Yonsei Med J.  2011 Jul;52(4):581-587. 10.3349/ymj.2011.52.4.581.

Apoptosis of T Lymphocytes Isolated from Peripheral Blood of Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease

• Lim SC
• Ju JY
• Chi SY
• Ban HJ
• Kwon YS
• Oh IJ
• Kim KS
• Kim YI
• Kim YC

• KMID: 1727989
• DOI: http://doi.org/10.3349/ymj.2011.52.4.581

Abstract

PURPOSE
Chronic obstructive pulmonary disease (COPD) is characterized by chronic inflammation of the airways and progressive destruction of lung parenchyma. Apoptosis is critical for the maintenance of normal tissue homeostasis and is in equilibrium with proliferation and differentiation. This study was undertaken to investigate relationship between apoptosis of peripheral blood lymphocytes during exacerbation of COPD and inflammatory response that characterizes this condition.
MATERIALS AND METHODS
Seventeen patients with COPD exacerbation, 21 stable COPD, and 12 control subjects were included. T lymphocytes were isolated from peripheral blood using MACS. Apoptosis of T lymphocytes was assessed with FACS using annexin V and 7-aminoactinomycin. Serum levels of interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-alpha were determined by an immunoassay technique.
RESULTS
There was significantly increased percentage of apoptotic lymphocytes, CD 4+, and CD 8+ T cells in the peripheral blood of patients with exacerbation of COPD compared with stable COPD. Serum levels of IL-6, IL-8, and TNF-alpha were significantly increased in patients with exacerbation of COPD compared with stable COPD. Only TNF-alpha presented a positive correlation with apoptotic lymphocytes in patients with exacerbation of COPD.
CONCLUSION
Increased apoptotic lymphocytes may be associated with upregulation of TNF-alpha in the peripheral blood of patients with acute exacerbation of COPD.

Keyword

Chronic obstructive pulmonary disease; apoptosis; lymphocyte; tumor necrosis factor-alpha

MeSH Terms

*Apoptosis
CD4-Positive T-Lymphocytes/pathology
CD8-Positive T-Lymphocytes/pathology
Flow Cytometry
Humans
Interleukin-6/blood
Interleukin-8/blood
Pulmonary Disease, Chronic Obstructive/blood/*pathology
T-Lymphocytes/*pathology
Tumor Necrosis Factor-alpha/blood

Figure

• 

  Fig. 1 Mean apoptosis percentage of peripheral blood T lymphocytes (A), CD 4+ T cells (B), and CD 8+ T cells (C) in control groups, stable COPD, and exacerbation of COPD measured by flow cytometry. COPD, chronic obstructive pulmonary disease.

• 

  Fig. 2 The serum levels of IL-6 (A), IL-8 (B), and TNF-α (C) in control groups, stable COPD, and exacerbation of COPD. Serum cytokine levels are expressed as the median value and interquartile range. •, outlier extending >1.5 box-lengths from the edge of the box. IL-6, interleukin-6; IL-8, interleukin-8; TNF-α, tumor necrosis factor-alpha. COPD, chronic obstructive pulmonary disease.

Reference

1. Pauwels RA, Rabe KF. Burden and clinical features of chronic obstructive pulmonary disease (COPD). Lancet. 2004. 364:613–620.

2. Barnes PJ, Shapiro SD, Pauwels RA. Chronic obstructive pulmonary disease: molecular and cellular mechanisms. Eur Respir J. 2003. 22:672–688.

3. Saetta M, Di Stefano A, Maestrelli P, Ferraresso A, Drigo R, Potena A, et al. Activated T-lymphocytes and macrophages in bronchial mucosa of subjects with chronic bronchitis. Am Rev Respir Dis. 1993. 147:301–306.

4. Pabst R, Binns RM. Lymphocytes migrate from the bronchoalveolar space to regional bronchial lymph nodes. Am J Respir Crit Care Med. 1995. 151:495–499.

5. Majo J, Ghezzo H, Cosio MG. Lymphocyte population and apoptosis in the lungs of smokers and their relation to emphysema. Eur Respir J. 2001. 17:946–953.

6. Segura-Valdez L, Pardo A, Gaxiola M, Uhal BD, Becerril C, Selman M. Upregulation of gelatinases A and B, collagenases 1 and 2, and increased parenchymal cell death in COPD. Chest. 2000. 117:684–694.

7. Saetta M, Di Stefano A, Turato G, Facchini FM, Corbino L, Mapp CE, et al. CD8+ T-lymphocytes in peripheral airways of smokers with chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 1998. 157:822–826.

8. Degterev A, Boyce M, Yuan J. A decade of caspases. Oncogene. 2003. 22:8543–8567.

9. Muzio M, Chinnaiyan AM, Kischkel FC, O'Rourke K, Shevchenko A, Ni J, et al. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death--inducing signaling complex. Cell. 1996. 85:817–827.

10. Hirata H, Takahashi A, Kobayashi S, Yonehara S, Sawai H, Okazaki T, et al. Caspases are activated in a branched protease cascade and control distinct downstream processes in Fas-induced apoptosis. J Exp Med. 1998. 187:587–600.

11. Gompertz S, O'Brien C, Bayley DL, Hill SL, Stockley RA. Changes in bronchial inflammation during acute exacerbations of chronic bronchitis. Eur Respir J. 2001. 17:1112–1119.

12. Song SH, Kim CH, Kwon SS, Kim YK, Kim KH, Moon HS, et al. Nuclear Factor-κB(NF-κB) activity and levels of IL-6, IL-8 and TNF-α in induced sputum in the exacerbation and recovery of COPD patients. Tuberc Respir Dis. 2005. 58:152–159.

13. Grutkoski PS, Graeber CT, Ayala A, Simms HH. Paracrine suppression of apoptosis by cytokine-stimulated neutrophils involves divergent regulation of NF-kappaB, Bcl-X(L), and Bak. Shock. 2002. 17:47–54.

14. Dunican AL, Leuenroth SJ, Grutkoski P, Ayala A, Simms HH. TNFalpha-induced suppression of PMN apoptosis is mediated through interleukin-8 production. Shock. 2000. 14:284–288.

15. Biffl WL, Moore EE, Moore FA, Barnett CC Jr. Interleukin-6 suppression of neutrophil apoptosis is neutrophil concentration dependent. J Leukoc Biol. 1995. 58:582–584.

16. Schmidt-Ioanas M, Pletz MW, de Roux A, Lode H. Apoptosis of peripheral blood neutrophils in COPD exacerbation does not correlate with serum cytokines. Respir Med. 2006. 100:639–647.

17. Rabe KF, Hurd S, Anzueto A, Barnes PJ, Buist SA, Calverley P, et al. Global strategy for the diagnosis, management, and prevention of chronic obstructive pulmonary disease: GOLD executive summary. Am J Respir Crit Care Med. 2007. 176:532–555.

18. Anthonisen NR, Manfreda J, Warren CP, Hershfield ES, Harding GK, Nelson NA. Antibiotic therapy in exacerbations of chronic obstructive pulmonary disease. Ann Intern Med. 1987. 106:196–204.

19. Kasahara Y, Tuder RM, Cool CD, Lynch DA, Flores SC, Voelkel NF. Endothelial cell death and decreased expression of vascular endothelial growth factor and vascular endothelial growth factor receptor 2 in emphysema. Am J Respir Crit Care Med. 2001. 163:737–744.

20. Hodge SJ, Hodge GL, Reynolds PN, Scicchitano R, Holmes M. Increased production of TGF-beta and apoptosis of T lymphocytes isolated from peripheral blood in COPD. Am J Physiol Lung Cell Mol Physiol. 2003. 285:L492–L499.

21. Hodge S, Hodge G, Holmes M, Reynolds PN. Increased airway epithelial and T-cell apoptosis in COPD remains despite smoking cessation. Eur Respir J. 2005. 25:447–454.

22. Bhowmik A, Seemungal TA, Sapsford RJ, Wedzicha JA. Relation of sputum inflammatory markers to symptoms and lung function changes in COPD exacerbations. Thorax. 2000. 55:114–120.

23. Crooks SW, Bayley DL, Hill SL, Stockley RA. Bronchial inflammation in acute bacterial exacerbations of chronic bronchitis: the role of leukotriene B4. Eur Respir J. 2000. 15:274–280.

24. Hurst JR, Donaldson GC, Perera WR, Wilkinson TM, Bilello JA, Hagan GW, et al. Use of plasma biomarkers at exacerbation of chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 2006. 174:867–874.

25. Prescott E, Lange P, Vestbo J. Chronic mucus hypersecretion in COPD and death from pulmonary infection. Eur Respir J. 1995. 8:1333–1338.

26. Agustí AG, Villaverde JM, Togores B, Bosch M. Serial measurements of exhaled nitric oxide during exacerbations of chronic obstructive pulmonary disease. Eur Respir J. 1999. 14:523–528.

27. Dekhuijzen PN, Aben KK, Dekker I, Aarts LP, Wielders PL, van Herwaarden CL, et al. Increased exhalation of hydrogen peroxide in patients with stable and unstable chronic obstructive pulmonary disease. Am J Respir Crit Care Med. 1996. 154:813–816.

28. 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.

29. Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY, Henson PM. Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF. J Clin Invest. 1998. 101:890–898.