non-Hodgkin Lymphoma Histologic Subtypes Research Articles

An Unexpected Diagnosis from ERCP for Ascending Cholangitis and Ampullary Mass: Extranodal Diffuse Large B-Cell Lymphoma

Diffuse large B cell lymphoma (DLBCL) is a histological subtype of non-Hodgkin lymphoma (NHL) that accounts for approximately 25% of all NHL cases. DLBCL usually presents as an enlarging mass, most commonly as nodal enlargement in the neck or axilla. Systemic B-symptoms are often reported, including severe night sweats, fever, and weight loss. Extranodal disease has been reported in up to 30 percent of cases, and the diagnosis of DLBCL is most commonly made with excisional lymph node biopsy. A 73 y/o F with a PMH of anemia presented with painless jaundice, abdominal pain, and weight loss. Physical was remarkable for jaundice and sclera icterus. Labs were positive for a WBC of 18.4, ALT of 73, AST of 57, ALKP of 786, total bilirubin of 3.7, direct bilirubin of 3.2, LDH of 247, and lipase of 859. US abdomen showed marked intra- and extra-hepatic biliary dilation, gallbladder distention, and abrupt termination of the CBD in the pancreatic head. CT scan confirmed biliary dilation with extensive peritoneal metastases or lymph nodes. A CA 19-9 level was 8894. An ERCP with cytology brushing and ampulla biopsy revealed an ampullary mass, a 1.5 cm dilated intrahepatic biliary tract, and a 5 cm distal and mid-common bile duct stricture beginning at the ampulla. A PET-CT scan was obtained, which showed abnormal uptake in posterior mediastinal, right axillary, right cardiophrenic, mesentery, and retroperitoneum lymph nodes, as well as in the right liver lobe and the pancreatic head and tail. Cytology brushing revealed atypical cells, and ampullary biopsy revealed DLBCL. This case of DLBCL was unique for multiple reasons. The initial symptoms of painless jaundice, abdominal pain, subjective fevers, and recent weight loss are not typical presenting symptoms of DLBCL; no rapidly enlargement mass or nodal enlargement of the neck and axilla was present. Her symptoms could have been explained solely by her ascending cholangitis. It was also remarkable how the clinical presentation resembled pancreatic adenocarcinoma but was actually due to a DLBCL with significant distant metastasis. This less common extranodal presentation would not have been expected given her complaints, as additional symptoms or a diagnosis would have presumably been previously suspected. Lastly, the majority of DLBCL diagnoses are made with excisional lymph node biopsy, but this case's diagnosis was interestingly made from an ampullary mass biopsy from an ERCP for ascending cholangitis.

Abstract 3838: Upregulation of DLEU1 significantly prolongs survival in a rituximab-treated DLEU1 knockout human Burkitt lymphoma (BL) xenograft NSG mouse model: DLEU1 may act as a tumor suppressor gene in pediatric BL

Abstract BACKGROUND: Burkitt lymphoma (BL) is the most common histological subtype of non-Hodgkin lymphoma (NHL) in children and adolescents (Cairo et al., Blood 2007; Miles/Cairo., BJHaem 2012). We identified that pediatric BL patients with chromosome 13q deletion, particularly 13q14.3, had significantly poorer outcome and inferior overall survival despite aggressive short, intensive multi-agent chemotherapy (Poirel/Cairo et al., Leuk 2009; Nelson/Cairo/Sanger et al., BJH 2009). Deleted in lymphocytic leukemia 1 (DLEU1) is a BL classifier gene in the 13q14.3 region (Dave et al., NEJM 2006) and interacts with C-MYC, TUBB2C and UBR1 (Stelzl et al., Cell 2005). We have previously reported a regulated DLEU1 expression significantly associated with changes in BL proliferation in vitro (Lee/Cairo et al., AACR 2015). We hypothesize that DLEU1 may have function as a tumor suppressor gene; however, functions and mechanisms underlying DLEU1 expression in the BL are poorly understood. OBJECTIVES: We hypothesize that DLEU1 may act as a tumor suppressor gene in BL and therefore examined whether up-regulation of DLEU1 results in changes in survival following targeted immunotherapy. METHODS: DLEU1 stably overexpressing Raji BL cells (Lee/Cairo et al., ASH 2013) were transfected with a firefly luciferase expression plasmid, kindly provided by Laurence Cooper MD, PhD. Four- to six- week-old female NSG (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ) mice from The Jackson Lab. Tumor burden and tumor progression were monitored by bioluminescence imaging system. Mice were treated with either rituximab (30 mg/kg) or cyclophosphamide (25 mg/kg) by intraperitoneal injection at 7 day intervals. Survival rates were analyzed by the Kaplan-Meier method using the Prism Version 6.0 software. RESULTS: We observed that DLEU1 stably overexpressing Raji BL cells xenografted (GFP-DLEU1) mice had significantly extended survival compared to GFP control mice following treatment rituximab (59 vs 51 days, p<0.05, n = 8), cyclophosphamide (50.5 vs 37.5 days, p<0.0001, n = 12) and in combination treatment (60.5 vs 53 days, p<0.05, n = 12) whereas there were no significant differences in survival between WT and DLEU1-KO mice with PBS treatment. We observed that there were no significant differences of luciferase intensity in between GFP and GFP-DLEU1 mice following rituximab, cyclophosphamide or in combination treatment. CONCLUSIONS: We demonstrated that the upregulation of DLEU1 expression significantly improved the survival in DLEU1 overexpressing BL cells xenografted NSG mice following rituximab, cyclophosphamide and in combination treatment. Therefore, the up-regulation of DLEU1 expression in BL may in part result in sensitizing to chemoimmunotherapy induced survival and may result in a consideration of potential therapeutic strategies Citation Format: Sanghoon Lee, Changhong Yin, Tishi Shah, Janet Ayello, Erin Morris, Carmella van de Ven, Mitchell S. Cairo. Upregulation of DLEU1 significantly prolongs survival in a rituximab-treated DLEU1 knockout human Burkitt lymphoma (BL) xenograft NSG mouse model: DLEU1 may act as a tumor suppressor gene in pediatric BL. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3838.

68. Unusual presentation of mediastinal lymphoma and role of cardiac MRI

Diffuse large B cell lymphoma (DLBCL) is the most common histologic subtype of non-Hodgkin lymphoma (NHL) accounting for approximately 25% of NHL cases. One of the common subtype of DLBCL is primary DLBCL of the mediastinum. Case report this 65 year old female known to have diabetes and hypertension presented to our emergency department with history of epigastric pain for last 2 h. Her electrocardiogram (ECG) showed right bundle block with left posterior hemi-block representing bi-fasicular block with minimal ST segment depression in leads V4–V6. Her initial routine laboratory results revealed normal renal function, blood counts and liver profile. Her cardiac bio-markers were elevated with Troponin I of 1.22 and CPK of 35. She was admitted by the cardiology team diagnosis of non-ST elevation myocardial infarction (NSTEMI). She was started with usual anti-ischemic. Next morning she had echocardiography which revealed a large mediastinal mass on the antero-lateral aspect of the left ventricle infiltrating the basal lateral and anterior wall. This mass was encasing the origin of the great vessels and also infiltrating the left atrium occluding the Left atrial appendage and left upper pulmonary vein. It was infiltrating the Right ventricular outflow tract causing obstruction to the flow with a gradient of 52 mmHg. Cardiac MRI showed multiple cardiac masses, the largest of which was originating from the anterior mediastinum and going posteriorly then infiltrating RV and within the right ventricular out-flow tract (RVOT) causing significant obstruction. The magnetic resonance characteristics of the intra cardiac and extra cardiac masses were same and with features of central necrosis was highly suggestive of lymphoma. The CT scan of the chest and abdomen showed the same cardiac findings as of cardiac MRI and multiple enlarged thoracic, retroperitoneal, left common iliac lymph nodes. Patient had CT guided Lymph node biopsy from the mediastinal lymph node and was reported as Diffuse Large B-Cell lymphoma of non-germinal center subtype with anaplastic features. Bone marrow biopsy was also performed as well PET oncology to complete staging process. The case was discussed in the multi-disciplinary oncology board and planned for chemotherapy with CHOP with no need for any cardiac intervention at this moment. The care of the patient was transferred to the hematology team with close follow-up by cardiology team. He received the chemotherapy for 4 cycles and had a repeat cardiac MRI which showed the complete resolution of the intra-cardiac mass. Patients with DLBCL typically present with a rapidly enlarging symptomatic mass, most usually nodal enlargement in the neck or abdomen, or, in the case of primary mediastinal large B cell lymphoma, the mediastinum, but may present as a mass lesion anywhere in the body. The prognosis is good as the rate of remission with chemotherapy is good. The cardiac MRI can assist in diagnosis.

Diffuse Large B-Cell Lymphoma Presenting as an Endobronchial Mass: A Case Report and Review of Literature

Diffuse large B-cell lymphoma (DLBCL) is the most common histological subtype of non-Hodgkin lymphoma (NHL). DLBCL normally presents with nodal involvement in the neck or the abdomen. Extranodal extramedullary disease is seen in up to 40% of cases. However, endobronchial involvement is very rare. Here we describe a 77-year-old woman with a history of bronchial asthma presenting with cough and shortness of breath who was initially treated for exacerbation of asthma. Persistent symptoms lead to further evaluation including CT scan and bronchoscopy, resulting in the diagnosis of DLBCL in this woman presenting as an endobronchial lesion. This case demonstrates the importance of considering lymphoma as a differential diagnosis of an isolated endobronchial lesion. J Hematol. 2016;5(3):106-109 doi: http://dx.doi.org/10.14740/jh283w

Downregulation of DLEU1 Significantly Shortened Survival in a Rituximab-Treated DLEU1 Knockout Human Burkitt Lymphoma (BL) Xenograft NSG Mouse Model: DLEU1 May Act As a Tumor Suppressor Gene in Pediatric BL

BACKGROUND: Burkitt lymphoma (BL) is the most common histological subtype of non-Hodgkin lymphoma (NHL) in children and adolescents (Cairo et al., Blood, 2007; Miles/Cairo, BJH, 2012). We have previously identified secondary chromosomal aberrations in 70% of pediatric BL patients (PBL) with a C-MYC gene rearrangement (Poirel/Cairo et al., Leukemia, 2009). Specifically, we identified significantly inferior event free survival (EFS) and overall survival in children and adolescents with a specific loss of the 13q14.3 locus (Poirel/Cairo et al., Leukemia, 2009; Nelson/Cairo/Sanger et al., Br. J. Haematol., 2009). In a multivariate analysis controlling for stage, lactate dehydrogenase levels, country of treatment, and group classification, children with BL who had a 13q deletion had significantly poorer EFS compared to the remainder of patients treated with the same French-American-British (FAB) chemotherapy regimen (Poirel/Cairo et al., Leukemia, 2009). Deleted in lymphocytic leukemia 1 (DLEU1) is a BL classifier gene in the 13q14.3 region (Dave et al., NEJM, 2006) and interacts with C-MYC, Tubulin beta-2C chain (TUBB2C), E3 ubiquitin-protein ligase (UBR1), cellular tumor antigen p53, and Ras association (RalGDS/AF-6) domain family member 1 (RASSF1) (Stelzl et al., Cell, 2005). The sequence-specific Transcription Activator-Like Effector Nucleases (TALENs) have been developed for targeted genome editing in in vitro and in vivo experiments with high efficiency as a new therapeutic tool (Sander et al., Nat. Biotechnol., 2011). We have previously reported a down-regulated DLEU1 mRNA expression significantly associated with an increase in BL proliferation in vitro (Lee/Cairo et al., AACR, 2015). We hypothesize that DLEU1 may have function as a tumor suppressor gene; however, the role of DLEU1 regulating programmed cell death in BL are poorly understood.OBJECTIVES: We hypothesize that DLEU1 may act as a tumor suppressor gene in BL and whether down-regulation of DLEU1 expression results in changes in BL survival following targeted immunotherapy.METHODS: TALENs induced DLEU1 knockout Raji cells were previously generated (Lee/Cairo et al., ASH, 2013) and DLEU1 knockout cells were stably transfected with a firefly luciferase expression plasmid (ffluc-zeo), kindly provided by Laurence Cooper MD, PhD. Four- to six- week-old female NSG (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) mice from The Jackson Laboratory were irradiated (2.5 Gy), and then mice were subcutaneously injected with 1x106 ffluc-zeo tumor cells. Tumor burden and tumor progression were monitored by bioluminescence imaging system using the Xenogen IVIS-200 (Caliper Life Sciences). Mice were treated with either PBS, IgG isotype control, rituximab (30 mg/kg) or cyclophosphamide (25 mg/kg) and in combination by intraperitoneal (i.p) injection at 7 day intervals. Survival rates were analyzed by the Kaplan-Meier method and differences evaluated by log-rank test using the Prism Version 6.0 software.RESULTS: There were significant increases of luciferase signal intensity in DLEU1 knockout mice (DLEU1-KO) compared to that in wild type (WT) mice on day 31 of rituximab (p<0.05), cyclophosphamide (p<0.05) or rituximab and cyclophosphamide (p<0.01) combined treatment. Consistent with these initial findings in tumor growth, we found that rituximab-treated DLEU1-KO mice (n=12 per group) had a significantly shortened survival time with a median of 42 days compared to that of WT mice (n=12 per group) (52 days, p<0.005) (Figure 1A). For the rituximab and cyclophosphamide combination treated group, WT mice (n=12 per group) had a significantly extended survival time compared to the DLEU1-KO group (55.5 days vs 48 days, p<0.05) (Figure 1B). There were no significant differences in survival between WT and DLEU1-KO mice with PBS and IgG isotope treatment.CONCLUSIONS: The down-regulation of DLEU1 expression significantly decreased the survival rate in DLEU1-KO xenografted NSG mice following rituximab and in combination with cyclophosphamide treatment. Therefore, the down-regulation of DLEU1 expression in BL may in part result in immunotherapy resistance and may result in a consideration of alternative therapeutic strategies. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Single Cell Mass Cytometry for Phenotypic Analysis of Diffuse Large B-Cell Lymphoma

Diffuse Large B-cell Lymphoma (DLBCL) is the most common histologic subtype of non-Hodgkin Lymphoma (NHL). Despite its improved outcome with R-CHOP chemotherapy, 40% of patients still suffer with relapsed or refractory disease. Further investigation is needed to understand the complexity of DLBCL. Time-of-flight mass cytometry (CyTOF) is a recently developed technology that combines traditional flow cytometry with mass spectrometry and supports analysis of up to 30-40 parameters simultaneously at the single cell level with minimal spectral overlap (Bendall et al, Science 2011). In this study, we sought to develop a CyTOF method for phenotypic analysis of DLBCL to obtain high resolution profiles of the malignant clone(s) represented in individual tumor samples and resolve any underlying population substructure that might be informative in understanding the clinical and biologic heterogeneity of this disease.We designed a two-tube assay for this study. Tube #1 contained 35 cell surface markers including CD45, CD19, CD20, CD22, CD79B, IgM, IgD, Ig Kappa, Ig Lambda, CD5, CD10, CD23, CD43, CD38, CD138, CD44, CD21, CD24, CD40, CD72, CD80, CD45RA, CD49D, CD49F, CD62L, CD25, CD27, CD30, CD127, CD184, CD194, CD200, CD34, HLA-DR, and CD3. Tube #2 contained 37 markers in total including 17 cell surface markers overlapping with Tube #1 plus an additional 20 intracellular markers (BCL2, BCL6, IRF4/MUM1, LMO2, MYC, MCL1, MEF2B, KAT3B/P300, CBP, FOXP1, RUNX1, Ikaros, EZH2, BMI1, NOTCH1, CARD11, IkBa, phospho-Rb, Ki67, and CyclinD2). Metal-conjugated antibodies were purchased from DVS Sciences or unconjugated antibodies labeled in-house using DVS MaxPar metal labeling kits. Data were acquired using a DVS CyTOF2 instrument. We examined both fresh and viably frozen single cell suspensions from diagnostic lymph node biopsy samples received for flow cytometric analysis at the BC Cancer Agency. We used SPADE (spanning-tree progression analysis of density-normalized events) for initial data analysis.We first performed preliminary validation studies including cross-comparison of mass cytometry (CyTOF2) vs. flow cytometry (BD Canto2) datasets and fresh vs. previously frozen cell suspension material. Although individual marker intensities using matched antibody clones were in general lower by CyTOF, eight-parameter surface staining results were qualitatively comparable between the two platforms. Also there were essentially no differences observed in CyTOF staining profiles between fresh and previously frozen samples. Preliminary clustering analysis of cell populations using SPADE revealed clear separation between normal and malignant B cell populations as well as apparent substructure to the malignant population in a subset of DLBCL samples. These findings suggest intratumoral heterogeneity can be resolved by high dimensional CyTOF analysis. Ongoing efforts will focus on determining if phenotypically defined subsets show enrichment for subclonal mutations. DisclosuresNo relevant conflicts of interest to declare.

A Prospective Analysis of Blood Donation History and Incidence of Non-Hodgkin Lymphoma

Background: The incidence rate of non-Hodgkin lymphoma (NHL) has more than doubled over the last few decades, but the reason for this increase has been largely unexplained. The long-term effect of blood donation on cancer incidence is not well investigated despite its potentially large impact on the population. Immune deficiency is associated with increased risk of NHL. Blood donation may influence subsequent NHL development via temporary immune system alterations following blood donation, which have been demonstrated in several human studies. A large nested case-control study within a cohort of blood donors in Sweden and Denmark showed an increased odds ratio (OR) of NHL among frequent plasma donors comparing to non-donors (OR 2.14, 95% confidence interval (CI) 1.22-3.74) [Edgren G et al., 2008]. Results of this and other studies have been difficult to interpret because of the possible influence of healthy donor effects or different baseline characteristics among blood donors compared to general population. To test the hypothesis that frequent blood donation is associated with an increased risk of NHL and its most common histologic subtypes in a general, non-immune compromised population, we conducted a prospective study in the Health Professional Follow-up Study (HPFS) cohort.Methods: We followed 36,580 men in the HPFS cohort, who in 1992 had no prior diagnosis of cancer and provided information on frequency of blood donation in the past 30 years. Throughout follow-up of the HPFS cohort, participants completed questionnaires about lifestyle and medical history every two years. Non-respondents were followed through the National Death Index. Identified diagnoses of cancer were confirmed by medical record review. The present analysis included all confirmed incident diagnoses of NHL through January 2008. Person-time accrued for each participant from January 1992 to the earliest among dates of NHL diagnosis, death, or January 2008. Cox proportional hazards regression was used to calculate hazards ratios (HR) and 95% CIs for the risk of all NHL and major NHL histologic subtypes associated with number of blood donations (1-5, 6-9, 10-19, or >20, vs. none), stratifying by age in months and follow-up cycle. Potential confounding by putative NHL risk factors (i.e. smoking, alcohol, BMI, height, physical activity, race) was evaluated by including these factors in multivariable models.Results: During 482,008 person-years of follow-up, 484 incident cases of NHL were confirmed. Participant characteristics were similar at baseline among blood donors and non-donors with respect to age, race, and selected lifestyle factors. In the age-adjusted model, there was no significant association between blood donation frequency and incidence of NHL (HR 1.12, 95% CI 0.82-1.55, comparing >20 donations vs. 0 donations over 30 years, p for trend = 0.44) or of any major NHL subtypes. Adjustment for potential confounders did not notably change the effect estimates for all NHL or any NHL subtypes.Conclusion: In this prospective study, there was no evidence that frequent blood donation is associated with incidence of all NHL or any major subtypes. DisclosuresNo relevant conflicts of interest to declare.

Autoimmune diseases associated with non-Hodgkin lymphoma: a nationwide cohort study

The cumulative risk of non-Hodgkin lymphoma (NHL) in Sweden by age 80 years has increased to 1.1 in women and 1.6% in men in 2011. Increased risk of NHL associated with personal histories of some autoimmune diseases (ADs) is known. It is unclear whether there are other NHL-related ADs and whether this association holds across different sex, age and year of diagnosis, or NHL histological subtypes. Over an average of 9.4-year (maximum 47 years) follow-up of 878 161 patients diagnosed in 1964-2010 with 33 different ADs, 3096 subsequent NHL were diagnosed (data: Swedish Cancer Registry). Of 33 studied ADs, 21 showed significantly increased risk of NHL; 6 of them tended to increase the risk and none significantly decreased it. The overall standardized incidence ratio (SIR) for NHL after ADs was 1.6 [novel findings: immune thrombocytopenic purpura (ITP) = 7.5, polymyositis/dermatomyositis = 4.1, primary biliary cirrhosis = 3.9, myasthenia gravis = 2.2, Behcet = 1.7, rheumatoid fever = 1.7, ulcerative colitis = 1.5, polymyalgia rheumatica = 1.4, and chronic rheumatic heart disease = 1.4; confirmatory findings: autoimmune hemolytic anemia = 27.2, Sjögren = 4.9, Celiac = 4.8, systemic lupus erythematosus = 4.4, polyarteritis nodosa = 2.9, discoid lupus erythematosus = 2.7, sarcoidosis = 2.6, Crohn = 2.1, systemic sclerosis = 2.1, rheumatoid arthritis = 2.0, and Hashimoto/hypothyroidism and psoriasis = 1.4]. SIR for NHL diagnosis before age 60 (2.2) was significantly higher than that in older ages (age ≥60: 1.5). The SIRs in women or men and in period 1993-2010 or 1964-1992 were similar. Risk of all common NHL histology subtypes significantly increased after ADs (cutaneous/peripheral T cell and anaplastic large T and null cell = 2.2; small B-cell lymphocytic = 1.7; diffuse large B cell = 1.6; follicular and mantel cell = 1.3). Many of 33 studied ADs (except for ankylosing spondylitis, diabetes type I graves/hyperthyroidism, multiple sclerosis, chorea minor, and pernicious anemia), especially when diagnosed at younger ages, were associated with higher risk of NHL. However, the absolute risk of NHL in many ADs is still small.

Abstract 5073: Serum sCD23 and sCD30 associated with non-Hodgkin lymphoma risk as far as 15 to 23 years after blood collection

Abstract Background: We and other investigators have recently reported associations between elevated serum concentrations of selected immune markers and subsequent risk of non-Hodgkin lymphoma (NHL). While these findings support a role for subtle immunologic effects in lymphomagenesis, the relatively short length of follow-up after phlebotomy in these studies (10-13 years) limits the ability to rule out reverse causation as an explanation for these findings. To clarify the temporal nature of these associations, we conducted a nested case-control study investigating serum levels of six immune markers and NHL diagnosed up to 23 years after phlebotomy within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study, a cohort with banked fasting serum from 29,133 male Finnish smokers. Methods: We selected 272 first-primary NHL cases diagnosed 2+ years after phlebotomy, identified through linkage with the Finnish Cancer Registry, and 325 controls individually matched to cases on baseline age, phlebotomy date, and number of previous specimen thaws. Serum from these individuals were tested for soluble CD23 (sCD23), sCD27, sCD30, B cell attracting chemokine-1 (BCA-1), soluble tumor necrosis factor receptor-2 (sTNFR-2), and soluble vascular endothelial growth factor receptor-2. Odds ratios (OR) and 95% confidence intervals (CI) relating categories of analyte concentration to NHL risk were computed through conditional logistic regression with adjustment for cigarette pack-years. Polytomous regression models were also fit to compute ORs for the two most common NHL histologic subtypes, chronic lymphocytic leukemia (CLL) and diffuse large B cell lymphoma (DLBCL). Results: We observed NHL associations with elevated pre-diagnostic concentrations of the B cell activation markers sCD23 (highest analyte category vs. lowest: OR 2.4, 95% CI 1.5-3.8, Ptrend = 0.0005) and sCD30 (OR 3.3, 95% CI 1.9-5.5, Ptrend &amp;lt; 0.0001). The associations with sCD23 and sCD30 were apparent for cases diagnosed as far as 15-23 years after blood collection (OR 5.1, 95% CI 1.8-14.6, Ptrend = 0.002 and OR 3.7, 95% CI 1.5-9.3, Ptrend = 0.01 respectively). In subtype-specific analyses, both sCD23 and sCD30 were significantly associated with DLBCL, both overall and in later follow-up periods. sCD23 was significantly associated with CLL, particularly for earlier follow-up periods, while sCD30 was not. Associations with NHL were observed for sCD27, BCA-1, and sTNFR-2 but became weaker or null for cases diagnosed &amp;gt;8 years post-phlebotomy. Conclusions: Our study provides strong evidence that elevated serum sCD23 and sCD30 are associated with future risk of NHL many years after blood collection. Our findings for these B cell activation markers suggest that B cell activation is an important mechanism in NHL development in the general population. Citation Format: Mark P. Purdue, Qing Lan, Troy J. Kemp, Allan Hildesheim, Stephanie J. Weinstein, Demetrius Albanes, Ligia A. Pinto, Nathaniel Rothman. Serum sCD23 and sCD30 associated with non-Hodgkin lymphoma risk as far as 15 to 23 years after blood collection. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5073. doi:10.1158/1538-7445.AM2014-5073

Association of XRCC1 genetic polymorphism with susceptibility to non-Hodgkin's lymphoma

The purpose of this study was to explore the association between X-ray repair cross-complementing group 1 (XRCC1)gene polymorphism and non-Hodgkin's lymphoma risk. A total of 282 non-Hodgkin's lymphoma (NHL) patients and 231 normal controls were used to investigate the effect of three XRCC1 gene polymorphisms (rs25487, rs25489, rs1799782) on susceptibility to non-Hodgkin's lymphoma. Genotyping was performed by using SNaPshot method. All statistical analyses were done with R software. Genotype and allele frequencies of XRCC1 were compared between the patients and controls by using the chi-square test. Crude and adjusted odd ratios and 95% confidence intervals were calculated by using logistic regression on the basis of genetic different models. For four kinds of NHL, subgroup analyses were also conducted. Combined genotype analyses of the three XRCC1 polymorphisms were also done by using logistic regression. The results showed that the variant genotype frequency was not significantly different between the controls and NHL or NHL subtype cases. Combined genotype analyses of XRCC1 399-280-194 results showed that the combined genotype was not associated with risk of NHL overall, but the VT-WT-WT combined genotype was associated with the decreased risk of T-NHL (OR: 0.21; 95%CI (0.06-0.8); P = 0.022), and the WT-VT-WT combined genotype was associated with the increased risk of FL(OR:15.23; 95%CI (1.69-137.39); P = 0.015). It is concluded that any studied polymorphism (rs25487, rs25489, rs1799782) alone was not shown to be rela-ted with the risk of NHL or each histologic subtype of NHL. The combined genotype with mutation of three SNP of XRCC1 was not related to the risk of NHL. However, further large-scale studies would be needed to confirm the association of decreased or increased risk for T-NHL and FL with the risk 3 combined SNP mutants of XRCC1 polymorphism.

Short Communication: Spectrum of Non-Hodgkin Lymphoma in an Urban Ryan White-Funded Clinic in the Established Antiretroviral Era

People living with HIV/AIDS (PLWHA) are at a higher risk of developing non-Hodgkin lymphoma (NHL). The influence of combined antiretrovirals (cART) on the presentation, treatment, and outcomes of HIV-associated NHL (HIV-NHL) warrants further investigation. We performed a retrospective analysis of PLWHA diagnosed with NHL who received care at the Infectious Diseases Ponce de Leon Center in Atlanta, Georgia, from January 1, 2004 to December 31, 2010. Thirty-five patients with HIV-NHL were identified. Among these patients, 7 had Burkitt lymphoma (BL), 20 had diffuse large B cell lymphoma (DLBCL), 7 had plasmablastic lymphoma (PL), and 1 had primary effusion lymphoma (PEL). The majority of patients (82.9%) presented with advanced disease, and 63% were not on ART at diagnosis. Despite having good performance status at presentation, the majority of patients presented with high International Prognostic Index (IPI) scores. There were differences between the histologic subtypes of NHL in regard to treatment, complications, and outcomes. The median CD4 lymphocyte count at diagnosis was 110 cells/mm(3) for patients with DLBCL [interquartile range (IQR): 66, 203], 165 cells/mm(3) for Burkitt lymphoma (IQR: 36, 199), and 98 cells/mm(3) for plasmablastic lymphoma (IQR: 34, 214). Overall, patients completed 67% of planned chemotherapy cycles. Common causes for chemotherapy termination were persistent myelosuppression (18.2%), social factors (22.7%), and disease progression (36.4%). Social factors included lack of transportation, substance abuse, unstable housing, and poor adherence. Two-year overall survival was 40% for all HIV-NHL. Half of the patients with DLBCL (n=10), 42% of patients with PL (n=3), and only 14.3% of patients with BL (n=1) were alive at 2 years. Among the overall survivors at 2 years, 85.7% had CD4 >200 cells/mm(3) and 78.6% had undetectable HIV viral loads (VL) at that time.

Predicting Cell Of Origin In Diffuse Large B-Cell Lymphoma By High Dimensional Flow Cytometry

IntroductionDiffuse Large B-cell Lymphoma (DLBCL) is the most common histologic subtype of non-Hodgkin Lymphoma (NHL). Patients with DLBCL can be divided into prognostic subgroups according to the cell of origin (COO) of the lymphomas that are characterized as the germinal centre B-cell-like (GCB) and the activated B-cell-like (ABC) subgroup (Alizadeh et al Nature 2000:403:503-511). While Gene Expression Profile (GEP) is the “gold standard” method for defining COO in DLBCL, to perform GEP routinely on clinical cases would incur both financial and logistical pressures that would be challenging to overcome in many centers. As a result, several groups have endeavored to devise immunohistochemical (IHC) staining panels that recapitulate GEP results, but have met with variable success. In contrast with the inherently subjective nature of IHC result interpretation and the limited number of IHC-stained cells that can be assessed visually, flow cytometry is a widely available,mature and robust methodology that achieves quantitative measurement of multiple simultaneous antibody staining parameters on tens to hundreds of thousands of individual cells in a matter of minutes. In this study, we sought to devise a flow cytometric method for determining COO in DLBCL from freshly disaggregated patient lymph node biopsy specimens. MethodsSix GCB cell lines (OCI-Ly1, SU-DHL-10, SU-DHL-4, SU-DHL-8, Karpas 422 and Pfeiffer) and 6 ABC cell lines (SU-DHL-9, NU-DUL-1, OCI-Ly3, HBL-1, MD903 and OCI-Ly10) were stained with eight cell surface markers (CD45, CD19, CD10, CD27, CD38, CD138, CD44 and CD34), two intracellular markers (BCL6 and IRF4), and a viability dye simultaneously using a single-tube, 11-color assay and analyzed on a 4-laser BD LSRFortessa flow cytometer. Antibodies were chosen to include known GCB (CD10, CD27, CD38 and BCL6) and ABC (CD138, CD44 and IRF4) markers based on prior studies. CD45 and CD19 were included to assist in identifying the malignant B cell population and CD34 was included as an investigational marker. FCS file data were analyzed using FlowJo software. ResultsEvaluation of mean fluorescence intensity (MFI) values and their corresponding standard deviations for 40,000-190,000 viable events for each sample revealed CD27 and BCL6 to be expressed at significantly higher levels in GCB cell lines, and CD44 and IRF4 to be significantly higher in ABC cell lines. CD10 and CD34 tended toward higher expression in GCB and ABC cell lines, respectively, but these differences did not reach statistical significance according to the Student's t-test. Analysis of all 10 fluorescence parameters simultaneously (i.e. 10-dimensional data projection) requiring more sophisticated bioinformatic tools is currently in progress. According to the serial univariate COO decision algorithm outlined by Hans et al (Blood 2004;103:275-282), our flow cytometry results correctly assigned 5 out of 6 GCB and 4 out of 6 ABC cell lines (cell line COO designations as defined by GEP). DiscussionIn this study, we assessed the performance of a flow cytometry-based assay for determining COO in DLBCL. Our results demonstrated that most GCB and ABC DLBCL cell lines can be correctly assigned with initial 10-color flow panel; however, some markers were highly informative while others were less or not informative at all in COO discrimination. Further development will be needed to improve the antibody panel design to the point they are able to replicate GEP results faithfully. Although flow cytometry cannot be performed on archival pathology material (i.e. formalin fixed, paraffin embedded tissue), most community and academic pathology departments have either their own flow cytometry instrumentation on site or established local referral centers and thus fresh tissue can usually be allocated for multidimensional flow analysis. Our results support the usefulness of flow cytometry for COO determination in DLBCL. However, further optimization of the panel design and testing on patient biopsy samples is needed. Disclosures:No relevant conflicts of interest to declare.

A Prospective Analysis of Body Size during Childhood, Adolescence, and Adulthood and Risk of Non-Hodgkin Lymphoma

The etiology of non-Hodgkin lymphoma (NHL) is poorly understood. Obesity is associated with inflammation, a cytokine milieu conducive to lymphocyte proliferation, and has been associated with NHL risk in some epidemiologic studies. To prospectively examine NHL risk in relation to adult and earlier life obesity, we documented 635 incident NHL diagnoses among 46,390 men in the Health Professionals Follow-up Study and 1,254 diagnoses among 116,794 women in the Nurses' Health Study over 22 to 32 years of follow-up. Using multivariable Cox proportional hazards models, we estimated cohort-specific incidence rate ratios (RR) and 95% confidence intervals (CI) for risk of NHL and major histologic subtypes associated with cumulative average middle and young adult (ages, 18-21 years) body mass index (BMI) and adolescent and childhood somatotype. NHL risk was modestly increased in men (but not women) with a cumulative average middle adult BMI ≥ 30 kg/m(2) (vs. 15-22.9 kg/m(2); RR, 1.28; 95% CI, 0.92-1.77; Ptrend = 0.05). In meta-analyses across cohorts, higher young adult BMI was associated with increased risk of all NHL (pooled RR per 5 kg/m(2), 1.19; 95% CI, 1.05-1.37), diffuse large B-cell lymphoma (DLBCL), and follicular lymphoma (all Ptrend ≤ 0.02). Adolescent somatotype was also positively associated with all NHL, DLBCL, and follicular lymphoma in pooled analyses (all Ptrend ≤ 0.03), whereas childhood somatotype was positively associated with NHL overall among women only (Ptrend < 0.01). These findings in two large prospective cohorts provide novel evidence that larger body size in childhood, adolescence, and young adulthood predicts increased risk of NHL, and particularly of DLBCL and follicular lymphoma.

Paediatric lymphoma in China: A clinicopathological study of 213 cases

This retrospective study was conducted to evaluate information on paediatric lymphoma in China. We reviewed the pathological files of patients less than 12 years of age with lymphoma in Shanghai Xinhua Hospital from January 1982 to June 2009. SPSS version 11.0 was used to analyse the results. Of the 213 subjects, 176 (82.6%) had non-Hodgkin's lymphoma (NHL) and 37 (17.4%) had Hodgkin's lymphoma (HL). All NHL cases had diffuse and high grade tumours, and 33.5% of these tumours primarily involved extra-nodal sites. Of the NHL cases, 56.6%, 43.3%, and 1.7% were derived from T, B, and null cells, respectively. Lymphoblastic lymphoma (LL, 50.6%), Burkitt's lymphoma (BL, 28.4%), and anaplastic large cell lymphoma (ALCL, 12.5%) comprised the majority of the NHL cases. A significant difference was found in the frequency of stage I/II cases between LL and ALCL. Paediatric HL resembled the disease in adults. Paediatric lymphoma in China is different from that in Western countries with respect to the incidence rate of HL and BL. The distribution pattern of NHL histological subtypes is more similar to that in Japan than that in Pakistan. These features suggest ethnic or geographic variations.

The Incidence, Characteristics, and Outcomes of Venous Thromboembolism in Patients with Follicular Lymphoma Compared to Diffuse Large B-Cell Lymphoma

Abstract 5090 Background:Previous studies have demonstrated a widely variable incidence of thromboembolism (VTE) in non-Hodgkin lymphoma (NHL) patients, ranging from 3% to 60%. This is in part because many previous studies have considered NHL as a single disease entity, combining a heterogeneous group of histological subtypes, each with a potentially different thrombogenic potential. We sought to clarify differences in VTE incidence between the indolent and intermediate grade forms of NHL, by looking at follicular lymphoma (FL) and diffuse large B-cell lymphoma (DLBCL), respectively. Furthermore, since chemotherapy is a major risk factor for VTE, we compared VTE incidence rate in the pre- and post-chemotherapy time periods. Patients/Methods:We assembled a retrospective cohort of the United States veterans diagnosed with DLBCL or FL between 1998 and 2009. VTE was identified by ICD-9 codes, while DLBCL or FL was identified by ICD-O3 codes in the Veterans Health Administration central cancer registry. Patients who did not receive any chemotherapy or had a history of VTE prior to the diagnosis of lymphoma were excluded. VTE rate was compared by Chi-square testing. Results:A total of 2094 DLBCL patients and 470 FL patients were included. In the period between lymphoma diagnosis and treatment initiation, VTE occurred at a rate of 18. 0 per 100 person-years in DLBCL and 2. 3 per 100 person-years in FL (p<0. 0001). Similarly, in the 6 months following treatment initiation, DLBCL patients had a VTE rate of 12. 3 per 100 person-years, compared to 7. 3 per 100 person-years in FL (p=0. 039). In DLBCL, patients had a higher risk of VTE between diagnosis and treatment compared to the 6 months after treatment initiation (p=0. 038). In FL, treatment was associated with a trend toward higher risk of VTE, although not statistically significant (p=0. 056). Conclusions:DLBCL is more “thrombogenic” than FL both before and during treatment. To our knowledge, this is the first report of direct comparison of VTE event rates between different histological subtypes of NHL, knowledge of which could influence decisions related to thromboprophylaxis. There are limitations of ICD-9 codes which should be acknowledged, and further research will be required to identify the optimal method to identify patients with VTE. The lower incidence of VTE after treatment initiation in DLBCL patients suggests that the reduced disease burden associated with treatment may actually decrease the overall risk of VTE despite the increased risk associated with chemotherapy. Conversely, in FL the risk of VTE appears to be higher in association with treatment. Disclosures:No relevant conflicts of interest to declare.

Occupational solvent exposure, genetic variation of DNA repair genes, and the risk of non-Hodgkin’s lymphoma

The main objective of this study was to test the hypothesis that genetic variations in DNA repair genes may modify the association between occupational exposure to solvents and the risk of non-Hodgkin's lymphoma (NHL). A population-based case-control study was conducted on Connecticut women including 518 histologically confirmed incident NHL cases and 597 controls. Unconditional logistic regression models were used to estimate the odds ratios and effect modification from the 30 single nucleotide polymorphisms in 16 DNA repair genes of the association between solvent exposure and the risk of NHL overall and subtypes. Single nucleotide polymorphisms in MGMT (rs12917) and NBS1 (rs1805794) significantly modified the association between exposure to chlorinated solvents and the risk of NHL (Pfor interaction=0.0003 and 0.0048, respectively). After stratification by major NHL histological subtypes, MGMT (rs12917) modified the association between chlorinated solvents and the risk of diffuse large B-cell lymphoma (Pfor interaction=0.0027) and follicular lymphoma (Pfor interaction=0.0024). A significant interaction was also observed between occupational exposure to benzene and BRCA2 (rs144848) for NHL overall (Pfor interaction=0.0042). Our study results suggest that genetic variations in DNA repair genes modify the association between occupational exposure to solvents and the risk of NHL.

Phytanic acid and the risk of non-Hodgkin lymphoma

Greater consumption of red meat, processed meat and dairy products has been associated with an increased risk of non-Hodgkin lymphoma (NHL) in several previous reports. Phytanic acid, a saturated fatty acid obtained primarily through the consumption of ruminant meat and dairy products, may offer a potential underlying mechanism for these associations. In a population-based case-control study of 336 cases and 460 controls conducted in Nebraska during 1999-2002, we examined whether phytanic acid-containing foods or total phytanic acid intake, estimated from a food frequency questionnaire and the published phytanic acid values of 151 food items, were associated with increased NHL risk. Unconditional logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals for overall NHL and the common NHL histologic subtypes. In multivariable models, higher intakes of density-adjusted beef [OR(T3 vs. T1) = 1.5 (1.1-2.2); P(trend) = 0.02], total dairy products [OR = 1.5 (1.1-2.2); P(trend) = 0.02) and milk [OR = 1.6 (1.1-2.3); P(trend) = 0.01] were associated with an increased risk of NHL. Intake of total phytanic acid was positively associated with NHL risk [OR = 1.5 (1.0-2.1); P(trend) = 0.04]. In analyses stratified by NHL subtype, greater consumption of beef was associated with an increased risk of diffuse large B-cell lymphoma, and greater consumption of milk was associated with an increased risk of follicular lymphoma (FL). Total phytanic acid intake was associated with an increased risk of FL and small lymphocytic lymphoma/chronic lymphocytic leukemia. Our results provide support that total phytanic acid and phytanic acid-containing foods may increase NHL risk.

WS12-3 - Marginal Zone Lymphoma, Clinical Features and Treatment

ABSTRACT Marginal zone lymphoma (MZL) is the second most frequent histologic subtype of non-Hodgkin lymphoma (NHL) following diffuse large B-cell lymphoma in Korea. In 2008 WHO classification, MZL comprise the splenic B-cell MZL (SMZL), the extranodal MZL of mucosa-associated lymphoid tissue (MALT lymphoma, ENMZL), and the nodal MZL (NMZL). In Korea, ENMZL account for 16.7% and NMZL accounts for 0.6% of all NHLs. In Korean MZL analysis, the principal clinical features were limited stage, small tumor burden, excellent performance status, and normal LDH. Patients with NMZL, however, are distinguished by male predominance, higher incidence of B symptoms, bone marrow involvement, poor performance status, and advanced stage. Because ENMZL of stomach is the most common and extensively studied MZL, this presentation will show non-gastric MZL studies conducted in Korea. For stage I–II non-gastric MZL patients, local treatment/radiotherapy should be considered for the principal treatment modality. For stage III–IV MZL, we have conducted a multicenter, phase II trial with rituximab plus CVP. In 40 patients with a total of 287 cycles R-CVP chemotherapy, overall response rate was 88% with 24 complete responses (60%). The median duration of response was 28.3 months. After a median follow-up of 38.2 months, the estimated 3-year progression-free survival and overall survival were 59% and 95%, respectively. There were 11% and 2% of grade 3 or 4 neutropenia and febrile neutropenia, respectively. The R-CVP regimen is the first recommended regimen for untreated advanced stage MZL in Korea. Further studies should explore the still unanswered strategies. The optimal first-line treatment of localized or advanced MZL, the usefulness of maintenance therapy for MZL, the proper salvage treatment modalities for relapsed cases, and so on.

Meat intake and risk of non-Hodgkin lymphoma

We conducted a population-based, case-control study to test the hypothesis that consumption of meat and meat-related mutagens increases the risk of non-Hodgkin lymphoma (NHL), and whether the associations are modified by N-acetyltransferase (NAT) 1 and 2. Participants (336 cases and 460 controls) completed a 117-item food frequency questionnaire. The risk of NHL was associated with a higher intake of red meat (OR=1.5; CI, 1.1-2.2), total fat (OR=1.4; CI, 1.0-2.1), and oleic acid (OR=1.5; CI, 1.0-2.2). NHL risk was also associated with a higher intake of very well-done pork (OR=2.5; 95% CI, 1.4-4.3) and the meat-related mutagen MeIQx (OR=1.6; 95% CI, 1.1-2.3). Analyses of the major NHL histologic subtypes showed a positive association between diffuse large B cell lymphoma (DLBCL) and higher intake of red meat (OR=2.1; 95% CI, 1.1-3.9) and the association was largely due to meat-related mutagens as a positive association was observed for higher intakes of both MeIQx (OR=2.4; 95% CI, 1.2-4.6) and DiMeIQx (OR=1.9; 95% CI, 1.0-3.5). Although the OR for follicular lymphoma (FL) was also increased with a higher red meat intake (OR=1.9; 95% CI, 1.1-3.3), the association appeared to be due to increased oleic acid (OR=1.7; 95% CI: 0.9-3.1). We found no evidence that polymorphisms in NAT1 or NAT2 modify the association between NHL and meat-related mutagens. Our results provide further evidence that red meat consumption is associated with an increase in NHL risk, and new evidence that the specific components of meat, namely fat and meat-related mutagens, may be impacting NHL subtype risk differently.

Meat Intake Is Not Associated with Risk of Non-Hodgkin Lymphoma in a Large Prospective Cohort of U.S. Men and Women

Meat intake has been inconsistently associated with risk of non-Hodgkin lymphoma (NHL), a heterogeneous group of malignancies of the lymphoid tissue etiologically linked to immunomodulatory factors. In a large U.S. cohort, we prospectively investigated several biologically plausible mechanisms related to meat intake, including meat-cooking and meat-processing compounds, in relation to NHL risk by histologic subtype. At baseline (1995–1996), participants of the NIH-AARP Diet and Health Study completed a diet and lifestyle questionnaire (n= 492,186), and a subcohort (n= 302,162) also completed a questionnaire on meat-cooking methods and doneness levels. Over a mean of 9 y of follow-up, we identified 3611 incident cases of NHL. In multivariable Cox proportional hazards regression models, we found no association between intake of red meat, processed meat, fish, poultry, heme iron, nitrite, nitrate, animal fat, or protein and NHL risk. MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline) and DiMeIQx (2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline), heterocyclic amines formed in meats cooked to well done at high temperatures, were inversely associated with chronic lymphocytic leukemia/small lymphocytic lymphoma [n= 979; HR (95% CI) for the highest vs. lowest quintile of intake: 0.73 (0.55, 0.96) and 0.77 (0.61, 0.98), respectively]. In this large U.S. cohort, meat intake was not associated with NHL or any histologic subtypes of NHL. Contrary to findings in animal models and other cancer sites, meat-cooking and -processing compounds did not increase NHL risk.