Neutrophil necrosis and annexin 1 degradation associated with airway inflammation in lung transplant recipients with cystic fibrosis

BMC Pulmonary Medicine

Keith C Meyer¹  Adnan Abbasi¹  Zhuzai Xiang¹  Francis H C Tsao¹ 

[1] Division of Pulmonary and Critical Care Medicine, Department of Medicine, Medical School, University of Wisconsin, K4/910 Clinical Science Center, 600 Highland Avenue, Madison, WI, 53792-9988, USA

关键词: Neutrophil;    Necrosis;    Lung transplant;    Cystic fibrosis;    Annexin 1;   

Others  :  1161075
DOI  :  10.1186/1471-2466-12-44

 received in 2012-03-07, accepted in 2012-07-28,  发布年份 2012

【 摘 要 】

Background

Neutrophils sequestered in lower respiratory tract secretions in the inflamed lung may undergo apoptosis and/or necrosis and release toxic cellular contents that can injure airways or parenchyma. This study examined the viability of neutrophils retrieved from the proximal airways of lung transplant recipients with bacterial tracheobronchitis.

Methods

Integrity and stability of intracellular proteins in neutrophils from proximal airways and peripheral blood from lung transplant recipients with bacterial tracheobronchitis were analyzed via Western blot analysis and determination of neutrophil viability by morphologic appearance and flow cytometry.

Results

Neutrophils in tracheobronchial secretions from lung transplant recipients with cystic fibrosis who had normal chest radiographic imaging but bronchoscopic evidence of purulent tracheobronchitis post-transplant were necrotic and associated with degradation of intracellular protein annexin 1. The neutrophil influx was compartmentalized to large airways and not detected in peripheral bronchoalveolar airspaces sampled via bronchoalveolar lavage. Peripheral blood neutrophils from healthy subjects cultured in vitro demonstrated that annexin 1 degradation, particularly to a 33 kDa annexin 1 breakdown product (A1-BP), was associated with neutrophil necrosis, but not apoptosis. Although annexin 1 degradation was not specific to neutrophil necrosis, it was a sensitive marker of intracellular protein degradation associated with neutrophil necrosis. Annexin 1 degradation to 33 kDa A1-BP was not observed in peripheral blood neutrophils from healthy subjects, but annexin 1 appeared to be degraded in peripheral blood neutrophils of lung transplant recipients despite a normal morphologic appearance of these cells.

Conclusions

Neutrophils were necrotic from the proximal airways of lung transplant recipients with bacterial tracheobronchitis, and this process may begin when neutrophils are still in the systemic circulation prior to sequestration in inflamed airways. Annexin 1 degradation to 33 kDa A1-BP may be useful as a sensitive marker to detect neutrophil necrosis.

【 授权许可】

   
2012 Tsao et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Bruce MC, Poncz L, Klinger JD, Stern RC, Tomashefski JF, Dearborn DG: Biochemical and pathological evidence for proteolytic destruction of lung connective tissue in cystic fibrosis. Am Rev Respir Dis 1985, 132:529-535.
• [2]Meyer KC, Zimmerman JJ: Neutrophil mediators, pseudomonas and pulmonary dysfunction in cystic fibrosis. J Lab Clin Med 1993, 121:654-661.
• [3]Ratjen F, Döring G: Cystic fibrosis. Lancet 2003, 361:681-689.
• [4]Chmiel JF, Konstan MW: Inflammation and anti-inflammatory therapies for cystic fibrosis. Clin Chest Med 2007, 28:331-346.
• [5]Haslett C: Granulocyte apoptosis and its role in the resolution and control of lung inflammation. Am J Respir Crit Care Med 1999, 160:S5-S11.
• [6]Liu CY, Liu YH, Lin SM, Yu CT, Wang CH, Lin HC, Lin CH, Kuo HP: Apoptotoc neutrophils undergoing secondary necrosis induce human lung epithelial cell detachment. J Biomed Sci 2003, 10:746-756.
• [7]Rydell-Tormanen K, Uller L, Erjefalt JS: Direct evidence of secondary necrosis of neutrophils during intense lung inflammation. Eur Respir J 2006, 28:268-274.
• [8]Naylor EJ, Bakstad D, Biffen M, Thong B, Calverley P, Scott S, Hart CA, Moots RJ, Edwards SW: Haemophilus influenzae induces neutrophil necrosis: a role in chronic obstructive pulmonary disease? Am J Respir Cell Mol Biol 2007, 37:135-143.
• [9]Minematsu N, Shapiro SD: To live and die in the LA (lung airway): mode of neutrophil death and progression of chronic obstructive pulmonary disease. Am J Respir Cell Mol Biol 2007, 37:129-134.
• [10]Meyer KC, Nunley DR, Dauber JH, Iacono AT, Keenan RJ, Cornwell RD, Love RB: Neutrophils, unopposed neutrophil elastase and alpha1-antiprotease defenses following human lung transplantation. Am J Respir Crit Care Med 2001, 164:97-102.
• [11]Tsao FHC, Meyer KC, Chen XM, Rosenthal NS, Hu JP: Degradation of annexin I in bronchoalveolar lavage fluid in patients with cystic fibrosis. Am J Respir Cell Mol Biol 1998, 18:120-128.
• [12]Perretti M: Lipocortin 1 and chemokine modulation of granulocyte and monocyte accumulation in exerimental inflammation. Gen Pharmacol 1998, 31:545-552.
• [13]Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 1951, 193:265-275.
• [14]Worlitzsch D, Tarran R, Ulrich M, Schwab U, Cekici A, Meyer KC, Birrer P, Bellon G, Berger J, Weiß T, Botzenhart K, Yankaskas JR, Randell S, Boucher RC, Döring G: Effects of reduced oxygen concentration in airway Pseudomonas infections of cystic fibrosis patients. J Clin Invest 2002, 109:317-325.
• [15]Nunley D, Dauber J, Iacono A, Keenan R, Zeevi A, Cornwell R, Love R, Meyer K: Unopposed neutrophil elastase in bronchoalveolar lavage fluid from transplant recipients with cystic fibrosis. Am J Respir Crit Care Med 1999, 159:258-261.
• [16]King MB, Campbell EJ, Gray BH, Hertz MI: The proteinase-antiproteinase balance in alpha-1-proteinase inhibitor–deficient lung transplant recipients. Am J Respir Crit Care Med 1994, 149:966-971.
• [17]Diep BA, Chan L, Tattevin P, Kajikawa O, Martin TR, Basuino L, Mai TT, Marbach H, Braughton KR, Whitney AR, Gardner DJ, Fan X, Tseng CW, Liu GY, Badiou C, Etienne J, Lina G, Matthay MA, DeLeo FR, Chambers HF: Polymorphonuclear leukocytes mediate Staphylococcus aureus Panton-Valentine leukocidin-induced lung inflammation and injury. Proc Natl Acad Sci U S A 2010, 107:5587-5592.
• [18]Watt AP, Courtney J, Moore J, Ennis M, Elborn JS: Neutrophil cell death, activation and bacterial infection in cystic fibrosis. Thorax 2005, 60:659-664.
• [19]Levada-Pires AC, Cury-Boaventura MF, Gorjão R, Hirabara SM, Puggina EF, Peres CM, Lambertucci RH, Curi R, Pithon-Curi TC: Neutrophil death induced by a triathlon competition in elite athletes. Med Sci Sports Exerc 2008, 40:1447-1454.
• [20]Papayannopoulos V, Metzler KD, Hakkim A, Zychlinsky A: Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular traps. J Cell Biol 2010, 191:677-691.
• [21]Moriceau S, Lenoir G, Witko-Sarsat V: In cystic fibrosis homozygotes and heterozygotes, neutrophil apoptosis is delayed and modulated by diamide or roscovitine: evidence for an innate neutrophil disturbance. J Innate Immun 2010, 2:260-266.
• [22]Carlsson M, Eriksson L, Pressler T, Kornfält R, Mared L, Meyer P, Wiik A, Wieslander J, Segelmark M: Autoantibody response to BPI predict disease severity and outcome in cystic fibrosis. J Cyst Fibros 2007, 6:228-233.
• [23]Aichele D, Schnare M, Saake M, Röllinghoff M, Gessner A: Expression and antimicrobial function of bactericidal permeability-increasing protein in cystic fibrosis patients. Infect Immun 2006, 74:4708-4714.24.
• [24]Hsu MJ, Lee SS, Lin WW: Polysaccharide purified from Ganoderma lucidum inhibits spontaneous and Fas-mediated apoptosis in human neutrophils through activation of the phosphatidylinositol 3 kinase/Akt signaling pathway. J Leuko Biol 2002, 72:207-216.
• [25]Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH: Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 1994, 84:1415-1420.
• [26]Homburg CH, de Haas M, von dem Borne AE, Verhoeven AJ, Reutelingsperger CP, Roos D: Human neutrophils lose their surface FcgRIII and acquire annexin V binding sites during apoptosis in vitro. Blood 1995, 85:532-540.
• [27]Verms I, Haanen C, Steffens-Nakken H, Reutelingsperger CP: A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods 1995, 184:39-51.
• [28]Takei H, Araki A, Watanabe H, Ichinose A, Sendo E: Rapid killing of human neutrophils by the potent activator phorbol 12-myristate 13-acetate (PMA) accompanied by changes different from typical apoptosis or necrosis. J Leukocyte Biol 1996, 59:229-240.
• [29]Suzuki K, Namiki H: Phorbol 12 myristate 13-acetate induced cell death of porcine peripheral blood polymorphonuclear leukocytes. Cell Struct Funct 1998, 23:367-372.