[1405] Identification of Novel Antigens To Identify the Neoplastic Cells of Nodular Lymphocyte Predominant Hodgkin Lymphoma Using a Lyoplate

Jonathan R Fromm, David Wu, Brent L Wood. University of Washington, Seattle, WA

Background: Nodular lymphocyte predominant Hodgkin's lymphoma (NLPHL) is usually confirmed by immunohistochemistry, but challenging cases exist in which this neoplasm can be difficult to differentiate from classical Hodgkin lymphoma (CHL) and B cell non-Hodgkin lymphomas.
Design: We sought to identify potentially useful novel antigens for the identification of L&H cells (neoplastic cells of NLPHL) using a Lyoplate™ (unconjugated antibodies to 242 cell surface antigens in 96-well plates). Antigen expression of a NLPHL cell line (DEV) was compared to a CHL cell line (L1236) and a Burkitt lymphoma cell line (Raji). Aliquots of the 3 cell lines were labeled with different fluorochromes (“barcoded”), processed with the antibody reagents, and evaluated by flow cytometry. Mean fluorescence intensity (MFI) of antigen expression by the different cell lines were measured and the MFI ratios calculated.
Results: Expression of antigens on DEV relative to the Raji and L1236 cells is shown in Table 1.

Table 1

↑ relative to Raji	↓ relative to Raji	↑ relative to L1236	↓ relative to L1236	↑ relative to Raji & L1236	↓ relative to Raji & L1236
CD30 CD39 CD44 CD59 CD62L CD79b CD87 CD106 CD108 CD146 CD171 CD273 CD275	CD10 CD21 CD27 CD32 CD38 CD58 CD72 CD85 CD49F CD97	CD19 CD20 CD22 CD37 CD39 CD45RB CD45RA CD50 CD53 CD62L CD75 CD79b CD71 CD81 CD87 CD102 CD146 CD171 CD268 CD275	CD15 CD26 CD32 CD31 CD58 CD97 CD227 CD274	CD39 CD87 CD275	CD32 CD97
	HLA-ABC B2M HLA-DR HLA DQ HLA-DP/DR/DQ		B2M HLA-DQ HLA-ABC DIS GD2 HLA-A2 HLA-DR/DQ/DP HLA-DR SSEA-1

DEV cells were then mixed with peripheral blood (PB) to evaluate the expression of CD39, CD87, CD275, CD32 and CD97 on DEV cells relative to PB leukocytes. CD39 was brightly expressed on DEV cells, moderately positive on macrophages, B cells, and granulocytes, but negative on T cells. CD87 was moderately positive on DEV cells, monocytes and granulocytes, but negative on B cells and T cells. CD275 was positive at an intermediate level on DEV cells (higher level than any other cell), weakly positive on B cells, and negative on other cells. CD32 was not expressed on DEV cells but was variably positive on T cells, and strongly positive on granulocytes, B cells and monocytes. Finally, CD97 was positive on DEV cells, monocytes and granulocytes, and T cells, but negative on B cells.
Conclusions: This study suggests that CD87 and CD275 (positive on L&H cells) and CD32 (negative on L&H cells) may be useful in diagnosing NLPHL. Further studies will required to evaluate the antigen expression of L&H cells from biopsies to validate these results.
Category: Hematopathology

Wednesday, March 21, 2012 9:30 AM

Poster Session V # 208, Wednesday Morning