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International Immunology, Vol. 15, No. 10, pp. 1183-1194, October 2003
© 2003 Japanese Society for Immunology

Immunogenicity of Peptide-25 of Ag85B in Th1 development: role of IFN-{gamma}

Ai Kariyone1, Toshiki Tamura1, Hideyuki Kano1, Yoichiro Iwakura2, Kiyoshi Takeda3, Shizuo Akira3 and Kiyoshi Takatsu1

1 Division of Immunology, Department of Microbiology and Immunology, and 2 Laboratory of Cell Biology, Center for Experimental Medicine, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan 3 Institute of Microbial Diseases, Osaka University, Osaka 565-0871, Japan

Correspondence to: K. Takatsu; E-mail, takatsuk{at}ims.u-tokyo.ac.jp
Transmitting editor: S. Koyasu

Ag85B (also known as {alpha} antigen or MPT59) is immunogenic, and induces expansion and differentiation of TCRVß11+CD4+ T cells to IFN-{gamma}-producing cells in C57BL/6 (I-Ab) mice. We reported that Peptide-25 (amino acids 240–254) of Ag85B is a major T cell epitope, and its amino acid residues at position 244, 247, 249 and 252 are I-Ab contact residues. Here we examined roles of IFN-{gamma} in the generation of Peptide-25-reactive CD4+ TCRVß11+ T cells and the efficacy of mutant peptides of Peptide-25 for Th1 development in mice other than C57BL/6 mice. Immunization of C57BL/6 mice with Peptide-25 included in incomplete Freund’s adjuvant led to preferential induction of CD4+ TCRVß11+ IFN-{gamma}- and tumor necrosis factor-{alpha}-producing T cells. Compared with other I-Ab-binding peptides such as Peptide-9 of Ag85B, 50V of pigeon cytochrome c and ovalbumin (OVA)265–280 peptide, only Peptide-25 was capable of inducing enormous expansion of TCRVß11+ IFN-{gamma}-producing T cells. Treatment of C57BL/6 mice with anti-Vß11 antibody before Peptide-25 immunization reduced the development of CD4+ IFN-{gamma}-producing T cells. Furthermore, B10.A(3R) mice, I-Ab-positive and TCRVß11– strain, showed remarkably lower response to Peptide-25 immunization than C57BL/6 mice. Peptide-25-primed IFN-{gamma}–/– cells showed significantly decreased expansion of CD4+ TCRVß11+ T cells as compared with wild-type cells. Interestingly, Peptide-25-primed cells from MyD88-deficient mice responded to Peptide-25 and differentiated into IFN-{gamma}-producing cells to a similar extent as wild-type mice, indicating Toll-like receptor-independent IFN-{gamma} production. These results imply that IFN-{gamma} plays important roles for the generation and expansion of CD4+ TCRVß11+ T cells in response to Peptide-25. Although Peptide-25 was non-immunogenic in C3H/HeN mice, a substituted mutant of Peptide-25, 244D247V, capable of binding to I-Ak, induced Th1 development. These results clearly demonstrate important roles of IFN-{gamma} in the expansion of CD4+ TCRVß11+ T cells, and will provide useful information for delineating the regulatory mechanisms of Th1-cell development and for analyzing mechanisms on Th1-dominant immune responses.

Keywords: Ag85B, mycobacterium, subunit vaccine, TCRVß11, tuberculosis


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