Effect of reovirus on primary human lymphoid neoplasms.

CLL indicates chronic lymphocytic leukemia; DLCBL, diffuse large B-cell lymphoma; BL, Burkitt lymphoma; and PBL, peripheral blood lymphocytes from healthy donors. (A) Reovirus replication in normal cells (lymphocytes, CD34⁺ cells) and primary human lymphoid neoplasms. Normal lymphocytes, normal bone marrow stem cells, and cell suspensions from primary lymphoma tumors were infected with reovirus. The cells were labeled with [³⁵S]-methionine from 24 to 48 hours after infection. The cells were then lysed, immunoprecipitated with antireovirus antibody, and analyzed by SDS-PAGE. (B) Flow cytometry analysis. Representative histograms of viable (7AAD⁻) CLL cells or Burkitt lymphoma cells after infection with 20 MOI of reovirus and staining with CD5-phycoerythrin (PE) and CD20–fluorescein isothiocyanate (FITC) (for CLL), or CD10-PE and CD20-FITC (for Burkitt lymphoma) are shown on the left. CLL cells coexpressing CD5 and CD20 were counted by means of Flow Count beads, and the results for 5 CLL samples are shown on the right. Burkitt lymphoma cells coexpressing CD10 and CD20 were counted similarly, and the results are shown. (C) Immunohistochemistry (IH) of reovirus antigens in virus-treated Raji and CLL cells. Cells were prepared and infected with reovirus as described in “Materials and methods.” Positive (brown) staining is observed in both Raji and CLL cells infected with reovirus, but not in the controls.