Non-Hogkin's Lymphoma Immunophenotype Detection And DNA Ploid Analysis By Flow Cytometry

Objective: To study the relation between NHL immunophenotype detection and DNA ploid analysis and NHL's diagnosis, typing, the malignant degree judging and prognosis. Methods: (1) To select 10 examples of RLN as control group. (2) To select 40 samples of NHL as test group.9 samples in NHL-1 stage, 11 samples in NHL-II stage, 12 samples in NHL-III stage and 12 samples in NHL-IV stage. (3) All NHL samples were tested by routine pathological morphology and immunohistochemistry according IWF standard to diagnose and type. (4) All samples' fresh lymph nodes tissue were obtained from lymph nodes biopsy or needle puncture and prepared suspended single cells by machine separation or enzyme separation. using CD1, cyCD3, CD3,CD5, CD7, CD10, CD19, CD20, CD23, CD25, CD43, CD56 monoclonal antibody labeling and PI staining method to analysis cell DNA content(DI SPF) and immunophenotype by Flow cytometry.Results: (1)3 samples of B-NHL and 1 sample of T-NHL in 40 lymph nodes,which failed to be diagnosed by routine methods, were determined by flow cytometric immunophenotype analysis. 1 sample was diagnosed B-NHL before, behaved multiclonal immunophenotype by FCM and was diagnosed RLN.Of 2 samples couldn' t be diagnosed before, 1 sample was diagnosed mantle cell lymphoma and 1 sample was diagnosed marginal zone lymphoma .1 sample of T-NHL was diagnosed NK\T NHL by FCM that can' t be typed according to IWF standard. (2) Analysis of cell DNA content in control group and test group:DL DNA heteroploid ratio and SPF were higher than that of control group (P<0.01) .DI and SPF analysis in different DNA ploid lymph nodes: DI .. SPF in ND cell and that in D cell were no notability differentiation (P>0.05) . DI , SPF in T, AN, M were higher than that of in D(P< 0.05). SPF in heteroploid was becoming higher and higher from ND→T→AN →M. (3) Analysis of DNA heteroploid ration , DI and SPF in different pathology groups and different clinical stages: DI , SPF in NHL- I , NHL- II and that in control group were no notability differentation (P>0.05 ) . DNA heteroploid ration , DI and SPF in NHL- III , NHL- IV and different pathology groups were notability differentation(P< 0.05).Conclusion: (1) Immunophenotype monoclona antibodys expressed differently in different NHL and improve the accuracy of diagnosis . (2) DI and SPF levels can reflect tumor proliferation pathology malignant degree and related to prognosis.