Expression of B-Cell Markers in Classical Hodgkin Lymphoma: A Tissue Microarray Analysis of 330 Cases

Abstract

Hodgkin and Reed-Sternberg cells of classical Hodgkin lymphoma arise from B-lymphocytes. However, classical markers of the B-cell phenotype, such as CD20, are present only in about 25% of the cases. The aim of the present study was to assess expression of the B-cell-related antigens CD20, CD79a, and CD138 in classical Hodgkin lymphoma using a tissue microarray consisting of 330 classical Hodgkin lymphoma cases. Expression of CD15, CD20, CD30, CD79a, CD138, and latent membrane protein 1 of Epstein-Barr virus was assessed by immunohistochemistry, and the methodology was validated by direct comparison of CD20 expression on the tissue microarray cores with corresponding large sections. The influence of the number of arrayed sample cores on the obtained expression levels of CD20 was analyzed by comparing the results from single, duplicate, and triplicate cores. Two-hundred fifty-three (77%) of the 330 cases were morphologically representative. CD20 was expressed in 84 cases (33%), CD79a in 26 (10%), and CD138 in 2 (1%), respectively. CD20 and CD79a were co-expressed in 16 cases (P <.005), and expression of CD20 correlated inversely with CD15 (P <.01). Comparing the tissue microarray results with those from conventional sections for expression of CD20 yielded a concordance of 94% (63/67). Examining one, two, and three cores from individual cases revealed positivity for CD20 at 24% (61/253), 32% (82/253), and 33% (84/253), respectively. We conclude that B-cell markers are expressed in 38% of classical Hodgkin lymphoma in the following rank order: CD20>CD79a>>CD138. The use of two cores per tissue sample renders the tissue microarray technology effectively representative and thus very useful for high-throughput evaluation of heterogeneously expressed markers in classical Hodgkin lymphoma.

Aberrant expression of cell cycle regulators in Hodgkin and Reed–Sternberg cells of classical Hodgkin's lymphoma

“…Several proteins that regulate the mitotic cycle, such as cyclin D1, D3, E, B1, cyclin-dependent kinases 1, 2 and 6, S-phase kinase associated protein-2, p16 INK4A , p18 INK4c , p21 CIP1 , p27 KIP1 , p53, the retinoblastoma protein and PCNA have been found to be deregulated in classical Hodgkin's lymphoma. [1][2][3][4]6,[20][21][22][23][24][25][26] In the present study, we used a previously validated classical Hodgkin's lymphoma tissue microarray with a cohort of clinically well-documented cases 6,17,27 to analyze the expression of the cyclin-dependent kinase inhibitors of the Cip/Kipfamily, p21 CIP1 and p27 KIP1 , as well as two proliferation markers, PCNA and cyclin A. Since Hodgkin and Reed-Sternberg cells are embedded within reactive normal lymphocytes, we used those lymphocytes as internal controls for our staining Aberrant cell cycle regulation in Hodgkin's lymphoma A Tzankov et al conditions.…”

“…6,17 Sections of the tissue microarray blocks, 4 mm thick, were transferred to an adhesive-coated glass slide system (Instrumedics Inc., Hackensack, NJ, USA) and stained with hematoxylin and eosin, Giemsa and with the periodic acid Schiff reaction. Only cases containing at least two morphologically unequivocal Hodgkin and Reed-Sternberg cells were analyzed.…”

Expression profiling of transcription factors Pax-5, Oct-1, Oct-2, BOB.1, and PU.1 in Hodgkin's and non-Hodgkin's lymphomas: a comparative study using high throughput tissue microarrays

“…1 In contrast, Reed-Sternberg and Hodgkin's cells in classical Hodgkin's lymphoma, despite their rearranged immunoglobulin genes, fail to express functional immunoglobulin mRNA and/or protein, and lack B-cell-specific antigens in most instances. [1][2][3][4] It has been speculated that the lack of immunoglobulin mRNA transcripts and immunoglobulin protein expression, as well as B-cell specific antigens may be due to either crippling mutations of the immunoglobulin genes found in approximately 25% of cases, 2,[5][6][7] or downregulation and disruption of the B-cell-specific transcription factors. [8][9][10] Recent studies have shown that transcription factors Pax-5, Oct-1, Oct-2, BOB.1, and PU.1 are expressed in the majority of B-cell lymphomas and nodular lymphocyte predominant Hodgkin's lymphoma, but are downregulated in Reed-Sternberg and Hodgkin's cells of classical Hodgkin's lymphoma.…”

“…In addition, in a significant proportion of classical Hodgkin's lymphoma, the Reed-Sternberg and Hodgkin's cells lack B-cellspecific antigens. [1][2][3][4] The lack of immunoglobulin mRNA transcripts and proteins as well as B-cell specific antigens have been speculated to be due to either crippling mutations of the immunoglobulin genes found in about 25% of cases, 2,5-7 or downregulation and disruption of the B-cell-specific transcription factors. [8][9][10] In contrast to classical Hodgkin's lymphoma, the neoplastic cells in nonHodgkin's lymphomas of B-lineage and nodular lymphocyte predominant Hodgkin's lymphoma express B-cell-specific antigens, immunoglobulin mRNA transcripts and their proteins.…”

“…[8][9][10] Recent studies have shown that transcription factors Pax-5, Oct-1, Oct-2, BOB.1, and PU.1 are expressed in the majority of B-cell lymphomas and nodular lymphocyte predominant Hodgkin's lymphoma, but are downregulated in Reed-Sternberg and Hodgkin's cells of classical Hodgkin's lymphoma. [1][2][3][8][9][10] Most studies report that the Pax-5 is expressed in 100% of non-Hodgkin's lymphomas of B-lineage and a significant proportion of classical Hodgkin's lymphoma. However, there is inconsistency in the reported incidence of expression of the Oct-2, Oct-1, BOB.1, and PU.1 in classical Hodgkin's and non-Hodgkin's lymphomas in the literature.…”