| Objective Epstein-Barr virus(EBV)is a carcinogenic herpes virus that has infected more than 90% of the world’s population.Most people infected by EBV have an asymptomatic and latent infection throughout their whole life,but in some infected people,EBV causes a series of diseases.There are dozens of diseases related to EBV infection,but the clinical manifestations of the diseases are diverse,the histological morphology is extensive,and the clinicopathological characteristics of different diseases overlap.The severity of the disease varies,and some diseases have a poor prognosis.The mechanism is still unclear,and current researches suggest one of its pathogenic mechanisms is that the imbalance between antiviral immunity and viral pathogenicity in the body.We retrospectively study the clinicopathological characteristics of EBV-related diseases in Southwest China,and statistically analyzed the relevant prognosis factors,as well as analyzed the degree of EBV infection and the immune microenvironment features of skin lesions with EBV-infected Hydroa vacciniforme-like lymphoproliferative disorder(HVLLPD).Methods This study consisted of two parts:1.Clinicopathological retrospective analysis of EBV-related diseases. EBER positive cases were collected from the First Affiliated Hospital and Affiliated Children’s Hospital of Chongqing Medical University from January 2013 to November 2020.All these cases were diagnosed according to WHO classification standards and had complete clinicopathological data;statistically analyzed the differences in the clinical characteristics between different diseases and analyzed prognostic factors.2.The degree of EBV infection and the characteristics of tissue immune microenvironment of skins with HVLLPD The infection of EBV in skin lesions with HVLLPD was analyzed by EBER in situ hybridization and the expression of viral latent membrane protein LMP1 was analyzed by immunohistochemistry.Immunohistochemical single staining and double staining were performed to analyze T cells(CD3+,CD5+,CD7+),T cell subtypes(CD4+,CD8+),na(?)ve CD4+T cell subtypes(Th1,Th2,Treg cells)and CD8+T cell differentiation subtypes(na(?)ve CD8+ T cells and terminally differentiated cytotoxic effector CD8+T cells),CD20+B cells,CD56+NK cells,CD68+ macrophages(M1 macrophages and M2 macrophages),cytotoxic granule-associated protein(Granzyme B and TIA-1),and the expression of immunomodulatory protein PD-1/PD-L1.Results 1.There were 313 EBV-infected patients,the ratio of male to female was 196 to 117,and the ratio of adults to children was 183 to 130.The median age of adult patients at diagnosis was 54 years(range 19-85years),of which NK/T cell lymphoma(39%,72/183)and nasopharyngeal carcinoma(28%,51/183)were the most common;The median age of children patients of EBV-associated disease at diagnosis was 7 years(range 1-17 years),of which Chronic active EBV infection of T/NK-cell type(CAEBV)(30.0%,39/130)and HVLLPD(23.8%,31/130)were the most common.Compared with the common symptoms in adults of EBV-associated neoplastic diseases(such as NK/T cell lymphoma and nasopharyngeal carcinoma),fever(P<0.001),lymphadenopathy(P<0.001),hepatosplenomegaly(P<0.001),three cell lines reduction(P<0.001),liver dysfunction(P<0.001)and LDH level elevated(P<0.001)were more common in children of EBV-positive diseases(such as CAEBV and HVLLPD)than adults.Histologically,the T/NK cell type(50%,156/313)was the most common,followed by the B cell type(28%,87/313),while the epithelial cell type(22%,70/313)was relatively rare.After follow-up,156 patients survived with a median time of 33months(range 1.3-129 months);104 patients died with a median time of8 months(range 0.5-122.5 months);53 patients were lost to follow-up.Univariate COX regression analysis revealed fever(HR 2.26,P <0.001),splenomegaly(HR 3.33,P <0.001),liver dysfunction(HR 2.23,P<0.001),elevated LDH levels(HR 2.30,P <0.001),anemia(HR 2.34,P<0.001),thrombocytopenia(HR 3.33,P <0.001)and leukopenia(HR 3.39,P <0.001)were poor prognostic factors;After diagnosis,patients who actively take corresponding radiotherapy,chemotherapy or surgery(HR0.40,P <0.001)have a better prognosis than those who only receive symptomatic treatment. 2.Thirty-four biopsies were obtained from 32 patients with HVLLPD.The number of EBER+ cells was higher than 100 cells/HPF in59%(20/34)biopsies,while the virus latent membrane protein LMP1 was usually(74%,25/34)negative.The proliferative lymphocytes in the lesions mainly were Pan-T cells expressed CD3,CD5,and CD7,while only a few scattered CD20+ B cells(12 cells/HPF),CD56+ NK cells(20cells/HPF),and CD68+ macrophages(62 cells/HPF)were observed.Among the CD4+ T cell,there were more Th1(CD4+T-bet+)than Th2(CD4+CMAF+)cells in 65%(24/34)of specimens.And only a few scattered FOXP3+ regulatory T cells were observed in most specimens.In 91%(31/34)of specimens,T-bet+ cytotoxic effector CD8+ T cells were more abundant than na(?)ve CD8+ T cells(T-bet-CD8+).Among macrophages,there were more M1(CD68+pstate1+)than CD206+ M2 and CD163+ M2 macrophages in 27 and 26 specimens(out of 34),respectively.Most specimens were diffusely positive for the cytotoxic granule-associated protein TIA-1,whereas granzyme B positive cells were rarely observed.In addition,only a few PD-1/PD-L1 positive cells were observed in only 1 and 6 biopsy tissues.Due to the progression of disease,2 patients underwent a second biopsy after 3 years and 1 year,respectively.Compared with the first biopsy,the number of EBER+ cells in the second biopsy increased by an average of 5.5 folds and the number of CD8+ T cells also increased.However,the terminally differentiated effector CD8+T cells decreased by an average of 21%;although the first biopsy showed scattered CD56+ and granzyme B+ cells,in the second biopsy,negative and only few CD56+ cells(13 cells/HPF)were observed,respectively,and the loss of granzyme B was occurred.Conclusion Among the 313 patients with EBV-related diseases,there were more males than females and mainly elderly and children.NK/T-cell lymphoma and nasopharyngeal carcinoma were more common in adults and CAEBV and HVLLPD were more common in children. EBV infection leads to neoplastic proliferation in adults.In children,CAEBV and HVLLPD often continuously progress,and the severe cases usually died of multiple organ failure or hemophagocytic syndrome.The prognosis of most patients is poor,about 40% of the follow-up cases died. In the skin tissues of Hydroa vacciniforme-like lymphoproliferative disorder,the number of infiltrating T/NK cells varied,showing a predominant infiltration of T cells,especially terminally differentiated effector CD8+T cells and Th1 cell.As the disease progresses,the loss of granzyme B was occurred. |
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