Richard S. Metzgar

Phenotypic characterization of cutaneous T-cell lymphoma. Use of monoclonal antibodies to compare with other malignant T cells

We studied the surface-antigen pattern of T cells in peripheral blood and cell lines from patients with advanced cutaneous T-cell lymphoma (CTCL) or T-cell acute lymphoblastic leukemia. The antigen patterns of cutaneous T-cell lymphoma cells from peripheral blood and established cell lines were nearly identical; the cells were negative for human thymus antigen (OKT6 and NA1/34), positive for pan-T-cell (OKT3, 17F12, 10.2, and 9.6) and helper-T-cell-subset (OKT4) antigens, and negative for T-cell-subset antigens 3A1 and OKT8. In contrast, the phenotypes of malignant T cells from patients with acute lymphoblastic leukemia were heterogeneous, with at least five patterns of reactivity. The T-cell-specific antibody 3A1 was the only monoclonal reagent that clearly distinguished the peripheral-blood T cells in CTCL (3A1-) from those in acute lymphoblastic leukemia (3A1+). Moreover, 3A1 was the most reliable T-cell marker in acute lymphoblastic leukemia. We conclude that circulating CTCL (Sézary) T cells are homogeneous in their antigen phenotype and are derived from a well-differentiated 3A1-, OKT4+, OKT8- helper-T-cell subset.

Antigens Specific for Human Lymphocytic and Myeloid Leukemia Cells: Detection by Nonhuman Primate Antiserums

Primate antiserums to human leukemia cells can detect antigens specific for lymphocytic leukemia cells or antigens present on certain myeloid leukemia cells. The antigen specific for lymphocytic leukemia cells is destroyed by treatment with neuraminidase or trypsin. Tryptic digests of lymphocytic leukemia cells contain the antigen, which has a high molecular weight.

Characterization and distribution of a 24,000-molecular weight antigen defined by a monoclonal antibody (DU-ALL-1) elicited to common acute lymphoblastic leukemia (cALL) cells

A monoclonal antibody (DU-ALL-1) was generated to common acute lymphoblastic leukemia (cALL) cells by microcytotoxicity and indirect immunofluorescence, DU-ALL-1 reacted only with cALL cell lines and not with the other hematopoietic cell lines tested. Peripheral blood lymphocytes, monocytes, granulocytes and mitogen-activated lymphocytes did not react significantly with this antibody. However, platelets (100%) and normal bone marrow cells (8.5%) reacted with DU-ALL-1. Microcytotoxicity testing of human leukemia cells showed that DU-ALL-1 reacted with cells from a majority of null and pre-B ALL patients (63/77) and with cells from some patients with acute myeloblastic leukemia (4/7) and T-ALL (4/20). DU-ALL-1 was generally non-reactive with cells from patients with B-cell leukemias (2/16) and chronic myelogenous leukemia in blast crisis (0/4). By an indirect immunoperoxidase technique, DU-ALL-1 reacted with a variety of non-hematopoietic tissues, including smooth and cardiac muscle and epithelia from several organs. The DU-ALL-1 antigen had an apparent mol. wt of 24,000 and did not bind to lectins or label with [3H]glucosamine. Thus, DU-ALL-1 defines a 24,000-mol. wt protein which is absent from most peripheral blood mononuclear cells, is expressed on normal platelets and several non-hematopoietic tissues, and may be a useful for subclassifying leukemias.

Embryonic development of human transplantation antigens.

SUMMARY Human fetal cells grown in tissue culture monolayers were tested using the technique of mixed agglutination and were found to possess some of the antigens controlled by the HL-A locus. Gestational specimens ranging from 6 weeks through maturity were studied. All fetal homogenates, as well as cells derived from numerous individual fetal organs, demonstrated similar patterns of reaction by mixed agglutination when tested with a panel of 22 human isoantisera. We have been unable to detect HL-A antigens on whole trophoblastic syncytia by direct mixed agglutination testing and have not demonstrated that this material is capable of absorbing out antibodies directed against HL-A specificities. Absorption studies utilizing human spermatozoa also indicate that spermatozoa, like the trophoblastic syncytia, either do not express their transplantation antigens or are lacking in these specificities.

Preparation and characterization of lymphocyte-activating factor (LAF) from acute monocytic and myelomonocytic leukemia cells

Leukemia cells from acute monocytic and myelomonocytic patients undergoing leukophoresis have been used to prepare large quantities of lymphocyte-activating factor (LAF). The chemical properties of normal leukocyte LAF and leukemia cell LAF from a single donor for industrial-scale purification. The human histiocytic lymphoma cell line U937 was also evaluated as a possible source of LAF. Although exhibiting several monocyte-specific properties, the U937 cell line failed to release LAF.

Characterization of clones of a human pancreatic adenocarcinoma cell line representing different stages of differentiation.

With a limiting dilution technique, clones have been established from the human pancreatic adenocarcinoma cell line, HPAF. Phenotypic analysis by a panel of murine monoclonal antibodies demonstrated distinct profiles of antigenic expression between the clones. However, identical isozyme patterns of different clones indicated their common origin from the parental HPAF cells. Two clones, CD11 and CD18, appeared to be arrested at different stages of secretory epithelial cell differentiation. CD11 cells demonstrated many characteristics of a well-differentiated state, including the formation of ductal structures with polarized, long columnar-shaped cells, the presence of secretory granules in the cytoplasm, high DU-PAN-2 antigen expression in nude mouse xenografts, and a longer doubling time (42 h) in tissue culture. In contrast, CD18 cells exhibited characteristics of a poorly differentiated state, including solid nests of isoprismatic cells without luminal spaces and cellular polarization, absence of secretory granules and DU-PAN-2 antigen expression in xenografts, and a shorter doubling time (26 h) in tissue culture. Since no culture systems of normal pancreatic ductal cells are currently available, these two pancreatic adenocarcinoma clones may provide a unique system to study genes and antigens related to pancreatic ductal cell differentiation.

Pediatric Oncology Group Utilization of Immunologic Markers in the Designation of Acute Lymphocytic Leukemia Subgroups: Influence on Treatment Responsea

The clinical application of blast cell immunophenotype testing is important in childhood ALL for the following reasons. (1) Knowledge of the immunologic group is important in predicting prognosis. Prognostic grouping may prove to be accomplished best by using a combination of traditional risk factors and immunologic phenotyping. However, definition of traditional risk factors may vary within the immunologic groups of ALL. (2) In assessing the relative effectiveness of different treatment regimens for children with ALL it is important to make comparisons among patients within the same major immunologic groups of ALL. (3) Identification of specific immunologic groups of patients within ALL may help in designing therapy for each group. The POG has already made preliminary attempts in this direction for T-ALL and B-ALL. However, leukemia species-specific therapy is still only a long-range goal. Laboratory research must endeavor to identify additional biologic characteristics peculiar to each major immunologic group of ALL. These characteristics may dictate therapeutic maneuvers in the future.

Immunologic and clinicopathologic features of common acute lymphoblastic leukemia antigen-positive childhood T-cell leukemia A pediatric oncology group study

The immunologic and clinicopathologic features of common acute lymphoblastic leukemia antigen (CALLA)‐positive and CALLA‐negative T‐acute lymphoblastic leukemia (ALL) and of CALLA‐positive non‐T, non‐B ALL (common ALL) of childhood were compared. Twenty‐seven percent of children with T‐ALL had blasts that expressed CALLA. This expression was not associated with a significantly different incidence of expression of sheep erythrocyte‐rosette receptors, glucocorticoid receptors, peanut agglutinin receptors, or T‐cell antigens. CALLA‐positive T‐cell blasts were more likely to express a p24 leukemia‐associated antigen (CD9, 50% versus 8%) and Ia antigens (39% versus 8%) than were CALLA‐negative blasts. Patients with CALLA‐positive and CALLA‐negative T‐ALL had similar clinicopathologic features at diagnosis. In contrast, compared to patients with common ALL, patients with CALLA‐positive T‐ALL were older, had higher leukocyte counts, and an increased incidence of splenomegaly, lymphadenopathy and mediastinal mass, similar to patients with CALLA‐negative T‐ALL. Patients with CALLA‐positive T‐ALL were more likely to achieve a complete remission (95% versus 83%, P = 0.055) and tended to have an increased duration of event‐free survival (P = 0.07) than did patients with CALLA‐negative T‐ALL. The expression of T‐cell antigens is more important than the expression of CALLA in defining biologically similar subgroups of childhood ALL. Preliminary evidence suggests that within T‐ALL the expression of CALLA may be prognostically important.

SPECIES DISTRIBUTION OF HUMAN TISSUE ISOANTIGENS. I. DETECTION OF HUMAN TISSUE ISOANTIGENS IN CHIMPANZEES

Human tissue isoantigens were detected on chimpanzee leukocytes and on a continuous heteroploid cell line of chimpanzee origin. The antisera used to detect these isoantigens were taken from multiparous women, multiparous chimpanzees, and chimpanzees immunized with human leukocytes and platelets. These sera were known to contain antibodies to human leukocyte isoantigens. The methods used to demonstrate the isoantigens were leukocyte agglutination and inhibition of leukocyte agglutination, as well as mixed agglutination and inhibition of this reaction.

Human thymus-leukemia related antigen(s): Detection by a nonhuman primate antiserum☆

Abstract A nonhuman primate antiserum to human thymus cells can detect an antigen which is specific for human thymus cells. In addition, this antiserum can also detect antigens which are cross-reactive between thymocytes and cells from patients with lymphocytic leukemia and perhaps some antigens which are present on actively dividing cell populations like cultured lymphoid cell lines derived from normal individuals. The relevance of these data to other human leukemia-associated antigens and murine TL antigens is discussed.