| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Research Abstract |
To identify the novel antigens that can effectively induce the immune response in order to develop an effective vaccine for the prevention of trypanosomiasis, neutral glycosphingolipids (GSLs) and gangliosdes were isolated from Trypanosoma brucei and analyzed by thin-layer chromatography (TLC) and TLC/secondary ion mass spectrometry (TLC/SIMS) or TLC immunostaining test. Three species of neutral GSLs, N-1, N-2 and N-3, were separated on TLC. The TLC/SIMS analysis of N-1 of the parasites revealed a series of (M-H)^- ions from m/z 698 to m/z 825 representing the molecular mass range of ceramide monohexoside (CMH) (GlcCer or galactosylceramide). On the other hand, the TLC/SIMS spectra of N-2 GSL revealed a series of (M-H)^- ions from m/z 944 to m/z 987 indicating the molecular mass range of LacCer. In the TLC/SIMS analysis of N-3 GSL, however, the characteristic molecular ions that can elucidate the structure of N-3 GSL were not obtained. Both GlcCer and LacCer were detected on the cell s
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urface of T. brucei by antibodies. In ganglioside fraction, four species of gangliosides, G-1, G-2, G-3 and G-4, were separated on TLC. G-1 ganglioside showed the reactivity to the anti-GM3 monoclonal antibody. G-2 was recognized by the anti-GM1 monoclonal antibody. G-3 gave reaction with the monoclonal antibody to GD1a G-4 had the reactivity to anti-GD1b monoclonal antibody. GM3, GM1, GD1a and GD1b were detected on the cell surface of T. brucei by using 4 kinds of monodonal antibodies. Furthermore, to investigate whether immune response against GSLs on T. brucei is effective against the inhibition of T. brucei infection, BALB/c mice were immunized with liposome containing ganglioside mixture (GM1, GD1a and GD1b) and challenged with T. brucei. All of control (non-immunized) mice died 6 days after challenge. However, none of mice immunized with ganglioside mixture-containing liposome died 40 days after challenge. In addition, no parasites were detectable in survived mice. These results suggest that induction of immune response against gangliosides on T. brucei would be effective for the prevention of T. brucei infection. Less
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