Measurement of ABO blood group antibody levels using renal cortical tubular epithelial cells of cynomolgus monkey / 中华器官移植杂志

ABSTRACT

Objective To investigate the possibility of measuring ABO blood group antibody levels using renal cortical tubular epithelial cells (RCTECs) of cynomolgus monkey.Method The primary RCTECs were isolated from cynomolgus monkey kidneys and identified by Western blotting and flow cytometry (FACS).FACS was applied to detect the expression of ABO histo-blood group antigens on RCTECs,the binding of blood group antibodies in human serum to RCTECs,and to compare the difference of measuring ABO blood group antibody levels between using human erythrocytes and RCTECs as target cells.Result The majority of cells derived from the kidney cortex were renal tubular epithelial cells.39.90％-73.80％ of RCTECs were found to express the ABO histoblood group antigens with the capability to bind the human blood group antibodies specifically.The expression level of ABO histo-blood group antigen on RCTECs was not influenced by long-term cryopreservation,and the percentage of B antigen positive cells was 39.90％ before frozen storage and 39.65％ after a 10-month cryopreservation respectively.The normal serum and allergic serum presensitized to synthetic A antigen-keyhole limpet hemocyanin (KLH-A) of cynomolgus monkey were used to compare the abilities of RCTECs and human erythrocytes to detect antibody titers.No agglutination was observed in RCTECs test group.The mean fluorescence intensity (MFI) values of antibody were highest when the dilution fold was lowest (1 ∶ 16) and gradually decreased with increased serum dilution in both serum groups.MFI fell towards baseline value at 1 ∶ 128 dilution in normal serum group while at 1∶8 192 dilution in allergic serum group.Between 1 ∶ 16 and 1∶8 192 dilutions,MFIs of normal serum group were all lower than those of allergic serum.In human red cells test group,obvious agglutination appeared at high concentrations of antibodies,and MFIs reached the peak at 1∶64 dilution in normal serum group and at 1∶32 dilution in allergic serum group,but fluctuated irregularly thereafter.Between 1∶64 and 1∶512 dilutions,MFIs of normal serum group were all higher than those of allergic serum.Data above showed that MFI values in RCTECs group could reflect the levels of blood group antibodies more exactly.Conclusion The RCTECs of cynomolgus monkey express ABO tissue-blood group antigens and would be more suitable to be used to measure ABO blood group antibody levels by FACS.