP of class 2 only in 8 sera and in 10 of class 1. This surprising result needs further investigation to clarify if the ratio IgE/IgG anti-RGP plays any role in the pathogenesis of pollinosis.(ABSTRACT TRUNCATED AT 400 WORDS)

Immunochemical properties of the antigens involved in the pigeon breeder's disease.

The pigeon breeder's disease is a form of extrinsic allergic alveolitis caused by the exposure to pigeon droppings. Chemical analysis of this antigen was carried out employing column fractionation techniques. Sephadex G-50 and DEAE cellulose were performed and several proteins and hexoses peaks were recorded. Molecular weights were determined by comparison with standardized marker proteins passed through a drop counting fraction collector. The whole extract revealed a molecular weight of 100 Kd while fraction 1 showed 66 Kd and fraction 2 only 30 Kd. A guinea pig experimental model was developed with the whole extract injected by the intradermal route, administered by an intragastric catheter or aerosolized in a glass chamber. Histopathological studies were carried out with the lungs, kidneys, liver and spleen obtained by the necropsy of the animals. The lungs and the kidneys showed the paramount changes in their structures with lymphomononuclear infiltrates and an Arthus-like phenomenon surrounding the vessels. Immunological techniques were applied to the sera and the lymphocytes obtained from the animals. Precipitin and hemagglutinating IgG antibodies were detected against the whole extract and the fractions obtained by column fractionation. Sensitized lymphocytes were also detected. This experimental model represents a guide in the approach to the human allergic alveolitis whose immunological findings will be presented in a forth-coming report.

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.

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.

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.

[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.

[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.

Asma polínica: estudio químico e inmunológico del polen de la gramínea Lolium perenne / Pollen asthma: chemical and immunologic study of the gramineous Lollium perenne pollen

La polinosis es una entidad clínica claramente establecida y su tratamiento hiposensibilizante provoca gratificantes resultados desde 1911 hasta la actualidad. Sin embargo, los autores han tendido siempre a emplear antígenos polínicos preparados con la mejor tecnología y estandarización, con el propósito de evitar substancias contaminantes cuyo destino se desconoce. Esta presentación aporta nuevas evidencias acerca de la composición química y la reactividad inmunológica del polen de Lolium perenne (Lp) frente a un antisuero de conejo específico y en humanos riníticos y asmáticos sensibilizados. Un extracto del polen de Lp con 10.000 UNP/nl fue pasado por columnas de Sephadex G-50 y de DEAE-celulosa. Los contenidos en proteínas y hexosas fueron registrados en un espectrofotómetro LKB Uvicord a 280 nm y 470 nm, respectivamente. Si bien se analizaron 400 tubos en cada fraccionamiento, se emplearon en las experiencias aquellos con los contenidos más altos en proteínas y en hexosas. Los pesos moleculares deteminados por columna, y comparados con marcadores especiales, fueron de 18.400, 25.500 y 68.000 daltons para las fracciones proteicas Nro. 33, 28 y 13 respectivamente. La determinación cuantitativa de proteínas arrojó valores significativos para las fracciones 33 (350 mcg/ml), 38 (250 mcg/ml) y 55 (200 mcg/ml). Por otro lado, conejos adultos fueron inmunizados con Lp más adyuvante de Freund completo, de acuerdo con el esquema clásico, durante 10 semanas. Este antisuero fue testificado con Lp y cada una de sus fracciones mediante el Ouchterlony y el Boyden. Cien pacientes afectados de rinitis estacional y asma con un IgE sérica total elevada y alta reactividad cutánea a Lp, fueron testificados con las 14 fracciones más significativas (7 proteínas y 7 hexosas) mediante pruebas cutáneas de lectura inmediata, destacándose los resultados encontrados con las fracciones proteicas Nro. 33, 38 y 359 y las hexosas Nro. 56, 192 y 360...

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...