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Background- High grade B-cell NHL's are 200 fold more common in seropositive as compared to seronegative patients. It includes diffuse large B cell lymphoma (DLBCL) (50%), Burkitt's (BL)(40%) and others such as plasmablastic lymphoma(PBL). They are biologically different from their seronegative counterparts with higher incidence of extranodal involvement, advanced stage and poor response to conventional therapy. In addition, they are at a higher risk of chemotherapy side effects and poorly tolerate dose intense regimens. To add to the complexity, social stigma and financial constraints decrease the compliance to therapy. R-EPOCH is designed to provide a balance between efficacy and safety. We report on our cohort of patients who were treated with this protocol. Methods- We retrospectively analysed HIV-associated de-novo high grade B-cell NHL patients who were treated with R EPOCH regimen at Tata Memorial Centre from 2011 till 2015. Patients aged ≥18 years who had received at least 1 cycle were included in the analysis. R EPOCH is an infusional regimen in which the dose of cyclophosphamide is adjusted depending on the baseline CD4+ T cell count and tolerance. The primary objective was the 3-year overall survival(OS), secondary objectives were response rates, the incidence of grade3/4 toxicities, dose intensity and correlation of OS with CD4 count, IPI, duration of HIV, Cyclophosphamide dose intensity (CDI). Demographic features, HIV related details (CD4 count at diagnosis, ART regimen), histological diagnosis, stage, bulky disease, extranodal sites, treatment details (number of cycles, dose administered), response, toxicity and status at last follow up were recorded. Descriptive statistics were summarised with median and range, survival outcomes were analysed with Kaplan meier method and impact of CD 4 count, CDI, IPI and duration of HIV on survival was assessed using log rank test. Results- A total of 40 patients(31males) with a median age-40 years (24-65 years) were treated. B symptoms were present in 19(48%). The cohort comprised of DLBCL-19 (48%), BL-16(40%), High grade B-Cell Lymphoma-Unclassifiable-4 (10%) and PBL 1 (2.5%). 16 (40%) patients had co-morbidities, including co-existent Hepatitis C in 5 and Hepatitis B in 2. HIV infection was detected at the time of lymphoma diagnosis in 18 (45%). The median CD4+ T cell count was 202 cells/mm3, 6 patients (15%)had count of Conclusions- R-EPOCH is a highly effective regimen in seropositive high-grade B-cell lymphoma even in the presence of adverse features (advanced stage, extranodal sites, CNS involvement, low CD4+ T cell count). The emphasis should now be on further reducing the toxicities. Figure Overall survival outcomes Figure. Overall survival outcomes Disclosures No relevant conflicts of interest to declare.

Introduction: MYC rearrangement in high-grade B-cell NHL(non-burkitt), either as single hit or double hit, has significant prognostic and therapeutic implications. There is remarkable paucity of data on frequency and clinico-pathological features of MYC-rearranged large B-cell lymphomas from developing world. Method: This study included de-novo high grade B-cell NHL cases (>15 years of age) registered at Tata Memorial Centre between January 2013- December 2014. Demography details, clinical features (stage at presentation, bone marrow involvement, presence and extent of extranodal involvement, B-symptoms, international prognostic index),original histopathological diagnosis, chemotherapy and radiotherapy details were recorded from electronic medical or paper case records. Response to therapy, relapses or death if any, status at last follow up, dates of relapse and/or death and last follow up were recorded. All cases were reviewed by expert hematopathologists to categorize the high grade B-cell NHL as per the WHO-2008 classification of hematopoietic malignancies. All cases with adequate formalin-fixed paraffin embedded (FFPE) blocks were evaluated by FISH for c-MYC , BCl-2 and BCl-6 using Vysis dual colour break apart probes. Cases which showed >10% cells with split signal were considered to harbor rearrangement. Result: A total of 114 cases of de-novo high-grade B-cell NHL with adequate FFPE blocks were evaluated. Based on WHO 2008 classification, 112 cases were classified as diffuse large B-cell lymphoma (DLBL)) and 1 each as Burkitt's (BL) and B- cell lymphoma unclassifiable -intermediate between DLBL and BL(BCLU). A total of 9/112 (8%) cases of DLBL showed MYC rearrangement. One of these 9 cases had both MYC and BCl-2 rearrangement. BCLU did not demonstrate MYC or BCl-2 rearrangement. The Ki-67 index was variable (40-95%)in MYC rearranged cases. The median age of the c-MYC rearranged cases was 55 year (range-23-79 years). 88% were males. Advanced stage disease, bulky mass and extranodal disease was seen in 67%, 44% and 55% of cases respectively. All but one patient had high LDH, however none of the patients had elevation more than 2XULN. These patients received rituximab based chemoimmunotherapy (RCHOP-4,RCEOP-4, REPOCH-1) and 77% achieved complete response. At median follow up of 5 months, 1 year-overall survival and progression free survivals were 80% and 75% respectively. There were no significant differences in clinical features (except higher proportion of males in the c-MYC rearranged subset), LDH levels, ki-67 index , response to therapy and survival between DLBL with or without c-MYC rearrangement Conclusion: In our study 8% of all DLBL cases showed c-MYC rearrangement. The frequency of double hit lymphoma was less than 1% (0.8%). Disclosures No relevant conflicts of interest to declare.

Introduction: Despite high initial success rates in the treatment of pediatric HL, nearly 20% of patients develop resistant / relapsed disease. While high-dose chemotherapy with autologous stem cell transplantation has improved outcomes, the best of these regimens still have limited success rates and also such therapies are not easily available for most patients in Low and Middle Income Countries (LMICs). Novel therapeutic agents are needed in this setting. Bendamustine is an alkylating agent with clinical activity against various adult lymphomas, including limited data supporting its use in heavily pretreated adult HL patients. There is no such data in pediatric HL patients. Here we retrospectively report the results of single agent bendamustine salvage regimen in pediatric patients with refractory/relapsed HL. Methods: Retrospective analysis of children with relapsed / refractory HL, who underwent treatment from January 2013 to February 2015, was performed. These patients, who were ineligible for autologous stem cell transplantation (ASCT) or had relapsed after ASCT (1), received bendamustine 120 mg/m2 as a 30 minutes infusion on days 1 and 2 every 28 days with growth factor support. Total 6 cycles were planned for each patient. Early response was evaluated using PET-CT following 2 cycles of bendamustine and best response was evaluated post completion of 6 cycles of bendamustine. Results: Of the 10 patients who were started on bendamustine, 8 received at least 2 cycles of bendamustine and were evaluable for response assessment. Of the 8, 7 were males and 1 female; median age was 13.5 years (range 5 to 15 years); the histology was mixed cellularity in 5 (62%) & nodular sclerosis in 3 (38%). Patients had received a median of 3 prior treatments (range 1 to 5). 5 had relapsed HL and 3 had primary progressive disease. On evaluation for early PET-CT response post 2 cycles of bendamustine, 4 patients (50%) had Complete Remission (CR) and 4 patients (50%) had Partial Remission (PR) with an Overall Response Rate (ORR) of 100%. One patient with PR post 2 cycles underwent haploidential allo-SCT and later died of sepsis. The remaining 7 patients went on to receive 6 cycles of bendamustine, 6 of them (85%) achieved CR and continue to be in CR. 1 patient who was in PR was started on lenalidomide plus celecoxib maintenance and died of sepsis at home post chicken pox infection at 7 months. The median follow-up for all 8 patients is 15 months (range 6 to 24 months). In general the treatment was well tolerated and toxicities, both hematological (grade II thrombocytopenia in 2 and febrile neutropenia in 1 patient) and extra-hematological were manageable. Conclusions: Within the limits of an observational retrospective study, these data indicates that bendamustine shows its efficacy in patients with relapsed/refractory HL, without any significant toxicity. It produces durable responses in patients with relapsed/refractory HL and may be an effective bridge to further therapeutic interventions. It may also be used earlier in the treatment strategy of HL and in combination with other active drugs. Disclosures Off Label Use: Bendamustine is used in treatment of relapsed/refractory Hodgkin Lymphoma, though it is not FDA approved for same at present..

Introduction: The role of FDG PET-CT in follicular lymphoma is limited to accurate assessment of disease extent in early stage patients and selection of biopsy site in cases of suspected high- grade transformation. Despite the known FDG avidity of follicular lymphoma, FDG PET-CT has not yet been included as part of standard staging procedures in these patients. FDG PET-CT has shown significant correlation with bone marrow biopsy in Hodgkin and diffuse large B-cell lymphomas. In this retrospective analysis we have assessed the correlation of PET-CT with that of bone marrow biopsy, the reference standard for assessment of bone marrow infiltration in follicular lymphoma. Methods: We retrospectively analyzed electronic medical records and database of patients with newly diagnosed follicular lymphoma registered at Tata Memorial Centre from July 2009 to Jun 2014, who underwent complete staging workup as per the current recommendations along with whole body 18FDG-PET/CT. The demographic features, performance status, stage, LDH, nodal sites, haemoglobin, follicular lymphoma international prognostic index (FLIPI), FDG PET-CT findings (bone marrow involvement, pattern of involvement- focal or diffuse, sites of marrow involvement, liver and spleen uptake, SUVmax of most FDG avid lesion) and bone marrow aspiration/biopsy (morphology, immunohistochemistry and immunophenotyping on aspirate, where available) findings were recorded. Focal uptake in marrow on baseline PET-CT was considered as marrow involvement if post therapy PET-CT showed resolution of these lesions. The sensitivity, specificity, negative and positive predictive value of PET-CT in detecting bone marrow infiltration was assessed taking bone marrow biopsy as gold standard. The factors responsible for discordant results were analyzed. Results: A total of 54 patients (males-38, females-16) were included in analysis with median age of 50 years, (range 22-73 years). At diagnosis 83% (45 patients) had stage III or IV disease and 57% patients had high-risk FLIPI score. Approximately 88% patients had good performance status (ECOG- Conclusion: This study shows that in patients with advanced stage follicular lymphoma bone marrow biopsy can be omitted if PET-CT shows focal or diffuse bone marrow uptake. Similarly, patients with early stage disease with no bone marrow uptake on PET-CT can be spared from bone marrow biopsy. Disclosures No relevant conflicts of interest to declare.

Poor prognosis DLBCL, including intermediate and high risk disease according to IPI accounts for approximately 20% of new cases of DLBCL. The addition of rituximab to conventional chemotherapy (CHOP) has improved the outcomes in this subset, with a 2-year overall survival (OS) of about 50%. However, 40-50% of these patients still have either primary refractory disease or relapse after an initial response. Rituximab-EPOCH (R-EPOCH), an infusional regimen has a dynamic dose adjustment strategy based on the hematopoietic nadir in previous cycle to achieve an optimal drug concentration. Phase II studies with R-EPOCH in untreated DLBCL with intermediate and high risk IPI have reported improved outcomes, with an estimated 2-year OS of 75% which appears superior to that of R-CHOP. Hence we analysed the outcomes of patients with de-novo, poor prognosis (intermediate and high risk IPI) DLBCL who received R-EPOCH and compared it to the historical cohort of patients who were treated with R CHOP at our centre. Methods Treatment-naïve patients of DLBCL with intermediate or high risk IPI, registered at our centre between November 2011 to June 2013, who received R-EPOCH regimen, were included for the analysis. Case records were reviewed for – demography, histology, stage, bulk of disease, extranodal sites, performance status, IPI, LDH, albumin, details of chemotherapy, grade ¾ toxicities (CTCAE version 4) and need for hospitalization. Responses were evaluated at mid and end of chemotherapy. Overall and progression free survival were calculated. Similar analysis was done for poor prognosis DLBCL patients treated with R-CHOP between Jan 2007 to December 2010. Results Baseline characteristics and treatment outcomes of 32 patients (males-24, females-8) treated with R-EPOCH were compared to 42 patients (males-28, females-14) who received R- CHOP. Median age in R- EPOCH group was 47 years (range-20-75 years) versus 55 years (23-72 years )in R- CHOP. Performance status≥ 2 was seen in 47% in R- EPOCH as compared to 28% in R-CHOP group. Significant proportion of patients in R-EPOCH had bulky disease(81% versus 16%) and stage III/IV disease (90% versus 81%) as compared to R-CHOP. Patients with IPI of two represented 8(25%), IPI of three, 11(34%), and IPI of four and five, 10(32%) on R- EPOCH compared to 21(50%), 19(45%) and 2(5%) on R-CHOP, respectively. Serum albumin Conclusion Our retrospective analysis indicates that treatment with R-EPOCH regimen resulted in similar results as with R-CHOP regimen. However patients treated with R-EPOCH had more adverse features in terms of disease bulk, poor performance status and high IPI score. A prospective randomized comparison is warranted between these two regimens. Disclosures: No relevant conflicts of interest to declare.

Introduction The two relevant questions that emerge and remains unanswered in the management of adult lymphoblastic lymphomas (LBL) are- how to select patients for mediastinal radiation and whom to subject for autologous transplant. Mediastinal radiation or intensification of therapy may reduce the mediastinal recurrence- a common site for relapse. It is also understood that not all patients with residual mediastinal mass relapse. The inability of computed tomography to differentiate between necrotic and viable disease in a residual mediastinal mass post induction therapy led us to explore PET-CT as modality to differentiate the same. In a retrospective analysis we evaluated the role of post-induction chemotherapy PET-CT in predicting the risk of relapse or progression in adult LBL patients. Methods In a single centre retrospective analysis we included newly diagnosed LBL patients (>15 years) who were treated with ALL- like therapy between January 2010 and February 2013. All patients who underwent PET-CT after induction chemotherapy were analysed. SUVmax was used to assess the response. Details of demographic variables, diagnosis, subtype, baseline LDH, bone marrow involvement, cerebrospinal fluid analysis, chemotherapy, radiotherapy, responses, relapse/progression, death and its cause and last follow up date were taken from electronic medical records. Results Twenty-two patients (17 males and 5 females) with median age 24.5 years (range, 16-44 years) were analysed. All patients had T- LBL. Thirteen patients (13/22) had stage IV disease (Ann Arbor stage) and all had bulky mediastinal mass at diagnosis. None of the patient had CSF or bone marrow involvement. Raised LDH and low albumin were present in 80% and 60% of patients respectively. Median WBC was 10,200/cumm (range, 4400-15300/cumm). All but 4 patients received BFM-90 ALL protocol based therapy. After induction 19 patients achieved complete remission on PET-CT. Patients who had residual mediastinal masses on PET-CT relapsed during later phase of therapy (all within 6 months) in mediastinum and died subsequently. Mediastinal radiation was given to one of these patients but it did not prevent a subsequent relapse. One patient who achieved complete response had bone marrow relapse after 16 months of therapy. Three patients had treatment related death (sepsis). At median follow up of 14 months 1-year overall survival was 69% and progression free survival was 78%. Conclusion Post induction therapy PET-CT seems to have prognostic role and may predict relapse for patients with residual disease activity. Such patients should be considered for treatment intensification. However, the role and timing of mediastinal radiation in such patients remains investigational. Disclosures: No relevant conflicts of interest to declare.

Abstract 2690 Background: Aggressive biology, tumor re-growth on treatment and poor tolerance to chemotherapy, are the factors responsible for dismal outcomes in patients with AIDS- related PBL. Treatment with CHOP, CHOP-like regimens or more intensive protocols (HyperCVAD, CODOX-M/IVAC) has failed to improve median survival beyond 15 months. To overcome the problems of tumor re-growth secondary to cancer cell resistance and treatment related toxicity, we devised a regimen in which drugs with proven anti-lymphoma activity were given in fractionated and continuous manner. Continuous daily dosing of oral etoposide for 2–3 weeks provides an effective concentration of drug for extended time periods. Anthracyclines were omitted to reduce the incidence of myelosuppression and mucositis. This regimen was tested prospectively to assess the efficacy and tolerability Methods: Between August 2007 to February 2011, consecutive patients with untreated AIDS-related PBL and age >18 years were counseled for treatment with the proposed regimen at our center. Diagnosis of PBL required absent or weak expression of CD20, expression of MUM-1 or CD 38 or CD138 to suggest plasma cell differentiation. Paraffin blocks were evaluated for expression of EBER, HHV-8 and Ki-67. All patients who were willing for treatment and follow up and did not have CNS involvement or concurrent infections were enrolled. Anti-retroviral therapy (ART) was started concurrently with chemotherapy if not received before. The 3-weekly regimen included cyclophosphamide 375 mg/m2 and vinblastine 4mg/m2 intravenously on day 1 and 8, oral etoposide 50 mg daily for 2 weeks and prednisolone 40 mg/m2//day for the first week of each cycle. Ten weekly doses of intrathecal methotrexate were given as CNS prophylaxis. Radiation was given to the bulky and extranodal sites. Mid and end of therapy responses were evaluated clinically and radiologically (CT or PET-CT). Results: Eighteen patients (males-11, females-7) with median age of 37.5 years (range, 22–51 years) were treated with the above mentioned regimen. Two-thirds of patients were not on ART at diagnosis. Ten patients had tuberculosis as an AIDS-defining illness. Median CD4 count at diagnosis was 175/μL (range 75–407/μL). Significant proportion of patients had adverse prognostic features like B symptoms (9/18), performance status (ECOG) ≥2 (11/18), stage III/IV disease (14/18), bulky disease (15/18), multiple extranodal sites (6/18), raised serum LDH (9/18). Extranodal disease was seen in 17/18 patients commonest being bone followed by anal canal. All except one patient received treatment with chemotherapy (median cycles-6, range 4–8). This patient did not follow up after the initial staging evaluation. Complete responses were seen in 15/17 patients after chemotherapy and 2 patients had partial response. Sixteen patients received radiation. All patients except one had complete response after RT. Patient who continued to have partial response at the end of radiation progressed after 7 months and died. At median follow up of 19 months (range, 3–48 months) both overall survival and event free survival are 87.7%. Median overall survival has not yet been achieved. Treatment was well tolerated with 6 episode of febrile neutropenia which were managed on outpatient basis. One patient was hospitalized for pneumonitis for 7 days. There were no treatment related deaths. On paraffin blocks expression of EBER was seen in 55%. None of them were positive for HHV-8. Six patients had Ki-67 of >90%. Conclusion: This is the first reported series from a single center which has shown improved response rates and survival for AIDS related PBL with a novel regimen as compared to available data till date. However the study has limitations of small size and short follow up. Disclosures: No relevant conflicts of interest to declare.

Abstract 4615 Diagnosis and subtyping of mature B cell Non-Hodgkin's lymphoma (NHL) in the bone marrow (BM) and peripheral blood in a leukemic phase may be challenging due to overlapping cell morphology and immunophenotypic features. This study aims to investigate the utility of CD200 and CD43 co-expression on lymphoid cells in differential diagnosis of mature B cell Non-Hodgkin's lymphoma by flow cytometry (FCM). CD200, known as OX-2 protein was first purified in 1982 as a type I membrane glycoprotein membrane of the Ig superfamily is postulated to play an immunoregulatory role in tumors. A prospective study of staging and diagnostic BM aspirates or peripheral blood samples for immunophenotyping from all consecutive cases of suspected Lymphomas referred to our center over a period of 3 months was done. Co-expression was determined by 3 color FCM. An additional tube with Fluorescein Isothiocyanate conjugated anti CD43, Phycoerythrin conjugated anti CD200 and Phycoerythrin cyanine5 conjugated CD19 was added to the routine panel of antibodies used for immunophenotyping in all the samples. Diagnostic utility of CD200 and CD43 co-expression was determined by comparison with Gold standard diagnosis made out of a combination of clinical features, morphology of tissue biopsies, FCM, immunohistochemistry and cytogenetics using Fluorescent insitu Hybridisation (FISH) for translocations involving IgH gene. Total 116 patients of suspected cases of Lymphomas were referred to our laboratory during the period of study. In addition to Bone marrow and peripheral blood FCM evaluation was done in ascitic fluid (n=1) and pleural fluid (n=3) and FNAC of lymph node (n=1) and retro-orbital mass FNAC (n=1). Out of these, 60 patients showed involvement by mature B cell NHL. Age range of the patients was 26 years to 86 years (Male:Female = 48:12). Chronic Lymphocytic Leukemia (CLL) was the commonest subtype (43.3%, 26/60) followed by follicular lymphoma (16.6%, 10/60), Diffuse Large B-cell Lymphoma (DLBCL) (13.3%, 8/60), Mantle cell lymphoma (MCL) (8.3%, 5/60), Splenic Marginal Zone Lymphoma (SMZL) (5.0%, 3/60), Hairy cell leukemia variant (HCLv) and Waldenstrom's Macroglobulinemia (3.3%, 2/60 each), one patient each (3.3%, 1/60) of Chronic Lymphocytic Leukemia/Prolymphocytic leukemia (CLL/PL), Prolymphocytic leukemia (PL) and Burkitt's Lymphoma. One patient was of unclassifiable low grade B-cell NHL presenting with splenomegaly and pancytopenia with bone marrow involvement and no lymphadenopathy and absence of any trans locations involving IgH gene.This patient had the immunophenotype of CD19, CD22, CD23, CD25, CD79b, CD200 and Kappa positive, with CD20 dim+ and CD5, CD11c, CD103, CD123 and CD43 negative. Annexin A1 was negative in the bone marrow biopsy. The detailed distribution of expression of CD200 and CD43 is given in Table 1.Table 1DiagnosisCD200 pos CD43 posCD200 pos CD43 negCD200 neg CD43 negCD200 neg CD43 posTOTALCLL2600026CLL/PL10001Follicular Lymphoma027110Splenic Marginal Zone Lymphoma02103Hairy cell leukemia variant00202Diffuse large B Cell Lymphoma12328Burkitts Lymphoma00101Small cell/Low grade B Cell Non-Hodgkins Lymphoma01001Mantle Cell Lymphoma00415Prolymphocytic Lymphoma10001Waldenstrom's macroglobulinemia02002Total29918460 Amongst all, CD200 and CD43 co-expression was noted in all cases of CLL, CLL/PL, PLL and only one case of DLBCL. This case of DLBCL was negative for CD5. The truth table for same is given in Table 2.Table 2CD43 and CD200CLL and related NHLNon CLL NHLTotalCoexpression present28129Coexpression absent03131Total283260 The sensitivity specificity data is given in Table 3.Table 3Specificity96.88%Sensitivity100%Positive predictive value96.55%Negative predictive value100% Conclusion: 1. Absence of CD200 and CD43 co-expression strongly rules out a diagnosis of chronic lymphocytic leukemia and related neoplasms. 2. In the differential diagnosis CD5 positive NHL, CD200 positivity strongly suggests the diagnosis of CLL/PLL or PLL. Disclosures: No relevant conflicts of interest to declare.