Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.
ADVANCED SEARCH
Log in
Top
BMC Pulmonary Medicine
Published in: BMC Pulmonary Medicine 1/2008

Open Access 01-12-2008 | Research article

Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide

Authors: Letícia Lintomen, Gilberto Franchi, Alexandre Nowill, Antonio Condino-Neto, Gilberto de Nucci, Angelina Zanesco, Edson Antunes

Published in: BMC Pulmonary Medicine | Issue 1/2008

Login to get access

Abstract

Background

Airway eosinophilia is considered a central event in the pathogenesis of asthma. The toxic components of eosinophils are thought to be important in inducing bronchial mucosal injury and dysfunction. Previous studies have suggested an interaction between nitric oxide (NO) and chemokines in modulating eosinophil functions, but this is still conflicting. In the present study, we have carried out functional assays (adhesion and degranulation) and flow cytometry analysis of adhesion molecules (VLA-4 and Mac-1 expression) to evaluate the interactions between NO and CC-chemokines (eotaxin and RANTES) in human eosinophils.

Methods

Eosinophils were purified using a percoll gradient followed by immunomagnetic cell separator. Cell adhesion and degranulation were evaluated by measuring eosinophil peroxidase (EPO) activity, whereas expression of Mac-1 and VLA-4 was detected using flow cytometry.

Results

At 4 h incubation, both eotaxin (100 ng/ml) and RANTES (1000 ng/ml) increased by 133% and 131% eosinophil adhesion, respectively. L-NAME alone (but not D-NAME) also increased the eosinophil adhesion, but the co-incubation of L-NAME with eotaxin or RANTES did not further affect the increased adhesion seen with chemokines alone. In addition, L-NAME alone (but not D-NAME) caused a significant cell degranulation, but it did not affect the CC-chemokine-induced cell degranulation. Incubation of eosinophils with eotaxin or RANTES, in absence or presence of L-NAME, did not affect the expression of VLA-4 and Mac-1 on eosinophil surface. Eotaxin and RANTES (100 ng/ml each) also failed to elevate the cyclic GMP levels above baseline in human eosinophils.

Conclusion

Eotaxin and RANTES increase the eosinophil adhesion to fibronectin-coated plates and promote cell degranulation by NO-independent mechanisms. The failure of CC-chemokines to affect VLA-4 and Mac-1 expression suggests that changes in integrin function (avidity or affinity) are rather involved in the enhanced adhesion.
Appendix
Available only for authorised users
Literature
1.
Spina D, Page CP: Asthma – a need for a rethink?. Trends Pharmacol Sci. 2002, 23: 311-315. 10.1016/S0165-6147(02)02022-9.CrossRefPubMed
2.
Gonglur U, Efeoglu T: Vascular adhesion and transendothelial migration of eosinophil leukocytes. Cell Tissue Res. 2004, 318: 473-482. 10.1007/s00441-004-0925-3.CrossRef
3.
Tachimoto H, Ebisawa M, Bochner BS: Cross-talk between integrins and chemokines that influences eosinophil adhesion and migration. Int Arch Allergy Immunol. 2002, 128: 18-20. 10.1159/000059414.CrossRefPubMed
4.
Lampinen M, Carlson M, Hakansson LD, Venge P: Cytokine-regulated accumulation of eosinophils in inflammatory disease. Allergy. 2004, 59: 793-805. 10.1111/j.1398-9995.2004.00469.x.CrossRefPubMed
5.
Chigaev A, Blenc AM, Braaten JV, Kumaraswamy N, Kepley CL, Andrews RP, Oliver JM, Edwards BS, Prossnitz ER, Larson RS, Sklar AS: Real time analysis of the affinity regulation of alpha 4-integrin. The physiologically activated receptor is intermediate in affinity between resting and Mn2+ or antibody activation. J Biol Chem. 2001, 276: 48670-48678. 10.1074/jbc.M103194200.CrossRefPubMed
6.
Horie S, Kita H: CD11b/CD18 (Mac-1) is required for degranulation of human eosinophils induced by human recombinant granulocyte-macrophage colony-stimulating factor and platelet-activating factor. J Immunol. 1994, 152: 5457-5467.PubMed
7.
Berends C, Dijkhuizen B, Monchy JGR, Dubois AEJ, Gerritsen J, Kauffman HF: Inhibition of PAF-induced expression of CD11b and shedding of L-selectin on human neutrophils and eosinophils by the type IV selective PDE inhibitor, rolipran. Eur Respir J. 1997, 10: 1000-1007. 10.1183/09031936.97.10051000.CrossRefPubMed
8.
Conran N, Ferreira HH, Lorand-Metze I, Thomazzi SM, Antunes E, De Nucci G: Nitric oxide regulates human eosinophil adhesion mechanisms in vitro by changing integrin expression and activity on the eosinophil cell surface. Br J Pharmacol. 2001, 134: 632-638. 10.1038/sj.bjp.0704295.CrossRefPubMedPubMedCentral
9.
Nagata M, Sedgwick JB, Bates ME, Kita H, Busse WW: Eosinophil adhesion to vascular cell adhesion molecule-1 activates superoxide anion generation. J Immunol. 1995, 155: 2194-2202.PubMed
10.
Higashimoto I, Chihara J, Kakazu T, Kawabata M, Nakajima S, Osame M: Regulation of eosinophil cell death by adhesion to fibronectin. Int Arch Allergy Immunol. 1996, 111: 66-69.CrossRefPubMed
11.
Sung K-LP, Yang L, Kim J, Ko D, Stachnick G, Castaneda D, Nayar J, Broide DH: Eotaxin induces a sustained reduction in the functional adhesive state of very late antigen 4 for the connecting segment 1 region of fibronectin. J Allergy Clin Immunol. 2000, 106: 933-940. 10.1067/mai.2000.110797.CrossRefPubMed
12.
Lamkhioued B, Renzi PM, Abi-Younes S, Garcia-Zepada EA, Allakhverdi Z, Ghaffar O, Rothenberg MD, Luster AD, Hamid Q: Increased expression of eotaxin in bronchoalveolar lavage and airways of asthmatics contributes to the chemotaxis of eosinophils to the site of inflammation. J Immunol. 1997, 159: 4593-4601.PubMed
13.
Minshall EM, Cameron L, Lavigne F, Leung DY, Hamilos D, Garcia-Zepada EA, Rothenberg M, Luster AD, Hamid Q: Eotaxin mRNA and protein expression in chronic sinusitis and allergen-induced nasal responses in seasonal allergic rhinitis. Am J Respir Cell Mol Biol. 1997, 17: 683-690.CrossRefPubMed
14.
Yamamoto K, Takanashi S, Hasegawa Y, Kanehira Y, Kaizuka M, Okumura K: Eotaxin level in induced sputum is increased in patients with bronchial asthma and in smokers. Respiration. 2003, 70: 600-605. 10.1159/000075205.CrossRefPubMed
15.
Tateno H, Nakamura H, Minematsu N, Nakajima T, Takahashi S, Nakamura M, Fukunaga K, Asano K, Lilly CM, Yamaguchi K: Plasma eotaxin level and severity of asthma treated with corticosteroid. Respir Med. 2004, 98: 782-790. 10.1016/j.rmed.2004.01.005.CrossRefPubMed
16.
Lilly CM, Nakamura H, Belostotsky OI, Haley KJ, Garcia-Zepeda EA, Luster AD, Israel E: Eotaxin expression after segmental allergen challenge in subjects with atopic asthma. Am J Respir Crit Care Med. 2001, 163: 1669-1675.CrossRefPubMed
17.
Ebisawa M, Yamada T, Bickel C, Klunk D, Schleimer RP: Eosinophil transendothelial migration induced by cytokines. Effect of the chemokine RANTES. J Immunol. 1994, 153: 2153-2160.PubMed
18.
Costa GG, Silva RM, Franco-Penteado CF, Antunes E, Ferreira HHA: Interactions between eotaxin and interleukin-5 in the chemotaxis of primed and non-primed human eosinophils. Eur J Pharmacol. 2007, 566: 200-205. 10.1016/j.ejphar.2006.09.075.CrossRefPubMed
19.
Ferreira HHA, Bevilacqua E, Gagioti SM, De Luca IM, Zanardo RC, Teixeira CE, Sannomiya P, Antunes E, De Nucci G: Nitric oxide modulates eosinophil infiltration in antigen-induced airway inflammation in rats. Eur J Pharmacol. 1998, 358: 253-259. 10.1016/S0014-2999(98)00575-5.CrossRefPubMed
20.
Zanardo RC, Costa E, Ferreira HHA, Antunes E, Martins AR, Murad F, De Nucci G: Pharmacological and immunohistochemical evidence for a functional nitric oxide synthase system in rat peritoneal eosinophils. Proc Natl Acad Sci USA. 1997, 94: 14111-14114. 10.1073/pnas.94.25.14111.CrossRefPubMedPubMedCentral
21.
Thomazzi S, Ferreira HHA, Conran N, De Nucci G, Antunes E: Role of nitric oxide on in vitro human eosinophil migration. Biochem Pharmacol. 2001, 62: 1417-1421. 10.1016/S0006-2952(01)00782-1.CrossRefPubMed
22.
Young SS, Ritacco G, Skeans S, Chapman RW: Eotaxin and nitric oxide production as markers of inflammation in allergic cynomolgus monkeys. Int Arch Allergy Immunol. 1999, 120: 209-217. 10.1159/000024269.CrossRefPubMed
23.
Hanasawa T, Antuni JD, Kharitonov SA, Barnes PJ: Intranasal administration of eotaxin increases nasal eosinophils and nitric oxide in patients with allergic rhinitis. J Allergy Clin Immunol. 1999, 105: 58-64. 10.1016/S0091-6749(00)90178-7.CrossRef
24.
Feder LS, Stelts D, Chapman RW, Manfra D, Crawley Y, Jones H, Minnicozzi M, Fernandez X, Paster P, Egan RW, Kreutner W, Kung TT: Role of nitric oxide on eosinophilic lung inflammation in allergic mice. Am J Respir Cell Mol Biol. 1997, 17: 436-442.CrossRefPubMed
25.
Iijima H, Duguet A, Eum S-Y, Hamid Q, Eidelman DH: Nitric oxide and protein nitration are eosinophil dependent in allergen-challenged mice. Am J Respir Crit Care Med. 2001, 163: 1233-1240.CrossRefPubMed
26.
Trifilieff A, Fujitani Y, Mentz F, Dugas B, Fuentes M, Bertrand C: Inducible nitric oxide synthase inhibitors suppress airway inflammation in mice through down-regulation of chemokine expression. J Immunol. 2000, 165: 1526-1533.CrossRefPubMed
27.
Frank S, Kampfer H, Wetzler C, Satllmeyer B, Pfeilschifter : Large induction of the chemotactic cytokine RANTES during cutaneous wound repair: a regulatory role for nitric oxide in keratinocyte-derived RANTES expression. Biochem J. 2000, 347: 265-273. 10.1042/0264-6021:3470265.CrossRefPubMedPubMedCentral
28.
Sato E, Simpson KL, Grisham MB, Koyama S, Robbins RA: Effects of reactive oxygen and nitrogen metabolites on eotaxin-induced eosinophil chemotactic activity in vitro. Am J Respir Cell Mol Biol. 2000, 22: 61-67.CrossRefPubMed
29.
Hansel TT, De Vries IJ, Iff T, Rihs S, Wandzilak M, Betz S, Blaser K, Walker C: An improved immunomagnetic procedure for the isolation of highly purified human blood eosinophils. J Immunol Meth. 1991, 145: 105-110. 10.1016/0022-1759(91)90315-7.CrossRef
30.
Anwar AR, Walsh GM, Cromwell O, Kay AB, Wardlaw AJ: Adhesion to fibronectin primes eosinophils via alpha 4 beta 1 (VLA-4). Immunology. 1994, 82: 222-228.PubMedPubMedCentral
31.
Kroegel C, Yukawa T, Dent G, Venge P, Chung KF, Barnes PJ: Stimulation of degranulation from human eosinophils by platelet-activating factor. J Immunol. 1989, 142: 3518-3526.PubMed
32.
Adamko DJ, We Y, Gleich GJ, Lacy P, Moqbel R: The induction of eosinophil peroxidase release: improved methods of measurement and stimulation. J Immunol Methods. 2004, 291: 101-108. 10.1016/j.jim.2004.05.003.CrossRefPubMed
33.
Ying S, Meng Q, Zeibecoglou K, Robinson DS, Macfarlane A, Humbert M, Kay AB: Eosinophil chemotactic chemokines (eotaxin, eotaxin-2, RANTES, monocyte chemoattractant protein-3 (MCP-3), and MCP-4), and C-C chemokine receptor 3 expression in bronchial biopsies from atopic and nonatopic (Intrinsic) asthmatics. J Immunol. 1999, 163: 6321-6329.PubMed
34.
Wardlaw AJ: Molecular basis for selective eosinophil trafficking in asthma: A multistep paradigm. Allergy Clin Immunol. 1999, 104: J917-926. 10.1016/S0091-6749(99)70069-2.CrossRef
35.
Ono SJ, Nakamura T, Ohbayashi M, Dawson M, Toda M: Chemokines: Role in leukocyte development, trafficking, and effector function. J Allergy Clin Immunol. 2003, 111: 1185-1199. 10.1067/mai.2003.1594.CrossRefPubMed
36.
Gale LM, McColl SR: Chemokines: extracellular messengers for all occasions?. Bioessays. 1999, 21: 17-28. 10.1002/(SICI)1521-1878(199901)21:1<17::AID-BIES3>3.0.CO;2-4.CrossRefPubMed
37.
Jia G-Q, Gonzalo J-A, Hidalgo A, Wagner D, Cybulsky M, Gutierrez-Ramos JC: Selective eosinophil transendothelial migration triggered by eotaxin via modulation of Mac-1/ICAM-1 and VLA-4/VCAM-1 interactions. Int Immunol. 1999, 11: 1-10. 10.1093/intimm/11.1.1.CrossRefPubMed
38.
Liu J, Munõz NM, Meliton AY, Zhu X, Lambertino AT, Xu C, Myo S, Myou S, Boetticher E, Johnson M, Leff AR: β2-Integrin adhesion caused by eotaxin but not IL-5 is blocked by PDE-4 inhibition and β2-adrenoceptor activation in human eosinophils. Pulm Pharmacol Ther. 2004, 17: 73-79. 10.1016/j.pupt.2003.10.005.CrossRefPubMed
39.
Kubes P, Suzuki M, Granger DN: Nitric oxide: an endogenous modulator of leukocyte adhesion. Proc Natl Acad Sci USA. 1991, 88: 4651-4655. 10.1073/pnas.88.11.4651.CrossRefPubMedPubMedCentral
40.
Giembycz MA, Lindsay MA: Pharmacology of the eosinophil. Pharmacol Rev. 1999, 51: 213-340.PubMed
41.
Del Pozo MA, Sanchez-Mateos P, Sanchez-Madrid F: Cellular polarization induced by chemokines: a mechanism for leukocyte recruitment?. Immunol Today. 1996, 17: 127-131. 10.1016/0167-5699(96)80604-9.CrossRefPubMed
42.
Sano H, Zhu X, Sano A, Boetticher EE, Shioya T, Jacobs B, Munoz NM, Leff AR: Extracellular signal-regulated kinase 1/2-mediated phosphorylation of cytosolic phospholipase A2 is essential for human eosinophil adhesion to fibronectin. J Immunol. 2001, 166: 3515-3521.CrossRefPubMed
43.
Conran N, Gambero A, Ferreira HH, Antunes E, de Nucci G: Nitric oxide has a role in regulating VLA-4-integrin expression on the human neutrophil cell surface. Biochem Pharmacol. 2003, 66: 43-50. 10.1016/S0006-2952(03)00243-0.CrossRefPubMed
44.
Canalli AA, Conran N, Fattori A, Saad ST, Costa FF: Increased adhesive properties of eosinophils in sickle cell disease. Exp Hematol. 2004, 32: 728-734. 10.1016/j.exphem.2004.04.010.CrossRefPubMed
45.
Sedgwick JB, Jansen KJ, Kennedy JD, Kita H, Busse WW: Effects of the very late adhesion molecule 4 antagonist WAY103 on human peripheral blood eosinophil vascular cell adhesion molecule 1-dependent functions. J Allergy Clin Immunol. 2005, 116: 812-819. 10.1016/j.jaci.2005.07.003.CrossRefPubMed
46.
Marcondes S, Cardoso MH, Morganti RP, Thomazzi SM, Lilla S, Murad F, De Nucci G, Antunes E: Cyclic GMP-independent mechanisms contribute to the inhibition of platelet adhesion by nitric oxide donor: a role for alpha-actinin nitration. Proc Natl Acad Sci USA. 2006, 103: 3434-3439. 10.1073/pnas.0509397103.CrossRefPubMedPubMedCentral
47.
Rothenberg ME, Hogan SP: The eosinophil. Annu Rev Immunol. 2006, 24: 147-174. 10.1146/annurev.immunol.24.021605.090720.CrossRefPubMed
48.
Filley WV, Holley KE, Kephart GM, Gleich GJ: Identification by immunofluorescence of eosinophil granule major basic protein in lung tissues of patients with bronchial asthma. Lancet. 1982, 2: 11-16. 10.1016/S0140-6736(82)91152-7.CrossRefPubMed
49.
Fujimoto K, Kubo K, Matsuzawa Y, Sekiguchi M: Eosinophil cationic protein levels in induced sputum correlate with the severity of bronchial asthma. Chest. 1997, 112: 1241-1247. 10.1378/chest.112.5.1241.CrossRefPubMed
50.
Horie S, Gleich GJ, Kita H: Cytokines directly induce degranulation and superoxide production from human eosinophils. J Allergy Clin Immunol. 1996, 98: 371-381. 10.1016/S0091-6749(96)70161-6.CrossRefPubMed
51.
Fujisawa T, Kato Y, Nagase H, Atsuta J, Terada A, Iguchi K, Kamiya H, Morita Y, Kitaura M, Kawasaki H, Yoshie O, Hirai K: Chemokines induce eosinophil degranulation through CCR3. J Allergy Clin Immunol. 2000, 106: 507-513. 10.1067/mai.2000.108311.CrossRefPubMed
52.
El-Shazly A, Masuyama K, Nakano K, Eura M, Samejima Y, Ishikawa T: Human eotaxin induces eosinophil-derived neurotoxin release from normal human eosinophils. Int Arch Allergy Immunol. 1998, 117: 55-58. 10.1159/000053573.CrossRefPubMed
53.
Badewa AP, Hudson CE, Heiman AS: Regulatory effects of eotaxin, eotaxin-2, and eotaxin-3 on eosinophil degranulation and superoxide anion generation. Exp Biol Med (Maywood). 2002, 227 (8): 645-651.
54.
Salvemini D, Masini E, Pistelli A, Mannaioni PF, Vane JR: Nitric oxide: a regulatory mediator of mast cell reactivity. J Cardiovasc Pharmacol. 1991, 17: S258-10.1097/00005344-199100001-00046.CrossRef
55.
Kimura M, Mitani H, Bandoh T, Totsuka T, Hayashi S: Mast cell degranulation in rat mesenteric venule: effects of L-NAME, methylene blue and ketotifen. Pharmacol Res. 1999, 39: 397-402. 10.1006/phrs.1999.0451.CrossRefPubMed
56.
Clark K, Simson L, Newcombe N, Koskinen AML, Mattes J, Lee NA, Lee JJ, Dent LA, Matthaei KI, Foster PS: Eosinophil degranulation in the allergic lung of mice primarily occurs in the airway lumen. J Leukoc Biol. 2004, 75: 1001-1009. 10.1189/jlb.0803391.CrossRefPubMed
Metadata
Title
Human eosinophil adhesion and degranulation stimulated with eotaxin and RANTES in vitro: Lack of interaction with nitric oxide
Authors
Letícia Lintomen
Gilberto Franchi
Alexandre Nowill
Antonio Condino-Neto
Gilberto de Nucci
Angelina Zanesco
Edson Antunes
Publication date
01-12-2008
Publisher
BioMed Central
Published in
BMC Pulmonary Medicine / Issue 1/2008
Electronic ISSN: 1471-2466
DOI
https://doi.org/10.1186/1471-2466-8-13

Other articles of this Issue 1/2008

BMC Pulmonary Medicine 1/2008 Go to the issue

Research article

Influence of the incremental step size in work rate on exercise response and gas exchange in patients with pulmonary hypertension

Research article

Exhaled and arterial levels of endothelin-1 are increased and correlate with pulmonary systolic pressure in COPD with pulmonary hypertension

Research article

Predictors of diagnostic yield in bronchoscopy: a retrospective cohort study comparing different combinations of sampling techniques

Research article

Do airway metallic stents for benign lesions confer too costly a benefit?

Research article

Improvement in symptoms and pulmonary function of asthmatic patients due to their treatment according to the Global Strategy for Asthma Management (GINA)

Research article

Predictors of long time survival after lung cancer surgery: A retrospective cohort study
Obesity Clinical Trial Summary

At a glance: The STEP trials

Read more

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits