Synthesis of interleukin-1α, interleukin-6, and interleukin-8 by cultured human nasal epithelial cells

doi:10.1016/S0091-6749(94)70055-9

Journal of Allergy and Clinical Immunology

Volume 93, Issue 6, June 1994, Pages 1060-1067

https://doi.org/10.1016/S0091-6749(94)70055-9 Get rights and content

Abstract

Nasal epithelium forms the initial barrier between the environment and the respiratory system and may be a potential source of proinflammatory interleukins, which contribute to the pathophysiology of allergic and nonallergic rhinitis. To explore this possibility, epithelium and cultured human nasal epithelial cells from nasal turbinates of patients undergoing surgery for treatment of upper airway obstruction were examined for the spontaneous expression of interleukin (IL)-1α, IL-1β, IL-6, and IL-8. Human nasal epithelial cell lysates and culture supernatants were assayed by two-site ELISAs specific for IL-1α, IL-1β, IL-6, or IL-8. Maximum concentrations of these cytokines in supernatants ranged from approximately 0.2 to 2 ng/ml for IL-1α, 1.5 to 7 ng/ml for IL-6, and 100 to 3000 ng/ml for IL-8. IL-1α was predominantly cell-associated, whereas most of the IL-8 and all of the IL-6 were detected in the supernatant. Little or no IL-1β was detected by ELISA in the supernatants or cell lysates. Whole tissue turbinates and isolated epithelium were also examined for IL-1β, IL-6, and IL-8 mRNA expression by Northern blot analysis. IL-6 and IL-8 mRNAs were detected, whereas IL-1β mRNA was not. Furthermore, IL-6 and IL-8 release from human nasal epithelial cell cultures was enhanced by addition to the cultures of lipopolysaccharide, and IL-6 release was inhibited by polymyxin B. Thus human nasal epithelium may be a major source of IL-1α, IL-6, and IL-8 in allergic and nonallergic rhinitis. Production of those proinflammatory cytokines by epithelial cells of the nasal and sinus mucosa may contribute to the pathologic and clinical events that occur in these diseases. (J ALLERGY CLIN IMMUNOL 1994;93:1060-7.)

Section snippets

Reagents

Dispase was from Boeringer-Mannheim (Indianapolis, Ind.). Dulbecco's modified Eagle's medium:nutrient mixture Ham's F-12 (DMEM:F-12) containing N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (HEPES) and L-glutamine, ethylenediaminetetraacetic acid, trypsin, phosphate-buffered saline without Ca + + and Mg + +, and penicillin-streptomycin were from M. A. Whitakker Bioproducts (Walkersville, Md.). NuSerum was from Collaborative Research, Inc. (Bedford, Mass.). Fungizone and pooled human AB

RESULTS

Production of IL-1α, IL-1β, IL-6, and IL-8 was first examined in the supernatants and lysates of 4-day HNEC cultures from four donors (Table I). IL-1α was detected in the cell lysates and supernatants of all cultures with median concentrations of 1612 and 170 pg/ml, respectively. In contrast, only one culture contained measurable levels of IL-1β, 337 pg/ml IL-1β in the cell supernatant and 45 pg/ml in the supernatant. This culture also had the highest level of expression of the other cytokines.

DISCUSSION

We have found, by means of ELISA and Northern blot analysis, that primary cultures of HNECs can spontaneously produce IL-1α, IL-6, and IL-8. IL-1α was at its highest levels within cells; this is most likely a reflection of the absence of a signal sequence within the IL-1α mRNA, which enhances protein secretion. ⁸ By contrast, IL-6 and IL-8, which have mRNAs that contain a signal sequence, 10, 12 were found predominately in supernatant (Table I).

Very little or no IL-1β production was detected

Acknowledgements

We thank Mariola Ilnicka for performing the Northern blot analysis, Drs. Elsie Eugui and Simon Lee for critical reading of the manuscript, and Dr. Anthony C. Allison for helpful discussions.

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^☆: From ^aInstitute of Immunology and Biological Sciences, Syntex Research, Palo Alto, Calif.; and ^bDepartment of Medicine, Division of Allergy and Infectious Diseases, University of Washington, Seattle.

^☆☆: Supported in part by National Institutes of Health grant NIH-NIEHS no. ES-05650.

^★: Reprint requests: Leonard C. Altman, MD, Division of Allergy and Infectious Diseases, Department of Medicine, SJ-10, University of Washington, Seattle, WA 98195.

^★★: 1/1/53426

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Synthesis of interleukin-1α, interleukin-6, and interleukin-8 by cultured human nasal epithelial cells☆,☆☆,★,★★

Abstract

Section snippets

Reagents

RESULTS

DISCUSSION

Acknowledgements

Immunol Allergy Clin North Am

J ALLERGY CLIN IMMUNOL

Immunol Allergy Clin North Am

Clin Immunol Immunopathol

Cytokine

Clin Immunol Immunopathol

Chest

J ALLERGY CLIN IMMUNOL

J Biol Chem

Kidney Int

J Biol Chem

Cytokine

Allergic rhinitis

N Engl J Med

The influx of inflammatory cells into nasal washings during the late response to antigen challenge: effect of systemic steroid pretreatment

Am Rev Respir Dis

Allergy

The regulation of allergic response by cytokine receptor networks

ACI News

Interleukin-1 and its biologically related cytokines

Interleukin-1: the first interleukin

Immunol Today

Interleukin-6: an overview

Annu Rev Immunol