Grading of follicular lymphoma: comparison of routine histology with immunohistochemistry

Follicular lymphoma (FL) grading is based on the average number of large transformed cells in 10 neoplastic follicles at x40 high-power field (x10-40 high-power field) examination (grade 1, 0-5 centroblasts per high-power field; grade 2, 6-15 centroblasts per high-power field; grade 3, >15 centroblasts per high-power field). Since there may be significant interobserver variability, we analyzed the usefulness of immunohistochemical stains in grading FLs more reliably. Forty-three FLs initially graded by World Health Organization criteria (grade 1, 12; grade 2, 18; grade 3, 13) were reviewed and stained with CD3, CD20, Ki-67, CD30, CD68, PAX-5, and BCL-6. Retrospective review was performed for the average number of large cells, of large lymphoid cells, of large cells staining with CD3, CD20, BCL-6 (40 cases), and PAX-5, and of all cells staining with CD68, Ki-67, and CD30. By histologic review, 8 of 43 FLs had a significant grade change (4 cases upgraded and 4 cases downgraded). CD3 and CD30 stained only 0 to 3 large cells and 0 to 3 cells, respectively, in neoplastic follicles. CD68+ cells represented the large nonlymphoid cells. Increasing FL grades demonstrated increases in Ki-67+ cells. The original grade showed substantial agreement with CD20 and moderate agreement with PAX-5 and BCL-6. The original histologic grade agreed with immunohistochemical-based grade using 2 or more antibodies in 5 of 8 discordant cases (4 by CD20 or BCL-6 and PAX-5; 1 by CD20, PAX-5, and BCL-6). Interobserver variability of histologic FL grading may be significant; we showed low-end "substantial agreement." Immunohistochemical stains (ie, CD20, PAX-5, and BCL-6) may more reliably determine the number of large transformed cells in neoplastic follicles; Ki-67 staining correlates with higher FL grades. Immunohistochemical stains may be evaluated in clinical trials of FL patients to determine prognostic significance.