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J M Pesando

Publications and source records attributed to J M Pesando.

At least 37 records · Page 2Linked to original sources
PubMed1986

Anti-CALLA antibodies identify unique antigens on lymphoid cells and granulocytes.

The common acute lymphoblastic leukemia antigen (CALLA) is a 100-kd surface glycoprotein that is present on normal and malignant lymphoid cells. It is a useful marker for distinguishing between clinically important types of acute leukemia. Anti-CALLA monoclonal antibodies (MoAb) also react with mature myeloid cells (granulocytes), where they identify an antigen having a similar molecular weight (mol wt). We now report that the antigens detected by anti-CALLA MoAb on human lymphoid and myeloid cells differ in their behavior and chemistry. Surface-labeling studies indicate that the antigen on lymphoid cells has a mol wt of approximately 100 kd v 110 kd for that on granulocytes. When cells are metabolically labeled with 35S-methionine, differences in the mol wt of these antigens are again observed. Unlike the lymphoid antigen, expression of that on purified granulocytes is not modulated by incubation with specific antibody. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of proteolytic digests of the two antigens fails to clarify their chemical relationship. Thus the antigens detected on these two cell types may share an epitope(s) but be chemically distinct, or CALLA may exist in distinct forms and behave differently on lymphoid cells and granulocytes.

Amino Acid Sequence↗
PubMed1984

Nonhuman primates express human leukemia-associated antigens.

Serologic studies using four murine monoclonal antibodies specific for the common acute lymphoblastic leukemia antigen (CALLA) and five monoclonal antibodies specific for the gp24 surface antigen indicate that these leukemia-associated antigens are present on cells of comparable tissues in man and in four nonhuman primates. As in man, adherent cell populations obtained from skin, lung, and bone marrow of Macaca fascicularis, M mulatta, M nemestrina, and Papio cynocephalus react with these antibodies. Similarly, granulocytes from both man and these nonhuman primates bind CALLA- and gp24-specific antibodies. Radioimmune precipitation experiments confirm the identity of these antigens. Our studies suggest that nonhuman primates can be used to screen serologic reagents to leukemia-associated antigens for potential toxic effects on normal tissues prior to their use in man. Similarly, nonhuman primates could be employed to assess the possible role of antigen-positive stromal cells in the reconstitution of bone marrow following transplantation.

Animals↗
PubMed1983

Distribution and modulation of a human leukemia-associated antigen (CALLA).

CALLA is a 100,000-dalton surface glycoprotein expressed by malignant cells of patients with clinically important subtypes of acute leukemia. Incubation of human leukemic cells expressing CALLA with specific monoclonal antibody (J5) at 37 degrees C causes rapid and selective internalization and degradation of this antigen (antigenic modulation). In these studies we show that CALLA-specific monoclonal antibodies also identify a cell surface glycoprotein having a m. w. of approximately 100,000 on 2 to 6% of nonmyeloid nucleated cells of normal adult bone marrow, on normal fibroblasts in tissue culture, and on cells of several nonhematopoietic human tumor cell lines. J5 antibody similarly modulates the surface expression of CALLA on nonleukemic cell populations, although the extent of modulation at a given concentration of antibody varied considerably. Modulation was almost complete for CALLA on cells of normal bone marrow, but was highly variable for cells of nonhematopoietic cell lines, possibly reflecting variability in antibody access to surface antigen. Using fluoresceinated or iodinated J5 antibody to modulate expression of CALLA on cells of leukemic cell lines, we show that antibody-antigen complexes undergo a temperature-dependent redistribution on the cell surface during modulation to form microaggregates. Antibody as well as antigen is then internalized. Studies of [35S]methionine-labeled cells indicate that synthesis of CALLA continues despite modulation of its surface expression by specific antibody, implying that the presence of CALLA on the cell surface reflects a dynamic equilibrium between the processes of surface expression of newly synthesized glycoprotein and its spontaneous and antibody-mediated clearance. The implications of these observations for immunotherapy are discussed.

Antibodies, Monoclonal↗
PubMed1981

Monoclonal antibody identifies a new Ia-like (p29,34) polymorphic system linked to the HLA-D/DR region.

The Ia antigens of the mouse are the basis for the genetic control of the immune response. The HLA-D/DR locus is considered to be the human counterpart of the Ia subregion of the murine major histocompatibility complex. The HLA-D/DR antigens are polymorphic, and eight well defined alleles have been identified using alloantisera. More recently, 'supertypic' antigens (MB and MT) have been defined which identify clusters of HLA-D/DR specificities. Little is known about the molecular basis for the cellular and serological polymorphism of the HLA-D/DR antigens, as alloantisera are usually of very low titre and heteroantisera frequently lack monospecificity. We present here the preparation and characterization of a monoclonal antibody which defines a new polymorphic system of the HLA-D/DR region. This and similar antisera should now begin to provide the reagents with which to correlate molecular structure with the functional repertoire of the human Ia-like antigens.

Antibodies↗
PubMed1981

Monoclonal antibodies defining serologically distinct HLA-D/DR related Ia-like antigens in man.

Four monoclonal antisera-identifying antigens with the identical tissue distribution and molecular weight of previously described Ia-like antigens were characterized. Two of these antisera, I-1 and I-2, identified antigens expressed on the HLA-D/DR positive cells from all HLA heterozygous individuals. Further characterization on homozygous typing cells (HTC's) demonstrated that I-2 was not reactive with most Dw7 and Dw11 HTC's. Monoclonal antisera, termed I-LR1 and I-LR2, defined polymorphic Ia-like antigens that demonstrated restricted expression on cells from HLA heterozygous individuals. Antigen I-LR1 was expressed on cells from 60% of HLA heterozygotes and its reactivity with HTC's did not conform to any previously described monotypic or supertypic HLA-D/DR pattern. In contrast, I-LR2 was expressed on 40% of HLA heterozygotes and identified only HLA-DR3, 5 and 6 HTC's. Studies of families with HLA recombinants permitted the demonstration that the I-LR1 and I-LR2 antigens are tightly linked to the HLA-D/DR locus. These experiments permit the direct demonstration by immunoprecipitation, linkage studies, and MHC recombinant families that the p29,34 complex in man is closely linked to or is within the HLA-D/DR locus. These studies suggest that the human Ia-like antigens are more heterogeneous than previously demonstrated and that monoclonal antisera will be useful in further defining the structural, genetic, and functional characteristics of these molecules.

Animals↗
PubMed1981

Human cell lines express multiple populations of Ia-like molecules.

Three monoclonal antibodies have been used to isolate Ia-like antigens from three human cell lines; two of which are thought to be homozygous at the HLA-D/DR locus. Complete extraction of the Ia antigens identified by one antibody leaves those recognized by the two remaining antibodies in three parallel sets of experiments, indicating that the antigenic determinants recognized by these antibodies are present on three different populations of Ia molecules from cells of single individuals. These three populations of Ia-like molecules may reflect serologic variants of the product of a single genetic locus or may represent the products of as many as three nonallelic genetic regions. Demonstration of the existence of these multiple populations of Ia-like molecules on presumed homozygous typing cells indicates that this antigenic system is much more complex than has been generally realized. Further study may clarify the relationship between HLA-D/DR type and susceptibility to a variety of diseases and ultimately lead to better matches and improved survival for allogenic transplants. Since the HLA-D region is intimately involved in regulation of the immune response and susceptibility to a variety of diseases, use of monoclonal antibodies specific for discrete Ia antigens, the only identified products of the HLA-D region, may facilitate dissection of its many biological functions.

Antibodies, Monoclonal↗
PubMed1981

A unique cell surface antigen identifying lymphoid malignancies of B cell origin.

A monoclonal antibody (anti-B1) specific for a unique B cell surface differentiation antigen was used to characterize the malignant cells from patients with leukemias or lymphomas. All tumor cells from patients with lymphomas or chronic lymphocytic leukemias, bearing either monoclonal kappa lambda light chain, expressed the B1 antigen. In contrast, tumor cells from T cell leukemias and lymphomas or acute myeloblastic leukemia were unreactive. Approximately 50% of acute lymphoblastic leukemias (ALL) of non-T origin and 50% of chronic myelocytic leukemia in blast crisis were also anti-B1 reactive. moreover, 21 of 28 patients with the common ALL antigen (CALLA) positive form of ALL were anti-B1 positive, whereas 0 of 13 patients with CALLA negative ALL were reactive. These observations demonstrate that an antigen present on normal B cells is expressed on the vast majority of B cell lymphomas and on approximately 75% of CALLA positive ALL, suggesting that these tumors may share a common B cell lineage.

Antibodies↗
PubMed1981

Serotherapy of acute lymphoblastic leukemia with monoclonal antibody.

We tested the efficacy of passive serotherapy in the treatment of acute lymphoblastic leukemia in four patients who had relapsed while receiving standard chemotherapeutic agents. Each patient received multiple intravenous infusions of J-5 monoclonal antibody specific for common acute lymphoblastic leukemia antigen (CALLA). In the three patients with circulating leukemic cells, there was a rapid decrease in circulating blasts that began immediately after antibody infusion, but not all leukemic cells were cleared, and remaining cells appeared to be resistant to further serotherapy. Although J-5 antibody was also demonstrable on bone marrow lymphoblasts immediately after antibody infusion in one patient, there was no change in bone marrow cellularity or differential during serotherapy. Analysis of the cell surface phenotype of leukemic cells during serotherapy and in vitro studies with patient cells suggests that resistance to serotherapy was mediated in part by antigenic modulation of CALLA in response to J-5 antibody.

Adolescent↗
PubMed1981

Fate of a common acute lymphoblastic leukemia antigen during modulation by monoclonal antibody.

Modulation of a human common acute lymphoblastic leukemia antigen (CALLA) by specific monoclonal antibody (J5) has been studied with the immune precipitation method to identify radiolabeled antigen. Surfaces of leukemic cells have been labeled using 125I both before and after modulation by J5 antibody for different time intervals. Leukemic cells have also been metabolically labeled with 35S-methionine before modulation. These studies indicate that the 100,000-dalton glycoprotein expressing CALLA (gp 100-CALLA) cannot be detected in cells that were modulated with J5 antibody before surface labeling but that it is easily detectable in cells that were surface labeled before modulation for 10 hr. At later time points, gp 100-CALLA is selectively lost from cells that were surface labeled before modulation. Gp 100-CALLA is not detected in the supernatants from cultures of these modulated cells. We conclude that gp 100-CALLA is rapidly internalized during modulation and that CALLA is degraded. Gp 100-CALLA is not shed into the culture media, nor does it remain on the cell surface in an altered form. Incubation of leukemic cells with antisera to beta2-microglobulin or IgM does not affect the expression of gp 100-CALLA.

Antibodies, Neoplasm↗
PubMed1981

Use of monoclonal antibodies as diagnostic and therapeutic reagents in acute lymphoblastic leukemia.

A monoclonal antibody specific for a common acute lymphoblastic leukemia (ALL) antigen has been generated and characterized. This antibody (J-5) is reactive with leukemic cells from most patients with non-T-cell ALL and some patients with chronic myelocytic leukemia in blast crisis. J-5 antibody is not reactive with leukemic cells from patients with either T-cell ALL, chronic lymphocytic leukemia, acute myeloblastic leukemia, or stable-phase chronic myelocytic leukemia. In addition to diagnostic applications, anti-common ALL antigen monoclonal antibody has been used to study the effect of specific serotherapy in the treatment of ALL. In the first patient with ALL treated with J-5 antibody, a 90% reduction in circulating lymphoblasts occurred within 1 hr of starting antibody infusion. Despite continued serotherapy, however, lymphoblasts persisted in peripheral blood and bone marrow. Analysis of cell surface markers during serotherapy suggested that resistance to antibody-mediated lysis in vivo may have been due to antigenic modulation of leukemic cells in response to J-5 monoclonal antibody.

Acute Disease↗
PubMed1981

Expression of common acute lymphoblastic leukemia antigen (CALLA) by lymphomas of B-cell and T-cell lineage.

Previous studies have demonstrated that the common acute lymphoblastic leukemia antigen (CALLA) is expressed by leukemic cells from approximately 80% of patients with non-T-cell ALL and 30%-50% of patients with chronic myelocytic leukemia in blast crisis. A small number of normal bone marrow and fetal liver cells also express CALLA, but the functional role of this molecule is unknown. In the present study, we have used a monoclonal antibody (J5) specific for CALLA to study the expression of this antigen in non-Hodgkin's lymphomas. Within the B-cell lymphomas, it was found the CALLA was expressed by almost all Burkitt's and nodular poorly differentiated lymphocytic lymphomas. Within the T-cell lymphomas, CALLA was expressed in 40% of patients with lymphoblastic lymphoma. Three of 3 Burkitt's lymphoma cell lines and three of eight T-lymphoblast cell lines were also found to express CALLA. Normal spleen, lymph node, and thymus cells were not reactive with J5 antibody. These findings indicate that expression of CALLA is not limited to relatively undifferentiated leukemic lymphoblasts but also occurs in more differentiated lymphoid malignancies. However, normal differentiated lymphoid cells in lymph node, spleen, and thymus, which have a phenotype similar to that of lymphoma cells, do not appear to express CALLA.

Antigens, Neoplasm↗
PubMed1980

A monoclonal antibody to human acute lymphoblastic leukaemia antigen.

Previous studies by Greaves and others have demonstrated the existence of an antigen associated with cells from many patients with acute lymphoblastic leukaemia (ALL) and some patients with chronic myelocytic leukamemia (CML) in blast crisis. Antisera to this common ALL antigen (CALLA) have been produced in rabbits and require extensive absorption which limits both the titre and quantity of antisera that can be generated and may result in variable specificity in different laboratories. The method for generation of specific antibody by somatic cell hybridisation introduced by kohler and Milstein has been successfully used to produce monoclonal antibodies against various normal human cell-surface proteins, including beta 2 microglobulin, histocompatibility antigens, thymocyte and peripheral T-cell antigens and Ia-like antigens. The present report describes the generation and characterisation of a monoclonal antibody specific for a common ALL antigen (CALLA) previously identified by conventional heteroantisera.

Animals↗
PubMed1980

Human leukemia-associated antigen: relation to a family of surface glycoproteins.

A heteroantiserum raised to leukemic cells of a patient with non-T cell acute lymphoblastic leukemia (ALL) has been extensively absorbed with cells from a leukemic T cell line and an autologous B lymphoblastoid cell line to produce a common ALL antiserum (CALLA). CALLA is specific for leukemic cells of most patients with non-T cell ALL and chronic myelogenous leukemia (CML) in lymphoid blast crisis. It has been extensively tested on a wide variety of normal cells and is unreactive with them. CALLA identifies a surface glycoprotein having a m.w. of approximately 100,000 on reactive cell populations. In contrast, partially absorbed anti-ALL sera detect a similar glycoprotein band on CALLA-negative B and T cell lines. The glycoprotein identified by CALLA has been isolated and used as an immunogen. This new antiserum (C129) detects a 100,000-dalton glycoprotein not only on CALLA-positive cell populations but also on most CALLA-negative normal and malignant hematopoietic cells and on B and T cell lines. We conclude that there exists a family of 100,000-dalton glycoproteins that are present on a variety of normal, transformed, and malignant cells and that possess shared as well as unique antigenic regions. The expression of at least one of these antigens, detected by CALLA, may be tumor specific.

Animals↗
PubMed1979

Ia determinants on human T-cell subsets defined by monoclonal antibody. Activation stimuli required for expression.

The nature of Ia antigens which appear on human T cells after activation and the stimuli required for their expression was examined utilizing a monoclonal antibody reactive with the Ia antigen framework. T cells were purified using monoclonal antibodies directed either at the entire T-cell population (OKT3) or the T-cell inducer subset (OKT4). By indirect immunofluorescence, it was shown that the human T-cell population contains no detectable Ia+ cells in the resting state. In contrast, in excess of 60% of the T-cell population expresses Ia antigen after alloactivation in the mixed lymphocyte culture. Moreover, these Ia antigens are expressed within both the OKT4+ and OKT4- subsets. Similarly, phytohemagglutinin and concanavalin A induced approximately 20% of peripheral T cells to express Ia antigen and the expression of these antigens is not restricted to either OKT4 subset. In contrast, only the inducer T-cell population which proliferates maximally to soluble antigen expresses Ia antigens after activation by tetanus toxoid. Thus, the expression of human Ia antigens on unique T-cell subsets depends upon the activation stimuli utilized and ability of the individual subset to respond to a given stimulus. Additional studies indicated that Ia antigens appear on previously Ia- T cells after activation and do not result from clonal expansion of a small subset of Ia+ T cells.

Antigen-Antibody Reactions↗
PubMed1979

Leukemia-associated antigens in ALL.

A cytotoxic common ALL antiserum (CALLA) specific for leukemic cells of most patients with non-T-cel- acute lymphoblastic leukemia (ALL) and of some patients with chronic myelogenous leukemia (CML) in blast crisis has been reproducibly prepared using cell lines for absorption. CALLA reacts with leukemic cells of 110 of 134 patients (82%) with non-T-cell ALL; 1 of 71 (1%) patients with acute myelogenous leukemia (AML); 2 of 7 patients (29%) with chronic myelogenous leukemia in blast crisis; 7 of 92 patients (8%) with other hematologic malignancies; and with the leukemic cell lines Laz 221 and NALM-1. It does not react with the normal hematopoietic cells, B- or T-cell lines, or cells from 26 patients with T-cell ALL that were tested. CALLA reactivity and periodic acid Schiff (PAS) staining correlate poorly, with CALLA reacting with cells from 86% (64 of 74) of patients with PAS-positive and 76% (29 of 38) of those with PAS-negative non-T-cell ALL. In these patients, CALLA reacts with cells from 89% of those under age 12 (78 of 88); 74% of those aged 12--20 (20 of 27); and 58% of those over 20 (11 of 19). Using only CALLA and antisera specific for Ia-like and T-cell antigens, we can now distinguish most cases of ALL from AML and other hematologic malignancies.

Absorption↗