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Ovalbumin-anti-CD-23 conjugates enhance ovalbumin-specific immune responses in a CD23-dependent manner

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Ovalbumin-anti-CD-23 conjugates enhance ovalbumin-specific immune responses in a CD23-dependent manner

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Samenvatting

A humoral immune response against blood-borne protein antigens is initiated in the white pulp of the spleen and requires activation of both B cells and helper T cells. Before a humoral immune response can be initiated, the antigen has to be transported to splenic follicles, because antigens have not access to the follicles by themselves. It has been shown that CD23+ B cells in vivo can transport IgE-antigen complexes into the follicles. CD23 is the low affinity receptor for IgE and is primarily expressed on B cells and follicular dendritic cells in mice. When mice are immunized with IgE-antigen complexes, an enhanced immune response can be seen. In the current study the transport function of CD23 on B cells was used to investigate whether ovalbumin conjugated to anti CD23 antibodies can facilitate antigen transport into the B cell follicles and enhance an antigen-specific immune response. Using CD23 as a transporting molecule, the ovalbumin conjugates can be efficiently transported to the follicles of the spleen.
First, ovalbumin was conjugated to anti-CD23 antibodies and to an isotype control. Subsequently, BALB/c and CD23 deficient mice were immunized with the ovalbumin conjugates and ovalbumin-specific IgG was measured at different time points. To analyze ovalbumin-specific T cell proliferation, BALB/c mice were adoptively transferred with DO11.10 cells, carrying a transgenic T cell receptor for a specific ovalbumin peptide, and immunized with ovalbumin conjugates. The number of transgenic CD4+ T cells was determined by flow cytometry of splenic cells. To analyze germinal center formation, spleens were isolated from conjugate immunized BALB/c mice and flow cytometry was performed to determine the number germinal center B cells. Also spleen sections were immunofluorescent stained and the percentage of germinal center containing follicles was determined by counting the germinal centers and follicles.
The primary ovalbumin-specific IgG antibody response in this study was significantly enhanced in mice immunized with ovalbumin-anti-CD23 conjugates compared to the mice immunized with the isotype conjugates or ovalbumin alone and compared to the immunized CD23 deficient mice. The proliferation of ovalbumin-specific CD4+ T cells was not enhanced in mice immunized with ovalbumin-anti-CD23 conjugates in comparison with the control groups. The ovalbumin-specific T cell response was even significantly lower than the mice immunized with isotype conjugates. The percentage of germinal center B cells in the mice immunized with ovalbumin-anti-CD23 conjugates was significantly higher than the percentage of the ovalbumin alone group. Also the percentage of germinal center containing follicles was higher in mice immunized with ovalbumin-anti-CD23 conjugates compared to the isotype control group and ovalbumin alone group. The increased germinal center formation in the mice immunized with ovalbumin-anti-CD23 conjugates is in correlation with the enhanced ovalbumin-specific antibody response.
In conclusion, ovalbumin-anti-CD23 conjugates immunized in mice enhance ovalbumin-specific immune responses in a CD23-dependent manner.

Toon meer
OrganisatieAvans Hogeschool
OpleidingBiologie en Medisch Laboratoriumonderzoek-Breda
AfdelingATGM Academie voor de technologie van Gezondheid en Milieu
PartnersUppsala University
Datum2013-04-26
TypeBachelor
TaalEngels

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