Double immunoenzymatic labeling of lymphomatous tissues for both immunologic phenotype and a malignancy-associated nucleolar antigen.

Defining cell lineage in the non-Hodgkin's lymphomas (NHL) is challenging for the immunopathologist. Cell surface marker techniques have made a major contribution to the understanding of the biology and classification of lymphoproliferative disorders by permitting the determination of the lymphoid (B- or T-cell) or monocytic lineage of the tumors. Because lymphoma cells often simulate the morphologic features and cell surface phenotype of their normal lymphocytic counterparts, it is difficult to discriminate normal from neoplastic lymphocytes. The authors have used representative monoclonal antibodies (MAb) to cell surface antigens to assess tumor cell surface antigens associated with various lymphoreticular cell lineages. Heteroantisera to the human malignancy-associated nucleolar antigen (HMNA) was utilized as a marker for neoplastic lymphoid cells as previously described. The use of double immunoenzymatic staining with both peroxidase and alkaline phosphatase allow us simultaneously to determine lymphoid lineage and malignancy on human lymphoma cells. In 101 cases of various cell types of NHL, the anti-HMNA antiserum reacted with nucleoli in the morphologically neoplastic lymphoma cells, but not with normal-appearing lymphoid and other cell types present in the lesions. Control specimens from normal and hyperplastic lymphoid tissue also failed to react with anti-HMNA antibodies.