Supplementary Figure S1 from Poor Concordance among Nine Immunohistochemistry Classifiers of Cell-of-Origin for Diffuse Large B-Cell Lymphoma: Implications for Therapeutic Strategies
Rita CoutinhoAndrew ClearAndrew OwenA. S. WilsonJanet MatthewsAbigail LeeRute AlvarezMaría Gomes da SilvaJosé CabeçadasMaria CalaminiciJohn G. Gribben
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<p>PDF file 265K, Supplementary Figure 1: Immunohistochemistry results for each antibody (A) BCL2; (B) CD10; (C) MUM1; (D) LMO2; (E) GCET1; (F) BCL6; (G) FOXP1. Original magnification: x20</p>Keywords:
Concordance
Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) account for nearly all pediatric nonlymphoblastic B-cell lymphomas. Because clinical behavior, prognosis, and response to therapy might differ, diagnostic accuracy is important. Morphologic examination often is sufficient, but occasionally, diagnostic ancillary studies are required. In adults, immunophenotyping is useful; however, pediatric data are limited. We characterized the immunohistochemical expression of 6 proteins (c-myc, CD10, bcl-6, bcl-2, CD138, and MIB-1) in pediatric BL (33 cases) and DLBCL (20 cases) with classic morphologic features. Significant differences in c-myc (BL, 30/33 [91%] vs DLBCL, 5/20 [25%]; P < .0001), bcl-2 (BL, 1/25 [4%] vs DLBCL, 7/19 [37%]; P < .02), and mean MIB-1 (BL, 99% vs DLBCL, 56%; P < .0001) expression were observed. There were no significant differences for CD10 (100% expression in BL and DLBCL), bcl-6 (BL, 23/33 [70%] vs DLBCL, 15/20 [75%]), or CD138 (no expression). Thus, pediatric BL and DLBCL have distinctive immunohistochemical profiles, and staining for c-myc, MIB-1, and bcl-2 might be useful in morphologically difficult cases.
Immunophenotyping
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T-Cell Rich-B cell Lymphoma (TCRBCL) is separately defined by World Health Organisation(WHO), it is a subtype of Diffuse Large B cell Lymphoma (DLBCL). It is an aggressive disorder treated same as that of DLBCL. But some times it may not respond to therapy and it has poor prognosis. It requires careful histopathological examination and immunohistochemistry (IHC) for confirmation. We reported a rare case of subcutaneous T cell rich B cell lymphoma, which is confirmed by IHC which shows CD20 and CD3 marker positive.
B-cell lymphoma
Large cell
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Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) account for nearly all pediatric nonlymphoblastic B-cell lymphomas. Because clinical behavior, prognosis, and response to therapy might differ, diagnostic accuracy is important. Morphologic examination often is sufficient, but occasionally, diagnostic ancillary studies are required. In adults, immunophenotyping is useful; however, pediatric data are limited. We characterized the immunohistochemical expression of 6 proteins (c-myc, CD10, bcl-6, bcl-2, CD138, and MIB-1) in pediatric BL (33 cases) and DLBCL (20 cases) with classic morphologic features. Significant differences in c-myc (BL, 30/33 [91%] vs DLBCL, 5/20 [25%]; P < .0001), bcl-2 (BL, 1/25 [4%] vs DLBCL, 7/19 [37%]; P < .02), and mean MIB-1 (BL, 99% vs DLBCL, 56%; P < .0001) expression were observed. There were no significant differences for CD10 (100% expression in BL and DLBCL), bcl-6 (BL, 23/33 [70%] vs DLBCL, 15/20 [75%]), or CD138 (no expression). Thus, pediatric BL and DLBCL have distinctive immunohistochemical profiles, and staining for c-myc, MIB-1, and bcl-2 might be useful in morphologically difficult cases.
Immunophenotyping
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Introduction: Diffuse large B-cell lymphoma (DLBCL) is the most common type of nonHodgkin lymphoma. The pathogenesis of DLBCL is complex because it involves at least two different pathways, a de novo pathway and a transformation pathway. MYC and BCL2 oncogenes are 2 key regulators implicated in the pathogenesis. DLBCL with concurrent expression of MYC and BCL2 has been shown to be clinically aggressive and confers a worse prognosis. MYC detection by immunohistochemistry is however not performed in a routine diagnostic work up of DLBCL cases. This study examined the presence of MYC and BCL2 proteins by immunohistochemistry in patients diagnosed to have DLBCL. Methods: This retrospective study involved patients diagnosed to have DLBCL at Tengku Ampuan Afzan Hospital, Kuantan, Pahang (Year 2009-2011) and Queen Elizabeth Hospital, Kota Kinabalu, Sabah Malaysia (Year 2012-2014). Immunohistochemistry for MCY and BCL2 were performed on sections of formalin fixed paraffin embedded tissue blocks. Results: There were 91 cases analyzed. Forty-nine cases (53.8%) exhibited concurrent expression of MYC and BCL2 proteins. In about one third of the cases, positivity was confined to BCL2. In 4 cases (4.4%) only MYC was expressed while in 9 cases (9.9%) both markers were negative. Overall about 60% and 85% of the cases were positive for MYC and BCL2 respectively. Conclusions: Approximately half of DLBCL case studied co-express MYC and BCL2. Prospective studies to look at the clinical significance and prognostic impact of this finding are advocated.
Pathogenesis
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Objective To investigate the relationship between BCL-6 and BCL-2 as well as the clinical features and prognosis of diffuse large B cell lymphoma(DLBCL). Methods Immunohistochemistry stain was used to determine the expression of BCL-6 and BCL-2. Results Positive rates of BCL-6 and BCL-2 were 60.9%(28/46) and 54.3%(25/46).BCL-6 positive and BCL-2 negative expression showed a trend to better overall survival(OS) and progression-free survival(PFS). Conclusion The results suggest significant correlationship among BCL-6,BCL-2 and prognosis.Moreover,BCL-6 and BCL-2 may be the useful prognostic indicators in diffuse large B cell lymphoma.
Stain
Clinical Significance
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Objective To study the expressions and significance of bcl-6 and Ki-67 in diffuse large B-cell lymphoma(DLBCL).Methods Immunohistochemistry technique was used to detect the expressions of bcl-6 and Ki-67 in the tissues of 90 DLBCL patients. 20 cases of reactive hyperplasia of lymph node (RH) were used as control. Results Positive expression rate of bcl-6 are 54.44 ％(49/90), 15.00 ％(3/20) in DLBCL or RH tissues respectively (x2=10.214,P=0.001). There were correlations between bcl-6 expression and Ann Arbor clinical stage, LDH, B symptoms, or Hans classification (x2=5.257,5.257,4.704,16.024 respectively,all P＜0.05).The high expression rate of Ki-67 were 80.00 ％(72/90),20.00 ％(4/20)in DLBCL or RH tissues respectively (x2=27.585,P=0.000). There were correlations between Ki-67 expression and Ann Arbor clinical stage,IPI or recent effect (x2=5.889,6.451,6.024 respectively,all P＜0.05).Conclusion There are significant correlations between the aberrant expression of bcl-6 or Ki-67 and Ann Arbor clinical stage,IPI or Hans classification. The expressions of bcl-6 and Ki-67 may provide important information for the clinical therapy and prognosis of DLBCL.
Key words:
Lymphoma, large B-cell, diffuse; bcl-6; Ki-67; Immunohistochemistry
Ki-67
Clinical Significance
B symptoms
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Diffuse large B-cell lymphomas (DLBCL) with aberrations of MYC probably represent a distinct clinicopathological entity following an aggressive course. Their incidence in unselected DLBCL collectives is debatable and the identification of such cases may be difficult. Therefore, the molecular epidemiology of MYC aberrations in DLBCL and whether they can be predicted by morphology and immunohistochemistry were studied on tissue microarrays containing 333 cases. Evaluation of MYC by fluorescence in situ hybridisation was successful in 220/333 (66%) cases. 9/220 (4%) cases showed MYC breaks. Re-evaluation of these tumours did not show any specific morphological and/or immunohistochemical features. The median survival time was 9 months for the respective patients, as opposed to 80 for patients without MYC breaks. The presence of MYC breaks in DLBCL cannot be reliably predicted by conventional methods. Since such patients might profit from different forms of treatment, routine testing of all DLBCL for MYC aberrations is suggested.
Tissue microarray
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Recent studies suggested that MYC and BCL2 protein co-expression is an independent indicator of poor prognosis in diffuse large B-cell lymphoma. However, the immunohistochemistry protocols for dual-expression staining and the scoring cut-offs vary by study. Sixty-nine cases of diffuse large B-cell lymphoma were evaluated for MYC and BCL2 protein expression using various cut-offs that have been recommended in prior studies. Independent of the International Prognostic Index risk group, cases with dual protein expression of BCL2 and MYC using ≥50%/40% cut-offs and ≥70%/40% had significantly shorter overall survival than cases without. It was verified in this patient population that the use of BCL2 and MYC immunohistochemistry, performed with available in vitro diagnostic-cleared antibodies, provides rapid prognostic information in patients with de novo diffuse large B-cell lymphoma. This study has practical implications for diagnostic laboratories and serves as a guide for implementation in the setting of future clinical trials.
International Prognostic Index
Clearance
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Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous group of lymphoid malignancies, which counts for more than a third of non-Hodgkin's lymphoma cases. The aim of the current study is to evaluate the prognostic role of several immunohistochemical (IHC) markers involved in the pathological process of DLBCL. This is a retrospective analysis of the 97 de novo DLBCL patients admitted between January 2007 and December 2016 in the Department of Hematology, "Filantropia" Municipal Hospital, Craiova, Romania. The expression of Bcl-2, Ki67, c-MYC and p53 was assessed by immunohistochemistry. A significant level of association was observed between high prognostic index values and Bcl-2, Ki67, c-MYC and p53 positive cases. Moreover, overall survival and disease-free survival were higher in patients with negative expression for these markers. Bcl-2, Ki67, c-MYC and p53 could make important diagnostic and therapeutic targets; therefore, their routine assessment should be mandatory.
Hematology
International Prognostic Index
Proliferation index
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Objective To explore the prognostic value of regulatory T cells (Tregs) and M2 macrophages in diffuse large B-cell lymphoma (DLBCL) tissues.Methods The expression of CD163 and Foxp3 was detected by immunohistochemistry in 92 cases of DLBCL,and it was
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