Establishment of tumor immunotherapy using class II

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In the first year, we tried to establish the system to delivery the recombinant hCIITA molecules into cancer cells. We first used VP22 tag derived from HSV as a candidate of deliverers, the VP22 interfered the function of hCIITA. Thus, we next used Tat tag derived from HIV and obtained the facts that Tat-hCIITA has enough function to induce MHC antigens in cancer cells and purified recombinant Tat-hCIITA protein in the culture media move into cancer cells and could make MHC induce. However, it was impossible to get enough amounts of recombinant Tat-hCIITA protein, since the protein formed inclusion body in E.coli.In the second year, we tried to establish the co-expression system of hCIITA and MHC haplotype-matched antigenicoligopeptides (Tb (k) and Tb (km) = I-A^k-haplotype matched oligopeptides, Tb (b) = I-A^b-haplotype matched oligopeptides) derived from Mycobacterium tuberculosis alpha antigen. We constructed the expression vectors, pZeo-hCIITA-HA-furin-Tb (k) -His, pZeo-hCIITA-HA furin-Tb (km) -His, and pZeo-hCIITA-HA-furin-Tb (b)-His and transfected into he C1300 (mouse neuroblastoma cell line derived from A/J mouse (I-A^k haplotype), MM102 (mouse breast cancer cell line derived from C3H/HeN mouse (I-A^k haptotype), and Lewis (mouse lung cancer cell line derived from C57BL/6 mouse (I-A^b haplotype). We have established stably hCIITA-and Tb (b)-coexpressed transfectants (Lewis-pZeo-hCIITA-HA-furin-Tb(b)-His and MM102-pZeo-hCIITA-HA furin-Tb (b)-His) and got the evidence that Lewis-pZeo-hCIITA-HA furin-Tb (b)-His generates well tumor immunity against tumor-laden hosts.