Abstract
Cellular inflammation of the nasal mucosa demonstrates a local immune response which plays an important role in allergic rhinitis. The aim of the present study was to characterize nasal mucosal lymphocytes regarding their activation and differentiation state by direct ex vivo flowcytometric analysis. Lymphocytes from the inferior turbinates were isolated by a mechanical method of preparation and, for comparison, from peripheral blood by Ficoll gradient centrifugation. Patients suffering from rhinitis or difficulty in nasal breathing were divided into an allergic (pollen-allergy, n = 13) and non-allergic group (n = 24). Expression of different T- and B-cell markers was determined by flowcytometric analysis. CD4+ T-cells from the nasal mucosa exhibited a memory phenotype (CD45RO+, 97%), were highly activated (CD69+, 43–73%), and showed low expression of the cutaneous lymphocyte antigen (CLA+, 5%). Nasal CD20+ B-lymphocytes expressed significantly higher levels of mIgE and lower levels of CD23 and CD80 than peripheral B-cells. Subsets of CD80+ (4%) and CD86+ (6%) CD20+ B-lymphocytes were identified in the nasal mucosa. No significant differences between allergic and non-allergic individuals were determined. As expected, the data show profound phenotypical differences between circulating peripheral blood and nasal mucosal lymphocytes. Activated memory lymphocytes are present in the nasal mucosa from allergic, but also non-allergic patients and may indicate to a significant role of a local inflammatory state without systemic criteria for allergy. In our study, we show that direct ex vivo isolation of lymphocytes is practicable method and offers a new technique to examine the local nasal allergic immune response using a multiparametric phenotypical analysis.
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Van Cauwenberge P, De Belder T, Vermeiren J, Kaplan A (2003) Global resources in allergy (GLORIA): allergic rhinitis and allergic conjunctivitis. Clin Exp Allergy Rev 3:46–50. doi:10.1046/j.1472-9725.2003.00062.x
Baraniuk JN (1997) Pathogenesis of allergic rhinitis. J Allergy Clin Immunol 99(2):S763–S772. doi:10.1016/S0091-6749(97)70125-8
Dykewicz MS (2003) Rhinitis and sinusitis. J Allergy Clin Immunol 111(Suppl 2):S520–S529. doi:10.1067/mai.2003.82
Durham SR (1998) Mechanisms of mucosal inflammation in the nose and lungs. Clin Exp Allergy 28(Suppl 2):11–16. doi:10.1046/j.1365-2222.1998.0280s6020.x
Bachert C, Prohaska P, Pipkorn U (1990) IgE-positive mast cells on the human nasal mucosal surface in response to allergen exposure. Rhinology 28(3):149–158
Otsuka H, Inaba M, Fujikura T, Kunitomo M (1995) Histochemical and functional characteristics of metachromatic cells in the nasal epithelium in allergic rhinitis: studies of nasal scrapings and their dispersed cells. J Allergy Clin Immunol 96(4):528–536. doi:10.1016/S0091-6749(95)70297-0
Lim MC, Taylor RM, Naclerio RM (1995) The histology of allergic rhinitis and its comparison to cellular changes in nasal lavage. Am J Respir Crit Care Med 151(1):136–144
Bascom R, Pipkorn U, Lichtenstein LM, Naclerio RM (1988) The influx of inflammatory cells into nasal washings during the late response to antigen challenge. Effect of systemic steroid pretreatment. Am Rev Respir Dis 138(2):406–412
Crobach M, Hermans J, Kaptein A, Ridderikhoff J, Mulder J (1996) Nasal smear eosinophilia for the diagnosis of allergic rhinitis and eosinophilic non-allergic rhinitis. Scand J Prim Health Care 14(2):116–121. doi:10.3109/02813439608997081
Hattori H, Okano M, Yoshino T et al (2001) Expression of costimulatory CD80/CD86-CD28/CD152 molecules in nasal mucosa of patients with perennial allergic rhinitis. Clin Exp Allergy 31(8):1242–1249. doi:10.1046/j.1365-2222.2001.01021.x
Smurthwaite L, Walker SN, Wilson DR et al (2001) Persistent IgE synthesis in the nasal mucosa of hay fever patients. Eur J Immunol 31(12):3422–3431. doi :10.1002/1521-4141(200112)31:12<3422::AID-IMMU3422>3.0.CO;2-T
Davidsson A, Karlsson MG, Hellquist HB (1994) Allergen-induced changes of B-cell phenotypes in patients with allergic rhinitis. Rhinology 32(4):184–190
Igarashi Y, Goldrich MS, Kaliner MA, Irani AM, Schwartz LB, White MV (1995) Quantitation of inflammatory cells in the nasal mucosa of patients with allergic rhinitis and normal subjects. J Allergy Clin Immunol 95(3):716–725. doi:10.1016/S0091-6749(95)70177-X
Calderon MA, Lozewicz S, Prior A, Jordan S, Trigg CJ, Davies RJ (1994) Lymphocyte infiltration and thickness of the nasal mucous membrane in perennial and seasonal allergic rhinitis. J Allergy Clin Immunol 93(3):635–643. doi:10.1016/S0091-6749(94)70075-3
Del Prete GF, De Carli M, D’Elios MM et al (1993) Allergen exposure induces the activation of allergen-specific Th2 cells in the airway mucosa of patients with allergic respiratory disorders. Eur J Immunol 23(7):1445–1449. doi:10.1002/eji.1830230707
KleinJan A, Vinke JG, Severijnen LW, Fokkens WJ (2000) Local production and detection of (specific) IgE in nasal B-cells and plasma cells of allergic rhinitis patients. Eur Respir J 15(3):491–497. doi:10.1034/j.1399-3003.2000.15.11.x
Powe DG, Huskisson RS, Carney AS et al (2004) Mucosal T-cell phenotypes in persistent atopic and nonatopic rhinitis show an association with mast cells. Allergy 59(2):204–212. doi:10.1046/j.1398-9995.2003.00315.x
Frotscher B, Anton K, Worm M (2002) Inhibition of IgE production by the imidazoquinoline resiquimod in nonallergic and allergic donors. J Invest Dermatol 119(5):1059–1064. doi:10.1046/j.1523-1747.2002.19531.x
Lukowsky A, Muche JM, Sterry W, Audring H (2000) Detection of expanded T cell clones in skin biopsy samples of patients with Lichen sclerosus et atrophicus by T cell receptor-gamma polymerase chain reaction assays. J Invest Dermatol 115:254–259. doi:10.1046/j.1523-1747.2000.00040.x
Testi R, Phillips JH, Lanier LL (1989) T cell activation via Leu-23 (CD69). J Immunol 143:1123–1128
Young JL, Ramage JM, Hill Gaston JS, Beverly PCL (1997) In vitro responses of human CD45RObrightRA- and CD45RA-RObright T cell subsets and their relationship to memory and naive T cells. Eur J Immunol 27:2383–2389. doi:10.1002/eji.1830270937
Fuhlbrigge RC, Kieffer JD, Armerding D, Kupper TS (1997) Cutaneous lymphocyte antigen is a specialized form of PSGL-1 expressed on skin homing T cells. Nature 389:978–981. doi:10.1038/40166
Rossiter H, Mudde GC, van Reijsen F et al (1994) Disease-related T cells from atopic skin express cutaneous lymphocyte antigen and sialyl Lewis X determinants, and bind to both E-selectin and P-selectin. Eur J Immunol 24:205–210. doi:10.1002/eji.1830240132
Tsicopoulos A, Joseph M (2000) The role of CD23 in allergic disease. Clin Exp Allergy 30:602–605. doi:10.1046/j.1365-2222.2000.00871.x
Corominas M, Mestre M, Bas J et al (1993) CD23 expression on B-lymphocytes and its modulation by cytokines in allergic patients. Clin Exp Allergy 23(7):612–617. doi:10.1111/j.1365-2222.1993.tb00902.x
Corominas M, Mestre M, Bas J, Buendia E (1998) Distinct modulation by interferon-gamma (INF-g) of CD23 expression on B and T lymphocytes of atopic subjects. Clin Exp Immunol 112:276–280. doi:10.1046/j.1365-2249.1998.00570.x
Gould HJ, Sutton BJ, Beavil AJ et al (2003) The biology of IgE and the basis of allergic disease. Annu Rev Immunol 21:579–628. doi:10.1146/annurev.immunol.21.120601.141103
Morikawa H, Nagashima S (2000) The role of costimulatory molecules (B7-1 and B7-2) on allergen-stimulated B cells in cedar pollinosis subjects. Clin Exp Allergy 30(3):383–392. doi:10.1046/j.1365-2222.2000.00713.x
Horiguchi S, Okamoto Y, Chazono H, Sakurai D, Kobayashi K (2005) Expression of membrane-bound CD23 in nasal mucosal B cells from patients with perennial allergic rhinitis. Ann Allergy Asthma Immunol 94(2):286–291
Pawankar RU, Okuda M, Okubo K, Ra C (1995) Lymphocyte subsets of the nasal mucosa in perennial allergic rhinitis. Am J Respir Crit Care Med 152(6 Pt 1):2049–2058
Pawankar RU, Okuda M, Suzuki K, Okumura K, Ra C (1996) Phenotypic and molecular characteristics of nasal mucosal gamma delta T cells in allergic and infectious rhinitis. Am J Respir Crit Care Med 153(5):1655–1665
Santamaria Babi LF, Picker LJ, Perez Soler MT et al (1995) Circulating allergen-reactive T cells from patients with atopic dermatitis and allergic contact dermatitis express the skin-selective homing receptor, the cutaneous lymphocyte-associated antigen. J Exp Med 181(5):1935–1940. doi:10.1084/jem.181.5.1935
Beyer K, Castro R, Feidel C, Sampson HA (2002) Milk-induced urticaria is associated with the expansion of T cells expressing cutaneous lymphocyte antigen. J Allergy Clin Immunol 109(4):688–693. doi:10.1067/mai.2002.123235
Campbell DE, Kemp AS (1999) Cutaneous lymphocyte-associated antigen expression in children with atopic dermatitis and non-atopic healthy children. Pediatr Allergy Immunol 10(4):253–257. doi:10.1034/j.1399-3038.1999.00042.x
Durham SR, Gould HJ, Hamid QA (1997) Local IgE production in nasal allergy. Int Arch Allergy Immunol 113(1–3):128–130
Cameron L, Hamid Q, Wright E et al (2000) Local synthesis of epsilon germline gene transcripts, IL-4, and IL-13 in allergic nasal mucosa after ex vivo allergen exposure. J Allergy Clin Immunol 106(1 Pt 1):46–52. doi:10.1067/mai.2000.107398
Cameron L, Gounni AS, Frenkiel S, Lavigne F, Vercelli D, Hamid Q (2003) S epsilon S mu and S epsilon S gamma switch circles in human nasal mucosa following ex vivo allergen challenge: evidence for direct as well as sequential class switch recombination. J Immunol 171(7):3816–3822
Takhar P, Smurthwaite L, Coker HA et al (2005) Allergen drives class switching to IgE in the nasal mucosa in allergic rhinitis. J Immunol 174(8):5024–5032
Karagiannis SN, Warrack JK, Jennings KH et al (2001) Endocytosis and recycling of the complex between CD23 and HLA-DR in human B cells. Immunology 103(3):319–331. doi:10.1046/j.1365-2567.2001.01238.x
Powe DG, Huskisson RS, Carney AS, Jenkins D, Jones NS (2001) Evidence for an inflammatory pathophysiology in idiopathic rhinitis. Clin Exp Allergy 31(6):864–872. doi:10.1046/j.1365-2222.2001.01106.x
Carney AS, Powe DG, Huskisson RS, Jones NS (2002) Atypical nasal challenges in patients with idiopathic rhinitis: more evidence for the existence of allergy in the absence of atopy? Clin Exp Allergy 32(10):1436–1440. doi:10.1046/j.1365-2745.2002.01465.x
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Christin Wolfram and Claudia Rasche contributed equally to this work.
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Wolfram, C., Rasche, C., Audring, H. et al. Isolation and phenotypic characterization of mucosal nasal lymphocytes by direct ex vivo analysis. Eur Arch Otorhinolaryngol 266, 677–683 (2009). https://doi.org/10.1007/s00405-008-0795-5
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DOI: https://doi.org/10.1007/s00405-008-0795-5