Biomedical subjects

I Mota

Publications and source records attributed to I Mota.

At least 19 recordsLinked to original sources

PubMed1995

The mast cell revisited.

The role of mast cells in allergic reactions is reviewed and the origin, distribution and properties of mast cells are reported. The characteristics of two phenotypically distinct mast cell populations are described. The function and properties of the IgE molecule as an antigen receptor on mast cells is discussed. The IgE receptor and its structure is updated. The participation of mast cells in the acute and late phase of the allergic reaction is pointed out. A double role for mast cells in allergic reactions is suggested: they may be responsible for the acute phase reaction through the release of mediators such as histamine and leukotrienes and for the late phase reaction through the release of pro-inflammatory cytokines.

Cytokines↗

PubMed1994

Adjuvant effect of Loxosceles gaucho (South American brown spider) venom.

Injection of L. gaucho venom and antigens (ovalbumin, ovomucoid and bovine gamma globulin) into rabbit skin induced an intense local inflammatory lesion and resulted in a significant increase in the level of IgG antibodies to the antigen in both the primary and secondary humoral immune response. The adjuvant activity of the venom was associated with its high mol. wt components, which are responsible for the inflammatory lesion. Rabbits rendered unresponsive to the venom and injected with venom plus antigen presented a very mild local inflammatory reaction and no increase in antibody formation. When venom and antigen were injected simultaneously but at different skin sites no adjuvant effect was induced. However, when antigen was injected 4 hr after venom injection but at the same skin site a significant adjuvant effect was produced. Furthermore, when venom plus antigen was injected intradermally into mice, a species in which the venom does not cause an inflammatory skin lesion, no adjuvant effect was detected. It is suggested that the adjuvant effect of L. gaucho venom in rabbits is probably due to its ability to cause a local severe inflammatory reaction.

Adjuvants, Immunologic↗

PubMed1994

The discovery of the relationship between mast cells, histamine and IgE.

Two years after graduating from medical school, Ivan Mota decided to spend a year doing postgraduate training. Little did he realise that he was embarking on a course that would see him lay the foundations for modern research into allergy and anaphylaxis. Here, he recounts his own tale.

Anaphylaxis↗

PubMed1994

Suppression of the antivenom antibody response by serum therapy.

1. The antivenom antibody response of mice injected with Bothrops jararaca venom and receiving specific serum therapy was studied under different experimental conditions. Balb/c mice (18-22 g) injected with venom (1.75 mg/kg) presented the clinical symptoms observed in patients bitten by B. jararaca and a high and long-lasting antivenom antibody response. 2. Injection of 0.1 ml of horse antiserum to venom 15 min after venom administration abolished the symptoms induced by the venom and induced an almost completely suppressed production of mouse antivenom antibodies. The extent of suppression of the antivenom antibody response depended on the dose of horse antiserum administered and was greater the sooner the serum therapy was applied after envenomation. 3. Injection of antiserum into envenomed mice that received an unrelated antigen (KLH) did not suppress the antibody response to KLH antigen though it inhibited production of antivenom antibodies. 4. Envenomed mice receiving an equivalent dose of F(ab')2 fragments obtained by pepsin digestion of horse antiserum presented the same extent of suppression of the antivenom antibody response as mice injected with the non-treated antiserum. 5. Mice whose antibody response was suppressed, when rechallenged with venom, presented a primary antibody response. 6. These results suggest that suppression of the antivenom antibody response presented by envenomed patients submitted to serum therapy is due to the masking of the venom epitopes by horse antibodies as well as to the rapid elimination of the venom epitopes.

Animals↗

PubMed1994

A rapid and efficient purification method for horse IgG(T) using a rat monoclonal antibody.

1. Louvain rats (IgK-1a) were immunized with horse IgG(T). To generate mAb to IgG(T), popliteal lymph node cells taken from the immunized animals were fused to a non-secreting LOU/C immunocytoma (IR983F). The hybridomas were cultured in HAT-containing medium and cloned under limiting dilution conditions. Supernatants from the growing hybrids were screened by ELISA using plates coated with horse IgG(T) or IgGa+b+c. 2. The anti-IgG(T) mAb obtained was named LO-HoGT-1 (LOU anti-horse IgG(T)). It is an IgG2a rat antibody whose light chain allotype is IgK-1a, and with an affinity constant of 2.9 x 10(10) M-1. 3. Ascites was induced in LOU (IgK-1b) rats by injecting the hybridoma cells and incomplete Freund's adjuvant ip. To obtain purified mAb, ascitic fluid was applied to a Sepharose anti-rat LOU IgK-1a chain column. 4. The purified mAb was then coupled to Sepharose. Immunoelectrophoretically pure IgG(T) was obtained by passage of horse serum through this column. The entire procedure took less than 30 min and resulted in a highly purified IgG(T).

Animals↗

PubMed1994

Detection and neutralization of B. jararaca venom in mice.

1. Bothrops jararaca venom was detected by ELISA at different times in the skin, muscle, blood, liver, lung, heart, kidney and spleen of mice injected with venom i.m. or i.d. 2. The results showed that even 10 min after i.m. injection the venom is detected mostly in skin rather than in the muscle of the venom injection site. A small amount of venom was detected in the kidney up to 12 h after im venom injection, and none was detected in tissues of lung, heart, liver or spleen. 3. However, in mice injected i.d., the venom could be detected in the skin up to 24 h after injection. Local necrosis and haemorrhage could be neutralized by antivenom injected by the i.d. or i.v. routes only if the antivenom was given a short time after venom injection, even when antivenom is administered in high concentration. 4. In contrast, experiments performed in mice receiving venom i.d. and treated by i.d. or i.v. routes with antivenom injected at different times after envenoming showed that the effect of venom on blood coagulation could be counteracted by antivenom administered by either route up to 2 h after venom injection. 5. We suggest that a feasible amount of antivenom administered i.d. could be given as a first aid measure after a snake bite accident. However, further experimental studies using the i.d. route for antivenom administration are essential to confirm this possibility.

Animals↗

PubMed1994

Role of platelets and complement in the clearance of epimastigote forms of Trypanosoma cruzi.

1. Trypanosoma cruzi epimastigote forms are very rapidly removed from the circulation of normal and C5-deficient mice. Depletion of C3 by cobra venom factor results in a significant delay in parasite clearance. 2. During parasite clearance there is a significant decrease in the number of circulating platelets and parasite clearance is considerably delayed in thrombocytopenic animals. 3. In vitro incubation of epimastigote forms with normal mouse serum leads to the formation of parasite clumps provided that platelets are present. Inactivation of factor B or depletion of C3 prevents this phenomenon. 4. When epimastigotes are incubated with normal mouse serum they absorb one or more factors required for their aggregation with platelets. 5. It is suggested that in mice T. cruzi epimastigote forms are removed from circulation by the alternative pathway of complement activation and that both C3 and platelets are required for parasite clearance.

Animals↗

PubMed1993

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.

Animals↗

PubMed1992

Neutralization of different activities of venoms from nine species of Bothrops snakes by Bothrops jararaca antivenom.

Antivenoms are the usual treatment in cases of systemic envenoming by Bothrops snakes. However, the neutralization of each venom component by the antivenom is not well established. Bothrops jararaca antivenom, produced in rabbits, recognizes the venoms of nine different Bothrops species with high ELISA antibody titres. Western blot analysis showed that almost all antigens present on both homologous and heterologous venoms are recognized. Neutralization tests were performed using whole antivenom or its IgG fraction. The antivenom was able to neutralize the haemorrhagic, coagulant and necrotizing activities of the heterologous venoms in the same antivenom/venom proportion as for the homologous venom. Myotoxic activity was only partially neutralized. Neutralization of the proteolytic activity of heterologous venoms required higher amounts of antivenom than for the homologous venom. Phospholipase and oedema-inducing activities were completely neutralized only in the homologous system.

Animals↗

PubMed1992

Toxic activities of venoms from nine Bothrops species and their correlation with lethality and necrosis.

The comparison of seven toxic activities contained in venoms from nine different species of Bothrops and the correlation of each activity with lethality and necrosis was the subject of this study. The haemorrhagic, coagulant, necrotizing, myotoxic, proteolytic and phospholipase activities were not equally distributed among the venoms studied except for the oedema-inducing activity which was almost equally distributed among them. The correlation coefficient was estimated for each activity in relation to lethality and necrosis induced by the venom. Lethality was significantly related to haemorrhagic and oedema-inducing activities, whereas the necrotizing activity showed significant correlation with phospholipase and coagulant activities. Proteolytic activity presented a significant inverse correlation with lethality.

Animals↗

PubMed1992

IgG antibodies to Loxosceles sp. spider venom in human envenoming.

The presence and specificity of IgG antibodies produced by patients with loxoscelism were studied. The loxoscelism diagnosis was supported mainly by clinical parameters. A search for IgG antibodies anti-Loxosceles gaucho venom in patients with loxoscelism submitted to serumtherapy showed antibodies in four out of 20 patients. The IgG antibodies were detected as early as 9 days and as late as 120 days after bite. The highest IgG antibody titer was 1:640 and the lowest was 1:80. Immunoblotting tests showed that human anti-L. gaucho IgG antibodies recognize preferentially the components responsible for the dermonecrotic and lethal activities of the venom. A comparison of the clinical picture, the level of serum IgG antibodies and the dose of antivenom administered suggest that there is no relationship between these parameters.

Antivenins↗

PubMed1992

Dermonecrotic and lethal components of Loxosceles gaucho spider venom.

Loxosceles gaucho spider venom causes a typical dermonecrotic lesion in bitten patients and rarely causes lethal systemic effects. Gel filtration on Sephadex G 100 of L. gaucho spider venom resulted in three fractions: fraction A, containing the higher mol. wt components (approximately 35,000); fraction B, containing lower mol. wt components (approximately 15,000); and fraction C, containing very low mol. wt components (probably small peptides). The dermonecrotic and lethal activities were detected exclusively in fraction A. The venom and fraction A produced large dermonecrotic lesions in rabbits with necrosis spreading by gravity to the skin of the lateral body wall. Analysis by SDS-PAGE showed that the proteins contained in fraction A are approximately 35,000 and 33,000 mol. wt. Immunoblotting analysis showed that the proteins responsible for the dermonecrotic and lethal activity are very immunogenic and the first to be detected by antibodies during the course of immunization.

Animals↗

PubMed1992

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.

Animals↗

PubMed1992

Effect of purified components of Ascaris suum extracts on IgE antibody response in guinea pigs.

The effect of components P530 and P29, isolated from Ascaris suum adult worm extract (ASC), on the heterologous IgE antibody response was studied in guinea pigs. Groups of 7 guinea pigs were immunized ip with 50 micrograms of ovalbumin (OA) alone or mixed with 200 micrograms of each component precipitated in an alum gel. The primary and secondary IgE antibody response was evaluated by passive cutaneous anaphylaxis reaction (PCA). Immunization of guinea pigs with P29 plus ovalbumin (OA) resulted in a significant increase in the level of serum IgE anti-OA antibodies, especially in the secondary response (almost 8-fold higher when compared with control group). This potentiated response was not observed when the animals received OA plus P530 or the crude extract. Indeed, the P530 component, as well as the crude extract, induced a depression of the anti-OA IgE antibody response (2-3 fold decrease when compared with OA-immunized animals). It was also shown that P29, but not P530 or ASC, was capable of eliciting a strong anti-ASC IgE antibody response. These results demonstrate that in guinea pigs these two Ascaris suum components have antagonistic biological effects, one inducing potentiation and the other suppression of the heterologous IgE antibody response.

Animals↗

PubMed1991

Neutralization of myotoxic activity of Bothrops venoms by antisera to purified myotoxins and to crude venoms.

A phospholipase myotoxin (MOO-1) and a non-phospholipase myotoxin (JSU-5) were studied for their antigenic cross-reactivity and neutralization by different antisera. Antisera against JSU-5 and MOO-1 reacted equally with both myotoxins in ELISA assays. The specificity of these antisera was also similar, recognizing the same 14,000-18,000 mol. wt components in the venoms of Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi and Bothrops pradoi. Using creatine kinase assays, JSU-5 myotoxicity was completely neutralized by B. jararacussu antivenom or anti-JSU-5 antibodies and partially neutralized by B. moojeni antivenom or anti-MOO-1 antibodies. MOO-1 myotoxicity was completely neutralized by antisera against JSU-5 and MOO-1 and B. jararacussu antivenom, and only partially neutralized by B. moojeni antivenom. B. jararacussu venom induced high titres of antibodies against purified myotoxins. This antiserum completely inhibited the myotoxicity of the homologous venom and significantly reduced the myotoxicity of the remaining myotoxin-containing venoms. It is suggested that B. jararacussu venom is a good immunogen to induce antibodies against myotoxins present in the venoms of the different species of Bothrops.

Animals↗

PubMed1991

Isolation of IgGT from hyperimmune horse anti-snake venom serum: its protective ability.

Hyperimmune horse anti-bothropic serum, used in serum therapy, was analyzed for its IgGT content and protective ability. IgGT was isolated through a combination of salt-mediated hydrophobic chromatography and protein A affinity chromatography. The chromatographic fractions obtained were analyzed with regard to their isotype content and protective ability. The results suggest that the protective ability of hyperimmune anti-venom serum is located mainly in the IgGT subclass.

Animals↗

PubMed1991

The possible mechanism of action of IgG antibodies and platelets protecting against Trypanosoma cruzi infection.

1. The role of IgG antibody and platelets in the mechanism of defense against Trypanosoma cruzi infection is reviewed. 2. Experimental data showing the participation of the different IgG subclasses in the immune lysis and immune clearance of the parasites are discussed. 3. The involvement of the platelets in the removal of the parasites from the circulation is considered. 4. It is suggested that IgG anti-T. cruzi antibodies interact with circulating parasites leading to formation of microaggregates, activation of C3 and deposition of C3b on the immune aggregates followed by adherence of platelets through C3b receptors. The immune aggregates would then be taken up by cells of the mononuclear phagocytic system.

Animals↗

PubMed1991

Isolation of horse IgG with protein A.

Horse immunoglobulins were obtained from normal serum defatted with dextran sulfate and precipitated with ammonium sulfate. Eight mg of this preparation was submitted to affinity chromatography with protein A-Sepharose CL-4B. Low temperature (4 degrees C) and a starting buffer at pH 8.0 were conditions required for all IgG subclasses to bind to protein A, even those with low affinity. The IgGs bound to protein A were eluted with glycine buffer at pH 2.8. The yield was about 90%. It is suggested that isolated IgG, instead of whole Igs, be used in serum therapy, reducing the amount of Igs and diminishing serum-related reactions.

Animals↗