Development of the new strategy in ABO blood group incompatible transplantation

abo血型不相容的实体器官移植的主要障碍是急性体液排斥反应，导致移植物功能障碍。与abo不相容的移植相关的体液性排斥反应是由与供体器官内皮结合的抗供体血型A/B抗体引发的。我们试图开发新的方法来中和血型抗原，以克服abo血型不相容的移植，这是通过(1)RNA干扰或(2)糖苷酶。(1)首先，构建ABO血型基因小干扰RNA。用小干扰RNA转染表达A或B抗原的癌细胞SW620、LoVo和SW48后，通过实时RT-PCR检测，ABO血型基因的信使RNA减少了70-80%。流式细胞术和westernblotting检测，ABO血型抗原的表达未被小干扰RNA抑制。(2) α- n -乙酰半乳糖胺酶不能完全降解A血型抗原。另一方面，α-半乳糖苷酶降解B抗原，将B组红细胞转化为B组红细胞，流式细胞术检测B组红细胞。与α-半乳糖苷酶反应的AB组红细胞转化为A组红细胞。α-半乳糖苷酶也将含有A和B抗原的癌细胞SW48转化为只含A抗原的细胞。我们成功地用糖苷酶中和了B型血抗原。

The major barrier to ABO-incompatible solid organ transplantation is the acute humoral rejection, leading to graft dysfunction. Humoral rejection associated with ABO-incompatible, transplantation is triggered by antidonor blood group A/B antibodies that bind to the endothelium of the donor's organ.We tried to develop the new methods of neutralizing the blood group antigen to overcome ABO-incompatible transplantation, which were performed by (1) RNA interference or (2) glycosidases.(1) Firstly, small interference RNA for the ABO blood group gene was created. When the cancer cell lines SW620, LoVo, and SW48 which express A or B antigen were transfected with the small interference RNA, the messenger RNA of the ABO blood group gene were reduced by 70-80%, detected by real time RT-PCR. However, expression of the ABO blood group antigen was not suppressed by small interference RNA, detected by flow cytometry and westernblotting.(2) Blood group A antigen was not completely degraded by α-N-acetylgalactosaminidase. On the other hand, B antigen was degraded by α-galactosidase, and group B RBCs were converted to group B RBCs, which were detected by flow cytometry. The group AB RBCs reacted with α-galactosidase were converted to group A RBCs. The cancer cell line SW48 which have A and B antigens were also converted to the cells with only A antigens by α-galactosidase.We succeeded in neutralizing the blood group B antigen by glycosidase.