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M S Macedo

Publications and source records attributed to M S Macedo.

28 records · Page 2Linked to original sources

Isolation of Ascaris suum components which suppress IgE antibody responses.

An extract from adult Ascaris suum was fractionated on a Sephadex G-200 column and individual elution fractions assessed for their ability to induce or suppress an IgE antibody response in mice immunized simultaneously with a heterologous antigen (ovalbumin). The results showed that, whilst fractions eluted in the first peak suppressed the anti-ovalbumin IgE antibody production, those eluted in the third peak induced a significant anti-Ascaris IgE response. The mixture of the two kinds of fractions resulted in suppression of anti-Ascaris IgE antibodies. The apparent molecular weight (MW) of the first and third peaks was 530,000 and 29,000 daltons, respectively. After delipidation, the extract still retained most of its biological activities. SDS-PAGE of this material showed selective loss of high-MW components when compared to the whole extract. No bands were observed in SDS-PAGE of peak I in contrast to peak III, which displayed several bands. Immunoblots of all these samples showed at least 3 bands above MW 150,000, which reacted with anti-peak I antiserum, in the unfractionated extracts and in peak I, but not in peak III. When the extract was fractionated by affinity chromatography using anti-peak I antibodies as ligands, the immunogenic components were present in the effluent, whereas the suppressive components were recovered in the eluate.

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Suppression of antibody response to an unrelated antigen in experimental murine paracoccidioidomycosis: effect of cyclophosphamide and indomethacin.

A suppressive effect of experimental paracoccidioidomycosis on the IgE antibody response to an unrelated antigen (ovalbumin) has been previously observed in mice. This effect was restricted to a short period, reaching maximum levels when OA was administered on the third day of Pb-infection. In order to study possible mechanisms involved in the establishment of this suppression, resistant (A/SN) and susceptible (B10.A) mice were treated with either a low dose of cyclophosphamide (CY) or indomethacin (INDO), a potent inhibitor of prostaglandin synthesis. While treatment with the first drug in A/SN mice induced only a recovery of the IgE anti-OA antibody response, in B10.A mice this effect was extended to IgG1, IgG2a and total levels of anti-OA antibodies. On the other hand, treatment with INDO reverted the anti-OA antibody suppression regarding each antibody class tested and in both strains of mice. These results suggest the participation of prostaglandins and of a cyclophosphamide-sensitive mechanism in the induction of the suppressive phenomenon in the human system.

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

The effect of X-irradiation on the suppression 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 24 h before immunization with 50 micrograms OVA and 200 micrograms ASC suppressive components abolished the damping effect on anti-OVA IgE antibody levels. The same effect was observed on the anti-ASC IgE antibody response obtained in mice injected with 200 micrograms ASC immunogenic plus 200 micrograms 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 suppresses both the homologous (anti-ASC) and heterologous (anti-OVA) antibody formation.

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PAF antagonists do not modify IgE antibody production in mice.

The effect of selective PAF antagonists on the in vivo production of IgE antibodies was investigated. The anti-ovalbumin IgE antibody content was estimated by passive cutaneous anaphylactic reaction (PCA) in the plasma of Balb/c mice 10 days after immunization with ovalbumin and alum. The PAF antagonists, BN 52021 (5 mg/kg, ip), BN 50730 (20 mg/kg, po), WEB 2086 (2 mg/kg, ip) and WEB 2170 (5 mg/kg, ip) were administered 1 h before immunization and twice a day for 8 days thereafter. The effect of the antagonists on the PAF-induced vasopermeability was also assayed. In the immunized mice the level of antiovalbumin IgE antibody, estimated by PCA titer, was 1/640. The treatment with the PAF antagonists did not change this level. At the concentrations employed, the antagonists BN 50730, WEB 2086 and WEB 2170 significantly reduced the PAF-induced vascular permeability. These results suggest that PAF does not seem to have a relevant effect on the production of IgE antibodies in vivo in the system used in the present study.

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Further characterization of Ascaris suum component(s) with suppressive activity on the IgE antibody response.

In order to characterize the component(s) of Ascaris suum responsible for damping of the IgE antibody production we demonstrated that the extract incubated in sodium acetate buffer, pH 4.5, maintained its suppressive effect and the same protein banding pattern by SDS-PAGE. Elimination of the lipoprotein components of the extract also left its damping properties unchanged. SDS-PAGE of the lipoprotein-free extract revealed practically the same pattern as shown by the whole extract, except for the high molecular weight polypeptides. These results indicate that the suppressive component(s) of A. suum did not precipitate and retained their activity at low pH. In addition, they appear not to be lipoproteins.

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Suppression of IgE antibody production against an unrelated antigen in experimental murine paracoccidioidomycosis.

A suppression of the IgE antibody response to ovalbumin was obtained in susceptible mice infected with Paracoccidioides brasiliensis yeast cells a few days prior to immunization with the former antigen plus adjuvant. A direct relationship between the number of injected fungi and the suppressive effect was established. When infection with a pathogenic isolate of P. brasiliensis (Pb 18) was compared to a non-pathogenic isolate (IVIC Pb267), the IgE anti-ovalbumin response was reduced by both. A similar effect was observed if mice were injected with dead yeast cells prior to immunization. Two strains of mice with completely opposite susceptibilities to infection with Pb18 cells (B10.A--susceptible and A/SN--resistant) both showed suppressed IgE anti-ovalbumin antibody production when infected 3 days prior to immunization. Injection of both strains of mice with P. brasiliensis antigen on the same day as immunization also had the same suppressive effect. These results suggest that the suppression of IgE response to an unrelated antigen in experimental murine paracoccidioidomycosis could be due to antigenic competition or to a suppressive component present in P. brasiliensis cells.

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Murine delayed type hypersensitivity is suppressed by Ascaris suum extract.

We studied the suppressive effect of an Ascaris suum extract on delayed-type hypersensitivity (DTH) reactions to ovalbumin (OVA) and to Bacillus Calmette-Guerin (BCG). The ability of mice to develop DTH reactions to both antigens was suppressed when an immunizing dose of the antigen was given subcutaneously together with the Ascaris extract. Partial or complete suppression of the response to OVA was obtained by the use of 400 or 1000 micrograms of Ascaris extract, respectively. The response to BCG, on the other hand, was totally suppressed by 400 micrograms of extract.

Animals↗

Suppression of IgE antibody production by Ascaris suum extract: characterization of suppressive component(s).

Partial characterization of the suppressive component(s) of A. suum extract that is (are) responsible for damping production of IgE antibody to ovalbumin was performed by physical and chemical methods. Digestion of the whole extract with trypsin and chymotrypsin completely abolished the suppressive activity. Oxidation with sodium metaperiodate or heating at 56 degrees C, however, had no effect. These results indicate that the integrity of heat-stable protein(s) present in the crude extract is essential for its suppressive effect. In addition, the carbohydrate moiety does not seem to play an important role in this effect.

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Suppressive effect of an Ascaris suum extract on IgE and IgG antibody responses in mice.

1. A suppressed cytotropic and agglutinating antibody response against ovalbumin was induced in mice immunized with this antigen and an Ascaris suum extract. 2. Suppression of IgE antibody production was abolished by administration of cyclophosphamide or X-irradiation before immunization. In contrast, suppression of homocytotropic IgG1 and heterocytotropic IgG2a antibody responses was considerably resistant to the same treatments. 3. The low levels of IgG agglutinating antibodies also remained unchanged after these treatments. 4. These results indicate that distinct regulatory mechanisms are the targets of suppression induced by A. suum extract in IgE and IgG responses.

Animals↗

Characterization of allergenic fractions from Drechslera monoceras.

Drechslera monoceras, a fungus of the Deuteromycota phylum, is fairly frequent in Brazil, and is spread through the atmosphere. In previous studies done in the city of Sao Paulo, it was found that in relation to 42 other fungi extracts, the crude extract of this fungi demonstrated a more intense cutaneous reaction in patients with respiratory allergies. Biochemical, antigenic and allergenic evaluations were carried out at various growth stages of this fungus. Based on these facts, the purpose of this research was the fractionation and allergenic characterization of the allergenic extract of D. monoceras to be used in diagnosis and immunotherapy in patients with positive cutaneous reaction to this fungus. In the city of Sao Paulo, 13 of 248 patients with respiratory allergy (asthma and/or rhinitis) showed positive reaction following cutaneous tests (skin prick tests). The crude extract of D. monoceras was fractionated by SDS-PAGE. The visible fractions were then separated by electroelution to be inoculated into BALB/c mice to evaluate the production of IgE antibody. The IgE content was detected by passive cutaneous anaphylaxis test in Wistar rats, and two fractions of approximate molecular weights of 14.4 and 36 KDa reacted to the test. The in vitro allergenic characterization was carried out by Western blotting, and three fractions of approximate molecular weights of 14.4, 36 and 60 KDa were positive. It was concluded that the extract of D. monoceras has at least three allergenic determinants, which can be used for diagnosis and immunotherapy in patients with respiratory allergy to this fungi.

Allergens↗