Analysis of a fuctor involving trnslocation of LT to the cell.

我们一直在分析除了GMI神经节苷脂受体外，是否存在与ETEC肠毒素(LT)对靶细胞的内吞作用有关的因素。将LT加入CHO细胞后，用抗LT抗体对细胞裂解物进行免疫印迹分析。除LT-B外，有一条相对分子质量为22,000的条带，可能使单体凝胶上部发生移动。然后我们考察了这一因素。然而，另一些检查者报告说，这种单体的移动是由细胞中LT-B的二聚体形成引起的。因此，我们研究了LT的内吞作用是否可能有其他因素参与，以制备一种阻断LT-B内吞的抗体。用NCTC 2071缺失GM1神经节苷脂制备的细胞裂解液免疫部分阻断LT内吞的CHO细胞的抗血清。然后，我们现在正在检查对这种抗血清起反应的一个因子。莫雷弗，当我们收到这笔赠款时，我们报告了以下数据：1.它也在染色体中编码，除了血浆。Arg 192附近A亚基缺失划痕位点的突变体对VZV、KKLH、卵清蛋白、BGG和麻疹病毒具有较强的佐剂作用。A亚基第192位附近的缺口位点不参与LT的某些生物活性。

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.