|Budget Amount *help
¥3,000,000 (Direct Cost : ¥3,000,000)
Fiscal Year 1999 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1998 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1997 : ¥1,200,000 (Direct Cost : ¥1,200,000)
We have been analyzing whether there might be a factor involving the endocytosis of ETEC enterotoxin (LT) to target cell except GMI ganglioside receptor. After LT was added to the CHO cell, Immunoprecipitates with anti-LT antibody to cell lysate was analyzed with Western blotting with anti-LT antibody. There was a band, of which molecular weight was 22,000 except the monomer of LT-B and might shift the monomer upper side of gel. Then we examined this factor. However, some other examiner reported that this shift of the monomer was caused by the formation of dimer of LT-B in the cell. Therefore, we examined whether other factor might be involved in the endocytosis of LT to make an antibody blocking the endocytosis of LT-B.
Some rabbits were immunized by cell lysate, which was prepared from NCTC 2071 defecting GM1 ganglioside and we can prepared the anti-serum partially blocking the internization of LT to CHO cells. Then, we are now examining a factor reacting to this antiserum.
Morecover, while we received this grant, we reported data as followed;
1. LT is also coded in the chromosome, except the plasmid.
2. The mutant defecting nicking site of A subunit near Arg 192 have strong adjuvant action to VZV, KKLH, Ovalbumin, BGG and measle virus.
3. The nicking site near Arg 192 of the A subnit is not involved in some biological activity of LT.