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Title: Phorbol esters alter the expression of lymphocyte membrane proteins

Abstract

T cell activation via the T cell receptor (T3-Ti complex) by OKT3 results in modulation of the T3-Ti complex, but does not affect T4, T8, or T11 antigen expression. To study the effect of other T cell activators on these T cell membrane antigens, the authors incubated mononuclear cells for 0-3 days with lectins or pharmacologic agents and stained with monoclonal antibodies to their antigens. The median fluorescence intensity (MFI) was measured with a fluorescence activated cell sorter. Activation of PBL with Con A, PHA, calcium ionophore A23187, or with dbcAMP, isoproterenol, or theophyllin had minimal effects on the MFI of T3, T4, T8, or T11. Phorbol myristate acetate (PMA), a protein kinase C activator which stimulates PBL though an alternate pathway, caused a 90-100% reduction of T3 and T4 MFI, a 25% reduction in T8 MFI, and a 400% increase in T11 MFI after 2 days. Addition of A23187 slightly increased these effects. PMA induced a 2-3-fold increase in cell diameter concomitant with the alterations in membrane antigens. These data suggest that T cell activation through pathways not directly linked to the T cell antigen receptor can result in surface antigen expression different from that which follows activation viamore » the T cell receptor.« less

Authors:
;
Publication Date:
Research Org.:
Univ. of Chicago, IL
OSTI Identifier:
5499363
Report Number(s):
CONF-8604222-
Journal ID: CODEN: FEPRA
Resource Type:
Conference
Resource Relation:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States); Journal Volume: 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; LYMPHOCYTES; RECEPTORS; PHORBOL ESTERS; BIOLOGICAL EFFECTS; AMP; ANTIGENS; CELL MEMBRANES; CONCANAVALIN; FLUORESCENCE; LECTINS; MONOCLONAL ANTIBODIES; PHYTOHEMAGGLUTININ; AGGLUTININS; ANIMAL CELLS; ANTIBODIES; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY FLUIDS; CARBOHYDRATES; CARCINOGENS; CELL CONSTITUENTS; CONNECTIVE TISSUE CELLS; ESTERS; HEMAGGLUTININS; LEUKOCYTES; LUMINESCENCE; MATERIALS; MEMBRANE PROTEINS; MEMBRANES; MITOGENS; MUCOPROTEINS; NUCLEOTIDES; ORGANIC COMPOUNDS; POLYSACCHARIDES; PROTEINS; SACCHARIDES; SOMATIC CELLS 560301* -- Chemicals Metabolism & Toxicology-- Cells-- (-1987)

Citation Formats

Reder, A.T., and Antel, J.P. Phorbol esters alter the expression of lymphocyte membrane proteins. United States: N. p., 1986. Web.
Reder, A.T., & Antel, J.P. Phorbol esters alter the expression of lymphocyte membrane proteins. United States.
Reder, A.T., and Antel, J.P. 1986. "Phorbol esters alter the expression of lymphocyte membrane proteins". United States. doi:.
@article{osti_5499363,
title = {Phorbol esters alter the expression of lymphocyte membrane proteins},
author = {Reder, A.T. and Antel, J.P.},
abstractNote = {T cell activation via the T cell receptor (T3-Ti complex) by OKT3 results in modulation of the T3-Ti complex, but does not affect T4, T8, or T11 antigen expression. To study the effect of other T cell activators on these T cell membrane antigens, the authors incubated mononuclear cells for 0-3 days with lectins or pharmacologic agents and stained with monoclonal antibodies to their antigens. The median fluorescence intensity (MFI) was measured with a fluorescence activated cell sorter. Activation of PBL with Con A, PHA, calcium ionophore A23187, or with dbcAMP, isoproterenol, or theophyllin had minimal effects on the MFI of T3, T4, T8, or T11. Phorbol myristate acetate (PMA), a protein kinase C activator which stimulates PBL though an alternate pathway, caused a 90-100% reduction of T3 and T4 MFI, a 25% reduction in T8 MFI, and a 400% increase in T11 MFI after 2 days. Addition of A23187 slightly increased these effects. PMA induced a 2-3-fold increase in cell diameter concomitant with the alterations in membrane antigens. These data suggest that T cell activation through pathways not directly linked to the T cell antigen receptor can result in surface antigen expression different from that which follows activation via the T cell receptor.},
doi = {},
journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:3,
place = {United States},
year = 1986,
month = 3
}

Conference:
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  • Activation of protein kinase C with phorbol ester modifies cyclic AMP production in several anterior pituitary cell systems. In the GH cell line from a rat pituitary tumor, exposure to phorbol 12-myristate 13-acetate (PMA: 100 nM) for 30 minutes significantly reduces vasoactive intestinal peptide (VIP: 100 nM) stimulated adenylate cyclase (AC) activity in subsequent membrane preparations to 62 + 4% of control (n = 6 independent studies). In contrast, these same membrane preparations respond to forskolin (1 ..mu..M) with significantly more activity, 130 +/- 6% of controls (n = 6 independent studies). Finally, phorbol ester does not block an inhibitorymore » hormone input into the AC system; somatostatin (100 nM) reduction of VIP-stimulated AC activity is not significantly different in membrane preparations from PMA treated and control cells (n = 3 independent studies). These other findings lead the authors to propose that protein kinase C can modify several sites in the AC complex in anterior pituitary cells.« less
  • The effect of phorbol esters on T cell adhesion to EC has been studied. The phorbol esters 12-0-tetradecanoyl-phorbol-13-acetate and 4-beta-phorbol-12-13-dibutyrate, but not the biologically inert 4-0-methyl-phorbol-12-13-didecanoate strongly increased the binding of /sup 51/Cr-labeled T cells to human umbilical vein EC monolayers in microtiter wells. Increase in binding was observed at 0.3 ng/ml with maximal enhancement at 50 ng/ml. Both unstimulated and phorbol ester activated T cells displayed a substantially greater binding affinity for EC than for fibroblasts or plastic. Binding enhancement occurred within one minute, with maximal increase after 15 min. Preincubation studies showed that binding enhancement was entirely attributablemore » to an effect on T cells, with no action on EC. Additive binding enhancement was seen when phorbol esters and reagents that increase adhesion by actions on EC (LPS, IL-1 and IFN-..gamma..) were used together. Increase in adhesion of activated T lymphocytes to EC may explain the greater emigration of activated T cells than small resting T cells into inflammatory foci in vivo. The rapid onset of the phorbol effect suggests that this may be an important mechanism for immediate localization of circulating T cells in the cellular immune response, activated, perhaps, at the endothelial blood-tissue interface.« less
  • Both insulin and phorbol esters are known to inhibit the transcription of the gene for P-enolpyruvate carboxykinase (GTP) (PEPCK). The promoter-regulatory region of the PEPCK gene was analyzed for responsive sequences, using a series of chimeric genes constructed by fusing segments of the 5'-flanking region of the PEPCK gene (containing a graded set of deletions) to the structural gene for Herpes virus thymidine kinase (TK). These chimeric genes were transfected into TK-deficient rat hepatoma cells and their level of expression and regulation by insulin, phorbol ester and calcium ionophore was tested. A segment of the PEPCK promoter-regulatory region spanning 2more » kb was shown to be sensitive to regulation by cAMP and glucocorticoids but not to insulin. In these same cells, insulin effectively inhibited the expression of the endogenous PEPCK gene. In a parallel set of experiments, they showed drastic alterations in the expression of the chimeric PEPCK-TK gene when cells were exposed to phorbol ester and/or calcium ionophore. These findings suggest that functional sequences required for insulin regulation of the PEPCK gene expression are 3' to the promoter-regulatory region of the gene.« less
  • The phorbol 12-myristate 13-acetate (PMA)-dependent down-regulation of immunoprecipitable protein kinase C was studied in human breast cancer cell lines that display different growth inhibitions toward the tumor promoter. PMA induces translocation of ({sup 35}S)methionine-prelabeled cytosolic protein kinase C to membranes, followed by complete degradation of the enzyme (t{sub {1/2}}, 2 hr). PMA does not affect the protein kinase C synthesis; 20-80% of total protein kinase C of control cells was still immunoprecipitable as membrane-bound 74- and 80-kDa protein kinase C-related polypeptides if cells were allowed to incorporate ({sup 35}S)methionine during PMA exposure for >6 hr. These two proteins lack proteinmore » kinase activity and phorbol ester binding but reveal V8 peptide patterns identical to the active forms of protein kinase C (77/80 kDa) of PMA-untreated cells. The amounts of the immunoprecipitable membrane-bound 80-kDa protein kinase C-related polypeptide synthesized during the prolonged PMA treatment appear to inversely correlate with the extent of PMA-mediated growth inhibition of the respective human breast cancer cell line. These data suggest that after homologous down-regulation, functional protein kinase C (77/80 kDa) is replaced by a population of membrane-associated but enzymatically inactive protein kinase C-related polypeptides (74/80 kDa).« less
  • The authors have studied the effect of phorbol esters on the induction of class I histocompatibility antigen (HLA) expression by interferons (IFNs) in the T-cell line MOLT-4 and in the MOLT-4 mutant YHHH. Addition of IFN-{alpha} to phorbol 12,13-dibutyrate-pretreated MOLT-4 cells causes a >20-fold increase in the expression of class I HLA, as compared to a 4- to 7-fold IFN-{alpha}-induced increase in control cells. Pretreatment with phorbol 12,13-dibutyrate does not alter the class I HLA response to IFN-{gamma} or the responses of other IFN-induced genes. This effect of phorbol 12,13-dibutyrate reproduces in MOLT-4 cells the phenotype of the mutant YHHH,more » which also displays a selective enhanced class I HLA response to IFN-{alpha}. Pretreatment of YHHH with phorbol 12,13-dibutyrate does not affect any of the responses induced by IFN. These findings suggest the existence of a phorbol ester-sensitive factor, inducible in MOLT-4 and constitutively expressed or modified in YHHH, which operates in the pathway of induction of class I HLA by IFN-{alpha} but not in the pathway used by IFN-{gamma}.« less