Anti cd10 56c6

Paraffin‐embedded sections of each sample were subjected to IHC staining, using anti‐CD10 (56C6; Leica Microsystems), anti‐CD20 (L‐26; DakoCytomation), anti‐B‐cell lymphoma 2 (BCL2) (124; DakoCytomation), anti‐BCL6 (P1F6; Leica Microsystems), anti‐MUM1 (MUM1p; DakoCytomation), and anti‐c‐MYC (Y69; Abcam) Abs. Each case was considered as positive if more than approximately 30% neoplastic cells were positive, except for BCL2 and MYC, for which each case was considered positive if more than approximately 50% and 40% neoplastic cells were positive, respectively.¹⁷ Tumors coexpressing the MYC and BCL2 proteins were considered as double expressor lymphomas (DELs).¹⁷ Immunohistochemical staining was also undertaken with the anti‐CD68 Ab (KP‐1; DakoCytomation) as the panmacrophage marker and the anti‐CD163 Ab (10D6; Leica Microsystems) for the M2 macrophage. The number of CD68‐ and CD163‐positive cells was evaluated in a representative high‐power field corresponding to that of SIRPα.

Tissue samples were processed as formalin‐fixed, paraffin‐embedded tissues according to standard institutional procedures. We created tissue microarrays from samples from 61 patients and undertook evaluations of IHC with antibodies using these microarrays. Antibodies (clones) used for IHC included anti‐CD20 (L‐26; DakoCytomation, Glostrup, Denmark), anti‐BCL2 (clone124; DakoCytomation), anti‐BCL6 (P1F6; Leica Microsystems, Wetzlar, Germany), anti‐Multiple myeloma oncogene ‐1 (MUM ‐1) (MUM1p; DakoCytomation), anti‐CD10 (56C6; Leica Microsystems), and anti‐c‐MYC (Y69) antibodies (Abcam, Cambridge, UK). Immunohistochemistry results were reviewed by two expert hematopathologists (H.M. and K.O.). Cut‐off points for MYC, BCL2, and BCL6 protein expression were defined as DLBCL with 30% or more, 1% or more, and 30% or more positive cells, respectively, as recommended in previous studies.12, 15, 34