Abstract
It is now clear that T-cell derived cytokines are important chemical mediators of inflammatory response in allergic diseases. According to their particular mRNA expression and cytokine secretion CD4 positive T lymphocytes were divided into T helper-1 (Thl) and T helper-2 (Th2) type cells (1). Thl type cell clones produce IL-2, IFNγ and lymphotoxin whereas Th2 type cell clones produce IL-4, IL-5, IL-6, IL-10 and IL-13. Both T cell subtypes produce IL-3, GM-CSF and TNFα. Thl and Th2 patterns of cytokine expression exhibit reciprocal inhibition via the release of IFNγ and IL-4 respectively. Due to their release of IL-4, Th2 cell clones are involved in immunoglobin production, specifically of the IgE subclass (2). Th2 cytokines are also instrumental in regulating eosinophil differentiation and survival in vitro via the actions of secreted IL-5, IL-3 and GM-CSF (3). Such effects in vivo may explain the increased tissue survival at the site of allergic inflammation and could also contribute to the inhibition of programmed cell death. It is now clear that T-cells are not the only source of cytokines; eosinophils, mast cells, basophils, macrophages and epithelial cells may also produce cytokines that could be involved in allergic inflammation. Whilst there is growing evidence to suggest that cytokines are playing an important role in initiating and maintaining inflammatory reactions associated with allergic disease in man, to confirm such a role it will be essential to identify the expression of cytokine genes and gene products and to localize cytokine receptors in vivo at the level of the tissue. During the last few years we have been using various methods to identify the in situ expression of cytokines in tissues obtained from asthmatics, allergic rhinitis and individuals with atopic dermatitis. The most widely used techniques to identify the expression of cytokines within tissues are in situ hybridization and immunocytochemistry. In this chapter, the techniques of in situ hybridization and immunocytochemistry will be described in the relation to the localization of cytokine genes and gene products. Examples of the application of these techniques to localize cytokines in tissue obtained from the sites of allergic inflammatory reactions will also be discussed.
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Mosmann TR, Cherwinski H, Bond MW, Gieldin MA & RL Coffman. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol 1986; 136:2348–2357.
Del Prete G, Maggi E, Parronchi P, Chretien I, Tiri A, Macchia D, Ricci M, Banchereau J, De Vries J & S Romagnani. IL-4 is an essential co-factor for the IgE synthesis induced in vitro by human T cell clones and their supernatants. J Immunol 1988; 140:4193–4198.
Clutterbuck EJ, Hirst EMA & CJ Sanderson. Human interleukin-5 (IL-5) regulates the production of eosi-nophils in human bone marrow cultures: comparison and interaction with IL-1, IL-3, IL-6 and GM-CSF. Blood 1989; 73:1504–1512.
Kay AB, Ying S, Varney V, Durham SR, Moqbel R, Wardlaw A & Q Hamid. Messenger RNA expression of the cytokine gene cluster, IL-3, IL-4, IL-5 and GM-CSF in allergen-induced late-phase cutaneous reactions in atopic subjects. J Exp Med 1991; 173:775–778.
Hamid Q, Azzawi M, Ying S, Moqbel R, Wardlaw A, Corrigan C, Bradley B, Durham SR, Collin J, Jeffery P, Quint D & AB Kay. Expression of mRNA for interleukin-5 in mucosal bronchial biopsies from asthma. J Clin Invest 1991; 87:1541–1546.
Robinson DS, Hamid Q, Ying S, Tsicopoulos A, Barkans J, Bentley AM, Corrigan C, Durham SR & AB Kay. Predominant TH2-like bronchoalveolar T-lymphocyte population in atopic asthma. New Engl J Med 1992; 326:298–304.
Pardue ML & JG Gall. Proc Natl Acad Sci USA 1969; 64:600.
Cox KH, DeLeon DV, Angerer LM, Angerer RC. Detection of mRNAs in sea wichin embryos by in situ hybridization using asymmetric RNA probes. Dev Biol 1984; 101:485–502.
Giaid A, Hamid Q, Adams C, Trenghi G & JM Polak. Non-isotopic RNA probes; comparison between different labels and detection systems. Histochemistry 1989; 93:191–196.
Ying S, Durham SR, Jacobson M, Masuyama M, AB Kay & Q Hamid. Phenotype of cells expressing inter-leukin-4 (IL-4), IL-5, IL-2 and interferon-gamma (IFNγ) mRNA in the nasal mucosa following allergen provocation. J Allergy Clin Immunol 1994; 93:270.
Hamid Q, Wharton J, Terenghi G, Hassall C, Aimi J, Taylor K, Nakazato H, Dixon J, Burnstock G & JM Polak. Localization of atrial natiuretic peptide mRNA and immunoreactivity in rat heart and human atrial appendage. Proc Natl Acad Sci USA 1987; 84:7315–7318.
Ploysongsang, M., Humbert, M, Ying S. Yasruel Z, Durham S, Kay, A.B., and Hamid, Q. Increased expression of interleuking-5 receptor gene in asthma. J Allergy Clin Immunol 1995; 95: 279.
Kita H, Ohnishi T, Okubo Y, Weiler D, Abrams JS & GJ Gleich. Granulocyte/macrophage colony-stimulating factor and interleukin-3 release from human peripheral blood esoinophils and neutrophils. J Exp Med 1991; 174:745–748.
Moqbel R, Hamid Q, Ying S, Barkans J, Hartneil A, Tsicopoulos A, Wardlaw AJ & AB Kay. Expression of mRNA and immunoreactivity for the granulocyte/macrophage colony-stimulating factor in activated human eosinophils. J Exp Med 1991; 174:749–752.
Hamid Q, Barkans J, Meng Q, Abrams J, Kay AB & R Moqbel. Human eosinophils synthesize and secrete interleukin-6 in vitro. Blood 1992; 80:1496–1501.
Broide DH, Paine MM & GS Firestein. Eosinophils express interleukin-5 and granulocyte macrophage colony-stimulating factor mRNA at sites of allergic inflammation in asthmatics. J Clin Invest 1992; 90:1414–1424.
Bradding P, Roberts JA, Britten KM, Montefort S, Djukanovic R, Mueller R, Heusser CH, Howarth PH & ST Holgate. Interleukin-4,-5 and-6 and tumor necrosis factor-α in normal and asthmatic airways: Evidence for the human mast cell as a source of these cytokines. Am J Respir Cell Mol Biol 1994;10:471–480.
Plaut M, Pierce JH, Watson CJ, Hanley-Hyde J, Nordan RP & WE Paul. Mast cell lines produce lymphoki-nes in response to cross-linkage of FcεRI or to calcium ionophores. Nature (London) 1989; 339:64–67.
Gordan JR & SJ Galli. Mast cells as a source of both preformed and immunologically inducible TNF-ε/cachetin. Nature (London) 1990; 346:274–276.
Cromwell O, Hamid Q, Corrigan C, Barkans J, Collins P & AB Kay. Expression and generation of IL-6, iL-8 and GM-CSF by human bronchial epithelial cells and enhancement by IL-lβ and TNF-α. Immunology 1992; 77:330–337.
Van Noorden S. Tissue preparation and immunostaining techniques for light microscopy. In: Immunocyto-chemistry. Modem methods and Applications. Eds, Polak JM & S Van Noorden. 1986. Chapter 3: pp26–53.
Frew AJ and Kay AB. The relationship between infiltrating CD4+ lymphocytes, activated eosinophils and the magnitude of the allergen-induced late-phase cutaneous reaction in man. J. Immunol. 1988; 141: 4158–4164.
Tsicopoulos, A., Hamid, Q., Varney V, Ying Sun, Moqbel, R., Durham, S.R., Kay, A.B. Preferential messenger RNA expression of Thl-type cells (IFN-γ+, IL-2+) in classical delayed-type (tuberculin) hypersensitiv-ity reactions in human skin. J. Immuno 1992; 148: 2058–2061.
Durham S, Ying S, Varney V, Jacobson M, Kay AB & Q Hamid. Cytokine messenger RNA expression for IL-3, IL-4, IL-5 and granulocyte/macrophage-colony stimulating factor in the nasal mucosa after local allergen provocation: Relationship to tissue eosinophilia. J Immunol 1992; 142:2390–2394.
Bentley AM, Meng Q, Robinson DS, Hamid Q, Kay AB & SR Durham. Increases in activated T lymphocytes, eosinophils and cytokine mRNA expression for interleukin-5, granulocyte/macrophage colony-stimulating factor in bronchial biopsies after allergen challenge in atopic asthmatics. Am J Respir Cell Mol Biol 1993; 8:35–42.
Robinson D, Hamid Q, Bentley A, Ying S, Kay AB & SR Durham. Activation of CD4+ T cells, increased Th2-type cytokine recruitment in bronchoalveolar lavage after allergen inhalation challenge in patients with atopic asthma. J Allergy Clin Immunol 1993; 92:313–324.
Robinson D, Hamid Q, Ying S, Bentley A, Assoufi B, Durham S & AB Kay. Prednisolone treatment in asthma is associated with modulation of bronchoalveolar lavage cell interleukin-4, interleukin-5 and inter-feron-γ cytokine gene expression. Am Rev Respir Dis 1993b; 148:401–406.
Leung DYM, Martin RJ, Szefler SJ, Sher ER, Ying S, Kay AB & Q Hamid. Dysregulation of interleukin-4, interleukin-5 and interferon γ gene expression in steroid-resistant asthma. J Exp Med 1995; 181:33–40.
Hamid Q, Boguniewicz M & DYM Leung. Differential in situ gene expression in acute versus chronic atopic dermatitis. J Clin Invest 1994; 94:870–876.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1996 Springer Science+Business Media New York
About this chapter
Cite this chapter
Hamid, Q.A., Minshall, E. (1996). In Situ Detection of Cytokines in Allergic Inflammation. In: Sehon, A., HayGlass, K.T., Kraft, D. (eds) New Horizons in Allergy Immunotherapy. Advances in Experimental Medicine and Biology, vol 409. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5855-2_46
Download citation
DOI: https://doi.org/10.1007/978-1-4615-5855-2_46
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7684-2
Online ISBN: 978-1-4615-5855-2
eBook Packages: Springer Book Archive