Carl H June - Storming Media

The Protein Tyrosine Kinase Inhibitor Herbimycin A, but not Genistein, Specifically Inhibits Signal Transduction by the T Cell Antigen Receptor

Jan 1992

12 pages

Authors: Martha Graber; Carl H June; Lawrence E Samelson; Arthur Weiss; NAVAL MEDICAL RESEARCH INST BETHESDA MD

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Several lines of evidence implicate a regulatory tyrosine phosphorylation in the activation of phospholipase C (PLC) by the T cell antigen receptor (TCR). These include studies using inhibitors of protein tyrosine kinases (PTKs). In Jurkat T cells expressing the heterologous human muscarinic receptor (HM1), PLC activity can be induced by either the TCR or HM1. HM1 activates PLC via a guanine nucleotide binding protein. We have studied the selectivity of ...

Expression of the Human Epidermal Growth Factor Receptor in a Murine T- Cell Hybridoma: A Transmembrane Protein Tyrosine Kinase Can Synergize with the T-Cell Antigen Receptor

Jan 1992

7 pages

Authors: Ian C Kennedy; Carl H June; John J O'Shea; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Variations in Thymocyte Susceptibility to Clonal Deletion During Ontogeny: Implications for Neonatal Tolerance

15-Jul-1991

7 pages

Authors: Charles M Zacharchuk; Mladen Mercep; Carl H June; Allan M Weissman; Jonathan D Ashwell; NAVAL MEDICAL RESEARCH INST BETHESDA MD

For mature T cells, the predominant outcomes of TCR occupancy include secretion of lymphokines and entry into the growth cycle. For many thymocytes, however, receptor occupancy leads to clonal inactivation. This process, known as negative selection, is fundamental to the establishment of immune self- tolerance. Two types of inactivation have been observed, clonal anergy and clonal deletion. Clonal anergy occurs when potentially autoreactive cells persist but fail to respond to ...

An Inducible Cytoplasmic Factor (AU-B) Binds Selectively to AUUUA multimers in the 3' Untranslated Region of Lymphokine mRNA.

Jun 1991

9 pages

Authors: Paul R Boujanen; Bronislawa Petryniak; Carl H June; Craig B Thompson; Tullia Lindstein; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Although the primary control of eukaryotic gene expression occurs at the transcriptional level, it has become clear that posttranscriptional mechanisms also play important roles in gene regulation. For instance, the efficiency of nuclear RNA splicing and processing has been suggested to play a regulatory role in the expression of some genes (21). Within the cytoplasm, selective mRNA compartmentalization, translation, and degradation, can each influence the final level of gene expression. ...

Bimodal Effect of Phorbol Ester on B Cell Activation.

05 Mar 1991

7 pages

Authors: James J Mond; Nili Feuerstein; Carl H June; Anil K Balapure; Robert I Glazer; NAVAL MEDICAL RESEARCH INST BETHESDA MD

The role of protein kinase C PKC in B cell activation is controversial. These studies were undertaken to determine whether protein kinase C has a stimulatory of inhibitory role in b cell activation. We found that treatment of B cells for a short period of time (30 min) with the PKC activator phorbol 12,13-dibutyrate (PDBU) primed the cells for enhanced proliferative responses to anti-immunoglobulin (anti-Ig) antibody whereas treatment for a ...

Signal Transduction in T Cells

Jan-1991

8 pages

Authors: Carl H June; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Rapid progress was during the past year in the delineation of the nature of the initial biochemical events triggered by the T-cell antigen receptor. Antigen-mediated activation of phospholipase C was demonstrated to require protein tyrosine phosphorylation and, most surprising, activation of the Ras family of signal transduction molecules was shown to closely follow simulation of the T-cell antigen receptor. Major controversy continues over which events are relevant to the various ...

Inhibition of T Cell Receptor Expression and Function in Immature CD4+CD8+ Cells by CD4

28 Sep 1990

5 pages

Authors: Toshinori Nakayama; Carl H June; Terry I Munitz; Michael Sheard; Susan A McCarthy; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Most immature CD4+CD8+ thymocytes express only a small number of T cell receptor (TCR) molecules on their surface, and the TCR molecules they do express are only marginally capable of transducing intracellular signals. TCR expression and function was not intrinsically low in immature CD4+CD8+ thymocytes, but was found to be actively inhibited by CD4-mediated signals. Indeed, release of CD4+CD8+ thymocytes from CD4-mediated signals resulted in significant increases in both TCR ...

The Geonomic Organization of the CD28 Gene. Implications for the Regulation of CD28 mRNA Expression and Heterogeneity

01 Jul 1990

11 pages

Authors: Kelvin P Lee; Carol Taylor; Bronislawa Petryniak; Laurence A Turka; Carl H June; NAVAL MEDICAL RESEARCH INST BETHESDA MD

CD28 is a 90-kDa homodimeric glycoprotein present on the surface of a large subset of T cells that appears to play an important role in the modulation of T cells activation. Although a number of physiologic effects associated with CD28 stimulation have been defined, relatively less is known about the structure and expression of the CD28 gene itself. We now show that CD28 is expressed in both Th cells and ...

Reciprocal Expression of Human ETS1 and ETS2 Genes during T-Cell Activation: Regulatory Role for the Protooncogene ETS1

May 1990

6 pages

Authors: Narayan K Bhat; Craig B Thompson; Tullia Lindsten; Carl H June; Shigeyoshi Fujiwara; NAVAL MEDICAL RESEARCH INST BETHESDA MD

The expression of the protooncogenes ETS1 and ETS2 has been studied in purified human T cells activated either by cross-linking of the T-cell receptor-CD3 complex on their cell surface or by direct stimulation with phorbol esters and ionomycin. Our results show that resting T cells express high levels of ETS1 mRNA and protein, while expression of ETS2 is undetectable. Upon T-cell activation, ETS2 mRNA and proteins are induced, while ETS1 ...

CD28 is an Inducible T Cell Surface Antigen that Transduces a Proliferative Signal in CD3(+) Mature Thymocytes

01 Mar 1990

9 pages

Authors: Laurence A Turka; Jeffrey A Ledbetter; Kelvin Lee; Carl H June; Craig B Thompson; NAVAL MEDICAL RESEARCH INST BETHESDA MD

T cells are important regulators of immune surveillance. The antigen specificity of individual T cells is determined by the TCR, a membrane-bound heterodimer that recognizes nominal Silver complexed to self-Major Histocompatibility Complex molecules. The alpha/beta subunits which comprise most TCR are expressed on the cell surface in close association with a series of five invariant proteins referred to as CD3 and together they form the TCR/CD3 complex. During intrathymic development, ...

Increases in Tyrosine Phosphorylation Are Detectable before Phospholipase C Activation after T Cell Receptor Stimulation

01 Mar 1990

10 pages

Authors: Carl H June; Mary C Fletcher; Jeffrey A Ledbetter; Lawrence E Samelson; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Antiphosphotyrosine immunoblots were used to characterize tyrosine phosphorylated proteins after stimulation of the human T Cell Receptor (TCR). Increased tyrosine phosphorylation was evident on at least 12 substrates within 2 min after ligation of the TCR with Monoclonal antibody. Analysis of the time course for increased tyrosine phosphorylation revealed distinct patterns. Increased phosphorylation of 135-kDa and 100-kDa substrates was evident within 5 s, whereas increased phosphorylation of the TCR-zeta-chain required ...

Monovalent Ligands of Complement Receptor 2 Inhibit Whereas Polyvalent Ligands Enhance Anti-Ig-Induced Human B Cell Intracytoplasmic Free Calcium Concentration

01 Mar 1990

7 pages

Authors: George C Tsokos; John D Lambris; Fred D Finkelman; Evangelos D Anastassiou; Carl H June; NAVAL MEDICAL RESEARCH INST BETHESDA MD

We have performed experiments to investigate the role of ligands for complement receptor 2 (CR2) in human B cell activation. Flow microfluorimetry was used to assess changes in free intracytoplasmic calcium concentration Ca(2+) in indo-loaded B cells, immediately after exposure to anti-micron antibody and to monovalent or polyvalent CR2 ligands. As monovalent ligands we used the C3d fragment and synthetic C3 peptides (peptides P14, residues 1201-1214, and P28, residues 1187-1214). ...

Activation of Human B Lymphocytes by Nanogram Concentrations of Anti- IgM-Dextran Conjugates

Jan 1990

7 pages

Authors: Gregory T Rehe; Ildy M Katona; Mark Brunswick; Larry M Wahl; Carl H June; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Surface immunoglobulin (sIg) cross-linking on B lymphocytes by high concentrations of anti-Ig antibody has been used to mimic antigen-stimulated B cell activation. In order to develop a system to study sIg-medicated T cell- independent B cell activation using low concentrations of antigen that are achieved under in vivo conditions, we conjugated monoclonal anti-human IgM antibody (anti-mu) to dextran (molecular weight 2 x 10 to the sixth power) thereby increasing its ...

Different Patterns of Inositol Polyphosphate Production Are Seen in B lymphocytes after Cross-Linking of sIg by Anti-Ig Antibody or by a Multivalent Anti-Ig Antibody Dextran Conjugate

01 Sep 1989

9 pages

Authors: Mark Brunswick; Carl H June; Fred D Finkelman; James J Mond; NAVAL MEDICAL RESEARCH INST BETHESDA MD

The current experiments were undertaken to determine whether the differences observed in the mitogenicity mediated by two agonists were also reflected in different patterns, kinetics and magnitudes of inositol polyphosphate production. Although both dextran conjugated and unconjugated anti-Ig antibodies induced rapid increases in level of I(1,4,5)P3, dextran conjugated anti-IG induced much greater levels of this isomer, which were sustained for significantly longer periods of time. Additional differences were found in ...

Mitogen-Induced Genes are Subject to Multiple Pathways of Regulation in the Initial Stages of T-Cell Activation

Mar 1989

8 pages

Authors: Steven G Irving; Carl H June; Peter F Ziffel; Ulrich Siebenlist; Kathleen Kelly; NAVAL MEDICAL RESEARCH INST BETHESDA MD

The delivery of a mitogenic signal to T cells via any one of several cell surface molecules elicits a variety of intracellular responses, some or all of which regulate subsequent gene expression events. The expression of nine novel mitogen-induced genes in response to various T-cell-activating agents was examined to evaluate the diversity of pathways which regulate such genes. The relative contribution of distinct secondary signals, individually or together, to mitogen-stimulated ...

Transcription of T Cell Antigen Receptor Genes is Induced by Protein Kinase C Activation

01 Sep 1988

7 pages

Authors: Tullia Lindsten; Carl H June; Craig B Thompson; NAVAL MEDICAL RESEARCH INST BETHESDA MD

The regulation of expression of the TCR-alpha and beta genes was stu died in the human T cell tumor line Jurkat. Treatment of the cells with PMA was shown to decrease the surface expression of the TCR-alpha/beta CD3 complex. Subsequent to PMA-induced modulation of the TCR/CD3 complex, increases in the MRNA levels of both the TCR-alpha and -beta genes were observed reaching a maximum 12 h after stimulation. Other T ...

Flow Cytometric Measurement of Cellular Ionized Calcium Concentration

Jan 1988

26 pages

Authors: Carl H June; Peter S Rabinovitch; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Considerable interest has been directed at measuring intracellular ionized calcium concentration (Ca2+)i) in living cells. Calcium has an important role in a number of cellular functions, and perhaps most interestingly, can function to transmit information from the cell membrane to regulate diverse cellular functions. Until the early 1980s it was not possible to measure (Ca2+)i in small intact cells, and attempts to measure cytosolic calcium were restricted mostly to large ...

Multiple Mechanisms Regulate c-myc Gene Expression during Normal T Cell Activation

Jan 1988

9 pages

Authors: Tullia Lindsten; Carl H June; Craig B Thompson; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Quiescent normal human T cells express low levels of steady-state c- myc mRNA as a result of low constitutive promoter utilization, a block to transcriptional elongation within the gene, and rapid degradation of c-myc mRNA in the cytoplasm. Following the activation of the T cell receptor (TCR)/CD3 complex, quiescent T cells are induced to express c-myc mRNA. Two intracellular pathways, one involving protein kinase C activation and the other mediated ...

Induction of Calcium Flux and Enhancement of Cytolytic Activity in Natural Killer Cells by Cross-Linking of the Sheep Erythrocyte Binding Protein (CD2) and the Fc-Receptor (CD16)

15 Sep 1987

9 pages

Authors: Claudio Anasetti; Paul J Martin; Carl H June; Karl E Hellstrom; Jeffrey A Ledbetter; Peter S Rabinovitch; Yoshihisa Morishita; Ingegerd Hellstrom; John A Hansen; NAVAL MEDICAL RESEARCH INST BETHESDA MD

Binding of the anti-cluster of differentiation (CD)2 monoclonal antibody 9-1 causes an increase in the concentration of cytoplasmic-free calcium ((Ca(2+)i) in cultured CD3(-)/CD16(+) natural killer (NK) cells. This response did not occur in cultured CD3(+)/CD16- cytotoxic T lymphocytes (CTL). Anti-CD16 antibodies could partially block the calcium response when NK cells were stimulated with intact antibody 9-1, and antigen-binding fragment F(ab')2 of antibody 9-1 did not produce a calcium response. Thus ...