ABSTRACT
One of the mechanisms by which adjuvants are believed to promote T-cell activation and prevent induction of oral tolerance is by up-regulating the expression of co-stimulatory molecules on antigen presenting cells. Mice treated orally with palmitoyl-ovalbumin conjugates become immunized, while those treated with native ovalbumin (Ova) become tolerant. Cells from the peritoneal cavity of B6D2F1 mice were cultured in the presence of 0.01, or 0.1 mg/100ml of either Ova, or palmitoyl-Ova and tested for the presence of cell markers. PE-conjugated anti-mouse CD80, CD86, and CD11b antibodies as well as biotin-PE were used to stain the antigen-activated peritoneal cells. A significant increase in the expression of CD86 and CD80 was observed following in vitro stimulation with palmitoyl-Ova; additionally, both Ova and palmitoyl-Ova induced the basal expression of CD11b. These findings could be related with the strong T-cell proliferative response induced by palmitoyl-Ova.
ABSTRACT
The aim of this work was to evaluate the effects of ethanol on the adsorption and tissue distribution of orally administered antigens in mice. Results showed that ethanol reduced the level of anti-ovalbumin IgA antibodies in intestinal fluid for the mice treated orally with a palmitoyl-ovalbumin conjugate. Ethanol was administered intragastrically to mice at 5 g/kg body weight for 14 days (chronic treatment), or 10 g/kg body weight every 7th day up to 14 days (acute treatment). Thereafter, 99m technetium-labeled antigens were administered and lymphoid tissues were collected. Ethanol interfered with the transport of ovalbumin to the liver. Moreover, the transport of palmitoyl-ovalbumin to mesenteric lymph nodes was reduced 6 h after the antigen administration. In conclusion, there was a relationship between the suppression of ethanol-mediated specific local IgA responses and the decreased transport of palmitoyl-ovalbumin to mesenteric lymph nodes.
ABSTRACT
The oral administration of proteic antigens, like ovalbumin, may result in the induction of oral tolerance or immunization. The aim of this work was to label a protein antigen with 99mTechnetium, encapsulate it in liposomes and investigate its absorption and tissue distribution after oral administration in mice. Ovalbumin was labeled with 99mTechnetium and encapsulated in small unilamellar vesicles. 99mTc-OVA encapsulated or not in liposomes was administrated to mice that were sacrificed after different times. The radioactivity was measured in various organs of the animals. Differences concerning the biodistribution of 99mTc-OVA were noticed. The technique may represent alternatives for the induction of immunization or oral tolerance.
A administração oral de antígenos protéicos pode levar a indução ou supressão da resposta imune. A indução da resposta imune é de grande importância para o desenvolvimento de vacinas orais. Já a supressão, denominada de tolerância oral, pode vir a representar uma solução terapêutica a numerosas enfermidades. O objetivo deste trabalho foi de compreender o comportamento in vivo de um antígeno modelo, a ovalbumina (OVA), marcado com um radioisótopo, o 99m tecnécio (99mTc), e administrado por via oral na sua forma livre e encapsulado em lipossomas de pequeno tamanho (SUV). As amostras foram administradas por gavagem à camundongos, sacrificados após 30, 60, 90, 120, 180 e 360 minutos. A radioatividade foi medida no estômago, intestinos, placas de Peyer, linfonodos mesentéricos, baço, fígado e sangue dos animais. Os resultados mostraram que a OVA livre ou encapsulada em lipossomas SUV apresenta biodistribuições distintas. As diferenças encontradas na biodistribuição da OVA livre ou encapsulada podem representar mecanismos diferentes para a indução ou não de tolerância oral.