Antimicrobial Lipids: Novel Innate Defense Molecules are Elevated in Sinus Secretions of Patients with Chronic Rhinosinusitis

Abstract

Background

Airway secretions possess intrinsic antimicrobial properties that contribute to the innate host defense of the respiratory tract. These microbicidal capabilities have largely been attributed to the presence of antibacterial polypeptides. However, recent investigation has indicated that host-derived lipids including cholesteryl esters also exhibit antimicrobial properties. The purpose of this study was to determine whether sinus secretions contain such antimicrobial lipids and to compare the lipid composition in patients with and without chronic rhinosinusitis (CRS).

Methods

Maxillary sinus fluid was obtained via antral lavage from subjects with (seven patients) and without (nine patients) a history of CRS. After specimen collection, total lipid was extracted according to Bligh and Dyer (Bligh EG and Dyer WJ, A rapid method of total lipid extraction and purification, Can J Biochem Physiol 37:911–918, 1959) and lipid profiles were obtained by reverse phase high-performance liquid chromatography on an amide-embedded C18 column. In addition, the neutrophil-specific antimicrobial peptides human neutrophil peptides 1–3 (HNP1–3) were quantified by Western immunoblotting.

Results

Lipids, including cholesteryl esters, were identified in the maxillary sinus secretions of patients with and without CRS. However, levels of lipid composition differed between the two groups with CRS patients exhibiting greater amounts of all classes of lipids, reaching over 10-fold higher concentration when compared with non-CRS patients. This increase was independent of HNP1–3 content.

Conclusion

Sinus secretions of patients with CRS appear to show elevated levels of antimicrobial lipids compared with controls independent from neutrophil influx. This up-regulation suggests that host-derived lipids act as mediators of mucosal immunity in CRS. Further study is necessary to determine if such antimicrobial lipids function alone or synergistically with antibacterial peptides in conferring such inherent microbicidal properties.

Keywords

Antimicrobial cholesteryl chronic esters immunity innate lipids rhinosinusitis

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References

Ganz

. Defensins: Antimicrobial peptides of innate immunity. Nature 3: 710–720, 2003.

Ganz

. Antimicrobial polypeptides. J Leukoc Biol 75: 34–38, 2004.

Medzhitov

, and Janeway

Jr.

Innate immunity.

N Engl J Med 343: 338–344, 2000.

Medzhitov

, and Janeway

Jr.

Innate immune recognition: Mechanisms and pathways.

Immunol Rev 173: 89–97, 2000.

Ooi

E.H.

, Wormald

P.J.

, and Tan

L.W.

. Innate immunity in the paranasal sinuses: A review of nasal host defenses. Am J Rhinol 22: 13–19, 2008.

McCormack

. New concepts in collectin-mediated host defense at the air-liquid interface of the lung. Respirology 11: s7–s10, 2006.

Vandermeer

, Sha

, Kane

A.P.

. Innate immunity of the sinonasal cavity: Expression of messenger RNA for complement cascade components and toll-like receptors. Arch Otolaryngol Head Neck Surg 130: 1374–1380, 2004.

Singh

P.K.

, Tack

B.F.

, McCray

P.B.

. Synergistic and additive killing by antimicrobial factors found in human airway surface liquid. Am J Physiol Lung Cell Mol Physiol 279: L799–L805, 2000.

Cole

A.M.

, Dewan

, and Ganz . Innate antimicrobial activity of nasal secretions. Infect Immun 67: 3267–3275, 1999.

10.

Cole

A.M.

, Liao

, Stuchlik

. Cationic polypeptides are required for antibacterial activity of human airway fluid. J Immunol 169: 6985–6991, 2002.

11.

Nijnik

, and Hancock

R.E.

. The roles of cathelicidin LL-37 in immune defenses and novel clinical applications. Curr Opin Hematol 16: 41–47, 2009.

12.

Herr

, Shaykhiev

, and Bals

. The role of cathelicidin and defensins in pulmonary inflammatory diseases. Expert Opin Biol Ther 7: 1449–1461, 2007.

13.

Thormar

, and Hilmarsson

. The role of microbicidal lipids in host defense against pathogens and their potential as therapeutic agents. Chem Phys Lipids 150: 1–11, 2007.

14.

T.Q.

, Moshkani

, Castillo

. Lipids including cholesteryl linoleate and cholesteryl arachidonate contribute to the inherent antibacterial activity of human nasal fluid. J Immunol 181: 4177–4187, 2008.

15.

Drake

D.R.

, Brogden

K.A.

, Dawson

D.V.

. Antimicrobial lipids at the skin surface. J Lipid Res 49: 4–11, 2008.

16.

Tollin

, Bergsson

, Kai-Larsen

. Vernix caseosa as a multi-component defense system based on polypeptides, lipids, and their interactions. Cell Mol Life Sci 62: 2390–2399, 2005.

17.

Thormar

, Isaacs

C.E.

, Brown

H.R.

. Inactivation of enveloped viruses and killing of cells by fatty acids and monoglycerides. Antimicrob Agents Chemother 31: 27–31.

18.

Isaccs

C.E.

, Kashyap

, Heird

W.C.

. Antiviral and antibacterial lipids in human milk and infant formula feeds. Arch Dis Child 65: 861–864, 1990.

19.

Isaacs

C.E.

, Litov

R.E.

, Marie

, and Thormar

. Addition of lipases to infant formulas produces antiviral and antibacterial activity. J Nutr Biochem 3: 304–308, 1992.

20.

Isaacs

C.E.

, Litov

R.E.

, and Thormar

. Antimicrobial activity of lipids added to human milk, infant formulas, and bovine mild. J Nutr Biochem 6: 362–366, 1995.

21.

Isaacs

C.E.

. Antimicrobial function of milk lipids. Adv Nutr Res 10: 271–285, 2001.

22.

Sun

C.Q.

, O'Connor

C.J.

, and Roberton

. The antimicrobial properties of milkfat after partial hydrolysis by calf pregastric lipase. Chem Biol Interact 140: 185–188, 2002.

23.

Sprong

R.C.

, Hulstein

M.F.

, and van der Meer

. Bactericidal activities of milk lipids. Antimicrob Agents Chemother 45: 1298–1301, 2001.

24.

Rosenfeld

, Andes

, Bhattacharyya

. Clinical practice guidelines: Adult sinusitis. Otolaryngol Head Neck Surg 137 (3 suppl): S1–S31, 2007.

25.

Bligh

E.G.

, and Dyer

W.J.

. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37: 911–918, 1959.

26.

Porter

, Yang

, Yavagal

. Distinct defensin profiles in Neisseria gonorrhoeae and Chlamydia trachomatis urethritis reveal novel epithelial cell-neutrophil interactions. Infect Immun 73: 4823–4833, 2005.

27.

Shen

, Porter

E.M.

, Reynoso

. Human defensin 5 expression in intestinal metaplasia of the upper gastrointestinal tract. J Clin Pathol 58: 687–694, 2005.

28.

Georgel

, Crozat

, Lauth

. A toll-like receptor 2- responsive lipid effector pathway protects mammals against skin infections with gram-positive bacteria. Infect Immun 73: 4512–4521, 2005.

29.

German

J.B.

, and Dillard

C.J.

. Composition, structure, and absorption of milk lipids: A source of energy, fat soluble nutrients and bioactive molecules. Crit Rev Food Sci Nutr 46: 57–92, 2006.

30.

Psaltis

A.J.

, Bruhn

M.A.

, Tan

L.W.

. Nasal mucosa expression of lactoferrin in patients with chronic rhinosinusitis. Laryngoscope 117: 2030–2035, 2007.

31.

Ooi

E.H.

. Wormald PJ, Carney AS, et al. Human cathelicidin antimicrobial peptide is upregulated in the eosinophilic mucus subgroup of chronic rhinosinusitis patients. Am J Rhinol 21: 395–401, 2007.

32.

Kim

S.T.

, Cha

H.E.

, Kim

D.Y.

. Antimicrobial peptide LL-37 is upregulated in chronic nasal inflammatory disease. Acta Otolaryngol 123: 81–85, 2003.

33.

Lee

S.H.

, Kim

J.E.

, Lim

H.H.

. Antimicrobial defensin peptides of the human nasal mucosa. Ann Otol Rhinol Laryngol 111: 135–141, 2002.

34.

Carothers

D.G.

, Graham

S.M.

, Jia

H.P.

. Production of beta defensin antimicrobial peptides by maxillary sinus mucosa. Am J Rhinol 15: 175–179, 2001.

35.

Ramanathan

Jr. , Lee

W.K.

, Dubin

M.G.

. Sinonasal epithelial cell expression of toll-like receptor 9 is decreased in chronic rhinosinusitis with polyps. Am J Rhinol 21: 110–116, 2007.

36.

Ramanathan

Jr. , Lee

W.K.

, Spannhake

E.W.

. Th2 cytokines associated with chronic rhinosinusitis with polyps down-regulate the antimicrobial immune function of human sinonasal epithelial cells. Am J Rhinol 22: 115–121, 2008.

37.

Lane

, Truong-Tran

Q.A.

, Schleimer

R.P.

. Altered expression of genes associated with innate immunity and inflammation in recalcitrant rhinosinusitis with polyps. Am J Rhinol 20: 138–144, 2006.

38.

Arikawa

, Ishibashi

, Kawashima

. Decreased levels of sphingosine, a natural antimicrobial agent, may be associated with vulnerability of the stratum corneum from patients with atopic dermatitis to colonization by Staphylococcus aureus. J Invest Dermatol 119: 433–439, 2002.

39.

Freedman

S.D.

, Blanco

P.G.

, Zaman

M.M.

. Association of cystic fibrosis with abnormalities in fatty acid metabolism. N Engl J Med 350: 560–569, 2004.

40.

Strandvik

, Gronowitz

, Enlund

. Essential fatty acid deficiency in relation to genotype in patients with cystic fibrosis. J Pediatr 139: 650–655, 2001.

41.

Keen

, Olin

A.C.

, Edentoft

. Airway nitric oxide in patients with cystic fibrosis is associated with pancreatic function, pseudomonas infection, and polyunsaturated fatty acids. Chest 131: 1857–1864, 2007.

42.

Coste

T.C.

, Armand

, Lebacq

. An overview of monitoring and supplementation of omega 3 fatty acids in cystic fibrosis. Clin Biochem 40: 511–520, 2007.

43.

Martinez

J.G.

, Waldon

, Huang

. Membrane-targeted synergistic activity of docosahexaenoic acid and lysozyme against pseudomonas aeruginosa. Biochem J 419: 193–200, 2009.