Basal gene expression by lung CD4+ T cells in chronic obstructive pulmonary disease identifies independent molecular correlates of airflow obstruction and emphysema extent
Christine M Freeman, Alexandra L McCubbrey, Sean Crudgington, Joshua Nelson, Fernando J Martinez, MeiLan K Han, George R Washko Jr, Stephen W Chensue, Douglas A Arenberg, Catherine A Meldrum, Lisa McCloskey, Jeffrey L Curtis, Christine M Freeman, Alexandra L McCubbrey, Sean Crudgington, Joshua Nelson, Fernando J Martinez, MeiLan K Han, George R Washko Jr, Stephen W Chensue, Douglas A Arenberg, Catherine A Meldrum, Lisa McCloskey, Jeffrey L Curtis
Abstract
Lung CD4+ T cells accumulate as chronic obstructive pulmonary disease (COPD) progresses, but their role in pathogenesis remains controversial. To address this controversy, we studied lung tissue from 53 subjects undergoing clinically-indicated resections, lung volume reduction, or transplant. Viable single-cell suspensions were analyzed by flow cytometry or underwent CD4+ T cell isolation, followed either by stimulation with anti-CD3 and cytokine/chemokine measurement, or by real-time PCR analysis. In lung CD4+ T cells of most COPD subjects, relative to lung CD4+ T cells in smokers with normal spirometry: (a) stimulation induced minimal IFN-γ or other inflammatory mediators, but many subjects produced more CCL2; (b) the T effector memory subset was less uniformly predominant, without correlation with decreased IFN-γ production. Analysis of unstimulated lung CD4+ T cells of all subjects identified a molecular phenotype, mainly in COPD, characterized by markedly reduced mRNA transcripts for the transcription factors controlling TH1, TH2, TH17 and FOXP3+ T regulatory subsets and their signature cytokines. This mRNA-defined CD4+ T cell phenotype did not result from global inability to elaborate mRNA; increased transcripts for inhibitory CD28 family members or markers of anergy; or reduced telomerase length. As a group, these subjects had significantly worse spirometry, but not DLCO, relative to subjects whose lung CD4+ T cells expressed a variety of transcripts. Analysis of mRNA transcripts of unstimulated lung CD4+ T cell among all subjects identified two distinct molecular correlates of classical COPD clinical phenotypes: basal IL-10 transcripts correlated independently and inversely with emphysema extent (but not spirometry); by contrast, unstimulated IFN-γ transcripts correlated independently and inversely with reduced spirometry (but not reduced DLCO or emphysema extent). Aberrant lung CD4+ T cells polarization appears to be common in advanced COPD, but also exists in some smokers with normal spirometry, and may contribute to development and progression of specific COPD phenotypes.
Trial registration: ClinicalTrials.gov as NCT00281229.
Conflict of interest statement
Competing Interests: The authors have declared that no competing interests exist.
Figures
References
- Mannino DM, Buist AS (2007) Global burden of COPD: risk factors, prevalence, and future trends. Lancet 370: 765–773.
- Cosio MG, Saetta M, Agusti A (2009) Immunologic aspects of chronic obstructive pulmonary disease. N Engl J Med 360: 2445–2454.
- Salvi SS, Barnes PJ (2009) Chronic obstructive pulmonary disease in non-smokers. Lancet 374: 733–743.
- Curtis JL, Freeman CM, Hogg JC (2007) The immunopathogenesis of chronic obstructive pulmonary disease: insights from recent research. Proc Am Thorac Soc 4: 512–521.
- Hogg JC, Chu F, Utokaparch S, Woods R, Elliott WM, et al. (2004) The nature of small-airway obstruction in chronic obstructive pulmonary disease. N Engl J Med 350: 2645–2653.
- Saetta M, Baraldo S, Corbino L, Turato G, Braccioni F, et al. (1999) CD8+ve cells in the lungs of smokers with chronic obstructive pulmonary disease. Am J Respir Crit Care Med 160: 711–717.
- Majori M, Corradi M, Caminati A, Cacciani G, Bertacco S, et al. (1999) Predominant Th1 cytokine pattern in peripheral blood from subjects with chronic obstructive pulmonary disease. J Allergy Clin Immunol 103: 458–462.
- Freeman CM, Curtis JL, Chensue SW (2007) CC chemokine receptor 5 and CXC chemokine receptor 6 expression by lung CD8+ cells correlates with chronic obstructive pulmonary disease severity. Am J Pathol 171: 767–776.
- Freeman CM, Han M-LK, Martinez FJ, Murray S, Liu LX, et al. (2010) Cytotoxic potential of lung CD8+ T cells in COPD is increased with disease severity and by in vitro stimulation with IL-18 and IL-15. J immunol 184: 6504–6513.
- Majo J, Ghezzo H, Cosio MG (2001) Lymphocyte population and apoptosis in the lungs of smokers and their relation to emphysema. Eur Respir J 17: 946–953.
- Lofdahl MJ, Roos-Engstrand E, Pourazar J, Bucht A, Dahlen B, et al. (2008) Increased intraepithelial T-cells in stable COPD. Respir Med 102: 1812–1818.
- Sullivan AK, Simonian PL, Falta MT, Mitchell JD, Cosgrove GP, et al. (2005) Oligoclonal CD4+ T cells in the lungs of patients with severe emphysema. Am J Respir Crit Care Med 172: 590–596.
- Freeman CM, Martinez FJ, Han MK, Ames TM, Chensue SW, et al. (2009) Lung dendritic cell expression of maturation molecules increases with worsening chronic obstructive pulmonary disease. Am J Respir Crit Care Med 180: 1179–1188.
- Lee SH, Goswami S, Grudo A, Song LZ, Bandi V, et al. (2007) Antielastin autoimmunity in tobacco smoking-induced emphysema. Nat Med 13: 567–569.
- Shan M, Cheng HF, Song LZ, Roberts L, Green L, et al. (2009) Lung myeloid dendritic cells coordinately induce TH1 and TH17 responses in human emphysema. Sci Transl Med 1: 4ra10.
- GOLD Executive Committee (2008) Global strategy for the diagnosis, management, and prevention of COPD (updated 2008).
- Freeman CM, Martinez FJ, Han MK, Washko GR Jr, McCubbrey AL, et al. (2013) Lung CD8+ T cells in COPD have increased expression of bacterial TLRs. Respir Res 14: 13.
- Cawthon RM (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res 30: e47.
- Savale L, Chaouat A, Bastuji-Garin S, Marcos E, Boyer L, et al. (2009) Shortened telomeres in circulating leukocytes of patients with chronic obstructive pulmonary disease. Am J Respir Crit Care Med 179: 566–571.
- Leopold JG, Gough J (1957) The centrilobular form of hypertrophic emphysema and its relation to chronic bronchitis. Thorax 12: 219–235.
- Thurlbeck WM (1963) A clinico-pathological study of emphysema in an American hospital. Thorax 18: 59–67.
- Auerbach O, Hammond EC, Garfinkel L, Benante C (1972) Relation of smoking and age to emphysema. Whole-lung section study. N Engl J Med 286: 853–857.
- Mets OM, van Hulst RA, Jacobs C, van Ginneken B, de Jong PA (2012) Normal range of emphysema and air trapping on CT in young men. AJR Am J Roentgenol 199: 336–340.
- Stephens R, Langhorne J (2010) Effector memory Th1 CD4 T cells are maintained in a mouse model of chronic malaria. PLoS Pathog 6: e1001208.
- Gebhardt T, Wakim LM, Eidsmo L, Reading PC, Heath WR, et al. (2009) Memory T cells in nonlymphoid tissue that provide enhanced local immunity during infection with herpes simplex virus. Nat Immunol 10: 524–530.
- Masopust D, Choo D, Vezys V, Wherry EJ, Duraiswamy J, et al. (2010) Dynamic T cell migration program provides resident memory within intestinal epithelium. J Exper Med 207: 553–564.
- Purwar R, Campbell J, Murphy G, Richards WG, Clark RA, et al. (2011) Resident memory T cells (T(RM)) are abundant in human lung: diversity, function, and antigen specificity. PLoS One 6: e16245.
- Rocha B, Tanchot C, Von Boehmer H (1993) Clonal anergy blocks in vivo growth of mature T cells and can be reversed in the absence of antigen. J Exp Med 177: 1517–1521.
- Singh NJ, Schwartz RH (2003) The strength of persistent antigenic stimulation modulates adaptive tolerance in peripheral CD4+ T cells. J Exp Med 198: 1107–1117.
- Gomez-Martin D, Diaz-Zamudio M, Galindo-Campos M, Alcocer-Varela J (2010) Early growth response transcription factors and the modulation of immune response: implications towards autoimmunity. Autoimmun Rev 9: 454–458.
- Paats MS, Bergen IM, Hoogsteden HC, van der Eerden MM, Hendriks RW (2012) Systemic CD4+ and CD8+ T-cell cytokine profiles correlate with GOLD stage in stable COPD. Eur Respir J 40: 330–337.
- Han MK, Agusti A, Calverley PM, Celli BR, Criner G, et al. (2010) Chronic obstructive pulmonary disease phenotypes: the future of COPD. Am J Respir Crit Care Med 182: 598–604.
- Manichaikul A, Hoffman EA, Smolonska J, Gao W, Cho MH, et al. (2014) Genome-wide study of percent emphysema on computed tomography in the general population. The Multi-Ethnic Study of Atherosclerosis Lung/SNP Health Association Resource Study. Am J Respir Crit Care Med 189: 408–418.
- Johannessen A, Skorge TD, Bottai M, Grydeland TB, Nilsen RM, et al. (2013) Mortality by level of emphysema and airway wall thickness. Am J Respir Crit Care Med 187: 602–608.
- Barcelo B, Pons J, Fuster A, Sauleda J, Noguera A, et al. (2006) Intracellular cytokine profile of T lymphocytes in patients with chronic obstructive pulmonary disease. Clin Exp Immunol 145: 474–479.
- Hogg JC, Macklem PT, Thurlbeck WM (1968) Site and nature of airway obstruction in chronic obstructive lung disease. N Engl J Med 278: 1355–1360.
- Brooks DG, Walsh KB, Elsaesser H, Oldstone MB (2010) IL-10 directly suppresses CD4 but not CD8 T cell effector and memory responses following acute viral infection. Proc Natl Acad Sci U S A 107: 3018–3023.
- Moore KW, de Waal Malefyt R, Coffman RL, O’Garra A (2001) Interleukin-10 and the interleukin-10 receptor. Annu Rev Immunol 19: 683–765.
- Burgess JL, Fierro MA, Lantz RC, Hysong TA, Fleming JE, et al. (2004) Longitudinal decline in lung function: evaluation of interleukin-10 genetic polymorphisms in firefighters. J Occup Environ Med 46: 1013–1022.
- Demeo DL, Campbell EJ, Barker AF, Brantly ML, Eden E, et al. (2008) IL10 polymorphisms are associated with airflow obstruction in severe alpha1-antitrypsin deficiency. Am J Respir Cell Mol Biol 38: 114–120.
- He JQ, Shumansky K, Zhang X, Connett JE, Anthonisen NR, et al. (2007) Polymorphisms of interleukin-10 and its receptor and lung function in COPD. Eur Respir J 29: 1120–1126.
- Barcelo B, Pons J, Ferrer JM, Sauleda J, Fuster A, et al. (2008) Phenotypic characterisation of T-lymphocytes in COPD: abnormal CD4+CD25+ regulatory T-lymphocyte response to tobacco smoking. Eur Respir J 31: 555–562.
- Smyth LJ, Starkey C, Vestbo J, Singh D (2007) CD4-regulatory cells in COPD patients. Chest 132: 156–163.
- Roos-Engstrand E, Ekstrand-Hammarstrom B, Pourazar J, Behndig AF, Bucht A, et al. (2009) Influence of smoking cessation on airway T lymphocyte subsets in COPD. COPD 6: 112–120.
- Plumb J, Smyth LJ, Adams HR, Vestbo J, Bentley A, et al. (2009) Increased T-regulatory cells within lymphocyte follicles in moderate COPD. Eur Respir J 34: 89–94.
- Sallusto F, Lenig D, Forster R, Lipp M, Lanzavecchia A (1999) Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature 401: 708–712.
- Ansel KM, McHeyzer-Williams LJ, Ngo VN, McHeyzer-Williams MG, Cyster JG (1999) In vivo-activated CD4 T cells upregulate CXC chemokine receptor 5 and reprogram their response to lymphoid chemokines. J Exp Med 190: 1123–1134.
- Fazilleau N, Mark L, McHeyzer-Williams LJ, McHeyzer-Williams MG (2009) Follicular helper T cells: lineage and location. Immunity 30: 324–335.
- Feghali-Bostwick CA, Gadgil AS, Otterbein LE, Pilewski JM, Stoner MW, et al. (2008) Autoantibodies in patients with chronic obstructive pulmonary disease. Am J Respir Crit Care Med 177: 156–163.
- Mueller SN, Gebhardt T, Carbone FR, Heath WR (2013) Memory T cell subsets, migration patterns, and tissue residence. Annu Rev Immunol 31: 137–161.
- Okada R, Kondo T, Matsuki F, Takata H, Takiguchi M (2008) Phenotypic classification of human CD4+ T cell subsets and their differentiation. Int Immunol 20: 1189–1199.
- Kapina MA, Shepelkova GS, Mischenko VV, Sayles P, Bogacheva P, et al. (2007) CD27low CD4 T lymphocytes that accumulate in the mouse lungs during mycobacterial infection differentiate from CD27 high precursors in situ, produce IFN-gamma, and protect the host against tuberculosis infection. J Immunol 178: 976–985.
- Di Stefano A, Caramori G, Gnemmi I, Contoli M, Vicari C, et al. (2009) T helper type 17-related cytokine expression is increased in the bronchial mucosa of stable chronic obstructive pulmonary disease patients. Clin Exp Immunol 157: 316–324.
- Bevan MJ (2004) Helping the CD8(+) T-cell response. Nat Rev Immunol 4: 595–602.
- Combe CL, Curiel TJ, Moretto MM, Khan IA (2005) NK cells help to induce CD8(+)-T-cell immunity against Toxoplasma gondii in the absence of CD4(+) T cells. Infect Immun 73: 4913–4921.
- Livingstone AM, Wilson EB, Ontiveros F, Wang JC (2009) Unravelling the mechanisms of help for CD8+ T cell responses. Immunol Res 45: 209–217.
- Dikopoulos N, Bertoletti A, Kroger A, Hauser H, Schirmbeck R, et al. (2005) Type I IFN negatively regulates CD8+ T cell responses through IL-10-producing CD4+ T regulatory 1 cells. J Immunol 174: 99–109.
Source: PubMed