Classical Hodgkin Lymphoma Cases Research Articles

Optimizing assessment of CD30 expression in Hodgkin lymphoma by controlling for low expression.

Since the approval of brentuximab vedotin (BV), assessment of CD30 status by immunohistochemistry gained increasing importance in the clinical management of patients diagnosed with CD30-expressing lymphomas, including classical Hodgkin lymphoma (CHL). Paradoxically, patients with low or no CD30 expression respond to BV. This discrepancy may be due to lack of standardization in CD30 staining methods. In this study, we examined 29 cases of CHL and 4 cases of nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) for CD30 expression using a staining protocol that was designed to detect low CD30 expression levels, and an evaluation system similar to the Allred scoring system used for breast cancer evaluation. For CHL, 10% of cases had low scores and 3% were CD30 negative, with 3 cases in which the majority of tumor cells showed very weak staining. Unexpectedly, one of four cases of NLPHL was positive. We demonstrate intra-patient heterogeneity in CD30 expression levels and staining patterns in tumor cells. Three CHL cases with weak staining may have been missed without the use of control tissue for low expression. Thus, standardization of CD30 immunohistochemical staining with use of known low-expressing controls may aid in proper CD30 assessment and subsequent therapeutic stratification of patients.

TMTV-Net: fully automated total metabolic tumor volume segmentation in lymphoma PET/CT images - a multi-center generalizability analysis.

Total metabolic tumor volume (TMTV) segmentation has significant value enabling quantitative imaging biomarkers for lymphoma management. In this work, we tackle the challenging task of automated tumor delineation in lymphoma from PET/CT scans using a cascaded approach. Our study included 1418 2-[18F]FDG PET/CT scans from four different centers. The dataset was dividedinto 900 scans for development/validation/testing phasesand 518 for multi-center external testing. The former consisted of 450 lymphoma, lung cancer, and melanoma scans, along with 450 negative scans, while the latter consisted of lymphoma patients from different centers with diffuse large B cell, primary mediastinal large B cell, and classic Hodgkin lymphoma cases. Our approach involves resampling PET/CT images into different voxel sizes in the first step, followed by training multi-resolution 3D U-Nets on each resampled dataset using a fivefold cross-validation scheme. The models trained on different data splits were ensemble. After applying soft voting to the predicted masks, in the second step, we input the probability-averaged predictions, along with the input imaging data, into another 3D U-Net. Models were trained with semi-supervised loss. We additionally considered the effectiveness of using test time augmentation (TTA) to improve the segmentation performance after training. In addition to quantitative analysis including Dice score (DSC) and TMTV comparisons, the qualitative evaluation was also conducted by nuclear medicine physicians. Our cascaded soft-voting guided approach resulted in performance with an average DSC of 0.68 ± 0.12 for the internal test data from developmental dataset, and an average DSC of 0.66 ± 0.18 on the multi-site external data (n = 518), significantly outperforming (p < 0.001) state-of-the-art (SOTA) approaches including nnU-Net and SWIN UNETR. While TTA yielded enhanced performance gains for some of the comparator methods, its impact on our cascaded approach was found to be negligible (DSC: 0.66 ± 0.16). Our approach reliably quantified TMTV, with a correlation of 0.89 with the ground truth (p < 0.001). Furthermore, in terms of visual assessment, concordance between quantitative evaluations and clinician feedback was observed in the majority of cases. The average relative error (ARE) and the absolute error (AE) in TMTV prediction on external multi-centric dataset were ARE = 0.43 ± 0.54 and AE = 157.32 ± 378.12 (mL) for all the external test data (n = 518), and ARE = 0.30 ± 0.22 and AE = 82.05 ± 99.78 (mL) when the 10% outliers (n = 53) were excluded. TMTV-Net demonstrates strong performance and generalizability in TMTV segmentation across multi-site external datasets, encompassing various lymphoma subtypes. A negligible reduction of 2% in overall performance during testing on external data highlights robust model generalizability across different centers and cancer types, likely attributable to its training with resampled inputs. Our model is publicly available, allowing easy multi-site evaluation and generalizability analysis on datasets from different institutions.

T-Cell-Rich Hodgkin Lymphoma With Features of Classic Hodgkin Lymphoma and Nodular Lymphocyte-Predominant Hodgkin Lymphoma: A Borderline Category With Overlapping Morphologic and Immunophenotypic Features.

It is known that a subset of cases of classic Hodgkin lymphoma (CHL) with B-cell-rich nodules (lymphocyte-rich CHL) exhibits morphologic and immunophenotypic features that overlap with nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL), raising diagnostic difficulties that can be resolved in most cases by performing an adequate battery of immunohistochemical studies. To fully characterize cases of T-cell-rich Hodgkin lymphoma where a specific diagnosis of NLPHL (ie, pattern D) or CHL could not be made even after complete immunophenotypic investigation. The clinical, immunomorphologic, and molecular (when applicable) presentation of 3 cases of T-cell-rich Hodgkin lymphoma was thoroughly investigated. These 3 cases harbored lymphocyte-predominant-like and Hodgkin and Reed-Sternberg-like cells that partially expressed B-cell and CHL markers and were negative for Epstein-Barr virus-encoded small RNA, in a T-cell-rich background with residual follicular dendritic cell meshworks; 1 case had frequent and the other 2 cases scant/absent eosinophils and plasma cells. Two patients with advanced-stage (III or IV) disease presented with axillary and supraclavicular lymphadenopathy, respectively, and without B symptoms. These patients underwent NLPHL-like therapeutic management with 6 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin hydrochloride [hydroxydaunorubicin], vincristine sulfate [Oncovin], and prednisone) chemotherapy; both are in complete remission 7 years posttherapy. One patient presented with stage I disease involving an internal mammary lymph node without B-symptoms and was treated with surgical excision alone; this patient is also in complete remission 1 year later. These cases illustrate overlapping features of T-cell-rich NLPHL and CHL with neoplastic cells expressing both B-cell program and CHL markers. This underrecognized overlap has not been fully illustrated in the literature, although it portrays a therapeutic challenge. These neoplasms may deserve in-depth investigation in the future that may bring up diagnostic or theragnostic implications.

Comparative Outcome Analysis of Nodular Lymphocyte-Predominant Hodgkin Lymphoma Vs Classical Hodgkin's Lymphoma - a SEER Based Study

Introduction Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) is a slower-growing variant of Hodgkin lymphoma that is uniquely characterized by its growth pattern and cellular appearance. While patients with NLPHL tend to demonstrate more promising prognoses than their counterparts with classical Hodgkin lymphoma (CHL), recent comparative studies remain limited. The objective of this study was to investigate the disparities in survival rates between NLPHL and CHL patients, accounting for variations in patient demographics and disease attributes. Methodology We utilized Surveillance, Epidemiology, and End Results (SEER) 17 registries database to identify patients diagnosed with NLPHL and CHL between 2000 - 2020. Univariate survival analysis was done using a Kaplan Meir plot with a log-rank test, and the Cox proportion hazard regression model was used for multivariate survival analysis. Results A total of 2,723 NLPHL cases and 42,581 CHL cases were included in the study. The median age of NLPHL patients stood at 44 years, whereas for CHL patients it was observed to be 38 years. Over an 83-month median follow-up, NLPHL demonstrated superior 5-year (92.2%) and 10-year (83.84%) survival rates compared to CHL (82.45% and 75.45%, respectively). When controlled for all available characteristics, NLPHL showed a significantly higher overall survival (HR: 2.373; 95% CI, 2.131 - 2.644; P&amp;lt;.001) than CHL. For those diagnosed with NLPHL, our data indicated that patients who underwent radiation therapy had better survival rate (HR: 1.98; 95% CI, 1.55 - 2.53; p&amp;lt;.001). Additionally, younger age and male gender were associated with a more favorable prognosis. Conclusion Our analysis highlights the superior survival outcomes for NLPHL patients as compared to those with CHL, even when accounting for available patient and disease characteristics. The further enhanced survival rates observed in NLPHL patients undergoing radiation therapy suggests potential for more tailored therapeutic approaches. Factors such as younger age and male gender were also identified as indicative of a more favorable prognosis, underscoring the need for further exploration of demographic and clinical influences on survival outcomes in Hodgkin lymphomas.

A Subset of Classical Hodgkin Lymphoma Cases Demonstrate CD19 Expression on Hodgkin Reed-Sternberg Cells By Immunohistochemistry

Background: The neoplastic Hodgkin Reed-Sternberg (HRS) cells in Classical Hodgkin Lymphoma (CHL) are thought to lose their B-cell phenotype and lack expression of most B-cell markers. Improvements in antibodies and detection methods have revealed that a subset of CHL cases may express some B-cell markers, including CD19 and CD20, with currently unclear significance. Introduction of CD19-directed therapies has changed the landscape of salvage treatment in CD19 expressing Non-Hodgkin Lymphomas. As such, we sought to characterize CD19 expression in HRS cells and its clinical implications and potential utility as a therapeutic target. Methods: We conducted an Institutional Review Board-approved retrospective review of records for CHL cases from 01/2015 to 07/2020 at a tertiary care center. 42 cases met the criteria and had sufficient tissue for analysis: 39 CHL, 2 CHL mixed with other B-cell lymphomas, and 1 Epstein Barr Virus Post-Transplant Lymphoproliferative Disease (EBV-PTLD)-CHL type. Cases were assessed for CD19 immunohistochemical (IHC) staining using the EP169 Rabbit Monoclonal Antibody. HRS cells were identified via morphology and characteristic immunoprofile, with all HRS cells expressing strong CD30 and dim PAX-5. CD19 staining intensity (0-3+) and pattern of expression (cytoplasmic, Golgi body, membranous) in HRS cells was independently evaluated by three hematopathologists. Based on their consensus, CD19 staining was categorized as negative, weak (variable across HRS cells), or positive. CD19 staining was compared with histologic subtype, other B-cell marker staining, EBV association (via EBER-ISH), and clinical data and outcomes, including, age, stage, treatment, response, overall survival, and demise. Results: Histologically, cases were composed of the following subtypes: 26 nodular sclerosis, 8 mixed cellularity, 3 lymphocyte-rich, 1 lymphocyte-depleted, 2 other (EBV-PTLD and malignant B-cell lymphoma) and 2 unclassifiable (due to treatment effect and biopsy size). Nine of 42 cases (21%) were EBV-associated by EBER-ISH. CD19 staining of any kind in HRS cells was found in 13/42 cases (31%), with 10/42 (24%) demonstrating positive staining and 3/42 (7%) demonstrating weak staining. Staining intensity and pattern varied across positive cases (Table 1 and Figure 1). CD19 staining was seen in 4/26 nodular sclerosis (15%), 6/8 mixed cellularity (75%), 1/3 lymphocyte-rich (33%), and 1/1 lymphocyte-depleted (100%) cases. The strongest correlation with other markers was seen with EBV, with 7/9 (78%) EBV+ cases staining with CD19. Five cases with CD19 staining also demonstrated predominantly patchy/dim CD20 staining. The clinical characteristics of CHL cases with CD19 expression varied, with an age range of 10-78 years old and variable clinical stage at time of tissue sampling. Following initial treatment, 4/13 patients failed to achieve remissions after frontline chemotherapy (mostly ABVD), with 3 of these patients now deceased due to disease. Conclusions: A significant subset of CHL cases demonstrate CD19 expression on HRS cells, and EBV-associated CHL has a high rate of CD19 expression, which may reflect its different underlying disease mechanism. Larger studies are necessary to characterize the staining pattern and intensity that may be most amenable to specific therapies and to better elucidate the prognostic implications of CD19 expression. Given the accessibility of CD19 IHC and the growing list of effective CD19-directed therapies, it is worthwhile assessing CD19 expression, particularly in EBV associated CHL, and further research into the utility of CD19 expression on HRS as a potential therapeutic target is warranted.

Specific Polo-Like Kinase 1 Expression in Nodular Lymphocyte-Predominant Hodgkin Lymphoma Suggests an Intact Immune Surveillance Program

Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) is a rare and relatively indolent B-cell lymphoma. Characteristically, the [lymphocyte-predominant (LP)] tumor cells are embedded in a microenvironment enriched in lymphocytes. More aggressive variants of mature B-cell and peripheral T-cell lymphomas exhibit nuclear expression of the polo-like kinase 1 (PLK1) protein, stabilizing MYC (alias c-myc) and associated with worse clinical outcomes. This study demonstrated expression of PLK1 in the LP cells in 100% of NLPHL cases (n=76). In contrast, <5% of classic Hodgkin lymphoma cases (n=70) showed PLK1 expression within the tumor cells. Loss-of-function approaches demonstrated that the expression of PLK1 promoted cell proliferation and increased MYC stability in NLPHL cell lines. Correlation with clinical parameters revealed that the increased expression of PLK1 was associated with advanced-stage disease in patients with NLPHL. A multiplex immunofluorescence panel coupled with artificial intelligence algorithms was used to correlate the composition of the tumor microenvironment with the proliferative stage of LP cells. The results showed that LP cells with PLK1 (high) expression were associated with increased numbers of cytotoxic and T-regulatory T cells. Overall, the findings demonstrate that PLK1 signaling increases NLPHL proliferation and constitutes a potential vulnerability that can be targeted with PLK1 inhibitors. An active immune surveillance program in NLPHL may be a critical mechanism limiting PLK1-dependent tumor growth.

The Association of Plasma Cell Density of the Microenvironment with Classical Hodgkin Lymphoma Stage

Background&#x0D; Plasma cells are important prognostic factors in various malignancies including Classical Hodgkin lymphoma (CHL). In the CHL microenvironment, plasma cells are the main prognostic factors, but research of relationship between plasma cell density in the microenvironment and CHL stage is still very limited. This study aims to determine the relationship between plasma cell density in the microenvironment and the CHL stage.&#x0D; &#x0D; Methods&#x0D; A case series procedure was conducted. The samples were paraffin blocks of CHL patients at Anatomic Pathology Department of Faculty of Medicine Universitas Sriwijaya/Mohammad Hoesin General Hospital. Among 47 cases of CHL diagnosed from 1st Januari 2018 to 30th November 2022, only 30 cases were included. Immunohistochemistry was performed by using specific antibody of plasma cells (anti-CD138 antibody). The number of plasma cells in each sample was calculated based on the expression of CD138 in the cell membrane and cytoplasm with weak-strong intensity. Then the cut-off point value was determined based on the ROC curve to classify the high and low plasma cell densities. The density of plasma cells was correlated to clinicopathological characteristics including age, sex, tumor location, CHL subtypes and stage. Statistical analysis was performed using Spearman test of SPSS with significant consideration if p&lt;0.05.&#x0D; &#x0D; Results&#x0D; A high proportion (&gt;10,9) of tumour-associated plasma cells of the tumor microenvironment was found in CHL advanced stage. Spearman test resulted in a significant correlation between density of plasma cell with advanced stage classical Hodgkin lymphoma (p=0.000).&#x0D; &#x0D; Conclusions&#x0D; There is significant relationship between high density of plasma cells with CHL stage.

ROLE OF GATA3 AND STAT6 IMMUNOHISTOCHEMISTRY IN CLASSIC HODGKIN LYMPHOMA (CHL) AND NODULAR LYMPHOCYTE PREDOMINANT HODGKIN LYMPHOMA (NLPHL)

Objective: The purpose of our study is to investigate the utility of immunohistochemical expression of GATA3 and STAT6 in differentiating Classic Hodgkin Lymphoma (CHL) from Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL). Material and Methods: We approached database of Shaukat Khanum Memorial Cancer Hospital and Research Centre and selected a total of 79 CHLs and 15 NLPHLs diagnosed either on Trucut biopsy or excision biopsy diagnosed during the time 2020-2021. One slide from each of these 94 cases was stained with GATA3 and STAT6 independently and pattern of staining (either nuclear or cytoplasmic or dual nuclear and cytoplasmic) was accessed. Results: 75/79(95%) cases of CHL were positive for GATA3 staining [60/79(75.9%) cases showed nuclear GATA3 staining, 15/79(18.9%) cases showed both nuclear and cytoplasmic staining] and 4/79(5%) CHL cases were negative for GATA3. 15/15(100%) cases of NLPHL were negative for GATA3[12/15(80%) cases showed no staining and 3/15(20%) cases showed only cytoplasmic blush]. 65/79(82%) cases of CHL were positive for STAT6[17/79(21.5%) cases showed nuclear STAT6 expression, 48/79(60.7%) cases showed both nuclear and cytoplasmic staining], and 14/79(18%) cases of CHL were negative for STAT6 expression [8/79(10%) cases showed only cytoplasmic staining and 6/79(8%) showed neither nuclear nor cytoplasmic staining]. 14/15(93.3%) cases of NLPHL were negative for STAT6[5/79(33.3%) cases showed only cytoplasmic staining and 9/15(60%) cases showed no staining] and 1/15(6.7%) case of NLPHL was positive for STAT6[showed nuclear STAT6 staining]. Conclusion: Nuclear staining of GATA3 and STAT6 either alone or in combination with cytoplasmic staining can be used to differentiate CHL from NLPHL. GATA3 effectively excludes NLPHL with 100% negative predictive value. Keywords: GATA3, STAT6, Classic Hodgkin lymphoma, Nodular lymphocyte predominant Hodgkin lymphoma

Diagnostic Utility of CD200 Immunohistochemistry in Distinguishing EBV-Positive Large B-Cell Lymphoma From Classic Hodgkin Lymphoma.

Epstein-Barr virus-positive large B-cell lymphoma (EBV+ LBCL) is a heterogeneous group of diseases that may resemble classic Hodgkin lymphoma (CHL) both morphologically and immunophenotypically. However, these diseases are treated with different therapies and carry distinct prognoses. We examined CD200 expression by immunohistochemistry in EBV+ LBCL and evaluated its diagnostic utility in the differential diagnosis with CHL. CD200 immunohistochemistry was performed on archival material from 20 cases of CHL (11 EBV+, 9 EBV-), 11 cases of EBV+ LBCL, and 10 cases of diffuse large B-cell lymphoma, not otherwise specified (DLBCL NOS). Staining pattern and intensity (0-3+ scale) were recorded. CD200 positivity was seen in Reed-Sternberg cells in 19 (95%) of 20 cases of CHL, predominantly in a strong (3+, 15/19) and diffuse (>50% of cells, 17/19) pattern. In contrast, CD200 was negative in 8 (73%) of 11 cases of EBV+ LBCL; the 3 positive cases showed 1 to 2+ staining in less than 50% of lesional cells. All cases of DLBCL NOS were negative for CD200. CD200 may be a useful immunophenotypic marker in differentiating EBV+ LBCL from CHL, with negative to partial/weak staining favoring a diagnosis of EBV+ LBCL and strong diffuse staining favoring a diagnosis of CHL.

CLINICO-PATHOLOGICAL EXAMINATION OF WALDEYER RING LYMPHOMAS

Introduction: Waldeyer's ring (WR) is the second most common site of extranodal lymphomas. Its clinical course may resemble those of nodal lymphomas. In this study, we evaluated the histopathological and clinical findings, the presence of Epstein-Barr virus (EBV), and overall survival of WR lymphomas diagnosed in our department. Materials and Methods: Retrospectively, 32 patients diagnosed with WR lymphoma between 2017-2022 were included. The demographic, clinical data of the patients and lymphoma types according to the latest classification of the World Health Organization, the presence of EBV (EBV early RNAs (EBER) by chromogenic in-situ hybridization (ISH) method) were examined. Descriptive statistics, categorical data were evaluated as percentage and frequency. Kaplan-Meier analysis was used for survival rate. A P value less than 0.05 was considered statistically significant. Results: The majority of 32 cases in our study were male. The mean age was 57.97±16.15 years. The clinical prediagnosis was carcinoma in 46.9% of the cases. Tonsil and nasopharynx had equal proportions as lymphoma origin. Only 3 (9.4%) cases are classic hodgkin lymphoma (CHL). Others were NHL, of which 16 (50.0%) were diffuse large B-cell lymphoma (DLBCL). EBER was positive only in tonsillar CHL cases. The survival of patients with lymphoma of nasopharyngeal origin was shorter than those of tonsillar origin (p=0.037). Conclusion: WR lymphomas are most often confused with carcinomas clinically. DLBCL is the most common type of lymphoma. EBER has been detected only in CHL. Survival is lower in lymphomas originating from the nasopharynx.&#x0D; Keywords: Non-Hodgkin Lymphoma, Nasopharynx, Tonsil, Hodgkin Lymphoma

PD-L1 immunohistochemistry assay optimization to provide more comprehensive pathological information in classic Hodgkin lymphoma

Overexpression of PD-L1 can be a predictive marker for anti-PD-1 therapeutic efficacy in classic Hodgkin lymphoma (CHL); however, harmonization of different IHC assays remains to be accomplished, and interpretations of PD-L1 immunostaining results remain controversial in CHL. In this study, we sought to optimize the PD-L1 immunohistochemistry (IHC) assay in CHL. All tests were performed on a tumour tissue microarray established from 54 CHL cases. Three IHC antibodies (405.9A11, SP142, 22C3) for detecting PD-L1 expression were compared semi quantitatively with the RNAscope assay (No. 310035, ACD), and the difference in the expression in background immune cells (ICs) between assays and the associations of expression levels with densities of TILs/TAMs were also analysed. 405.9A11 demonstrated best specificity in HRS cells and best sensitivity in ICs. Positive expression of PD-L1 was more frequent in ICs (85.2%) than in HRS cells (48.1%). Different subgroups of background ICs, including tumour-associated macrophages (TAMs), were assessed and scored for CD4, CD8, FOXP3, and CD163 expression. PD-L1 expression on ICs was the factor most associated with the density of TAMs. 405.9A11 provided the most convincing PD-L1 expression results. Pathologists should report PD-L1 expression in a combined manner, including both the status of HRS cells and the percentage of PD-L1-positive ICs.

Hodgkin Lenfomada B Hücre Transkripsiyon Faktörlerinin İmmünohistokimyasal İfadesi ve Ayırıcı Tanıda Kullanımı

Aim: Classical Hodgkin lymphoma is common, it is one of the lymphomas whose differential diagnosis can be difficult. It is thought that Hodgkin cell may originate from the germinal center. Our aim in this study was to determine the germinal center transformation markers OCT-2, BOB.1, BCL-6, PAX-5, CD20 and MUM-1 in Classic Hodgkin Lymphoma (CHL), Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL) and Diffuse Large B-cell Lymphoma (DLBCL) to evaluate the expressions of by immunohistochemical method and chromogenic in-situ hybridization (ISH) of EBV early RNAs (EBER).&#x0D; Material and methods: 49 biopsies diagnosed with Hodgkin lymphoma (HL), 5 with NLPHL and 19 with DLBCL were evaluated for CD30, PAX-5, OCT-2, BOB.1, MUM-1, BCL-6, and CD20, and EBER positivity. SPSS 18 was used for statistical analysis.&#x0D; Results: 73 lymphoma cases were included in the study, 61.6% males and 38.4% females. The median age of patients was 50 years. CHL (67.1%) was the most common lymphoma type, and mixed cellular Hodgkin lymphoma (MSHL) was the most common subtype. There was a statistically significant difference in CD30, OCT-2, BOB.1, MUM-1, PAX-5, CD20, BCL-6, EBER expression between CHL and DLBCL cases (p

Aberrant synaptophysin expression in classic Hodgkin lymphoma

BackgroundSynaptophysin is an immunohistochemical marker for neuroendocrine differentiation and is widely used in pathologic diagnosis. Its expression in malignant lymphoma has not yet been described. However, we experienced an index case of classic Hodgkin lymphoma with synaptophysin expression. This experience prompted us to investigate synaptophysin expression in classic Hodgkin lymphoma.MethodImmunohistochemical staining of synaptophysin was performed in 59 diagnosed cases of classic Hodgkin lymphoma, 10 anaplastic large cell lymphomas, 16 diffuse large B-cell lymphomas, and 5 extranodal marginal zone lymphoma of the mucosa-associated tissue. Synaptophysin-positive cases were stained for both chromogranin and CD56a.ResultOf 59 classic Hodgkin lymphoma cases, 11 (19%) were positive for synaptophysin. None of the anaplastic large cell lymphomas expressed synaptophysin. Synaptophysin showed weak but specific expression in the cytoplasm of the Hodgkin lymphoma tumor cells. Other background inflammatory cells (such as macrophages, B-, and T-lymphocytes) were all negative for synaptophysin expression. Chromogranin and CD56a were not expressed in the synaptophysin-positive classic Hodgkin lymphomas.ConclusionsSynaptophysin is an integral glycoprotein present in presynaptic vesicles of neurons and neuroendocrine cells. It is a diagnostic marker for neuroendocrine tumors. Aberrant synaptophysin expression has been reported in non-neuroendocrine tumors but not in lymphoma or leukemia. To the best of our knowledge, synaptophysin positivity has only been reported in a single case of precursor T-lymphoblastic leukemia/lymphoma to date. Our study showed that aberrant synaptophysin expression in classic Hodgkin lymphoma is an unexpectedly frequent finding. The mechanism underlying, and prognostic significance of, such aberrant expression is unclear. Thus, in a small biopsy, aberrant synaptophysin expression could be a diagnostic pitfall and should be carefully avoided.

A Unusual Case of Classic Hodgkin Lymphoma (cHL) with Aberrant T-Antigens Expression

Abstract Introduction/Objective T-cell-associated antigens (TCA) are rarely expressed by Hodgkin/Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (cHL) and is associated with uncertain clinical significance. Methods/Case Report We present a very recent case of a 68 year-old female with no significant past medical history presented with a large right groin mass for 1 month. She denied any fever, night sweats, fatigue or weight loss. CT abdomen/pelvis showed inguinal/pelvic lymphadenopathy. Resection showed effaced nodal architecture with a vague nodular appearance and with large, atypical nucleolated cells in a background of small, mature lymphocytes and histiocytes. Eosinophils are conspicuously absent in the background. The large cells were polylobated lymphocyte and histiocyte rich (L &amp; H) like, classic HRS cells as well as multinucleated in appearance. The morphologic features overlapped between Classic Hodgkin Lymphoma, and Nodular Lymphocyte Predominant Hodgkin Lymphoma. Given the immunohistochemical (IHC) features of the neoplastic cells [CD45 negative, PAX5+ (weak), CD30+ (strong), CD15+ (majority) and MUM1 +] the diagnosis was most compatible with CHL. Interestingly, a subset of the Hodgkin cells (10-20%) aberrantly expressed a number of T-cell markers (CD2, CD3, CD4, and CD8). However, the background smaller T-cells did not show cytologic atypia or aberrancies of T-cell antigen markers. The large cells were negative for CD43 and ALK1. TCR gamma gene rearrangement was also negative for clonal T-lymphocytes, thus excluding the possibility of peripheral T-cell lymphoma with Hodgkin-like cells. Based on the morphology and IHC profile, this case proves to be CHL with multiple TCA expression including CD2, CD3, CD4 and CD8. She was planned for chemotherapy. Four months after the initial diagnosis and after completion of 2 cycles of chemotherapy, her PET CT scan showed improvement with right inguinal adenopathy being smaller in size and decreased avidity, indicating a response to treatment. She is due for 2 more cycles. Till todate she is doing well with no further symptoms. Results (if a Case Study enter NA) NA. Conclusion So far reported cases of classic Hodgkin lymphoma (CHL) with aberrant T-cell antigen (TCA) expression were associated with decreased event-free survival and overall survival when compared with TCA-negative CHLs. Our finding contributes to the literature and improves our knowledge of the disease.

HL-048 Clinicopathological, Immunohistochemical Characteristics, and Outcomes of Hodgkin Lymphoma Patients in Erbil City, Iraq

Hodgkin lymphoma (HL) has unique epidemiological features with diverse pathologies and exhibits considerable clinicopathological variations in different parts of the world. In this study, we aimed to assess the clinicopathological features, immunohistochemistry, and outcomes of HL patients treated in Erbil, northern Iraq. This was a retrospective study conducted in Nanakaly Hospital for Blood Diseases and Oncology in Erbil, Iraq. A total of 125 patients diagnosed between January 2012 and December 2016 were assessed for their clinical characteristics, histopathology, immunophenotype, and outcome. The median age was 28 years (range: 18-71 years); 55% were male and 41% had Stage II HL. The most common histological type was nodular sclerosis (51.2%) followed by mixed cellularity (43.2%). CD30 was positive in nearly all cases of classical HL. CD15 and CD20 were positive in 98.7% and 75% of patients with the nodular lymphocyte predominant subtype, respectively. Most of the patients received adriamycin, bleomycin, vinblastine, and dacarbazine chemotherapy, and the 5-year overall survival in our study is 70%. Advanced stage (IV), high lactate dehydrogenase levels, low hemoglobin, and splenomegaly are significant predictors of poor survival. Our patients exhibited outcomes that were lower than those reported in developed countries.

The Association of Programmed Death Ligand 1 (PD-L1) and Cluster of Differentiation 95 Ligand (CD95L) Immunoexpression with Chemotherapy Response in Classical Hodgkin Lymphoma

Background: Programmed Death Ligand (PD-L1) and Cluster of Differentiation 95 (CD95L) are influenced by oncogenes and function in the anti-apoptosis process which is thought to play a role in chemotherapy resistance. This study aimed to analyze the association of PD-L1 and CD95L immunoexpression with chemotherapy response in Classical Hodgkin Lymphoma (CHL).Method: This study involved 40 cases of histopathologically diagnosed CHL treated with doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) chemotherapy. PD-L1 and CD95L immunohistochemical staining were performed in selected paraffin-embedded tissue blocks of all cases. The chemotherapy response status of the patients was taken from the medical recordResults: High PDL-1 immunoexpression was evident in 19 (47.5%) cases while positive CD95L immunoexpression was found in 14 (35%) CHL cases. High PD-L1 immunoexpression was significantly associated with the Non-Responsive (NR) group (78.9%) with p-value = 0.0001. Positive CD95L immunoexpression was greater in the NR group (71.4%) with p-value = 0.37.Conclusion: : High PD-L1 immunoexpression indicated an unfavorable response to ABVD chemotherapy in CHL. CD95L immunoexpression was not associated with ABVD chemotherapy response in CHL.

IRF8 as a Novel Marker to Differentiate Between CD30-Positive Large Cell Lymphomas.

Interferon regulatory factor 8 (IRF8) is a new biomarker shown to be positive in monocytic leukemias as well as in B cells. As a transcription factor, it plays a critical role in pre-B-cell differentiation and induction of tolerance pathways, among other functions. Given the frequent diagnostic dilemma in CD30-positive large cell lymphomas that could resemble both Hodgkin lymphoma and anaplastic large cell lymphoma (ALCL), we sought to determine whether IRF8 can be useful in distinguishing between these neoplasms that require different treatment strategies. In this retrospective study, 74 cases of classic Hodgkin lymphoma (CHL) and 7 cases of nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) on a tissue microarray (TMA), as well as 15 individual cases of ALK-negative ALCL and 4 cases of ALK-positive ALCL, were stained for IRF8. Paired box 5 (PAX5) immunostaining of the TMA was also performed and compared alongside since that marker is occasionally the only marker to help clinically differentiate between T- and B-cell lymphomas with anaplastic/Hodgkin-like cytology. None (0%) of the ALCLs were positive for IRF8 while all (100%) of the NLPHLs and 85% of the CHLs were positive for IRF8. Six (8%) cases of CHL were PAX5 negative but IRF8 positive. Conversely, seven (10%) cases of CHL were PAX5 positive but IRF8 negative. Four (6%) cases of CHL were negative for both PAX5 and IRF8. There is significant morphologic and immunophenotypic (CD30 positive and CD45 and CD20 negative) overlap between CHL and ALCL. Since many ALCLs show downregulation of lineage-specific T-cell markers or are "null cell" type, only PAX5 has been a reliable marker to differentiate between borderline cases. This is further confounded by positivity of PAX5 in some ALCLs due to amplification of PAX5. On the basis of recent discoveries of IRF8 function as well as performance as an immunostain, we tested this marker in human lymphoma samples and found that it aids in the discrimination between these tumors.

The characterization and frequency of diagnosis of EBV-positive variants with a lymphoid predominance in the structure of Hodgkin lymphoma

Introduction. Epstein — Barr virus (EBV) plays an important role in the pathogenesis of lymphoid tumors, in particular Hodgkin lymphoma. The frequency of expression of the EBV varies in different histological variants of classical Hodgkin lymphoma and is rarely observed in nodular lymphocyte predominant Hodgkin lymphoma.Aim — to study the pathomorphological features of the histological variants of Hodgkin lymphoma with lymphoid predominance associated with the EBV, as well as the frequency of their diagnosis in the structure of Hodgkin lymphoma.Materials and methods. The retrospective study included 794 patients with a verified diagnosis of Hodgkin lymphoma using histological and immunohistochemical methods on biopsy material for the period 2018–2019 (age range — 18–91 years old; median — 34 years old; men : women — 1.1 : 1). The presence of EBV in biopsies was assessed by immunohistochemical reaction with antibodies to EBV (clone LMP1), or by chromogenic in situ hybridization with probes for EBV-encoded small RNAs.Results. Classical Hodgkin lymphoma was diagnosed in 91 % (725/794) cases, nodular lymphocyte predominant Hodgkin lymphoma — in 9 % (69/794) cases. EBV-positive Hodgkin lymphoma accounted for 11 % (82/725) of all cases of classical Hodgkin lymphoma, (age range — 18–81 years old, median — 45 years old; men : women — 2.5 : 1). All cases of nodular lymphocyte predominant Hodgkin lymphoma were EBV-negative. Lymphocyte-rich classical Hodgkin lymphoma was found in 14 patients (14/725, 2 %), 4 patients showed intermediate morphoimmunohistochemical features with nodular lymphocyte predominant Hodgkin lymphoma, which were statistically significantly different from classical Hodgkin lymphoma by the presence of B-zones in the form of large nodules (p = 0.0157) and expression CD20 by tumor cells (p = 0.0404).Conclusion. Nodular lymphocyte predominant Hodgkin lymphoma is not characterized by a connection with EBV infection, unlike classical variant — lymphocyte-rich classical Hodgkin lymphoma. The obtained data support the concept of the existence of a transient form of Hodgkin lymphoma, which has the features of nodular lymphocyte predominant Hodgkin lymphoma and classical Hodgkin lymphoma, in the pathogenesis of which the Epstein — Barr virus likely plays a role.

Accuracy of Pathologic Diagnosis in Patients With Lymphoma and Survival: A Prospective Analysis From Botswana.

PURPOSEWith intense HIV epidemics, southern African countries have a high burden of classic Hodgkin lymphoma (CHL) and non-Hodgkin lymphoma (NHL). However, suboptimal access to pathology resources limits subtype classification. We sought to assess the diagnostic accuracy of specimens classified as lymphoma and to determine association between discordant pathologic diagnosis and overall survival.METHODSSeventy patients with CHL or NHL and treated at three Botswana hospitals from 2010 to 2016 were analyzed. Local pathologic assessment relied primarily on morphology. All cases underwent secondary US hematopathology review, which is considered gold standard.RESULTSThe median follow-up was 58 months. The overall reclassification rate was 20 of 70 cases (29%). All 20 CHL cases were correctly classified in Botswana, and mixed cellularity was the most common subtype, diagnosed in 11 (55%) cases. Of 47 confirmed NHL cases, diffuse large B-cell lymphoma was the final US diagnosis in 28 cases (60%), another aggressive B-cell NHL in nine (19%), an indolent B-cell NHL in six (13%), and T-cell NHL in four (9%). Common types of diagnostic discordance included NHL subtype reclassification (11 of 20, 55%) and CHL reclassified as NHL (7 of 20, 35%). Concordant versus discordant diagnosis after secondary review was associated with improved 5-year overall survival (60.1% v 26.3%, P = .0066). Discordant diagnosis was independently associated with increased risk of death (adjusted hazard ratio 2.733; 95% CI, 1.102 to 6.775; P = .0300) even after stratifying results by CHL versus NHL.CONCLUSIONIn this single prospective cohort, discordant pathologic diagnosis was associated with a nearly three-fold increased risk of death. Limited access to relatively basic diagnostic techniques impairs treatment decisions and leads to poor patient outcomes in low-resource countries.

Downregulation of the Tumor Suppressor SAMHD1 in Classical Hodgkin Lymphoma

Introduction: The SAM domain and HD domain 1 (SAMHD1) protein is a deoxynucleoside triphosphate (dNTP) triphosphohydrolase, which has been initially described to restrict human immunodeficiency virus type 1 (HIV-1) in certain cell types through depletion of intracellular dNTP substrates for HIV-1 reverse transcription. Mutations of SAMHD1 gene have been linked to Aicardi-Goutières syndrome (AGS) and have been identified as putative drivers of chronic lymphocytic leukemia resulting in decreased SAMHD1 mRNA and protein levels. Therefore, a line of evidence suggests that SAMHD1 may play a role in oncogenesis as a tumour suppressor. In addition, SAMHD1 may confer resistance to nucleoside-based chemotherapies by hydrolysing their active triphosphate metabolites, and thus, it may lead to resistance to certain chemotherapeutic agents with cytarabine in acute myeloid leukemia being a recent example (Herold et al, Nat Med 2017; 23(2):256-263). The expression patterns and the potential role of SAMHD1 in the pathogenesis of classical Hodgkin lymphoma (HL) are not yet known.Methods: SAMHD1 protein levels were assessed by Western blot analysis in 20 human lymphoma and leukemia cell lines including 5 classical HL (L1236, L428, HDML2, HDMyZ, MDA-V) and 3 diffuse large B-cell lymphoma (DLBCL) (RC-K8, OCI-Ly3, U2932) cell lines. As our findings from in silico analysis revealed potential binding sites for MYC on the SAMHD1 gene promoter, HL cell lines were treated with selective MYC inhibitors to assess possible MYC-associated regulation of SAMHD1 expression. The patient cohort included 212 cases of classical HL diagnosed and treated at the University of Texas MD Anderson Cancer Center (USA), University of Athens (Greece) and University of Crete (Greece). A control group of 4 reactive lymph nodes and 76 B-cell non-Hodgkin lymphomas (58 DLBCLs) was included in the study for comparison. All tissue samples were obtained prior to therapy. SAMHD1 expression was assessed by immunohistochemistry performed on TMAs with duplicate tumor cores from each case or full tissue sections using a monoclonal antibody (Bethyl Laboratories, San Antonio, TX). An arbitrary 10% cutoff was used to define SAMHD1 positivity in the neoplastic cells. Overall survival (OS) was defined as time from diagnosis to death or last follow-up. Event-free survival (EFS) was defined as time from diagnosis to relapse, death, or last follow-up. Survival analyses were performed using the Kaplan-Meier method (log-rank test) and Cox regression models.Results: In 4 out of 5 HL cell lines, SAMHD1 protein levels were dramatically reduced as compared to all 3 DLBCL and other lymphoma and leukemia cell lines tested. Inhibition of MYC activity resulted in a substantial increase in the SAMHD1 protein level suggesting a possible MYC-associated gene regulation. Similarly, the neoplastic Hodgkin-Reed Sternberg (HRS) cells of classical HL were SAMHD1-negative in 132 of 212 (62.3%) patients. Among the SAMHD1-positive cases, the percentage of positive HRS cells ranged from 10 to 95% with a median 40%. By contrast, SAMHD1 was negative in only 15% of the control group of DLBCLs. In addition, most of the germinal center (GC) cells of the reactive lymph nodes were SAMHD1-positive with consistently higher intensity of immunostaining as compared to HRS cells of classical HL. No significant associations between SAMHD1 levels and clinicopathologic parameters were observed in the HL cohort. In a subset of 56 HL patients with long follow up period (median 195 months), FFS at 10 years was 79% vs. 71% for patients with SAMHD1-negative vs. SAMHD1-positive HL, respectively. Similarly, OS at 15 years was 71% vs. 63% for SAMHD1-negative vs. SAMHD1-positive HL, respectively. These differences did not reach statistical significance (p>0.05, log-rank) likely because of the relatively small number of patients included in this preliminary survival analysis.Conclusions: SAMHD1 is shown for the first time to be downregulated in HRS cells of classical HL further supporting its role as a tumor suppressor and its regulation may involve MYC. Preliminary survival analysis shows a statistical trend towards inferior FFS and OS for patients with SAMHD1-positive HRS cells, however, analysis of the entire cohort of HL patients is underway to draw definite conclusions. DisclosuresKhoury:Pfizer: Research Funding; Angle: Research Funding; Kiromics: Research Funding; Stemline Therapeutics: Research Funding.