Trial of recombinant MSP-1 vaccine combined with T lymphocyte triggering factor(TLTF)

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We examined the basic research of malaria vaccicne development using African trypanosome (Trypanosoma brucei). Outlines of the results are as follows.1 : The trypanosome expression vectors were constructed with MSP-1 gene (msp-1). These vectors were transfected into T. brucei procyclic form, and expressions were analysed. However, we could not confirm the expression of MSP-1.It is suggested that msp-1 or MSP-1 may have some responsibility in molecular folding or conformation, for example, because the drug resistance gene included in the vectors were functioned and the other reporter genes constructed into the vectors were expressed as expected.Then, the policy of this project was converted to malaria vaccine development using the adjuvant effect of variable surface glycoproteins (VSG) of the T brucei bloodstream form.2 : The fusion proteins of [Histidine TagIVSG N-terminal conserved domain/MSP-1/VSG C-terminal conserved domain] (VSG-MSP) and [Histidine Tag/MSP-1] were tried to express using vaculovirusinsect cell system. The recombinant fusion proteins were solubilized with guanidine, and were purified using the Ni-agarose affinity column.3 : Mice were immunized by subcutaneous injection of recombinant proteins and were subsequently boosted 3 weeks and 6 weeks after the initial immunization. Three weeks after the final inoculation, all mice were challenged with the parasitized 106 erythrocytes by intraperitoneal injection. As the result, slight preventive effects on 9arasitemia were obserbed when mie were immunized with VSGMSP, though they could not survived the infection.