The Effects of Inhaled Interferon Gamma in Normal Human Airways

Abstract The physiological role of the multidrug resistance P-glycoprotein (P- gp), which is expressed by normal human T lymphocytes, is still largely unknown. To investigate whether or not P-gp is involved in the transport of cytokines, peripheral blood lymphocytes were stimulated with phytohemagglutinin (PHA) in the absence or presence of P-gp inhibitors, and concentrations of cytokines (interleukin-2 [IL-2], IL- 4, IL-6, interferon-gamma [IFN-gamma]) in the supernatants of these cultures were quantitated by enzyme-linked immunosorbent assay. P-gp inhibitors included verapamil (Ver), tamoxifen (Tmx), and the P-gp specific monoclonal antibody UIC2. Release of IL-2 was significantly suppressed by these inhibitors at concentrations that were also effective in blocking efflux of Rhodamine-123 from normal T lymphocytes. IL-2 mRNA expression in lymphocytes was not different between PHA control and the cultures with P-gp inhibitors. Ver and Tmx did not interfere with T-cell activation as determined by CD25 and CD69 expression. In a nonhematological model, the P-gp expressing HCT-8 adenocarcinoma cell line, exogenously added IL-2 was shown to exert an inhibitory effect on P-gp mediated Rhodamine-123 efflux. In addition, transepithelial transport of IL-2 by electrophysiologically tight and polarized HCT-8 monolayers was examined. A time-dependent flux of IL-2 across dense monolayers, which was partially inhibited by Ver, was observed. We also investigated whether or not P-gp inhibitors suppressed release of other cytokines produced by activated T cells (IL- 4, IL-6, IFN-gamma). Release of IL-4 and IFN-gamma was significantly inhibited by Ver, Tmx, and UIC2; however, release of IL-6 remained unaffected. These data show P-gp mediated transmembrane flux of IL-2 in T lymphocytes and HCT-8 cells. We conclude that P-gp participates in the transport of cytokines (IL-2, IL-4, and IFN-gamma) in normal peripheral T lymphocytes.

Download Full-text

The Effect of In Vivo Interferon-Gamma on the Distribution of LFA-1 and ICAM-1 in Normal Human Skin

Journal of Investigative Dermatology ◽

10.1111/1523-1747.ep12284016 ◽

1989 ◽

Vol 93 (4) ◽

pp. 439-442 ◽

Cited By ~ 86

Author(s):

Jonathan N.W.N. Barker ◽

Michael H Allen ◽

Donald M MacDonald

Keyword(s):

Human Skin ◽

Interferon Gamma ◽

Normal Human Skin ◽

Normal Human

Download Full-text

Plasma Antiviral Activity and Interferon-gamma Production by Superantigen-stimulated Lymphocytes during Normal Human Pregnancy

American Journal Of Reproductive Immunology ◽

10.1111/j.8755-8920.2001.450404.x ◽

2001 ◽

Vol 45 (4) ◽

pp. 217-225 ◽

Cited By ~ 1

Author(s):

GIOVANNI GRASSO ◽

LAURETTA MASSAI ◽

PIERLUIGI MIGLIACCIO ◽

ENRICO PICCIOLINI ◽

MICHELA MUSCETTOLA

Keyword(s):

Antiviral Activity ◽

Interferon Gamma ◽

Human Pregnancy ◽

Normal Human

Download Full-text

Immunohistochemical and ultrastructural studies of basal cells, Clara cells and bronchiolar cuboidal cells in normal human airways

Pathology International ◽

10.1111/j.1440-1827.1998.tb03865.x ◽

1998 ◽

Vol 48 (12) ◽

pp. 944-953 ◽

Cited By ~ 47

Author(s):

Yichiko Nakajima ◽

Oichi Kawanami ◽

Enjing Jin ◽

Mohammad Ghazizadeh ◽

Mitsuyoshi Honda ◽

...

Keyword(s):

Basal Cells ◽

Clara Cells ◽

Ultrastructural Studies ◽

Normal Human ◽

Human Airways

Download Full-text

Characterization of mucins from cultured normal human tracheobronchial epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.2000.278.6.l1118 ◽

2000 ◽

Vol 278 (6) ◽

pp. L1118-L1128 ◽

Cited By ~ 53

Author(s):

David J. Thornton ◽

Thomas Gray ◽

Paul Nettesheim ◽

Marj Howard ◽

Ja Seok Koo ◽

...

Keyword(s):

Charge Density ◽

Gradient Centrifugation ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Mrna Levels ◽

Early Passage ◽

Respiratory Mucin ◽

Normal Human ◽

Human Airways

Early-passage normal human tracheobronchial epithelial (NHTBE) cells grown in air-liquid interface cultures in medium containing retinoids differentiate into a mucociliary epithelium over a 2- to 3-wk period and express increasing mRNA levels of the airway mucin genes MUC5AC and MUC5Bas the cultures age; the levels of MUC2 mRNA were very low throughout the study. Using specific antibodies to MUC5AC and MUC5B mucins, we noted a gradual increase in these two mucins in the intracellular and apically secreted pools as a function of time. A low level of MUC2 mucin was detected, which did not change with time. The intracellular and apically secreted mucins isolated from day 14and day 21 cultures by density gradient centrifugation were similar in density to those previously isolated from human respiratory mucus secretions. The sedimentation rate of the apically secreted mucins indicated that they were highly oligomerized, polydisperse macromolecules similar to those previously documented from in vivo secretions. In contrast, the cell-associated mucins from the cultured NHTBE cells were much smaller, possibly only monomers and dimers. Anion-exchange chromatography detected no differences in charge density between the reduced and carboxymethylated cell-associated and secreted forms of the MUC5AC and MUC5B mucins. The MUC5AC mucin was of similar charge density to its in vivo counterpart; however, MUC5B was more homogeneous than that found in vivo. Finally, evidence is presented for an intracellular NH2-terminal cleavage of the MUC5B mucins. These studies indicate that the mucins produced by cultured NHTBE cells are similar to those found in human airways, suggesting that this cell culture model is suited for studies of respiratory mucin biosynthesis, processing, and assembly.

Download Full-text

MEASUREMENT OF NORMAL HUMAN AIRWAYS RESPONSE TO β-ADRENOCEPTOR STIMULATION USING A FORCED OSCILLATION TECHNIQUE

Journal of Clinical Pharmacy and Therapeutics ◽

10.1111/j.1365-2710.1991.tb00302.x ◽

1991 ◽

Vol 16 (3) ◽

pp. 187-191 ◽

Cited By ~ 2

Author(s):

B.J. Lipworth ◽

R. A. Clark ◽

D. G. McDevitt

Keyword(s):

Forced Oscillation ◽

Forced Oscillation Technique ◽

Normal Human ◽

Human Airways

Download Full-text

Involvement of P-glycoprotein in the transmembrane transport of interleukin-2 (IL-2), IL-4, and interferon-gamma in normal human T lymphocytes

Blood ◽

10.1182/blood.v88.5.1747.bloodjournal8851747 ◽

1996 ◽

Vol 88 (5) ◽

pp. 1747-1754 ◽

Cited By ~ 1

Author(s):

J Drach ◽

A Gsur ◽

G Hamilton ◽

S Zhao ◽

J Angerler ◽

...

Keyword(s):

T Lymphocytes ◽

Interferon Gamma ◽

T Cell Activation ◽

Interleukin 2 ◽

Rhodamine 123 ◽

Enzyme Linked Immunosorbent Assay ◽

Ifn Gamma ◽

Human T Lymphocytes ◽

P Glycoprotein ◽

Normal Human

The physiological role of the multidrug resistance P-glycoprotein (P- gp), which is expressed by normal human T lymphocytes, is still largely unknown. To investigate whether or not P-gp is involved in the transport of cytokines, peripheral blood lymphocytes were stimulated with phytohemagglutinin (PHA) in the absence or presence of P-gp inhibitors, and concentrations of cytokines (interleukin-2 [IL-2], IL- 4, IL-6, interferon-gamma [IFN-gamma]) in the supernatants of these cultures were quantitated by enzyme-linked immunosorbent assay. P-gp inhibitors included verapamil (Ver), tamoxifen (Tmx), and the P-gp specific monoclonal antibody UIC2. Release of IL-2 was significantly suppressed by these inhibitors at concentrations that were also effective in blocking efflux of Rhodamine-123 from normal T lymphocytes. IL-2 mRNA expression in lymphocytes was not different between PHA control and the cultures with P-gp inhibitors. Ver and Tmx did not interfere with T-cell activation as determined by CD25 and CD69 expression. In a nonhematological model, the P-gp expressing HCT-8 adenocarcinoma cell line, exogenously added IL-2 was shown to exert an inhibitory effect on P-gp mediated Rhodamine-123 efflux. In addition, transepithelial transport of IL-2 by electrophysiologically tight and polarized HCT-8 monolayers was examined. A time-dependent flux of IL-2 across dense monolayers, which was partially inhibited by Ver, was observed. We also investigated whether or not P-gp inhibitors suppressed release of other cytokines produced by activated T cells (IL- 4, IL-6, IFN-gamma). Release of IL-4 and IFN-gamma was significantly inhibited by Ver, Tmx, and UIC2; however, release of IL-6 remained unaffected. These data show P-gp mediated transmembrane flux of IL-2 in T lymphocytes and HCT-8 cells. We conclude that P-gp participates in the transport of cytokines (IL-2, IL-4, and IFN-gamma) in normal peripheral T lymphocytes.

Download Full-text