Paraffin-resistant Antigens Detectable by Antibodies L26 and Polyclonal CD3 Predict the B- or T-Cell Lineage of 95% of Diffuse Aggressive Non-Hodgkin’s Lymphomas

Abstract

The reactivity of eight preferential B-cell (L26, 4KB5, and KiB3) and T-cell (polyclonal CD3, Leu22, MT-1, UCHL-1, and OPD4) antibodies which detect paraffin-resistant antigens was examined by a threestep immunoperoxidase technique in 111 formalin-fixed, paraffin-embedded diffuse aggressive non-Hodgkin's lymphomas (NHLs) to determine the optimal panel for accurate lineage assignment. L26 (CD20) and polyclonal CD3 (CD3) were the most sensitive (> 95%) and specific (100%) antibodies. They identified the B-or T-cell lineage correctly in 106 (95%) cases. The five L26-negative, polyclonal CD3-negative cases included all three precursor B lymphoblastic NHLs and two (one B and one T) diffuse large-cell NHLs. Immunostaining with the second most sensitive preferential B-cell (4KB5) and T-cell (Leu22) antibodies correctly identified the lineage in two addiThe recent advent of a variety of monoclonal and polyclonal antibodies reactive with paraffin-resistant B-and T-cell-associated antigens has markedly increased our ability to analyze lymphoid proliferations in routinely processed tissues. However, the immunophenotypic analysis of non-Hodgkin's lymphomas (NHLs) with these antibodies in paraffin-embedded tissue sections has often been considered inferior to the immunophenotypic analysis of viable cells in suspension and frozen tissue sections. In some cases, however, the use of routinely processed, paraffin-embedded tissue is preferable to frozen tissue-for instance, when accurate morphologic identification of the malignant cells is necessary or when it is important to preserve unfixed tissue for other studies. Furthermore, because specimens often arrive in the pathology laboratory already in fixative, fresh tissue may not be available for immunophenotyping. In addition to the histologic subtype and stage, the B-or T-cell lineage of an NHL often is clinically important because tional NHLs, but one false-positive result occurred. Preferential B-cell antibody KiB3 reacted with two precursor B lymphoblastic NHLs. Use of additional paraffin-reactive antibodies did not increase the number of NHLs assigned to the correct cell lineage. In conclusion, it appears that a two-tiered approach, with a first-line panel consisting of L26 and polyclonal CD3, followed by 4KB5 and Leu22 in nonlymphoblastic NHLs and by KiB3 in lymphoblastic NHLs, represents the most efficient method of correctly identifying the B-or T-cell lineage of diffuse aggressive NHLs by paraffin tissue section immunohistochemistry.

Immunoreactivity of B-Cell Markers (CD79a, L26) in Rare Cases of Extranodal Cytotoxic Peripheral T- (NK/T-) Cell Lymphomas

“…The monoclonal antibody L26 recognizes an intracellular epitope of CD20 and has been shown to be a valuable reagent for the diagnosis of B-cell lymphomas in routinely processed tissues (3)(4)(5)(6)(7)(8)24). A few cases of CD20-positive (L26) nodal PTLs, however, have been reported (5,10,11).…”

“…The report on the revised European-American classification of lymphoid neoplasms includes a record of the most characteristic immunophenotype for each lymphoma entity, and the majority of antigens listed can be detected on paraffin sections by commercially available antibodies (1,2). Some antibodies, such as those that recognize CD20 and CD79a, have gained widespread acceptance because they are considered to be largely B-lineage restricted and therefore useful for differential diagnosis of B-cell versus T-cell lymphoma (3)(4)(5)(6)(7)(8)(9). Although occasional examples of CD20-positive T-cell lymphomas have been reported (5,10,11), to our knowledge no case of CD79a-positive peripheral T-cell lymphoma (PTL) has been published.…”

Staging bone marrow in diffuse large B-cell lymphoma: the role of ancillary investigations

“…It is recommended that at least two B-cell and Tcell markers be used to prevent false results relating to the antibody, with CD20 and CD3 being reported as the most sensitive in assigning lineage in diffuse aggressive NHL. 45 The other options in B-cell markers include CD79a, PAX5, 4KB5, and KiB3, and often the antibody of choice is the one the laboratory has validated and is most comfortable with using. These antibodies may be of particular relevance in cases where the primary tumour is CD20 negative.…”

Role of Immunohistochemistry in Staging Diffuse Large B-cell Lymphoma (DLBCL)

“…We used two B-cell and two T-cell markers to corroborate our findings. CD20 and CD3 have been shown to be most sensitive at assigning lineage in diffuse aggressive NHL (Chadburn and Knowles 1994) and are the most commonly used antibodies in our laboratory. The two additional antibodies, CD79a and CD45RO, were selected to maintain consistency because these are the antibodies used in diagnostic tissue sections.…”

“…The streptavidin-biotin method was used to immunostain sections using the following monoclonal antibodies: CD3 (Dako clone CD3, DakoCytomation Inc, Carpenteria, CA; 1:200 dilution), CD45RO (Novacastra clone UCLH-1, Novacastra Laboratories Ltd, Newcastle, UK; 1:1000 dilution), CD20 (Zymed clone L26, Zymed Laboratories, Invitrogen Immunodetection Corp, Carlsbad, CA; 1:50 dilution), CD79a (Dako clone JCB117, 1:500 dilution), k (Novacastra clone kp-53, 1:750 dilution), and l (Novacastra clone Hp-6054, 1:750 dilution). CD20 and CD3 are known to be most sensitive at assigning lineage in diffuse aggressive NHL (Chadburn and Knowles 1994) and are the most commonly used antibodies in our laboratory. The two additional antibodies, CD79a and CD45RO, were selected to maintain consistency because these are the antibodies used for diagnostic tissue sections in our laboratory.…”