Lymphocyte Biology Research Articles

Efficient Gene Transfer Into Primary Murine Lymphocytes Obviating the Need for Drug Selection

The efficient introduction of exogenous genes into primary lymphocytes is potentially important both for somatic cell gene therapy and for studying lymphocyte biology. We describe the use of retroviral vectors to efficiently introduce exogenous genes into primary, mature murine lymph node T and B cells, and primary, immature murine CD4- CD8- double-negative (DN) thymocytes. Efficient infection of primary cells was achieved by cocultivation of target cells with lethally irradiated helper cells that produce high titers of retroviral vectors containing either the neomycin phosphotransferase II (neo) gene, or both the neo and the human adenosine deaminase (ADA) genes, in the presence of lymphokines and/or mitogens. Two days postinfection, without neomycin selection, one to five copies of the exogenous genes per cell were detected by Southern blot analysis. Expression of the exogenous human ADA protein was detected at levels comparable to the endogenous murine ADA protein in the mature T and B lymphocytes, and was somewhat lower for the immature DN thymocytes.

Efficient gene transfer into primary murine lymphocytes obviating the need for drug selection

The efficient introduction of exogenous genes into primary lymphocytes is potentially important both for somatic cell gene therapy and for studying lymphocyte biology. We describe the use of retroviral vectors to efficiently introduce exogenous genes into primary, mature murine lymph node T and B cells, and primary, immature murine CD4- CD8- double- negative (DN) thymocytes. Efficient infection of primary cells was achieved by cocultivation of target cells with lethally irradiated helper cells that produce high titers of retroviral vectors containing either the neomycin phosphotransferase II (neo) gene, or both the neo and the human adenosine deaminase (ADA) genes, in the presence of lymphokines and/or mitogens. Two days postinfection, without neomycin selection, one to five copies of the exogenous genes per cell were detected by Southern blot analysis. Expression of the exogenous human ADA protein was detected at levels comparable to the endogenous murine ADA protein in the mature T and B lymphocytes, and was somewhat lower for the immature DN thymocytes.

Signal transduction through the T-cell antigen receptor

The mechanism by which ligand binding to the T-cell antigen receptor triggers the T-cell activation program has long been one of the most fascinating questions in lymphocyte biology. Here, we review recent insights into the transmembrane signaling functions of this multisubunit receptor complex.

Lymphocyte biology in lupus

Lymphocyte biology in lupus

Interaction of histidine-rich glycoprotein with human T lymphocytes

Histidine-rich glycoprotein (HRGP), a human plasma and platelet protein, interacts with multiple ligands in vitro, including heparin, plasminogen, thrombospondin, and fibrinogen/fibrin. In this study, the binding of HRGP to human T lymphocytes was characterized. The binding was specific, concentration-dependent, saturable, and reversible. Scatchard plot analysis revealed two classes of binding sites: the high affinity class had an apparent dissociation constant (Kd) of 1.92 X 10(-8) M, with 0.92 X 10(4) sites/cell, and the low affinity class had a Kd of 4.97 X 10(-7) M, with 3.7 X 10(4) sites/cell. HRGP binding to T cells in the presence of HRGP-depleted serum was comparable to that observed in buffer. Dot-blot analysis showed that HRGP bound to specific T cell proteins. Using both HRGP affinity chromatography and immunoprecipitation with affinity-purified anti-HRGP IgG, a major 56-kDa HRGP-binding protein in surface labeled T cell lysates was demonstrated. The 56-kDa protein was shown not to be related to the CD2 molecule on T cells. The binding characteristics of HRGP to T lymphocytes indicate a specific ligand-receptor interaction. This is the first demonstration of HRGP binding to a cell surface, and its binding to human T cells may play an important role in T lymphocyte biology.

Report on the 13th Meeting of the European Federation of Immunological Societies, Rome, 13–16 September 1988.

Knowledge of the interactions between T lymphocytes and other immunocompetent cells are central to understanding the pathogenesis of allergic disease. In recent years immunologists have made considerable progress towards delineating the mechanisms which regulate the production of IgE and which modulate the activity of non‐specific effector cells (eosinophils, neutrophils and monocytes). Papers on this subject were presented in several sessions at the recent EFIS meeting (held in September in Rome). In this report, Tony Frew summarizes the news from this meeting and explains why researchers in the field of allergy need to keep abreast of developments in T lymphocyte biology.

The effect of platelet activating factor and BN52021, a platelet activating antagonist, on the biology of lymphocytes

The effect of platelet activating factor and BN52021, a platelet activating antagonist, on the biology of lymphocytes

Thymic lymphocyte differentiation and thymic leukemogenesis

Henry Kaplan helped establish the fields of lymphocyte biology and viral leukemogenesis by his early and continuing studies on radiation leukemogenesis. As one of Henry's students I carried on these dual preoccupations with thymic lymphocytopoiesis and thymic lymphomagenesis. This communication demonstrates that 1) thymic lymphocytes are derived from bone marrow precursors which lack any T cell markers; 2) these bone marrow cells (or their clonogenic subsets) can give rise to either thymic cortical plus medullary progeny, or medullary progeny alone; 3) thymic lymphocytes mature in contact with 3–5 classes of nonlymphoid cells (thymic nurse cells, cortical dendritic epithelial cells, medullary epithelial cells, dendritic reticular cells, and macrophages), and one of these subsets, cortical dendritic epithelial cells, express an unusual distribution of MHC antigen (perhaps utilized in the maturation of T cell MHC restriction); 4) the population of cells which are poised to emigrate from the thymus are a unique subset of cortical cells which possess peripheral lymphoid organ homing receptors; and 5) the thymic target cells for retrovirus lymphomagenesis express highly specific retrovirus receptors that are analogous (and perhaps synonymous) with antigen-specific T cell receptors.

Multiparameter analysis of human lymphocyte subpopulations using flow cytometry.

This review has emphasized several recent advances in our understanding of human lymphocyte biology made possible by mAbs that recognize specific functional subpopulations of lymphocytes, coupled with multiparameter flow-cytometric analysis. In the last 3 years, both the T4+ and T8+ subsets of T cells have been further divided according to function and appearance of specific cell surface antigens. In addition, subpopulations of NK cells with different maturation states and B cells with distinct functional abilities have been discovered. Multiparameter flow-cytometric analysis of lymphocyte subpopulations can provide clues to the pathogenetic mechanisms of immune disorders, as has been demonstrated in AIDS patients and children with JRA. The use of multiparameter flow-cytometric analysis to study human immune responses in vitro has enabled a clearer understanding of the complex lymphoid interactions and, as we believe, may play an important role in the analysis of immune-associated disease processes.

T and B lymphocytes: Two different cell populations for the “first defence barrier” at cutaneous and mucosal levels?

Data on lymphocyte biology indicate that the humoral system of B-cells is specifically active with precise analogies at all mucosal levels. Might there be a corresponding function for T-lymphocytes at the cutaneous structure level? Certain physiological findings appear to suggest that the two main lymphocyte populations share the role of “first defence barrier”.

The pathogenesis of autoimmunity in New Zealand mice

In the past few years there has been an explosion of information in the area of NZ mouse research. This has reflected a general increase in knowledge and technology in cellular immunology and lymphocyte biology. Continuing research in many specific areas promises to expand this increasing data base. The numerous abnormalities of CMI in NZ mice chronicled in the past are currently being analyzed in terms of cell synergism rather than as simple effector mechanisms. Such analysis is indicating more selective rather than general impairment of CMI in aging NZ mice. Similarly, the allogeneic H-2 identical cytotoxic activity expressed by NZ mice and its relevance to autoimmune disease are being explored. In the area of suppressor functions in NZ mice, continued refinement and standardization of suppressor assays must be achieved before adequate interpretation of the present information can be made. Suppressor activity in aging NZ mice has been described as depressed, normal, and elevated. This is an area of acute interest. While the primary etiologic significance of T cell regulatory defects is currently being minimized, their possible secondary role in pathogenesis of autoimmune disease in NZ mice is being investigated. The recent discovery of a significant B cell abnormality present from birth in all SLE-prone murine strains has increased the probability that intrinsic B cell hyperactivity is a primary etiologic factor in murine autoimmunity. The hypersecretion of IgM and the increased incidence of B cell colony-forming units observed in NZ mice is being subjected to further genetic analysis in many laboratories. Similarly, the continued evaluation of the (CBA/N × NZB) hybrid promises to be enlightening, particularly the development of congeneic mice with the CBZ/N X chromosome on the NZB background. In addition, the development of H-2 congeneic NZB strains and immunoglobulin allotype congeneic NZB strains are proceeding. These strains will provide information on the possible roles of MHC and V H regions in autoimmune pathogenesis. The NZB.ch congeneic strain has already proven to be of great value. The continued development and further characterization of congenitally immunologic mutant NZ mice has been and will continue to be useful in elucidating the mechanism of autoimmune development in these animals. While NZ mice express a variety of host-viral interactions, recent genetic analysis suggests that viral infection is not a primary etiologic agent in the autoimmune disease of NZ mice. A well-defined clinical entity and a range of abnormalities have been delineated in NZ mice. The present goal is to define the cellular basis of autoimmunity through continued immunologic and genetic analysis of this important animal model of human autoimmune disease.

B lymphocyte biology studied with anti-Ig antibodies.

B lymphocyte biology studied with anti-Ig antibodies.

Activation of leopard frog ( Rana pipiens) spleen lymphocytes by concanavalin A and phytohemagglutinin

Activation times for concanavalin A (Con A)- and phytohemagglutinin (PHA)-induced lymphocyte mitogenesis in leopard frogs were determined by competitive inhibition analysis using α-methyl- d-mannopyranoside and N-acetyl- d-galactosamine. Con A activation required approximately 18–20 hr whereas PHA was required for at least the first 24 hr. Temporal events for mammalian lymphocytes are similar suggesting that activation parameters of lymphocyte biology have remained unchanged since the first vertebrate tetrapods evolved.

Fine structure of leopard frog (Rana pipiens) lymphocytes after in vitro transformation by phytohemagglutinin

Stimulation of leopard frog (Rana pipiens) spleen lymphocytes in vitro with phytohemagglutinin (PHA) caused a gradual change from small or medium sized lymphocytes to lymphoblasts. Lymphocytes cultured with PHA formed cell clusters with uropods oriented towards the center, giving them a rosette-like appearance. Mitotic figures were observed within lymphocyte clusters. At the ultrastructural level, unstimulated lymphocytes had sparse cytoplasm containing a few mitochondria, scattered free ribosomes and nuclei containing heavy clumps of heterochromatin. In contrast, after PHA stimulation, lymphocytes possessed the typical ultrastructure of lymphoblasts. The nucleus contained loosely packed euchromatin and prominent nucleoli. Ribosomes were aggregated to form polysomes, rough endoplasmic reticulum was sparse and mitochondria numerous. Uropods contained a well developed Golgi apparatus, centrioles with associated microtubules, various membrane bound bodies and abundant mitochondria and polysomes. The fine structural features of transformed frog lymphocytes are similar to those observed in several mammalian species, suggesting that this facet of lymphocyte biology has been conserved during evolution.

Cutaneous T-Cell Lymphomas: Clues Of A Skin Thymus Interaction

The common cellular denominator of the neoplastic lymphocytes of the cutaneous lymphomas is the presence of membrane markers of T-cell identity. For this reason these disorders are grouped together as "cutaneous T cell lymphomas". These neoplastic T-cells have a characteristic tissue distribution (preferentially infiltrating the skin and sparing the bone marrow). The abnormal T-cells of the leukemic phase of these disorders produce large amounts of macrophage migration inhibitory factor, which may adversely affect macrophage mobilization, and also stimulate differentiation of B-cells into plasma cells. The antigenic properties, the usual slow rate of replication, and the circulatory route of these cells may be exploitable in the development of more specific therapeutic approaches to the management of affected patients. Although these T-cells have an affinity for the skin, the role of this organ in the proliferation and differentiation of these cells is as yet not established. Localization of the primary site(s) of proliferation awaits completion of in vivo kinetic studies. The neoplastic T-cells from such patients provide important cellular reagents for the study of diverse aspects of lymphocyte biology. They have already been used to investigate mechanisms of lymphocyte triggering, isolate histocompatibility antigens, and characterize anti-T-cell immunoglobulin from patients with systemic lupus erythematosus.

The Lymphatic System

In this handsome volume, Battezzati and Donini have contributed significantly to the complex matter of lymphocyte biology. The inclusion of this material illuminates the subject of immunological functions of the lymphocyte, and makes this more comprehensive than most books on the lymphatic system. The basic physiology of the lymphatics has been well delineated by Courtice and Yoffey in past years, but the present chapters give a good review of both the old and more recent literature on this subject. The pertinent references are numerous; unfortunately some significant writings, and some quoted authors, are omitted. Nevertheless, the material on lymphocyte physiology should have particular value to workers in this field. The anatomy and pathology of lymph nodes and lymphatics are covered in some detail. The sections on lymphangiography are extensive and well done, with interesting presentations of pathology demonstrated by supplemental techniques of simultaneous venography, tomography of visualized nodes, and scintillation

The Production of a Monocyte Chemotactic Factor by Agammaglobulinemic Chicken Spleen Cells

Abstract The classification of lymphocytes into bursal derived (B)2 and thymic derived (T) subpopulations has been a significant advance in our understanding of lymphocyte biology. A variety of immune functions have been attributed to each of these subpopulations, while other functions depend on a cooperation between both cell types (1). It has been suggested that lymphokines, the biologically active substances produced in vitro by activated lymphocytes (2, 3) are produced by T lymphocytes (2). However, little definitive experimental evidence to substantiate this conclusion has been reported. The agammaglobulinemic (aγ) chicken has been shown to be largely depleted of B cells (4, 5) and represents an excellent model in which to investigate the contributions of B and T cells to various aspects of the immune response (6). In the present investigation we have adapted a recently described assay of mononuclear leukocyte (MNL) chemotaxis (7) to study lymphokine production by mitogen-stimulated normal and aγ chicken spleen cells.