Breaking the Non‐Responsiveness of C57BL/6 Mice to the Malarial Antigen EB200 – The Role of Carrier and Adjuvant Molecules

Abstract

Poor immunogenicity and major histocompatibility complex (MHC) restriction of immune responses to certain recombinant proteins or synthetic peptides impose problems in developing effective vaccines. EB200 is one of the vaccine candidate antigens from Plasmodium falciparum, which induces MHC-restricted immune responses in mice of different haplotypes. A way of overcoming this problem is to conjugate the antigen to an immunogenic protein carrier and to use optimal adjuvant substances. We have investigated the carrier effect of glutathione-S-transferase (GST) in CBA and C57BL/6 mice which are high and low responder to EB200, respectively. Our results reveal that the MHC restriction in C57BL/6 mice was broken by the use of GST as a carrier. Studies on the B-cell repertoires in both strains of mice immunized with GST-EB200 by preparing hybridoma cell lines indicate that the B-cell repertoires were similar in both CBA and C57BL/6 mice. However, the antibody affinity and the magnitude of the response were still lower in the low-responder C57BL/6 mice compared with that in CBA even when cholera toxin (CT) was used as adjuvant. To improve the response, the efficacy of various adjuvant substances like alum and Hsp 70 from Trypanosoma cruzi and the combination of various adjuvants was analysed. CT and Hsp 70 together act synergistically and markedly improve the immunogenicity of EB200 by increasing antibody affinity and the magnitude of the responses in C57BL/6 mice, which may be explained by the complementary effect of adjuvants. These results are of importance in the design of efficient vaccines.

Monocyte‐derived dendritic cells from chagasic patients vs healthy donors secrete differential levels of IL‐10 and IL‐12 when stimulated with a protein fragment of Trypanosoma cruzi heat‐shock protein‐70

“…It has been shown that HSP70 from T. cruzi , Plasmodium falciparum , Mycobacterium tuberculosis and Leishmania braziliensis may function as adjuvants in the induction of the immune response in animal models 24 , 25 , 26 . However, studies on the interaction of DC and T. cruzi demonstrated that the parasite may infect such cells and even proliferate intracellularly, altering their maturation program by inhibiting the production of cytokines, downregulating co‐stimulatory molecules, diminishing the expression of class I and class II histocompatibility complex molecules, and therefore altering the capacity to present antigenic peptides to T lymphocytes 27 , 28 .…”