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.

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).

Bat feces as an indoor allergen.

We have demonstrated in an animal model (specific IgG) as well as in atopic patients suffering from rhinitis/asthma (specific IgE) that bat feces have antigenic properties. A single peak with high glycoprotein content was obtained by chromatography, while SDS-PAGE revealed several proteins between 29 and 116 kDa which showed immune serum blotting at 43.6 and 66 kDa. Positive specific skin tests with bat feces and IgE-RAST anti-bat feces were detected in atopic patients living in tall buildings and old houses in Buenos Aires. As bat feces did not cross-react with bat epithelium, studies evaluating rat serum and epithelium and pigeon feces in order to analyze the role of bat serum proteins, such as IgA, are encouraged.

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.

IgE antibodies against snake venoms.

A similar event was detected in the clinical records of a small group of atopic patients living in the northern provinces of Argentina, i.e., they were bitten by a snake of the Bothrops species (or yarará) during their rural activities (woodcutters, cattle-drivers and farmers). Those who were bitten twice suffered an acute episode of hives and angioedema within 15 minutes after the snake bite. The presence of specific antibodies against Bothrops alternata (Ba) extract was detected by means of RAST for IgE and Ouchterlony and Boyden for IgG. The Ouchterlony also demonstrated crossreactivity among the venoms of the Bothrops species and the positivity of the six fractions obtained by DEAE-cellulose column fractionation against the horse anti-Ba serum. The Ba antigen induced a definite inhibition of the RAST. We presume that hives and angioedema in atopic patients immediately after a second snake bite could be attributed to the presence of a specific IgE antibody against the venom, and must not be misinterpreted with the toxic effects that appear later.

Interstitial pneumonitis induced in guinea pigs by Triatoma infestans antigens.

Data concerning the experimental induction of hypersensitivity pneumonitis in guinea pigs with a Triatoma infestans antigen are presented. Glycoproteins obtained from the chitinous structures of T. infestans (79 kd + 11 kd) were aerosolized daily to guinea pigs during 7 weeks. The presence of specific antibodies (IgG and IgE) was detected by serological techniques; histopathological studies of the lungs showed interstitial infiltrates of macrophages and T-cells. Single non-necrotizing granulomas were seen at the seventh week 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 T. infestans.

A citrus fruit-exclusion diet in sensitive patients and its influence on specific antibodies.

The presence and kinetics of specific IgE and IgG antibodies against the oranges Citrus aurantium sinensis (CAS) and Citrus silension (CS) were studied in 41 atopic and 20 non-atopic children aged 8-12 years. Diagnostic procedures such as intracutaneous skin tests, IgE PRIST and anti-CAS and anti-CS IgE and IgG RAST were performed in both groups. A citrus fruit-exclusion diet was maintained for 180 days. The comparison of the results before and after the diet showed no significant changes in skin reactivity and RAST values, and even a slight increase was recorded in the latter. We have attempted to explain these puzzling findings.

Immunochemical properties of the antigens of Triatoma infestans.

Taxonomically related to the cockroach Periplaneta americana (Pa), Triatoma infestans (Ti) is an insect that parasitizes two-thirds of Argentina and acts as a vector for American trypanosomiasis. The glycoproteins obtained by a Sephadex G-150 column fractionation from the whole-body extract prepared with the chitinous structures revealed antigenicity in rabbits, which developed specific IgG after 8 weeks of immunization, as detected by several serological techniques. Atopic patients living in the northern provinces of Argentina--Santiago del Estero and Chaco (endemic area for Ti)--with rhinitis/asthma showed immediate positive skin tests to Ti and its fractions, as well as an IgE-RAST-anti-Ti in 30-36% of them. Healthy nonatopic adults of the same area and other atopics and nonatopics from Buenos Aires (non-endemic area) were selected as controls and showed negative type I skin tests, as well as specific RAST. The existence of common epitopes between Ti and Pa was confirmed by absorption techniques, RAST inhibition, IEF and STIF-RAST. These findings have clinical and epidemiological connotations.

IgE antibodies against Neisseria flavescens.

Bacterial allergy is still a matter of controversy. We sustain that this name should be employed only in the presence of a specific IgE against antigens from bacteria. In 100 atopic patients and 100 healthy controls with Neisseria flavescens in their pharyngeal exudates, we performed type I immediate skin tests with Neisseria flavescens and IgE-RAST throughout 1 year. Positive wheal and flare reactions were elicited in 8 of 100 atopic patients as well as in 3 of 100 nonatopic subjects. IgE-RAST/anti-Neisseria flavescens was found in 6 of the former group and in 1 of the latter. Neither late-phase nor Arthus-like reactions were recorded. Neisseria flavescens is a non-pathogenic commensal of the oropharynx with scarce antigenic properties and seems not to play an important role in these conditions (rhinitis/asthma). Bacterial immunotherapy should be considered only in the presence of specific IgE antibodies with careful selection of the bacteria or their antigens.

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.

Evaluation of the high density lipoprotein cholesterol protective effect against atherogenesis in rabbits fed cholesterol supplemented diets

El colesterol de las lipoproteínas de alta densidad (C-HDL) plasmático fue evaluado en 15 conejos alimentados con dietas suplementadas con colesterol respecto a su efecto protector en el proceso aterogénico. Partiendo de un nivel basal de C-HDL de 29 ñ 11 mg/dl (promedio ñ DS), sólo se logró como máximo duplicar dicho valor en tres conejos, mientras que el colesterol total (CT) aumentó hasta 20 veces. El índice plasmático CT/HDL aumentó 80 veces su valor basal (2,4 ñ 0,9) y fue el parámetro que detectó la mejor correlación con el colesterol acumulado en la aorta y con los scores de los estudios patológicos (r:0,956, p < 0,001 y r:0,805, p < 0,003, respectivamente). El contenido de CT aórtico aumentó 10 veces y la relación colesterol libre/colesterol esterificado disminuyó 20 veces. Los resultados patológico demostraron scores de lesiones aórticas que se elevaron desde 0 a 4. se concluye que las altas correlaciones obtendias cuando el índice CT/C-HDL fue graficado contra la acumulación del colesterol aórtico o contra los scores de las lesiones, avalan la teoría del transporte reverso del colesterol y la efectividad de dicho índice para predecir el grado del proceso aterogénico. Por otra parte, la escasa respuesta del C-HDL en este modelo experimental alienta a emprender futuras investigaciones con fármacos con el objeto de elevar este parámetro para normalizar el índice CT/C-HDL y evitar las lesiones

Evaluation of the high density lipoprotein cholesterol protective effect against atherogenesis in rabbits fed cholesterol supplemented diets

El colesterol de las lipoproteínas de alta densidad (C-HDL) plasmático fue evaluado en 15 conejos alimentados con dietas suplementadas con colesterol respecto a su efecto protector en el proceso aterogénico. Partiendo de un nivel basal de C-HDL de 29 ñ 11 mg/dl (promedio ñ DS), sólo se logró como máximo duplicar dicho valor en tres conejos, mientras que el colesterol total (CT) aumentó hasta 20 veces. El índice plasmático CT/HDL aumentó 80 veces su valor basal (2,4 ñ 0,9) y fue el parámetro que detectó la mejor correlación con el colesterol acumulado en la aorta y con los scores de los estudios patológicos (r:0,956, p < 0,001 y r:0,805, p < 0,003, respectivamente). El contenido de CT aórtico aumentó 10 veces y la relación colesterol libre/colesterol esterificado disminuyó 20 veces. Los resultados patológico demostraron scores de lesiones aórticas que se elevaron desde 0 a 4. se concluye que las altas correlaciones obtendias cuando el índice CT/C-HDL fue graficado contra la acumulación del colesterol aórtico o contra los scores de las lesiones, avalan la teoría del transporte reverso del colesterol y la efectividad de dicho índice para predecir el grado del proceso aterogénico. Por otra parte, la escasa respuesta del C-HDL en este modelo experimental alienta a emprender futuras investigaciones con fármacos con el objeto de elevar este parámetro para normalizar el índice CT/C-HDL y evitar las lesiones (AU)

Evaluation of the high density lipoprotein cholesterol protective effect against atherogenesis in rabbits fed cholesterol supplemented diets.

Plasma high density lipoprotein cholesterol (HDL-C) was evaluated in 15 rabbits fed cholesterol supplemented diets to assess its protective effect on the atherogenic process. From a baseline level of 29 +/- 11 mg/dl (mean +/- SD) the maximum attained for HDL-C was twofold in only three rabbits, whereas total cholesterol (TC) increased 20 fold. Plasma TC/HDL-C ratio rose 80 fold from the baseline (2.4 +/- 0.9) and it was the best parameter that correlated with aortic cholesterol accumulation and pathological scores. Aortic TC content increased 10 fold and free cholesterol/cholesterol esters ratio decreased 20 fold. Pathological studies showed that aortic lesion scores rose from 0 to 4. It can be concluded that the high correlations obtained when TC/HDL-C ratio was plotted against both aortic cholesterol deposition and lesion scores, support the theory of the reverse cholesterol transport and the effectiveness of this index to predict the degree of the atherogenic process. On the other hand, the poor response of HDL-C in this model encourages future research using drugs to increase this parameter in order to normalize TC/HDL-C ratio and avoid lesions.

Evaluation of the high density lipoprotein cholesterol protective effect against atherogenesis in rabbits fed cholesterol supplemented diets.

Plasma high density lipoprotein cholesterol (HDL-C) was evaluated in 15 rabbits fed cholesterol supplemented diets to assess its protective effect on the atherogenic process. From a baseline level of 29 +/- 11 mg/dl (mean +/- SD) the maximum attained for HDL-C was twofold in only three rabbits, whereas total cholesterol (TC) increased 20 fold. Plasma TC/HDL-C ratio rose 80 fold from the baseline (2.4 +/- 0.9) and it was the best parameter that correlated with aortic cholesterol accumulation and pathological scores. Aortic TC content increased 10 fold and free cholesterol/cholesterol esters ratio decreased 20 fold. Pathological studies showed that aortic lesion scores rose from 0 to 4. It can be concluded that the high correlations obtained when TC/HDL-C ratio was plotted against both aortic cholesterol deposition and lesion scores, support the theory of the reverse cholesterol transport and the effectiveness of this index to predict the degree of the atherogenic process. On the other hand, the poor response of HDL-C in this model encourages future research using drugs to increase this parameter in order to normalize TC/HDL-C ratio and avoid lesions.

Induction of anti-orange antibodies in rabbits.

The capacity of certain foods to cause allergic reactions is well known. The four types of mechanisms that Gell and Coombs described in 1968, are involved in these reactions, although the reaginic antibodies retain the paramount attention of the immunologists. The physiochemical composition of the allergen molecule is the goal of investigators with the purpose to clarify the intrinsic kinetics of antibody synthesis. This paper contributes to the conflicting data about orange allergens especially those obtained from Citrus Aurantium Sinensis and Citrus Silension (CAS and CS, respectively). Glycoproteins were separated by gel filtration through a Sephadex G-50 column. A definite protein peak was obtained meanwhile several hexoses appeared throughout the fractionation procedure. These molecules have adequate physiochemical properties that make them able to trigger the immunological response (molecular weight, definite chemical composition and glycoprotein content). Although CAS and CS have a similar chemical composition a slight inverse proportion of proteins and hexoses was demonstrated between the two classes. Molecular weights were different for CAS (51.500) and for CS (37.000) in comparison with well established protein makers. Ouchterlony revealed two precipitin lines in the CAS-anti-CAS system but none in the CS-anti-CS one. The Boyden technique showed a titre of 1/256 in the first case and only of 1/64 in the second of specific anti-orange antibodies. All the eluted fractions gave negative results although they were concentrated ten times by pre-evaporation. This animal model reinforced the statement that after a long and continuous exposure to orange antigens it was possible to develop specific antibodies. It is assumed that this phenomenon happens in atopic children with it's diagnostic and therapeutic importance.

Immunological findings in human beings with autoimmune uveitis.

Thirty-four adult patients suffering from uveitis of unknown origin were submitted to an immunological protocol where specific and non-specific parameters were checked. Serum IgA, IgG and IgM appeared within normal values meanwhile IgE-PRIST was undoubtedly increased in 27 out of 34 patients. The presence of autoantibodies such as antimitochondrial and antismooth muscle were detected in a few patients with Harada's syndrome; high serum levels of immune complexes were also detected in this little group of patients with posterior uveitis. Serum complement fractions showed no significant changes. Antirretinal IgG antibodies were present in almost 40% of the patients especially in those who also have autoantibodies and immunocomplexes. The histoimmunofluorescent technique revealed positive images at the retinal membrane agreeing with the histopathological lesions obtained in the experimental model. The positive cross reactivity between bovine tissue sections and the sera from patients with uveitis reinforced the idea of the presence of common epitopes in the S-antigen of both origins. LT-CD4 and LT-CD8 serum lymphocytes showed contradictory results though both subpopulations appeared diminished. The ratio between them kept within the normal range or slightly increased (2.3-2.8). Sensitized lymphocytes to the S-antigen were detected by the MIF technique in one-third of the patients. The presence of IgG antiretinal antibodies led us to speculate about the possibility that an ADCC mechanism could also participate in the etiopathogenesis of the autoimmune uveitis. On the other hand the presence of a specific antiretinal IgE antibody must play a paramount role in increasing the vascular permeability and thus enhancing the humoral and cellular events in the inflammatory focus.(ABSTRACT TRUNCATED AT 250 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.

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.