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ABSTRACT Introduction: Post-transplant lymphoproliferative disorders (PTLDs) are a heterogeneous group of lymphoid proliferations occurring after solid organ or bone marrow transplantation. The primary aims of our study were to characterize cumulative incidence of PTLDs, clinical and pathological features according to the Epstein-Barr virus (EBV) status and survival. Methods: This was a retrospective cohort study on adult and pediatric patients, from January 2001 to December 2017. The cumulative incidence of PTLD was calculated by analyzing all the patients transplanted at our hospital, based on the database of the Organ Donation and Ablation Authority of Argentina (INCUCAI). The Kaplan-Meier method was used to plot the survival. Results: Fifty-eight cases of biopsy-confirmed PTLD were identified and 12 cases of clinical data were incomplete and these patients were excluded. The median age at the time of the PTLD diagnosis was 17.5 years (interquartile range [IQR] 9 - 57). The median interval between transplant and PTLD diagnosis was 39 months (IQR 9 - 113). The most commonly transplanted organ was the liver (24 cases, 52.2%), followed by kidney (20 cases, 43.5%). The Epstein-Barr encoding region in situ hybridization (EBER ISH) was positive in 29 (69.8%) of the 43 evaluable biopsies. The PTLD cumulative incidence was 1.84% (95%CI 1.77 - 1.91) for solid organ and 0.84% (95%CI 0.48 - 1.2) for bone marrow transplant patients. The overall survival rate at 5 years was 0.77 (95%CI 0.61 - 0.87). Subgroups by the EBV EBER status, transplant type, PTLD subtype and age group (adult vs. pediatric) showed no statistically significant association with the overall survival. Conclusion: The PTLD incidence was similar to that of previous series and the EBER did not appear as a relevant factor in our patient survival.
Subject(s)
Humans , Child , Adolescent , Adult , Transplants , Lymphoproliferative Disorders , Adjustment Disorders , Herpesvirus 4, Human , Epstein-Barr Virus InfectionsABSTRACT
OBJECTIVE@#To establish the technique that take the advantages of flow cytometry combined fluorescence in situ hybridization (Flow-FISH) to identify the Epstein-Barr virus(EBV) infected lymphocyte subtypies in patients' peripheral blood sample.@*METHODS@#Peripheral Blood monocyte from 9 patients with EBV infection enrolled at Children's Hospital in Chongqing Medical University were isolated by Ficoll-paque centrifugal separation. The expressions of EBER1, EBER2 in cell were detected by qRT-PCR. The surface markers of cell were detected by Flow cytometry after staining with their antibodies. The cell was treated Fix-Permeabilization Buffer before hybridization with fluorescent labeled probe at 37 ℃ overnight. The cell status, surface markers and targeted mRNA are detected by flow cytometry and fluorescence microscope.@*RESULTS@#It was optimized that the Fix-Permeabilization Buffer and recipe with 0.2% Tween-20 were picked out as providing a good cell integrity and high resolution of surface markers. Hybridization with 20% formamide and 7% dextran sulfate at 37 ℃ overnight is the optimal hybridization condition as a good hybridization effect, a detectable cell integrity and a high resolution of cell markers under flow cytometry detection. Finally, upon the established Flow-FISH method, lymphocyte subpopulations of the EBV+ cells from cell lines and blood samples of patients were identified successfully.@*CONCLUSION@#A Flow-FISH technology is established, which can be applied in the identification of EBV infected cell subtypes. This research provides a foundmental for its application in clinical test in EBV+ related proliferative diseases.
Subject(s)
Humans , Epstein-Barr Virus Infections , Flow Cytometry/methods , Herpesvirus 4, Human , In Situ Hybridization, Fluorescence/methods , Lymphocyte SubsetsABSTRACT
Objective: To report the first case of lymphomatoid gastropathy in China, and to demonstrate the clinical characteristics, diagnostic approach, treatment and prognosis in this kind of patients. Methods: One patient was diagnosed as lymphomatoid gastropathy at Peking Union Medical College Hospital, and her clinical characteristics, lab data, treatment and follow-up outcomes were reviewed. Results: A case of a 51-year-old female was presented, who underwent esophagogastroduodenoscopy (EGD) due to slight epigastric discomfort. EGD revealed multiple ulcers and erosions. Biopsies showed atypical lymphocytes infiltration with CD3(+), CD56(+), CD20(-), CD8(-), TIA(+), Granzyme B(-) and Ki-67 (75%). Epstein-Barr virus-encoded RNA in situ hybridization was negative. Four months later, repeated EGD examination showed regression of the lesions without specific treatment. Conclusion: Lymphomatoid gastropathy was a unique disease entity mimicking NK/T-cell lymphomas in pathology, with the quite different profile of treatment and prognosis. It's important to consider this issue during the differential diagnosis to avoid any excessive treatment.
Subject(s)
Female , Humans , Middle Aged , Biopsy , China , Immunophenotyping , In Situ Hybridization , Lymphatic Diseases , Lymphoma, T-Cell, Peripheral , Stomach DiseasesABSTRACT
Objective@#To report the first case of lymphomatoid gastropathy in China, and to demonstrate the clinical characteristics, diagnostic approach, treatment and prognosis in this kind of patients.@*Methods@#One patient was diagnosed as lymphomatoid gastropathy at Peking Union Medical College Hospital, and her clinical characteristics, lab data, treatment and follow-up outcomes were reviewed.@*Results@#A case of a 51-year-old female was presented, who underwent esophagogastroduodenoscopy (EGD) due to slight epigastric discomfort. EGD revealed multiple ulcers and erosions. Biopsies showed atypical lymphocytes infiltration with CD3(+), CD56(+), CD20(-), CD8(-), TIA(+), Granzyme B(-) and Ki-67 (75%). Epstein-Barr virus-encoded RNA in situ hybridization was negative. Four months later, repeated EGD examination showed regression of the lesions without specific treatment.@*Conclusion@#Lymphomatoid gastropathy was a unique disease entity mimicking NK/T-cell lymphomas in pathology, with the quite different profile of treatment and prognosis. It’s important to consider this issue during the differential diagnosis to avoid any excessive treatment.
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Objective:To explore the clinical characteristics and laboratory data of aggressive non-Hodgkin's lymphoma(NHL)compli-cated by autoimmune hemolytic anemia(AIHA).Methods:Data of six patients with aggressive NHL complicated by AIHA treated at the Second Affiliated Hospital of Suzhou University between September 2013 and July 2016 were reviewed retrospectively. The onset symptoms,disease progression,therapy,and prognostic factors were analyzed.Results:From September 2013 to July 2016,155 pa-tients with aggressive NHL were treated in our hospital.Six of them were complicated by AIHA(3.9%),with three males and three fe-males,aged from 62 to 74.The median age was 67 years.The first clinical symptoms included the following:five presented with lymph-adenectasis,three had fever,and one presented with multiple bone destruction and bone pain,all complicated by progressive hemo-globin decrease.Histological examination of the six patients revealed three cases with diffuse large B-cell lymphoma(DLBCL),including one case with positive Bcl-2,Bcl-6,and C-myc,and one case with positive CD5;one case was peripheral T-cell lymphoma-not other-wise specified(PTCL-NOS);and two cases with angioimmunoblastic T-cell lymphoma(AITCL).Epstein-Barr virus(EBV)-mRNA(EBER) was detected by chromogenic in situ hybridization(CISH).Up to 100%(5/5)cases were EBER-positive.Eastern Cooperative Oncology Group scores of the six patients were three to four.All six cases were in the Ann-Arbor stagesⅢ-Ⅳand International Prognostic Index score 4-5.All cases belonged to the high risk group.At the time when lymphoma was confirmed by pathology,the median level of he-moglobin was 56(34-79)g/L.The median ratio of reticulocytes was 6.7(0.2-21.0)%.The positive rate of Coombs test was 100%.All cas-es showed autoantibodies against C3(1:64-1:2 048).Four cases showed antoantibodies against G antigen(±~1:16).The plasma con-centration of EBV DNA of three patients was detected and all increased.Except that 1 case gave up treatment,five patients received the chemotherapy with CHOP or R-CHOP.Four patients received prednisone between chemotherapy intermittent period.Two cases showed sustained complete response(CR).The overall survival(OS)was 20 and 14 months.Another patient with DLBCL and one pa-tient with PTCL-NOS died from secondary severe pulmonary infection and heart failure during the myelosuppression.The OS times were 1.5 and 2 months,respectively.One patient with AITCL died in the disease progression after four cycles of chemotherapy.The OS was 4.5 months.Conclusion:Aggressive NHL complicated by AIHA is common in older patients with poor prognosis.The incidence rate of EBV infection was high,and hemolysis is rapid and serious.The patients'tolerance to chemotherapy was poor.Early diagnosis and effective chemotherapy may improve patients'prognosis.
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Objective:To explore the clinical characteristics and laboratory data of aggressive non-Hodgkin's lymphoma(NHL)compli-cated by autoimmune hemolytic anemia(AIHA).Methods:Data of six patients with aggressive NHL complicated by AIHA treated at the Second Affiliated Hospital of Suzhou University between September 2013 and July 2016 were reviewed retrospectively. The onset symptoms,disease progression,therapy,and prognostic factors were analyzed.Results:From September 2013 to July 2016,155 pa-tients with aggressive NHL were treated in our hospital.Six of them were complicated by AIHA(3.9%),with three males and three fe-males,aged from 62 to 74.The median age was 67 years.The first clinical symptoms included the following:five presented with lymph-adenectasis,three had fever,and one presented with multiple bone destruction and bone pain,all complicated by progressive hemo-globin decrease.Histological examination of the six patients revealed three cases with diffuse large B-cell lymphoma(DLBCL),including one case with positive Bcl-2,Bcl-6,and C-myc,and one case with positive CD5;one case was peripheral T-cell lymphoma-not other-wise specified(PTCL-NOS);and two cases with angioimmunoblastic T-cell lymphoma(AITCL).Epstein-Barr virus(EBV)-mRNA(EBER) was detected by chromogenic in situ hybridization(CISH).Up to 100%(5/5)cases were EBER-positive.Eastern Cooperative Oncology Group scores of the six patients were three to four.All six cases were in the Ann-Arbor stagesⅢ-Ⅳand International Prognostic Index score 4-5.All cases belonged to the high risk group.At the time when lymphoma was confirmed by pathology,the median level of he-moglobin was 56(34-79)g/L.The median ratio of reticulocytes was 6.7(0.2-21.0)%.The positive rate of Coombs test was 100%.All cas-es showed autoantibodies against C3(1:64-1:2 048).Four cases showed antoantibodies against G antigen(±~1:16).The plasma con-centration of EBV DNA of three patients was detected and all increased.Except that 1 case gave up treatment,five patients received the chemotherapy with CHOP or R-CHOP.Four patients received prednisone between chemotherapy intermittent period.Two cases showed sustained complete response(CR).The overall survival(OS)was 20 and 14 months.Another patient with DLBCL and one pa-tient with PTCL-NOS died from secondary severe pulmonary infection and heart failure during the myelosuppression.The OS times were 1.5 and 2 months,respectively.One patient with AITCL died in the disease progression after four cycles of chemotherapy.The OS was 4.5 months.Conclusion:Aggressive NHL complicated by AIHA is common in older patients with poor prognosis.The incidence rate of EBV infection was high,and hemolysis is rapid and serious.The patients'tolerance to chemotherapy was poor.Early diagnosis and effective chemotherapy may improve patients'prognosis.
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Objective To investigate the EB virus (EBV) infection and its clinical significance in angioimmunoblastic T-cell lymphoma(AITCL).Methods 62 patients diagnosed as AITCL between 2008 Jan and 2011 Dec were retrospective analyzed.All cases were re-confirmed and classified with the histology and immunology according to 2008 WHO Classification.In situ hybridization of EBV encoded RNA (EBV-EBER) was performed.Clinical characteristics and follow-up data of patients were collected.Results 42 ％ (26/62) AITCL cases were EBER-positive.EBV infection was found to be significant correlation with age over 60 years and poor response to therapy (P =0.025,P =0.049,respectively).However,EBV infection had no relationship with the overall survival,the presence of B symptom and the Ann Arbor stage.Conclusion In AITCL,EBV infection seems not to be associated with the overall survival,B symptom and Ann Arbor stage,but it may have impacts on theraputic response.
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ObjectiveTo investigate the diagnostic significance of different pathology techniques (Immunohistochemistry and ISH)to detect EBV on bone marrow biopsy tissues of hemophagocytic syndrome. Methods Histological,immunohistochemicalandinsituhybridizationwereusedtostudythe hemophagocytic features and expression of LMP-1,EBER and other markers.Results25 out of 51 cases (49.0 ％)showed an active proliferation growth pattern while other 12 cases(23.5 ％)demonstrated a deterioration morphological character compared with their same age group. The left 14 cases (27.5 ％) showed a normal myeloproliferative pattern. 91.4 ％ (32/35) expressed CD68/KP-1 and 89.5 ％ (17/19) cases expressed CD68/PG-M1, which showed an abnormal increase of monocytes. 26 out of 51 cases (51.0 ％) were positive for EBER,while EBER and immunohistochemistry on LMP-1 were detected simultaneously in 41 cases.The EBER were positive in 19 cases(46.3 ％)but none for LMP-1(P =0.000).ConclusionHypoplasia or hemophagocytic features could be detected on bone marrow biopsy tissues of hemophagocytic syndrome, which might be slightly difficult to diagnose. So immunohistochemistry would be essential. Meanwhile, the detection of EBER is more helpful than LMP-1 for diagnosis.
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Forty eight cases of nasopharyngeal carcinoma patients who had been treated with radiotherapy were analysed for the expression of EBER in the nucleus of the tumor cells by in situ hybridization technique. EBER was expressed in 69% of the cases. Histologic specimens positive for EBER were then examined for the expression of EBV-LMP1 by immunohistochemistry, which showed 68% positivity. Clinical staging of 48 cases and radiation response of 32 cases were collected. Data of up to 2 years of survival of 16 cases were obtained through postal and phone communication. Statistical analysis showed that the Working Formulation histologic classification of NPC correlated significantly with radiation response. The expression of EBV-LMP1 was correlated significantly with the survival of the patients. Thus EBV-LMP1 which has been shown to possess influence on lymphoid and epithelial cells, clinicopathologically proved to have a function as prognostic indicator.
Subject(s)
Nasopharyngeal Neoplasms , Viral Matrix ProteinsABSTRACT
Epstein-Barr virus (EBV) has been linked to a spectrum of neoplastic conditions, including Burkitt's lymphoma, nasopharyngeal carcinoma, Hodgkin's disease, lymphoepithelioma-like carcinomas and malignant lymphomas in immunocompromised state. To determine the prevalence and the subtype of EBV in gatrointestinal malignancies, fifty cases of adenocarcinomas and seventeen cases of malignant lymphomas were analyzed by EBERs in situ hybridization and polymerase chain reaction using primers for EBNA-1, EBNA-2A and EBNA-2B, on the paraffin sections. In addition, immunohistochemical stain for p53 protein was performed to investigate the potential role of EBV infection on tumor suppressor gene, p53, during tumorigenesis. EBER was detected in 6 of 26 gastric adenocarcinomas, 2 of 24 colon adenocarcinomas, and 8 of 17 malignant lymphomas. EBER was more prevalent in malignant lymphoma arising in the intestine (6/6) than in the stomach (2/11), and was detected in both B and T cell phenotypes. EBNA-1 was positive in 11 of 16 EBER positive cases and the subtyping was possible in 8; both type 1 and 2 were detected in gastric cancers, whereas only type 2 was found in intestinal neoplasms. In adenocarcinomas the high rate of p53 protein overexpression was found in both EBER positive (8/8) and negative cases (32/42), whereas the positive rate was higher in EBER positive cases (7/8) than in EBER negative cases (4/9) of malignant lymphomas. From the results, it can be concluded that EBV infection and the p53 tumor suppressor gene are independently associated in a significant portion of the gastrointestinal malignancies, but the mechanism of action remains to be elucidated.
Subject(s)
Adenocarcinoma , Burkitt Lymphoma , Carcinogenesis , Colon , Epstein-Barr Virus Infections , Gastrointestinal Tract , Genes, Tumor Suppressor , Herpesvirus 4, Human , Hodgkin Disease , In Situ Hybridization , Intestinal Neoplasms , Intestines , Lymphoma , Paraffin , Phenotype , Polymerase Chain Reaction , Prevalence , Stomach , Stomach NeoplasmsABSTRACT
To determine the prevalence of Epstein-Barr virus infection in lymphoid neoplasms of various histology and location, the paraffin tissues of 74 non-Hodgkin's lymphomas and 13 Hodgkin's diseases were studied by EBER and BHLF RNA in situ hybridization as well as immunostaining using LMP-1, EBNA-2, and ZEBRA. As a control, non-neoplastic lymphoid tissues from the nasal cavity(10), lymph node(38) and Waldeyer's ring(12) were investigated. In non-neoplastic control, EBV genome was detected in none of 10 nasal mucosa, 6 of 38 lymph node, and 1 of 12 Waldeyer's ring. EBV-positive non-neoplastic lymphocytes expressed CD45RO in 2 cases and CD20 in 4 cases. Non-Hodgkin's lymphoma was positive for EBV in 37.8% of the cases in which T-cell lymphoma showed higher rate(56%) than B-cell lymphoma(15%), especially in nasal lymphoma(80%) and angiocentric lymphoma(63.6%). Hodgkin's disease was EBV positive in 38.4% of the cases. EBV genome in tumor tissue existed in latent form as well as in lytic form. LMP-1 was positive in 80% of Hodgkin's disease and 39% of non-Hodgkin's lymphoma in which EBV genome was detected. EBNA-2 was expressed in 3 cases of non-Hodgkin's lymphoma. On the basis of protein expression, most lymphomas belonged to type II latency. These results support that EBV is associated with pathogenesis of malignant lymphoma although its mechanism still awaits to be clarified.
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Epstein-Barr virus(EBV) is associated with a wide spectrum of benign and malignant disorders including leukoplakia, Hodgkin's lymphoma, central nervous system lymphoma, peripheral T cell lymphoma and nasopharyngeal undifferentiated carcinoma. There are several distinctive aspects of biology of the virus that are important in investigation of virus in clinical specimens. The abundant expression of the EBER mRNA transcripts makes possible the sensitive detection of latent expression in EBV-associated tumors. Although there has been a dramatic increased interest in the direct characterization of EBV in clinical specimens, there have been few studies about the effective and reliable positive controls in performing in situ hybridization technique for EBV, especially on paraffin-embedded tissue. We applied Burkitts lymphoma cell line as positive control in EBV in hydridization using Oncor Kit. The cell block of Burkitt lymphoma cell line(CCL85 EB-3) showed strong and specific positivity for EBER in situ in nuclei of EBV infected cells.
Subject(s)
Biology , Burkitt Lymphoma , Carcinoma , Cell Line , Central Nervous System , Herpesvirus 4, Human , Hodgkin Disease , In Situ Hybridization , Leukoplakia , Lymphoma , Lymphoma, T-Cell, Peripheral , RNA, MessengerABSTRACT
AIM: To study the expression of mic2/CD99 protein and their correlation with Eber-1/LMP-1 in Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma. METHODS: Immunohistochemical staining, in situ hybrization and tissue microarry technique were used to detect the expressions of mic2/CD99 and Eber-1/LMP-1 of H/RS cells in 43 cases of cHL and 16 cases of NHL. RESULTS: The positive rate of CD99 protein expression in 43 cases of cHL was 2.3% (1/43) , mic2 was 55.8% (24/43), LMP1 was 58.1% (25/43) and Eber-1 was 53.5% (23/43). The expressions of CD99 and mic2 in the NHL group were higher than those in cHL group (P0.05). There was a negative correlation between the expression of CD99 protein and LMP1 in H/RS cells (P0.05). There was a significant correlation between the high expression of LMP1 and a low expression of CD99 in the young patients (P0.05). CONCLUSION: There is a negative correlation between the expression of LMP1 and CD99 in Hodgkin and Reed-Sternberg cells of classical Hodgkin's lymphoma.