Ultrastructural identification of human tonsil T-lymphocytes by peroxidase-conjugated anti-HTLA serum.

A horse anti-serum rendered specific for human T-lymphocytes was conjugated with peroxidase and used for ultrastructural identification of human tonsil T-lymphocytes. With T- and B-enriched suspensions, virtually all T-lymphocytes were labelled with Po-anti-HTLA, whereas no B-cells were stained with this conjugate. The labelling HTLA conjugate confirm the ultrastructural characteristics of thymus-dependant cells.     

Cytotoxicity of human peripheral blood T-lymphocyte clones activated by hepatitis B virus surface antigen

Summary The present studies examined the cytotoxic activities of peripheral blood lymphocytes (PBL) from volunteers with (sero-positive) and without (sero-negative) circulating antibodies to hepatitis B virus surface antigen before and 30 days after vaccination with hepatitis B virus surface antigen (HBsAg). Long-term culture of monospecific hepatitis B surface (HBsAg)-responsive T-lymphocytes were isolated and grown in large numbers. The mechanism of T-cell mediated cytolysis, and the identification of the carbohydrate determinants on the surface of these effector cells responsible for the killing effect, are being examined.     

Cytotoxicity of human peripheral blood T-lymphocyte clones activated by hepatitis B virus surface antigen

The present studies examined the cytotoxic activities of peripheral blood lymphocytes (PBL) from volunteers with (sero-positive) and without (sero-negative) circulating antibodies to hepatitis B virus surface antigen before and 30 days after vaccination with hepatitis B virus surface antigen (HBsAg). Long-term culture of monospecific hepatitis B surface (HBsAg)-responsive T-lymphocytes were isolated and grown in large numbers. The mechanism of T-cell mediated cytolysis, and the identification of the carbohydrate determinants on the surface of these effector cells responsible for the killing effect, are being examined.     

Ultrastructural identification of human tonsil T-lymphocytes by peroxidase-conjugated anti-HTLA serum

Summary A horse anti-serum rendered specific for human T-lymphocytes was conjugated with peroxidase and used for ultrastructural identification of human tonsil T-lymhocytes. With T- and B-enriched suspensions, virtually all T-lymphocytes were labelled with Po-anti-HTLA, whereas no B-cells were stained with this conjugate. The labelling was found to be irregularly distributed on the plasma membrane of T-cells. Direct identification with specific Po-anti-HTLA conjugate confirm the ultrastructural characteristics of thymus-dependant cells.This work was supported by I.N.S.E.R.M. grants No. 74-5-027-01 and A.T.P. No. 73-16.Acknowlegments. We are indebted toMartine Blanc who prepared the cell suspensions and toDanielle Germain who did the preparations for electron microscopic examination.     

Surface receptors and functional interactions of human natural killer cells: from bench to the clinic

The past 10years have witnessed dramatic progress in our understanding of how natural killer (NK) cells function and their role in innate immunity. Thanks to an array of inhibitory receptors specific for different HLA class I molecules, human NK cells can sense the decrease or loss of even single alleles at the cell surface. This represents a typical condition of a potential danger, i.e. the presence of tumor or virally infected cells. NK cell triggering and lysis of these cells is mediated by several activating receptors and coreceptors that have recently been identified and cloned. While normal cells are usually resistant to NK-mediated attack, a remarkable exception is represented by dendritic cells (DCs). In their immature form they are susceptible to NK-mediated lysis because of the expression of low levels of surface HLA class I molecules. The process of DC maturation (mDCs) is characterized by the surface expression of high levels of HLA class I molecules. Accordingly, mDCs become resistant to NK cells. A recent major breakthrough highlighted the role played by donor NK cells in allogenic bone marrow transplantation to cure acute myeloid leukemias. Alloreactive NK cells derived from donor hematopoietic precursors not only prevented leukemic relapses, but also prevented graft rejection and graft-versus-host disease.Received 12 March 2003; received after revision 18 April 2003; accepted 30 April 2003     

Detection of varicella zoster virus DNA in human tissue by standard and nested polymerase chain reaction

Results and conclusion The sensitivity of the PCR assay was determined with cloned VZV DNA. About 200 copies of the target sequence were necessary for detectable amplification by standard PCR and less than 20 copies by nested PCR. Out of 24 human trigeminal ganglia five tested positive for VZV DNA by standard PCR (21%), in eleven cases VZV DNA was detectable using nested PCR (46%). Sequences specific for VZV could be detected in PMBC from children with acute varicella up to six days after the onset of rash by standard (one child) or nested (three children) PCR. This confirms that at the time of haematogenous spread before and during the rash viral DNA can be found within the mononuclear cells. Thus the use of nested primers enhances the sensitivity of the assay and allows the detection of only a few genomic copies of viruses in human tissues.     

Rosette formation by human T and B lymphocytes in the presence of adrenergic and cholinergic drugs

Summary It was shown that adrenergic drugs, which increase the intracellular levels of cAMP, inhibit the rosette formation by T-lymphocytes, but stimulate the rosettes produced by B-lymphocytes. Cholinergic drugs, which increase the levels of cGMP, on the contrary, stimulate the formation of rosettes by T-lymphocytes but inhibit those produced by B-lymphocytes.Acknowledgments. This investigation was supported by Grant from National Council for Scientific and Technological Development (CNPq), Rio de Janeiro, Brazil.     

Possible target of Abelson virus phosphokinase in cell transformation

By fusing interphase cells to cells undergoing mitosis, the interphase chromosomes can be visualized. When analyzed in this way, chromosomes of normal mouse cells show characteristic undercondensed centromeric regions. We have found that the centromeric regions of chromosomes from Abelson virus-transformed cells are fully condensed. Abelson virus transforms mouse cells by introducing into them a virally encoded phosphokinase that is expressed constitutively. Thus, we propose that the condensation of centromeric chromatin is a result of overphosphorylation by the Abelson virus phosphokinase, and that the centromeric region is the relevant target of overphosphorylation in transformed cell growth.     

Detection, by direct lymphocyte immunocytotoxicity, of antigens in cells transformed by Rous virus]

The hamster cells transformed by the Rous Sarcoma Virus (V.S.R.) evidenced surface antigenic alterations that were detected by a method of cellular mediated immunocytotoxicity. Immune hamster lymphocytes were added to tritiated proline prelabeled target cells. These lymphocytes were able to lyse specifically the V.S.R. transformed cells.     

Endothelial cells as antigen-presenting cells: role in human transplant rejection

The immunological properties of human endothelial cells suggest they perform a pivotal role in acute and chronic rejection following solid organ transplantation. In this review the basic features of acute and chronic rejection are described as are the cellular and molecular requirements for antigen presentation. Traditionally, antigen-presenting cells are considered to be bone marrow-derived cells. However, these conclusions have been derived from rodent models of allograft rejection where bone marrow-derived passenger leukocytes are the only source of donor major histocompatibility complex (MHC) class II in the grafted organ. In contrast, in humans, virtually all the microvascular and small vessel endothelial cells are ‘constitutively’ positive for MHC class II antigens. The phenotypic properties of human endothelial cells, their response to cytokines and their ability to stimulate resting T cells are described. Unlike bone marrow-derived antigen presenting cells (APCs), which utilise B7/CD28 interactions, human endothelial cells utilise lymphocyte function antigen 3 (LFA3)/CD2 pathways to stimulate T cells. They activate a CD45RO + B7-independent subpopulation of T cells. Their effect on allogeneic T cells is compared with other non-bone marrow-derived cells such as fibroblasts, epithelial cells and smooth muscle cells, which are unable to stimulate resting T cells. Evidence is presented suggesting that release of MHC and non-human leukocyte antigens (HLA) from endothelial cells stimulates an alloantibody and autoimmune response leading to chronic rejection. Received 30 March 1998; received after revision 4 May 1998; accepted 4 May 1998     

Molecular pathogenesis of virus infections

Although a very wide range of viral diseases exists in vertebrates, certain generalizations can be made regarding pathogenetic pathways on the molecular level. The presentation will focus on interactions of virions and their components with target cells. Using coronaviruses as examples the changes in virulence have been traced back to single mutational events; recombination, however, is likely to be an alternative mechanism by which virus-host interactions (e.g. the cell-, organ- or animal species-spectrum) can dramatically change. Receptor molecules are essential for the early interactions during infection and some of these have been identified. Events in the target cell and the host organism are discussed, and wherever possible, aspects of virus evolution and cooperation between infectious agents are highlighted.     

Rosette formation by human T and B lymphocytes in the presence of adrenergic and cholinergic drugs.

It was shown that adrenergic drugs, which increase the intracellular levels of cAMP, inhibit the rosette formation by T-lymphocytes, but stimulate the rosettes produced by B-lymphocytes. Cholinergic drugs, which increase the levels of cGMP, on the contrary, stimulate the formation of rosettes by T-lymphocytes but inhibit those produced by B-lymphocytes.     

The importance of HLA-G expression in embryos,trophoblast cells,and embryonic stem cells

The nonclassical HLA-G molecule is a trophoblast-specific molecule present in almost every pregnancy. It differs from classical HLA class I molecules by the low degree of allelic variants and the high diversity of protein structures. HLA-G is reported to be a tolerogenic molecule that acts on cells of both innate and adaptive immunity. At the maternal–fetal interface HLA-G seems to be responsible largely for the reprogramming of local maternal immune response. This review will focus on the HLA-G gene expression profile in pregnancy, in preimplantation embryos, and in human embryonic stem cells with emphasis on the structural diversity of the HLA-G protein and its potential functional and diagnostic implications.     

Isolation and partial characterization of receptor sites of normal human lymphocytes for lectins]

Glycoproteins of the lymphocyte surface are involved in many membrane mediated events. Their specific carbohydrate determinants might interact with lectins. Purification of macromolecules released from normal human lymphocytes by trypsin was performed using gel filtration and affinity properties for Ricinus sanguineus agglutinin. Some structural and biochemical characteristics are given. Relation to HLA determinants and surface immunoglobulins is discussed.     

Cellular therapy to target neuroinflammation in amyotrophic lateral sclerosis

Neurodegenerative disorders are characterized by the selective vulnerability and progressive loss of discrete neuronal populations. Non-neuronal cells appear to significantly contribute to neuronal loss in diseases such as amyotrophic lateral sclerosis (ALS), Parkinson, and Alzheimer’s disease. In ALS, there is deterioration of motor neurons in the cortex, brainstem, and spinal cord, which control voluntary muscle groups. This results in muscle wasting, paralysis, and death. Neuroinflammation, characterized by the appearance of reactive astrocytes and microglia as well as macrophage and T-lymphocyte infiltration, appears to be highly involved in the disease pathogenesis, highlighting the involvement of non-neuronal cells in neurodegeneration. There appears to be cross-talk between motor neurons, astrocytes, and immune cells, including microglia and T-lymphocytes, which are subsequently activated. Currently, effective therapies for ALS are lacking; however, the non-cell autonomous nature of ALS may indicate potential therapeutic targets. Here, we review the mechanisms of action of astrocytes, microglia, and T-lymphocytes in the nervous system in health and during the pathogenesis of ALS. We also evaluate the therapeutic potential of these cellular populations, after transplantation into ALS patients and animal models of the disease, in modulating the environment surrounding motor neurons from pro-inflammatory to neuroprotective. We also thoroughly discuss the recent advances made in the field and caveats that need to be overcome for clinical translation of cell therapies aimed at modulating non-cell autonomous events to preserve remaining motor neurons in patients.     

Genetic control by the HLA region of the immune response to factor VIII in hemophilic patients]

In 10 sibships of haemophilic A patients, every one including 1 haemophilic brother with an antibody to factor VIII and 1 haemophilic brother without antibody, pattern of HLA segregation was analysed. All sibships were HLA haploidentical or different and not HLA identical. This fact suggested a linkage between the major histocompatibility complex and an "Ir" gene to factor VIII.     

Inhibition of neutrophil-mediated cytotoxicity by alpha 2 macroglobulin

Highly purified human alpha 2 macroglobulin (1.2--10 mg/ml) was shown to inhibit phytohemagglutinin-induced or antibody-induced lysis of chicken erythrocytes by polymorphonuclear neutrophils. Inhibition was not associated with impaired contact between effector and target cells but rather with the antiprotease activity of alpha 2 macroglobulin.     

Oncogenic protein tyrosine kinases

Signals through Kit receptor tyrosine kinase are essential for development of erythrocytes, melanocytes, germ cells, mast cells and interstitial cells of Cajal (ICCs). Mice and rats with a double gene dose of loss-of-function mutations of Kit show depletion of these cells. Although human homozygotes with loss-of-function mutations of Kit have not been reported, gain-of-function mutations of Kit result in development of tumors from mast cells, germ cells and ICCs in humans. The ICC tumors are called gastrointestinal stromal tumors (GISTs), and GISTs are a good target for the Kit inhibitor imatinib mesylate. The interrelationship between the type of Kit gain-of-function mutations and the therapeutic effect of imatinib mesylate has been well characterized in GISTs. Kit is interesting from both a biological and clinical view-point.     

Cell-mediated cytotoxicity by natural killer and killer cells, lipid peroxidation and glutathione

In the course of spontaneous cell-mediated cytotoxicity (SCMC) and antibody-dependent cell-mediated cytotoxicity (ADCC) with human peripheral lymphocytes as effector cells, no lipid peroxidation occurred as measured by the production of ethane and thiobarbituric acid-reactive material. Furthermore, impairment of major cellular defense systems of target cells (K562 cells for SCMC, Chang liver cells for ADCC), by decreasing their glutathione content, had no effect on either lipid peroxidation or the cytotoxic response. These findings indicate that peroxidative damage is not a mechanism of NK and K cell-mediated cytotoxicity.