Boosting the suppressive effects of Ascaris suum components in IFN-γ-deficient mice / Potencialização do efeito supressor de componentes do Ascaris suum em camundongos deficientes em IFN-γ

High molecular weight components from Ascaris suum extract suppress ovalbumin-specific immunity in mice. In IFN-γ-deficient mice, ovalbumin-specific delayed-type hypersensitivity reactions are more strongly downregulated by these suppressive components. Here, the cellularity of the delayed-type hypersensitivity reaction in IFN-γ-deficient mice and the increased downregulation induced by Ascaris suum components were analyzed. IL-12p40-dependent neutrophilic influx was predominant. Suboptimal doses of the suppressive fraction from this nematode completely inhibited the hypersensitivity reaction, thus indicating intensification of the immunosuppression under conditions of intense recruitment of IFN-γ-independent neutrophils.

Componentes de alto peso molecular do extrato de Ascaris suum suprimem a imunidade específica à ovalbumina em camundongos. Em camundongos geneticamente deficientes de IFN-γ a reação de hipersensibilidade tardia específica para ovalbumina foi mais fortemente prejudicada por estes componentes supressivos. Aqui, a celularidade da reação de hipersensibilidade tardia em camundongos deficientes de IFN-γ e o incremento na supressão induzida por componentes do Ascaris suum foram analisados. Influxo neutrofílico, dependente de IL-12p40, foi predominante. Dose sub-ótima da fração supressiva do nematódeo inibiu completamente a reação de hipersensibilidade, indicando uma intensificação da imunossupressão em condições de recrutamento intenso de neutrófilos independente de IFN-γ.

Suppressive effect of strychnos nux-vomica on induction of ovalbumin-specific IgE antibody response in mice.

Strychnos nux-vomica Linn. (SNV; Loganiaceae), a medicinal plant has been used as folk medicine for alleviating inflammation, joint pains and allergic symptoms. In the present study, we examined its possible immunomodulatory effect on induction of ovalbumin (OVA)-specific IgE antibody response in a murine model, as evaluated by passive cutaneous anaphylaxis (PCA). The OVA-specific IgE antibody response was significantly suppressed in BALB/c mice (H-2d), following intraperitoneal administration of aqueous stem extract of the plant along with OVA. Furthermore, the different doses of SNV extract were found to significantly suppress the induction of OVA-specific IgE antibody response. The anti-OVA IgE antibody response was suppressed in different haplotypes of mice viz., C57BL/6 (H-2b) and SWR/J (H-29). However, preliminary findings revealed no significant change in the total IgG antibody response against OVA, as evaluated by ELISA. These results confirm the suppressive activity of S. nux-vomica on allergen-specific IgE antibody response and suggest its possible application in allergic conditions. Keywords: Strychnlos nux-vomica, Immunomodulation, Immunosuppression, IgE antibody response, Passive cutaneous anaphylaxis, ELISA

A study of allergen-specific IgE antibodies in Indian patients of atopic dermatitis.

BACKGROUND: Atopic dermatitis (AD) is a chronic, relapsing, pruritic dermatitis frequently associated with the hyperproduction of IgE to various allergens. Identification of these allergens is possible by various laboratory investigations. AIM: The present study was designed to assess these allergen-specific antibodies in the diagnosis of AD in the Indian context. METHODS: This prospective study comprised 50 patients of AD. The diagnosis was made clinically after satisfying Hanifin and Rajka's criteria. Serum IgE levels were estimated and specific IgE antibodies were measured for 20 food allergens and aeroallergens. RESULTS: Serum IgE was elevated in 88% of the patients. The highest elevation of mean IgE levels was seen in the 10-20 years age group. Sixty five percent of the children under the age of ten years were positive to one or more food allergens. Food allergens were more often positive in the < or = 10 years age group and specific antibodies to inhalants were seen more frequently in the older age groups. Specific antibodies to apples were found in all age groups. CONCLUSION: Antibodies against apples and hazelnuts were the more commonly seen specific antibodies in children. Incidence of positivity was much higher in children when compared to earlier studies. Identification of food allergens can be an important factor in the diagnosis of AD in children in India. Positivity to inhalant allergens in the older age groups was lower in this study. The allergen profile with regard to inhalants in Indian patients was similar to that of earlier studies.

Identification of allergens in Indian fishes: hilsa and pomfret exemplified by ELISA and immunoblotting.

Enzymed-linked immunosorbent assay of hilsa and pomfret muscle extracts showed specific IgE binding to ten allergic patients' sera, the results corroborated to that of skin prick test. Comparison of allergen profiles of the two fish extracts by immunoblotting revealed a common antigenic protein of 50 kDa and some high molecular weight fish allergens instead of low molecular weight parvalbumin found in several fishes. Purified and well characterized fish allergens are always considered better than crude fish extracts for diagnostic use.

The Inverse Association Between the Presence of Antibody to Hepatitis B Surface Antigen and Atopy in Young Adults

BACKGROUND: Some bacterial and viral infections may reduce the risk of atopy, and this is based on the concept of their ability to divert the immune system towards the Th1 responses. Most of the hepatitis B virus (HBV) infections occur in the developing countries and this is where atopic disorders are least prevalent. Th1 responses are important for the viral clearance of HBV and also for antibody production. The aim of the study is to investigate whether the presence of antibodies to the hepatitis B surface antigen (anti-HBs) is inversely associated with atopy in adults. METHODS: A random sample of 358 subjects, who were without hepatitis B surface antigen, was recruited; they were aged from 18 to 79 years (105 young adults aged 40 years). Determinations of the anti-HBs and skin prick tests using aeroallergens were performed. Those subjects with one or more positive skin reactions (a mean wheal diameter > or=3 mm) were considered atopic. RESULTS: The prevalence rate of atopy (p=0.035) or the sensitization to Dermatophagoides farinae (p=0.01) was significantly lower in the subjects with anti-HBs than in those subjects without anti-HBs for the young adults, but not for the older adults. The logistic regression analysis that was done on the young adults showed that the presence of anti-HBs was associated with a significantly lower risk of atopy (the odds ratio adjusted for confounding variables=0.40 [95% CI 0.16-0.98], p=0.046) or with the sensitization to D. farinae (0.20 [0.06-0.65], p=0.008). CONCLUSION: The presence of anti-HBs produced by a natural HBV infection or vaccination might be inversely associated with atopy in young adults.

Somatropin promotes dermatophagoides farinae-specific IgE generation independently of IL-4 and IL-10.

Interleukin (IL)-10 accelerates the IgE production of anti-CD40- and IL-4-stimulated PBMC by enhancing the IL-6 production of T lymphocytes or antigen-primed spleen cells, in addition to its role as a regulator of the inflammatory responses. To further investigate the mechanisms enhancing IgE synthesis, we determined the effect of somatropin as well as IL-10 on the secretion of Dermatophagoides farinae (Df)-specific IgE by K7 cells, which originate from an EBV-immortalized cell line. Df-pulsed autologous T cells, as well as the supernatants of these cultures, increased the synthesis of Df-specific IgE. Antigen-specific IgE was also enhanced when K7 cells were treated with anti-CD40 antibody and with both IL-4 and IL-10, or with IL-4 and IL-10 without anti-CD40 antibody. The treatment of K7 cells with anti-CD40 antibody and IL-4, or anti-CD40 antibody and IL-10 did not increase IgE production. The Df-specific IgE activity of the supernatants of K7 cells treated with somatropin alone was increased significantly although somatropin did not show any additive effect on the IgE production of anti-CD40 antibody-treated cells. The results indicate that IL-10, a Th2-type cytokine, directly affects the mature B cells that produce IgE, and that the secretion of IgE is increased by treatment with IL-10 in cells that are stimulated with anti-CD40 and IL-4 at the level of the EBV-immortalized cell line, which has already switched to IgE production. Somatropin similarly stimulates activated mature B cells to enhance their production of antigen-specific IgE without class switching, independently of IL-4 and IL-10.

Intranasal sensitization with Blomia Tropicalis antigens induces allergic responses in mice characterized by elevated antigen-specific and non-specific serum ige and peripheral blood eosinophil counts

Para avaliar a capacidade alergizante do antígeno da Blomia tropicalis (Bt) a produção de IgE específica e não específica a antígeno Bt foi monitorada em camundongos BALB/c após exposição ao antígeno por via nasal. Foi evidenciado que Bt contem um alérgeno funcional em seus componentes. Os componentes alergênicos entretanto, quando administrados por via intra-nasal, sem qualquer adjuvante, não induzem resposta IgE durante um pequeno período. Por outro lado, a inoculação intra-nasal de antígenos Bt aumentou a resposta sérica de IgE em camundongos pré-tratados por uma injeção inicial sensibilizante sub-cutânea aos mesmos antígenos. A inoculação do antígeno Bt sem as injeções sensibilizantes iniciais induziu a produção de anticorpos IgE somente quando o antígeno foi administrado de maneira contínua, por um período longo de mais de 24 semanas. Mesmo quando as injeções sensibilizantes iniciais foram ausentes, o antígeno Bt inoculado com a toxina de cólera (CT) como adjuvante mucoso também aumentou de maneira significante a resposta IgE antígeno específica do Bt dependendo da dose de CT administrada conjuntamente. O presente estudo também demonstrou que camundongos inoculados com antígeno Bt/CT mostram aumento do nível IgE não específico no soro e médias de eosinófilos no sangue periférico sem qualquer elevação da contagem total de leucócitos. A análise por Immunoblot demonstrou cinco principais componentes antigênicos reativos aos anticorpos IgE induzidos. Estes componentes na posição 44-64 kilodaltons foram considerados importantes antígenos-candidatos para o diagnóstico da alergia relacionada ao ácaro.

Airway Inflammation and Allergen Specific IgE Production May Persist Longer Than Airway Hyperresponsiveness in Mice

During the preclinical study of new therapeutic modality, we evaluate whether the treatment can reverse the established asthma phenotypes in animal model. However, few have reported on the long term persistence of asthma phenotypes upon re-challenge with allergen (secondary challenge) in animal model. We evaluated the persistence of asthma phenotypes by secondary challenge at different times in previously challenged murine asthma model. BALB/c mice sensitized by intraperitoneal injections of 20 microgram of ovalbumin and 1 mg of alum on days 1 and 14 were challenged initially by the inhalation of 1% ovalbumin for 30 min on days 21, 22, and 23. Each group of mice was rechallenged at 5, 7, 9, or 12 weeks after the initial challenge. Airway hyperresponsiveness, BAL fluid, airway histology and serum ovalbumin-specific IgE level were evaluated. Airway eosinophilia, airway inflammation and serum ovalbumin-specific IgE production persisted upon secondary allergen challenges at least 12 weeks after the initial challenge. However, airway hyperresponsiveness persisted only until mice were rechallenged 7 weeks after the initial challenge. Airway inflammation and allergen specific IgE production may persist longer than airway hyperresponsiveness in a mouse asthma model of secondary allergen challenge.

Production and characterization of a monoclonal antibody against an Ascaris suum allergenic component

Ascaris suum allergenic components (PIII) separated by gel filtration chromatography of an adult worm extract were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells using polyethylene glycol (MW 1450) as fusogen. The hybridomas were cultured in HAT-containing medium and cloned at limiting dilutions. Supernatants from the growing hybrids were screened by ELISA using plates coated with PIII or the A. suum crude extract. The monoclonal antibody obtained, named MAC-3 (mouse anti-A. suum allergenic component), is an IgG1 kappa mouse immunoglobulin that specifically recognizes a 29,000 molecular weight protein (called allergenic protein) with an affinity constant of 1.7 x 10(9) M-1. The A. suum components recognized by MAC-3 induce specific IgE antibody production in immunized BALB/c mice. Ascitic fluid induced in Swiss mice by injecting ip the hybridoma cells and incomplete Freund's adjuvant was purified by affinity chromatography using a protein A-Sepharose column. The purified monoclonal antibody was then coupled to activated Sepharose beads in order to isolate the A. suum allergenic component from the whole extract by affinity chromatography

Diverse profiles of specific IgE response to toluene diisocyanate (TDI)-human serum albumin conjugate in TDI-induced asthma patients

The prevalence studies on specific IgE to toluene diisocyanate (TDI)-human serum albumin (HSA) conjugate in TDI-induced asthma have shown variable results. In this study, we attempted to compare specific IgE bindings to TDI-HSA conjugate and its specificity using 3 different conjugates. Sera were collected from 20 TDI-induced asthma and 10 controls. Specific IgE were measured by ELISA using three TDI-HSA conjugates; two from Carnegie Mellon (CM; 98 and 99 CM conjugates) and one from Ajou University. To evaluate specificity and cross-reactivity, ELISA inhibition tests were applied. Positive and negative predictive values between Ajou conjugate and 98 CM conjugate were 75% and 100%. Those between Ajou and 99 CM were 100% and 93.8%. One patient showed an isolated positive response to the Ajou with negative responses to the other two conjugates. ELISA inhibition test using this patient's serum revealed the significant inhibitions by the Ajou and minimal inhibitions by the others. On the other hand, another patient showed an isolated positive response to 99 CM with negative responses to the others, and ELISA inhibition test showed significant inhibition by 99 CM with minimal inhibitions by the others. These results suggest that specific IgE bindings to a new antigenic determinant of TDI-HSA conjugate can be heterogeneous and differ from one individual to another.

modulatory effects of interleukin-4 and interferon-gamma production by T-helper cells on IgE synthesis in children with atopic disease

Interleukin-4 [IL-4] is produced by T-helper cells type 2 [TH2] and induces IgE synthesis. T-helper cells type 1 [TH1] produce interferongamma [IFN-gamma] which suppresses TH2 and reduces IL-4 induced IgE production. In this study, we demonstrated that the levels of specific IgE in the serum of atopic children [n = 20] were elevated while IL-4 production was increased and IFN-gamma secretion was reduced, compared to those of control group. Interleukin -4-induced IGE synthesis by peripheral blood mononuclear cells of atopic children in vitro was blocked in the presence of IFN-gamma. In addition, levels of soluble CD23-which is specifically induced by IL-4, were significantly elevated in our atopic patients. The data indicate that enhanced production of IL-4 and lowered IFN-gamma secretion by T-helper cells correlate with the elevated specific IGE levels in the serum of atopic children

Participación de los linfocitos Th2 en la enfermedad alérgica / Participation of lymphocites Th2 in allergic disease

Los padecimientos alérgicos son determinados genéticamente y afectan al 20 a 30 por ciento de la población de los países desarrollados. En la última década se ha observado un incremento en la prevalencia de los mismos, caracterizado por un aumento en la capacidad de los linfocitos B para producir anticuerpos inmunoglobulinas tipo E (IgE). Esta síntesis de la IgE humana resulta de la colaboración entre los subtipos de la células T auxiliadoras (Th) CD4+ y las células B. En años recientes se han estudiado intensamente las propiedades funcionales de la células Th, particularmente las tipo 2 en el mecanismo de unión de las células B productoras de IgE, células cebadas o basófilos y eosinófilos en reacciones alérgicas. Estos conceptos son conocidos en la actualidad e alergia como la teoría Th2

Biology of basophils and their role in allergic inflammation.

Chronic allergic inflammatory reactions involve the infiltration and participation of many different cell types. Although it has been evident for many years that tissue specific mast cells have a primary role in the early stages of these reactions, recent studies indicated that, in addition to mast cells response, a later reaction which selectively recruits the circulating lymphocytes, eosinophils and basophils to the site of inflammation, is the hallmark for the progression of allergic diseases.

IgE production from the nasal polyp tissue: comparison between atopic and non-atopic subjects

OBJECTIVES AND METHODS: To confirm the local production of IgE antibody from the nasal polyp tissue, and to evaluate the difference between atopics and non-atopics, nasal polyp tissues were taken from both 10 atopic and 10 non-atopic subjects. The tissue total IgE (tlgE) level was measured by enzyme-linked immunosorbent assay (ELISA) and serum tlgE level by radio-immunoassay. The tissue albumin level was measured by nephelometry, and serum albumin level by Bromocresol green method. RESULTS: The polyp tissue tlgE/albumin as well as serum tlgE/albumin ratio were significantly higher in atopics than in non-atopics (p 0.05). Three non-atopic subjects had high polyp tissue tlgE/albumin (> 10). A significant correlation was noted between serum tlgE/albumin and polyp tlgE/albumin (r = 0.46, p = 0.04). The ratio of polyp tlgE/albumin to serum tlgE/albumin was greater than 1 in all of the non-atopic subjects and 7 of 10 atopic subjects. CONCLUSION: These findings support the hypothesis that IgE antibody could be locally produced from the nasal polyp tissue of non-atopic subjects as well as atopic subjects. The possibility of an isolated local production of IgE antibody was suggested.

Skin reactivity and specific IgE antibody to two nonbiting midges in Korean respiratory allergy patients

To evaluate the significance of chironomid as a respiratory allergen, we performed skin prick tests with Chironomus plumosus (CP) and Tokunagayusurika akamusi (TA) extracts on 475 respiratory allergy patients, and their specific IgE antibodies were detected by enzyme-linked immunosorbent assay (ELISA) in 106 positive reactors to skin prick test and 30 negative controls. Ninety-seven (20.4%) showed more than 2+ of allergen to histamine ratio to CP and 98 (20.6%) to TA on skin prick test. Seventy-one (73.2%) of 97 positive reactors had increased specific IgE to CP, and 34 (34.7%) of 98 positive reactors, to TA. CP-specific IgE was detected in 14 (14.4%) non-atopic asthmatics and 6 (6.2%) non-allergic rhinitis patients. TA-specific IgE was detected in 17 (17.4%) non-atopic asthmatics and 6 (6.1%) non-allergic rhinitis patients. No association was noted between skin reactivity to Dermatophagoides farinae and the prevalence of specific IgE to CP or TA (p > 0.05). The correlation between total IgE level and specific IgE level to CP and TA was poor (r = 0.07, 0.04). ELISA inhibition test suggested specificity of IgE binding and cross-allergenicity between CP and TA. It is suggested that CP and TA can induce IgE-mediated reaction in exposed patients and should be considered as important causative allergens in respiratory allergy patients in Korea.

Decrease in passive cutaneous anaphylaxis reactions in rats secondary to stress

Anaphylaxis was assayed by the passive cutaneous anaphylaxis (PCA) test in male Wistar rats (250 g body weight). Three experimental groups were used: animals restrained in an electric chamber and submitted to electric shock immediately after sensitization and 24 h before anaphylaxis (31 animals), animals restrained in the electric chamber for the same time but receiving no electric shock (23 animals), and non-manipulated, home-cage control animals (24 animals). The frequency of PCA reactions was decreased in the group of animals submitted to restraint when compared with the home-cage control group. However, the group of animals submitted to both restraint and electric shock showed no decrease in the frequency of PCA reactions. It is suggested that, in rats, stress induced by restraint decreases PCA reactions and that this decrease is counteracted by a simultaneous stress induced by electric shock

Suppression of the IgE antibody response by Ascaris suum components: effect of x-irradiation

The effect of X-irradiation on the supperession of IgE antibody responses induced by some of the Ascaris suum (ASC) components was analyzed in mice (7-week old A?Sn females). Treatment with 300 R 24h before immunization with 50 *g OVA and 200 *g ASC suppressive components abolished the damping effect on ati-OVA IgE antibody levels. The same effect was observed on the anti-ASC IgE antibody response obtained in mice injected with 200 *g ASC immunogenic plus 200 *g ASC suppressive components. Moreover, the failure of suppressive components to induce an IGE anti-ASC antibody response on their own was also abolished by X-irradiation. These results indicate that the suppressive components are able to elicit an IgE antibody response, but simultaneously activate a regulatory mechanism which suppressive both the homologous (anti-ASC) and heterologous (anti-OVA) antibody formation