Rituximab is a chimeric monoclonal antibody that recognizes the CD20 antigen. It has been used to treat B-cell non-Hodgkin lymphoma (B-NHL), but recently rituximab resistance has been a cause for concern. We examined histological and immunohistochemical changes in 59 patients with B-NHL after rituximab therapy. The patients comprised 32 men and 27 women with a median age of 59 years. Pre-rituximab specimens comprised 34 follicular lymphomas (FL), 11 diffuse large B-cell lymphomas (DLBCL), 10 mantle cell lymphomas, two marginal zone B-cell lymphomas (MZBCL), and two chronic lymphocytic leukemias (CLL). CD20 expression in lymphoma cells was evaluated by immunohistochemistry or flow cytometry. Postrituximab materials were taken a median of 6 months (4 days to 59 months) after rituximab therapy. Sixteen cases ( R ituximab is a chimeric monoclonal antibody that has recently been incorporated into the treatment of B-cell non-Hodgkin lymphoma (B-NHL). It recognizes the CD20 antigen, a pan-B-cell marker, binds to it, and induces apoptosis of CD20-positive cells.(1-4) Rituximab can be used as a monotherapy or in combination with conventional chemotherapy for treatment of low-and high-grade, untreated, relapsed, or refractory CD20-positive B-NHL, achieving a high response rate with a low toxicity.Recent studies have reported that B-NHL show CD20-negative relapse after rituximab therapy.(5-17) Transformation of follicular lymphoma (FL) to CD20-negative diffuse large B-cell lymphoma (DLBCL), (15) proliferation of CD20-negative plasmacytoid tumor cells of marginal zone B-cell lymphoma (MZBCL) (16) or lymphoplasmacytic lymphoma,transformation of FL to classical Hodgkin's lymphoma, (17) and progression of nodular lymphocyte-predominant Hodgkin lymphoma to CD20-negative T-cell-rich B-cell lymphoma have also been reported. (18) Several mechanisms of resistance to rituximab have been suggested, including selection of a CD20-negative clone as a consequence of rituximab exposure, masking of CD20 epitopes by rituximab itself, or true loss of CD20 antigen by genetic and epigenetic changes. (12,13,15,(19)(20)(21)(22)(23)(24) In the present study we carried out retrospective analyses of histological and immunophenotypic changes and outcome in 59 patients with B-NHL after rituximab-containing therapy, to explore the effect of rituximab on CD20 expression and morphology in B-NHL.
Materials and MethodsPatient selection. We reviewed the pathology archives of the National Cancer Center Hospital, Tokyo, Japan, for the period 2002 to 2007. Fifty-nine consecutive cases of CD20-positive B-NHL treated with rituximab, with or without chemotherapy, for which pre-and post-rituximab specimens were available, were included in our study. Rituximab (Zenyaku Kogyo, Tokyo, Japan) was used at a standard dose of 375 mg/m 2 once a week for rituximab monotherapy and once every 3 weeks for the rituximab-cyclophosphamide, doxorubicin, vincristine and prednisone (CHOP) regimen. Clinical information was extracted from the medical records, and the Ann Arbor syste...