| Objective To prepare a rabbit monoclonal antibody GCET2 and investigate its function in DLBCL diagnosis. Methods Rabbit was immunized by GCET2 recombination protein, and the rabbit spleen cells with higher serum titer were fused with the 240E-W3 rabbit myeloma cells. The GCET2 rabbit monoclonal antibody was developed by using RabMAb(?) technology, and its specificity was confirmed by ELISA, WB, ICC, IHC and flow cytometry. IHC was used to detect a panel of markers including GCET2, CD10、 Bc16、 MUM1、GCET1、FOXP1、Ki67 and c-myc in 81 cases of DLBCLs,5 cases of follicular lymphoma (FL) and 2 cases of Burkitt’s lymphoma tissues. Then GCET2 was added to Hans and Choi algorithms to compare their sensitivity of diagnosing GCB-DLBCLs. Result We developed a rabbit monoclonal GCET2 antibody (clone EP316), which specifically expressed in normal germinal center B-cells (GCB) and GCB derivation lymphomas. The proportions of GCB-DLBCL cases in the original Hans and Choi algorithms were 21% (17/81) and 23%(19/81), respectively. With the inclusion of GCET2, the frequencies of GCB-DLBCL cases were increased to 54% (44/81) and 52% (42/81), respectively. Furthermore, in 81 DLBCLs, the proportions of high c-myc expression in GCET2 negative and positive groups were 15.2% (7/46) and 2.8% (1/35), respectively. Conclusion:Based on results from this study, we conclude that GCET2 is a sensitive marker for GCB derivation lymphomas. Add GCET2 into Hans and Choi algorithms can improve their sensitivity to GCB-DLBCL. |
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