Research The Clonality Of Ebv-associated T Cell Lymphoproliferative Disorder And Study On The Role Of Eb Virus In The Pathogenesis In T Cell Lymphoproliferative Disorder

EBV-associated lymphoproliferative disorder includes chronic active EBV infection、hemophagocytic syndrome and infantile fulminant EBV-associated T LPD, the proliferative cell are B or T/NK cell. EBV-associated T cell lymphoproliferative disorder(T-LPD) is a rare disease, it occurrs mainly in Asia and South America. EBV-associated T LPD is a systemic disease, it can impair lymph node, liver, spleen, skin, gastrointestinal tract. The clinical feature is fever and multi-organ disfunction. Some patients die of severe complications or lymphoma. In the reactive appearance, the lymph nodes morphologically show hyperplasia of the paracortical area and lymph node follicles with a mild increase in transformed lymphocytes in the paracortex and diffuse proliferation of atypical small to large lymphoid cells. The clonality of cell include polyclone oligoclone and monoclone. The categorization of EBV-T LPD as follow:category A1, polymorphic LPD without clonal prolifeartion of EBV-infected cells; category A2, polymorphic LPD with clonality; category A3, monomorphic LPD with clonality; category B, monomorphic LPD with clonality and fulminant course. We study on the clonality of EBV-associated T LPD and research EBV genome polymorphism immunohistochemistry contrast with nasopharyngeal carcinoma NK/T cell lymphoma to find the role of EBV in the pathogenesis in T LPD.The paper includes three parts as follows:The first partResearch the clonality of T cell receptor gene rearrangement in EBV-associated T cell lymphoproliferative disorder[Objectives]To collect EBV-associated T LPD in the first hospital of Guangxi Medical University from2008august to2011march and to investigate T cell receptor gene rearrangement in EBV-associated T cell lymphoproliferative disorder in Guangxi and research the feature of rearrangement in EBV-T LPD.[Materials and methods]EBV-T LPD were selected by clinical feature, organizational form, immunohistochemistry and in situ hybridization(ISH) for EBER positive,Use BIOMED-2PCR and heteroduplex analysis to research TCRβ (TCR B) and TCRy (TCR G) gene rearrangement.[Results]TCR β and TCR Y of10cases EBV-T LPD were polyclonal rearrangements[Conclusions]To investigate the lymphocyte whether it was monoclonal rearrangement were a best way to distinguish hyperplasia and neoplastic hyperplasia.10cases T LPD all were polyclonal rearrangements, suggesting diseases did not achieve lymphoma; Use BIOMED-2PCR and heteroduplex analysis to research TCRβ and TCRy gene rearrangement was an objective method to distinguish T LPD and T cell lymphoma. The patients with polyclonal rearrangements can also have severe clinical manifestations, suggesting the clonality of TCR were not necessarily bound up with clinical manifestations. The second partEBV subtypes and variants of EBV-T LPD NK/T cell lymphoma and nasopharyngeal carcinoma[Objectives]To investigate the EBV subtypes and variants of EBV-associated T cell lymphoproliferative disorder(T LPD) NK/T cell lymphoma and nasopharyngeal carcinoma(NPC) in Guangxi, and to clarify whether has the special relations between EBV variants and three diseases, inorder to clarify the pathogenic roles of EBV. [Materials and methods]EBV isolated from10cases T LPD16cases NK/T cell lymphoma and54cases NPC were investigated for EBV subtypes and variants. Five regions of the EBV gene were amplified by polymerase chain reaction (PCR). The resultant PCR products were then purified and subjected to restriction fragment length polymorphism(RFLP) analysis and DNA sequencing.[Results](1)Amplification of the five regions of EBV gene was performed successfully in10cases T LPD16cases NK/T cell lymphoma and54cases NPC.(2)The predominant type in T LPD were A(10/10,100%)、F (7/10,70%)、I (10/10,100%)、XhoI-(10/10,100%)、del-LMP1(10/10,100%);(3)The predominant type in NK/T cell lymphoma were A (11/16,68.8%)、F (13/16,81.3%)、I (13/16,81.3%)、XhoI-(15/16,93.7%)、del-LMP1(9/16,56.3%);(4)The predominant type in NPC were A (54/54,100%), f (44/54,81.5%)、1(54/54,100%)、XhoI-(54/54,100%)、del-LMP1(52/54,96.3%);(5)The proportion of type A has significant difference among three groups diseases, P=0.000. The proportion of type A in NPC was higher than in NK/T cell lymphoma,P=0.000.(6)The proportion of type F has significant difference among three groups diseases, P=0.000. The proportion of type F in T-LPD was higher than in NPC,P=0.003. The proportion of type F in NK/T cell lymphoma was higher than in NPC,P=0.000.(7)The proportion of type I has significant difference among three groups diseases, P=0.006. The proportion of type I in NPC was higher than in NK/T cell lymphoma,P=0.011.(8)The proportion of type Xhol-has no significant difference among three groups diseases, P=0.195.(9)The proportion of type del-LMP1has significant difference among three groups diseases, P=0.000. The proportion of type del-LMP1in T LPD was higher than in NK/T cell lymphoma,P=0.046; The proportion of type del-LMP1in NPC was higher than in NK/T cell lymphoma,P=0.000.[Conclusions](1)Type A EBV is predominant type in three diseases and NK/T cell lymphoma has some significant relationship with type B EBV.(2)The predominant type in Guangxi were type I and XhoI-, suggesting type I and XhoI-were associated with region.(3)The predominant type in NK/T cell lymphoma and T LPD were type F EBV, The predominant type in NPC were type f EBV, suggesting the mechanism and pathway of EBV in NK/T cell lymphoma and NPC were different. Type F EBV has close relationship with lymphocyte diseases and type f EBV has close relationship with nasopharyngeal epithelial cell.(4)Type del-LMP1EBV has significant relationship with T LPD and NPC, NK/T cell lymphoma has significant relationship with both type del-LMP1and wt-LMP1EBV.(5)Type F EBV and type f EBV were related with different cell types, the EBV types of EBV-T LPD were similar to NPC but different from NK/T cell lymphoma.Special EBV types were closely related to some neoplasm and diseases. The three partExpression and significance of NF-κB bcl-2and survivin in EBV-T LPD NK/T cell lymphoma and nasopharyngeal carcinoma[Objectives]To investigate the expression and clinical significance of NF-κB bcl-2and survivin in EBV-T LPD NK/T cell lymphoma and nasopharyngeal carcinoma and to analyse relations among them.[Materials and methods]18cases T LPD24cases NK/T cell lymphoma and74cases NPC were selected by in situ hybridization(ISH) for EBER positive; the expression of NF-κB bcl-2and survivin was detected by immunohistochemistry method in three groups cases.[Results](1)18cases T LPD24cases NK/T cell lymphoma and74cases NPC were EBER positive;(2)The positive expression rates of NF-κB bcl-2and survivin in T LPD were61.1%(11/18)、44.4%(8/18)、16.7%(4/18);(3)The positive expression rates of NF-κB bcl-2and survivin in NK/T cell lymphoma were54.2%(13/24)、37.5%(9/24)、50%(12/24);(4)The positive expression rates of NF-κB bcl-2and survivin in NPC were74.3%(55/74)、81.1%(60/74)、58.1%(43/74);(5)The expression of bcl-2has significant difference among three groups diseases, P=0.000. The expression of bcl-2in NPC was higher than in NK/T cell lymphoma,P=0.000; The expression of bcl-2in NPC was higher than in T LPD, P=0.004; The expression of survivin has significant difference among three groups diseases, P=0.024. The expression of survivin in NPC was higher than in T LPD, P=0.006;(6)There was significant expression relationship between NF-κB and bcl-2in NPC, P=0.049,r=0.192. There was significant expression relationship between NF-κB and survivin in NK/T cell lymphoma, P=0.000,r=0.553.[Conclusions] The expression of bcl-2and survivin is low in T-LPD, suggesting T-LPD had not activate inhibite apoptosis.The expression of bcl-2in NPC was higher than in NK/T cell lymphoma and T LPD, there was significant expression relationship between NF-κB and bcl-2in NPC, suggesting NF-κB was upstream regulation factors and up-regulated the expression of bcl-2to inhibite apoptosis and promote cell proliferation for NPC occurrence and development. On the other hand, there was significant expression relationship between NF-κB and survivin in NK/T cell lymphoma, suggesting both NPC and NK/T cell lymphoma were related to inhibite apoptosis but the mechanism and pathway were different.