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1.
Parasite Immunol ; 39(9)2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28703913

RESUMEN

Helminth infections have the ability to modulate host's immune response through mechanisms that allow the chronic persistence of the worms in the host. Here, we investigated the mechanisms involved on the suppressive effect of Ascaris suum infection using a murine experimental model of LPS-induced inflammation. We found that infection with A. suum markedly inhibited leucocyte influx induced by LPS into air pouches, suppressed secretion of pro-inflammatory cytokines (IL-1ß, TNF-α and IL-6) and induced high levels of IL-10 and TGF-ß. Augmented frequency of CD4+ CD25high Foxp3+ T cells was observed in the mesenteric lymph nodes of infected mice. Adoptive transfer of purified CD4+ CD25+ T cells to recipient uninfected mice demonstrated that these cells were able to induce a suppressive effect in the LPS-induced inflammation in air pouch model. In addition, adoptive transfer of CD4+ CD25+ T cells derived from IL-10 knockout mice suggests that this suppressive effect of A. suum infection involves IL-10 cytokine. In conclusion, our results demonstrated that A. suum experimental infection was capable of suppressing LPS-induced inflammation by mechanisms, which seem to be dependent on responses of CD4+ CD25+ T cells and secretion of IL-10 cytokine.


Asunto(s)
Ascariasis/inmunología , Ascaris suum/inmunología , Traslado Adoptivo , Animales , Ascariasis/parasitología , Antígenos CD4/metabolismo , Citocinas/metabolismo , Femenino , Factores de Transcripción Forkhead/metabolismo , Inflamación/inducido químicamente , Interleucina-10/metabolismo , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Interleucina-6/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Linfocitos T/inmunología , Factor de Crecimiento Transformador beta/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo
2.
J Helminthol ; 89(3): 367-74, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-24703095

RESUMEN

Helminths use several strategies to evade and/or modify the host immune response, including suppression or inactivation of the host antigen-specific response. Several helminth immunomodulatory molecules have been identified. Our studies have focused on immunosuppression induced by the roundworm Ascaris suum and an A. suum-derived protein named protein 1 from A. suum (PAS-1). Here we assessed whether PAS-1 is an excretory/secretory (E/S) protein and whether it can suppress lipopolysaccharide-induced inflammation. Larvae from infective eggs were cultured in unsupplemented Dulbecco's modified Eagle medium (DMEM) for 2 weeks. PAS-1 was then measured in the culture supernatants and in adult A. suum body fluid at different time points by enzyme-linked immunosorbent assay (ELISA) with the monoclonal antibody MAIP-1. Secreted PAS-1 was detected in both larval culture supernatant and adult body fluid. It suppressed lipopolysaccharide (LPS)-induced leucocyte migration and pro-inflammatory cytokine production, and stimulated interleukin (IL)-10 secretion, indicating that larval and adult secreted PAS-1 suppresses inflammation in this model. Moreover, the anti-inflammatory activity of PAS-1 was abolished by treatment with MAIP-1, a PAS-1-specific monoclonal antibody, confirming the crucial role of PAS-1 in suppressing LPS-induced inflammation. These findings demonstrate that PAS-1 is an E/S protein with anti-inflammatory properties likely to be attributable to IL-10 production.


Asunto(s)
Ascaris suum/fisiología , Proteínas del Helminto/inmunología , Proteínas del Helminto/metabolismo , Tolerancia Inmunológica , Inmunosupresores/metabolismo , Animales , Ascaris suum/química , Ascaris suum/inmunología , Movimiento Celular/efectos de los fármacos , Medios de Cultivo/química , Citocinas/metabolismo , Ensayo de Inmunoadsorción Enzimática , Interacciones Huésped-Patógeno , Larva/química , Larva/inmunología , Larva/fisiología , Leucocitos/metabolismo , Leucocitos/fisiología , Factores de Tiempo
3.
Scand J Immunol ; 72(6): 491-503, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21044123

RESUMEN

We have previously demonstrated that PAS-1, a 200 kDa protein from Ascaris suum, has a potent immunomodulatory effect on humoral and cell-mediated responses induced by APAS-3 (an allergenic protein from A. suum) or unrelated antigens. In this study, we investigated the mechanisms by which PAS-1 is able to induce this effect on an allergic airway inflammation induced by OVA in mice. C57BL/6 mice were adoptively transferred on day 0 with seven different PAS-1-primed cell populations: PAS-1-primed CD19(+) or B220(+) or CD3(+) or CD4(+) or CD8(+) or CD4(+) CD25⁻ or CD4(+) CD25(+) lymphocytes. These mice were immunized twice with OVA and alum by intraperitoneal route (days 0 and 7) and challenged twice by intranasal route (days 14 and 21). Two days after the last challenge, the airway inflammation was evaluated by antibody levels, cellular migration, eosinophil peroxidase levels, cytokine and eotaxin production, and pulmonary mechanical parameters. Among the adoptively transferred primed lymphocytes, only CD4(+) CD25(+) , CD8(+) or the combination of both T cells impaired the production of total IgE and OVA-specific IgE and IgG1 antibodies, eosinophilic airway inflammation, Th2-type cytokines (IL-4, IL-5 and IL-13), eotaxin release and airway hyperreactivity. Moreover, airway recruited cells from CD4(+) CD25(+) and CD8(+) T-cell recipient secreted more IL-10/TGF-ß and IFN-γ, respectively. Moreover, we found that PAS-1 expands significantly the number of CD4(+) CD25(+) FoxP3(+) and CD8(+) γδTCR(+) cells. In conclusion, these findings demonstrate that the immunomodulatory effect of PAS-1 is mediated by these T-cell subsets.


Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Proteínas del Helminto/inmunología , Inmunomodulación , Receptores de Antígenos de Linfocitos T gamma-delta/fisiología , Hipersensibilidad Respiratoria/inmunología , Traslado Adoptivo , Animales , Linfocitos T CD4-Positivos/patología , Linfocitos T CD8-positivos/patología , Factores de Transcripción Forkhead/biosíntesis , Inflamación/inmunología , Subunidad alfa del Receptor de Interleucina-2/biosíntesis , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratas , Ratas Wistar , Hipersensibilidad Respiratoria/patología
4.
Cytokine ; 44(3): 335-41, 2008 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19008120

RESUMEN

Helminths and their products have a profound immunomodulatory effect upon the inductive and effector phases of inflammatory responses, including allergy. We have demonstrated that PAS-1, a protein isolated from Ascaris suum worms, has an inhibitory effect on lung allergic inflammation due to its ability to down-regulate eosinophilic inflammation, Th2 cytokine release and IgE antibody production. Here, we investigated the role of IL-12, IFN-gamma and IL-10 in the PAS-1-induced inhibitory mechanism using a murine model of asthma. Wild type C57BL/6, IL-12(-/-), IFN-gamma(-/-) and IL-10(-/-) mice were immunized with PAS-1 and/or OVA and challenged with the same antigens intranasally. The suppressive effect of PAS-1 was demonstrated on the cellular influx into airways, with reduction of eosinophil number and eosinophil peroxidase activity in OVA+PAS-1-immunized wild type mice. This effect well correlated with a significant reduction in the levels of IL-4, IL-5, IL-13 and eotaxin in BAL fluid. Levels of IgE and IgG1 antibodies were also impaired in serum from these mice. The inhibitory activity of PAS-1 was also observed in IL-12(-/-) mice, but not in IFN-gamma(-/-) and IL-10(-/-) animals. These data show that IFN-gamma and IL-10, but not IL-12, play an important role in the PAS-1 modulatory effect.


Asunto(s)
Ascaris suum/inmunología , Hiperreactividad Bronquial/inmunología , Proteínas del Helminto/metabolismo , Interferón gamma/inmunología , Interleucina-10/inmunología , Animales , Formación de Anticuerpos/inmunología , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/metabolismo , Movimiento Celular/inmunología , Inmunoglobulina G/inmunología , Interferón gamma/deficiencia , Interferón gamma/genética , Interferón gamma/metabolismo , Interleucina-10/deficiencia , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-12/deficiencia , Interleucina-12/genética , Interleucina-12/inmunología , Interleucina-12/metabolismo , Leucocitos/citología , Leucocitos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados
5.
Parasite Immunol ; 28(9): 453-61, 2006 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-16916369

RESUMEN

The inflammatory and functional changes that occur in murine lung after infection with 2500 infective Ascaris suum eggs were studied in this work. A sequential influx of neutrophils, mononuclear cells and eosinophils occurred into airways concomitantly with migration of larvae from liver to the lungs. Histological analysis of the lung showed a severe intra-alveolar haemorrhage at the peak of larval migration (day 8) and the most intense inflammatory cell infiltrate on day 14. Ascaris L3 were found in alveolar spaces and inside bronchioles on day 8. The number of eosinophils was elevated in the blood on days 8 and 14. The peak of eosinophil influx into the lung was at day 14, as indicated by the high levels of eosinophil peroxidase activity, followed by their migration into the airways. The antibody response against egg and larval antigens consisted mainly of IgG1 and IgM, and also of IgE and anaphylactic IgG1, that cross-reacted with adult worm antigens. Total IgE levels were substantially elevated during the infection. Measurement of lung mechanical parameters showed airway hyperreactivity in infected mice. In conclusion, the murine model of A. suum infection mimics the Th2-induced parameters observed in pigs and humans and can be used to analyse the immunoregulatory properties of this helminth.


Asunto(s)
Ascaris suum/inmunología , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/parasitología , Enfermedades Pulmonares Parasitarias/inmunología , Enfermedades Pulmonares Parasitarias/parasitología , Anafilaxia/inmunología , Animales , Anticuerpos Antihelmínticos/sangre , Anticuerpos Antihelmínticos/inmunología , Especificidad de Anticuerpos , Ascaris suum/crecimiento & desarrollo , Lavado Broncoalveolar/métodos , Peroxidasa del Eosinófilo/metabolismo , Eosinofilia/inmunología , Eosinofilia/parasitología , Femenino , Enfermedades Pulmonares Parasitarias/enzimología , Ratones , Ratones Endogámicos BALB C , Ratas , Ratas Wistar , Regulación hacia Arriba
6.
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
7.
Inflamm Res ; 54(1): 17-21, 2005 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-15723200

RESUMEN

BACKGROUND: Recently, we identified a 200 kDa protein (PAS-1) from Ascaris suum worms, that suppresses the humoral immune response. Here, the effect of PAS-1 on inflammatory leukocyte migration induced by bacterial lipopolysaccharide (LPS) was investigated. METHODS: Cellular migration and cytokine release, stimulated by LPS or LPS+PAS-1, were analyzed in air pouches induced in the shaved back of BALB/c mice. Cytokines were determined by ELISA and RT-PCR on air pouch exudates and in vitro stimulated peritoneal macrophages. RESULTS: The significant cellular influx induced by LPS, consisting predominantly of neutrophils, was highly suppressed in the presence of PAS-1, but not a non-related protein. PAS-1 led also to a marked reduction of TNF-alpha, IL-1 beta and IL-6 levels in both LPS-stimulated air pouches and peritoneal macrophage cultures. In contrast, PAS-1 induced a significant increase of IL-10 and TGF-beta production. CONCLUSIONS: These results demonstrate that PAS-1 has a potent anti-inflammatory activity, probably due to the stimulation of regulatory cytokines in macrophages, thus leading to the inhibition of pro-inflammatory cytokine production.


Asunto(s)
Antiinflamatorios/farmacología , Ascaris suum/química , Proteínas del Helminto/farmacología , Animales , Antiinflamatorios/aislamiento & purificación , Células Cultivadas , Quimiotaxis de Leucocito/efectos de los fármacos , Citocinas/biosíntesis , Citocinas/genética , Citocinas/metabolismo , Femenino , Proteínas del Helminto/aislamiento & purificación , Leucocitos/citología , Leucocitos/efectos de los fármacos , Lipopolisacáridos/farmacología , Ratones , Ratones Endogámicos BALB C
8.
Braz J Med Biol Res ; 37(2): 223-6, 2004 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-14762577

RESUMEN

The extract of Ascaris suum suppresses the humoral and cellular immune responses to unrelated antigens in the mouse. In order to further characterize the suppressive components of A. suum, we produced specific monoclonal antibodies which can provide an important tool for the identification of these proteins. The A. suum immunosuppressive fractions isolated by gel filtration from an extract of adult worms were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells and the cloned hybrid cells obtained were screened to determine the specificity of secreted antibodies. Three monoclonal antibodies named MAIP-1, MAIP-2 and MAIP-3 were selected and were shown to react with different epitopes of high molecular weight proteins from the A. suum extract. All antibody molecules have kappa-type light chains but differ in heavy chain isotype. MAIP-1 is a mouse IgM, MAIP-2 is an IgA immunoglobulin and MAIP-3 is an IgG1 immunoglobulin and they recognize the antigen with affinity constants of 1.3 x 10(10) M-1, 7.1 x 10(9) M-1 and 3.8 x 10(7) M-1, respectively. The proteins recognized by these monoclonal antibodies (PAS-1, PAS-2 and PAS-3) were purified from the crude extract by affinity chromatography and injected with ovalbumin in BALB/c mice in order to determine their suppressive activity on heterologous antibody production. It was demonstrated that these three proteins are able to significantly suppress anti-ovalbumin antibody secretion, with PAS-1 being more efficient than the others.


Asunto(s)
Anticuerpos Antihelmínticos/inmunología , Anticuerpos Monoclonales/inmunología , Ascaris suum/química , Proteínas del Helminto/inmunología , Inmunosupresores/inmunología , Animales , Anticuerpos Antihelmínticos/biosíntesis , Anticuerpos Monoclonales/biosíntesis , Especificidad de Anticuerpos/inmunología , Ascaris suum/inmunología , Western Blotting , Cromatografía de Afinidad , Electroforesis en Gel de Poliacrilamida , Proteínas del Helminto/aislamiento & purificación , Inmunosupresores/aislamiento & purificación , Ratones , Ratones Endogámicos BALB C
9.
Braz. j. med. biol. res ; 37(2): 223-226, Feb. 2004. ilus
Artículo en Inglés | LILACS, SES-SP | ID: lil-354183

RESUMEN

The extract of Ascaris suum suppresses the humoral and cellular immune responses to unrelated antigens in the mouse. In order to further characterize the suppressive components of A. suum, we produced specific monoclonal antibodies which can provide an important tool for the identification of these proteins. The A. suum immunosuppressive fractions isolated by gel filtration from an extract of adult worms were used to immunize BALB/c mice. Popliteal lymph node cells taken from the immunized animals were fused with SP2/O myeloma cells and the cloned hybrid cells obtained were screened to determine the specificity of secreted antibodies. Three monoclonal antibodies named MAIP-1, MAIP-2 and MAIP-3 were selected and were shown to react with different epitopes of high molecular weight proteins from the A. suum extract. All antibody molecules have kappa-type light chains but differ in heavy chain isotype. MAIP-1 is a mouse IgM, MAIP-2 is an IgA immunoglobulin and MAIP-3 is an IgG1 immunoglobulin and they recognize the antigen with affinity constants of 1.3 x 10(10) M-1, 7.1 x 10(9) M-1 and 3.8 x 10(7) M-1, respectively. The proteins recognized by these monoclonal antibodies (PAS-1, PAS-2 and PAS-3) were purified from the crude extract by affinity chromatography and injected with ovalbumin in BALB/c mice in order to determine their suppressive activity on heterologous antibody production. It was demonstrated that these three proteins are able to significantly suppress anti-ovalbumin antibody secretion, with PAS-1 being more efficient than the others.


Asunto(s)
Animales , Ratones , Anticuerpos Antihelmínticos , Proteínas del Helminto , Ascaris suum , Inmunosupresores , Anticuerpos Monoclonales , Anticuerpos Antihelmínticos , Proteínas del Helminto , Western Blotting , Ascaris suum , Electroforesis en Gel de Poliacrilamida , Inmunosupresores , Ratones Endogámicos BALB C , Anticuerpos Monoclonales , Especificidad de Anticuerpos
10.
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
11.
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
12.
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
13.
Eur J Pharmacol ; 421(2): 133-40, 2001 Jun 08.
Artículo en Inglés | MEDLINE | ID: mdl-11399269

RESUMEN

The effect of neonatal capsaicin (8 methyl-N-vanillyl-6-nonenamide) treatment on the leucocyte infiltration into the airways and pleural cavity was investigated in rats actively sensitized with ovalbumin. The animals were neonatally injected with either capsaicin (50 mg/kg, s.c., 2nd day of life) or vehicle (10% ethanol and 10% Tween 80). At adult ages, the animals were actively sensitized with ovalbumin (200 microg, s.c.) and 14 days later they were intratracheally (or intrapleurally) challenged with ovalbumin. The substance P level in bronchoalveolar lavage fluid of the capsaicin group was reduced by >90% compared to control group (vehicle), confirming the efficacy of capsaicin treatment. In the capsaicin group, the number of neutrophils (but not of eosinophils and mononuclear cells) in bronchoalveolar lavage fluid of sensitized animals was significantly higher than the control group. Intrapleural injection of ovalbumin in sensitized rats caused a significant neutrophil influx at 6 h that was markedly increased in the capsaicin-pretreated animals compared to control group. The counts of eosinophils and mononuclear cells in the pleural exudates did not differ significantly between capsaicin and control groups. The increased levels of immunoglobulin (Ig)E, IgG1 and IgG2a anti-ovalbumin antibodies in serum of sensitized rats did not differ between capsaicin and control groups. In conclusion, the exacerbated pulmonary neutrophil recruitment caused by the capsaicin neonatal treatment is unrelated to increase in serum immunoglobulin antibodies, and suggests a protective role for C-fibers in attenuating the allergic neutrophil infiltration.


Asunto(s)
Líquido del Lavado Bronquioalveolar/citología , Capsaicina/farmacología , Leucocitos/efectos de los fármacos , Fibras Nerviosas/fisiología , Pleura/efectos de los fármacos , Animales , Animales Recién Nacidos , Anticuerpos/sangre , Anticuerpos/efectos de los fármacos , Líquido del Lavado Bronquioalveolar/química , Recuento de Células , Femenino , Inmunoglobulina E/sangre , Inmunoglobulina E/efectos de los fármacos , Inmunoglobulina G/sangre , Inmunoglobulina G/efectos de los fármacos , Leucocitos/citología , Masculino , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Pleura/citología , Pleuresia/inducido químicamente , Pleuresia/fisiopatología , Ratas , Ratas Wistar , Sustancia P/metabolismo
14.
J Allergy Clin Immunol ; 104(3 Pt 1): 595-600, 1999 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-10482833

RESUMEN

BACKGROUND: Hormones play a modulating role in allergic inflammation. Hyperthyroidism may increase the severity of asthma, and hypothyroidism may ameliorate coexistent asthma. The mechanisms regulating this interaction are not completely understood. OBJECTIVE: The purpose of this study was to test the hypothesis that thyroid hormones influence the development of allergic airway inflammation after antigen challenge in rats. METHODS: The experimental design included either sensitized or nonsensitized surgically thyroidectomized and sham-operated rats. Experiments were performed 50 days after surgery. Thyroidectomized rats and sham-operated controls were sensitized by subcutaneous injection of ovalbumin (OVA) and Al(OH)(3) and challenged 14 days later by OVA inhalation. Bronchoalveolar lavages were performed 24 hours after challenge. RESULTS: Compared with controls, thyroidectomized animals presented markedly decreased cell yields from bronchoalveolar lavage fluid after OVA challenge. The impaired response was not related to changes in the number of circulating leukocytes. Determination of antibody serum concentrations indicated that thyroidectomized rats presented a marked reduction in the level of anti-OVA IgE compared with controls, without significant differences in IgG(1) and IgG(2a) serum concentrations. Reversal of the impaired responses was attained by 16-day treatment of hypothyroid animals with thyroxine, but not by 1- or 3-day treatment. CONCLUSION: The data presented suggest that the continuing deficiency of thyroid hormones influences the development of the inflammatory component of asthma. This is due, at least in part, to a decrease in the production of IgE.


Asunto(s)
Hipersensibilidad Respiratoria/etiología , Hormonas Tiroideas/fisiología , Animales , Formación de Anticuerpos , Líquido del Lavado Bronquioalveolar/citología , Hipotiroidismo/complicaciones , Recuento de Leucocitos , Masculino , Ovalbúmina/inmunología , Ratas , Ratas Wistar , Hipersensibilidad Respiratoria/inmunología , Tiroidectomía
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