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1.
Clin Exp Allergy ; 35(7): 873-9, 2005 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-16008672

RESUMEN

BACKGROUND: We have recently isolated two distinct components from Ascaris suum adult worms with different effects on the immune system: the allergenic protein of A. suum (APAS-3), which induces IgE antibody production, and suppressive protein of A. suum (PAS-1), which inhibits humoral and cellular immune responses induced by unrelated antigens. In this study, we investigated the immunomodulatory effect of PAS-1 on a murine model of asthma induced by APAS-3. METHODS: BALB/c mice were immunized twice with APAS-3 or APAS-3 plus PAS-1 by the intraperitoneal and subcutaneous route (on days 0 and 7) and challenged twice with the same antigens intranasally (days 14 and 21). Two days after the last challenge, the allergic airway inflammation was evaluated by cellular migration, eosinophil peroxidase (EPO) activity, cytokine and chemokine production and pulmonary mechanical parameters. RESULTS: The allergenic properties of APAS-3 were confirmed by the stimulation of anaphylactic IgE and IgG1 antibody production and eosinophilic airway inflammation and hyper-responsiveness. On the other hand, PAS-1-treated mice showed a marked suppression of cellular migration and EPO activity that correlated well with a significant reduction in the levels of IL-4, IL-5, eotaxin and RANTES in the bronchoalveolar lavage (BAL) fluid. In contrast, considerable amounts of IL-10 were observed in the BAL fluid of PAS-1-treated mice. Airway hyper-responsiveness was obtained in APAS-3-immunized mice, but the conductance of the respiratory system was restored to normal values in the presence of PAS-1. CONCLUSION: These results indicate that A. suum allergenic protein APAS-3 induces a T helper 2-type immune response and, consequently, eosinophilic airway inflammation and hyper-responsiveness. Moreover, the modulatory protein PAS-1 has a marked suppressive effect on this response, and the inhibition of cytokine (IL-4, IL-5) and chemokine (eotaxin and RANTES) release, probably because of the presence of IL-10, may contribute to this effect.


Asunto(s)
Ascaris suum/inmunología , Asma/inmunología , Proteínas del Helminto/inmunología , Alérgenos/inmunología , Anafilaxia/inmunología , Animales , Líquido del Lavado Bronquioalveolar/inmunología , Inhibición de Migración Celular , Quimiocina CCL11 , Quimiocina CCL5/inmunología , Quimiocinas CC/inmunología , Factores Quimiotácticos Eosinófilos/inmunología , Modelos Animales de Enfermedad , Peroxidasa del Eosinófilo/inmunología , Eosinófilos/inmunología , Femenino , Inmunoglobulina E/inmunología , Inmunoglobulina G/inmunología , Interleucinas/inmunología , Pulmón/inmunología , Ratones , Ratones Endogámicos BALB C , Células Th2/inmunología
2.
Allergy ; 58(11): 1117-24, 2003 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-14616121

RESUMEN

The aim of this study was to investigate the role of immunoglobulin E (IgE) in the late phase reaction (LPR) of murine experimental asthma. Our model consisted of an implant of DNP-conjugated, heat-coagulated hen's egg white (DNP-EWI), followed 14 days later by an intratracheal challenge with aggregated DNP-ovalbumin. Airway inflammation was analyzed 48 h after challenge and compared with a similarly immunized group of mice with highly suppressed humoral response due to anti-micro and anti-delta antibody treatment. Total number of cells in the bronchoalveolar lavage (BAL) (with predominance of eosinophils) and EPO activity in the lung homogenate were increased in the DNP-EWI-immunized group compared with immunosuppressed or nonimmunized mice. However, the cellular infiltration and EPO activity observed in the immunosuppressed group were still significantly above those obtained in the nonimmunized group, indicating that inhibition of antibody production did not completely prevent the inflammatory manifestations in BAL and lung. Airway hyperresponsiveness to methacoline was obtained in DNP-EWI-immunized mice, but the respiratory mechanical parameters returned to normal levels in the immunosuppressed group. When these mice were reconstituted with monoclonal anti-DNP antibodies, only IgE, but not IgG1, restored lung inflammation and decreased the conductance of the respiratory system, therefore, increasing hyperresponsiveness. These results indicate that antibodies are not essential for induction of LPR in the lung. However, IgE enhances pulmonary inflammation and hyperresponsiveness.


Asunto(s)
Formación de Anticuerpos , Asma/fisiopatología , Hiperreactividad Bronquial/inmunología , Inmunoglobulina E/inmunología , Animales , Asma/inmunología , Asma/patología , Hiperreactividad Bronquial/fisiopatología , Líquido del Lavado Bronquioalveolar/citología , Recuento de Células , Dinitrofenoles/inmunología , Clara de Huevo , Peroxidasa del Eosinófilo , Eosinófilos/enzimología , Eosinófilos/patología , Inmunoglobulinas/sangre , Inflamación , Masculino , Ratones , Ratones Endogámicos BALB C , Ovalbúmina/inmunología , Peroxidasas/metabolismo
3.
Braz. j. med. biol. res ; 34(8): 1033-1036, Aug. 2001. ilus
Artículo en Inglés | LILACS, SES-SP | ID: lil-290152

RESUMEN

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


Asunto(s)
Animales , Ratones , Anticuerpos Antihelmínticos/biosíntesis , Alérgenos/inmunología , Ascaris suum/inmunología , Anticuerpos Monoclonales/biosíntesis , Inmunoglobulina E/biosíntesis , Ensayo de Inmunoadsorción Enzimática , Alérgenos/aislamiento & purificación , Proteínas del Helminto/inmunología , Cromatografía de Afinidad , Ratones Endogámicos BALB C
4.
Braz J Med Biol Res ; 34(8): 1033-6, 2001 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-11471042

RESUMEN

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.


Asunto(s)
Alérgenos/inmunología , Anticuerpos Antihelmínticos/biosíntesis , Anticuerpos Monoclonales/biosíntesis , Ascaris suum/inmunología , Alérgenos/aislamiento & purificación , Animales , Cromatografía de Afinidad , Ensayo de Inmunoadsorción Enzimática , Proteínas del Helminto/inmunología , Inmunoglobulina E/biosíntesis , Ratones , Ratones Endogámicos BALB C
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