Establish an in Vitro Immune Rejection Response for the Receptor to Realize the Transplantation of NK Cells from Different Species

Abstract

Healthy adult grass turtle as the donor, draw 10ml of blood, prepare PBMC, isolate and extract purified grass turtle NK cell suspension.

Healthy male Big dog is a receptor. Under anesthesia, 200 milliliters of venous blood is extracted out of the body, enters the blood cell separator, extracts plasma and lymphocyte components, a total of 115 ml, and puts a blood collection bag with heparin sodium. After preserving 10ml plasma, divide the lymphocyte plasma into 3 bags of 35 ml each. Storage.

Take 1 bag of Big Dog Lymphocyte Plasma 35ml, dilute isovolume physiological saline, inject 15ml of grass turtle NK cell suspension, produce a mixed lymphocyte plasma between Turtle and Big Dog, and put it in the incubator. On the third day after mixing, the immune rejection began, and the grass turtle NK cells accounted for 76% of the dead cells. On the fifth day, the response peaked. Grass turtle NK cells accounted for 92% of the dead cells. On the 10th day, the immune rejection response was stationary, and 28% of the grass turtle NK cells survived, achieving a non-rejective peacekeeping coexistence state with the receptor Big Dog lymphocytes. When this group of grass turtle NK cells that have undergone the immune rejection test of the receptor Big dog in vitro are imported into the Big dog body with the Big dog lymphocytes, they are no longer regarded as heteroactive antigens by the receptor Big dog immune system and are not attacked by immune rejection. It became an integral part of the lymphocytes in the Big dog, and the experimental plan for the transplantation of xenogeneic NK cells from grass turtles into Big dog has been completed.‌

During the natural survival period of grass turtle NK cells, we can select the blood at a different time to prepare the peripheral blood mononuclear cell PBMC of the Big dog. According to the principle of autologous NK cell retransfusion, we can isolate and cultured mixed NK cells of grass turtle and Big dog in vitro, and retransfuse them into the Beagle. Through staining comparison, grass turtle NK cells can also be specially selected from the mixed NK cells of grass turtle and Big dog for isolation and culture in vitro, and returned to the Big dog. Both methods belong to autologous NK cell retransfusion therapy.

The article expounds the clinical significance of establishing an in vitro immune rejection for the receptor, and points out that establishing an in vitro immune rejection for the receptor is a routine test that must be carried out by allogeneic immune cell transplantation and the problem of disparity between the in vivo and in vitro survival environments of immune cells.