Late diagnosis of lung cancer is still the main reason for high mortality rates in lung cancer. Lung cancer is a heterogeneous disease which induces an immune response to different tumor antigens. Several methods for searching autoantibodies have been described that are based on known purified antigen panels. The aim of our study is to find evidence that parts of the antigen-binding-domain of antibodies are shared among lung cancer patients. This was investigated by a novel approach based on sequencing antigen-binding-fragments (Fab) of immunoglobulins using proteomic techniques without the need of previously known antigen panels. From serum of 93 participants of the NELSON trial IgG was isolated and subsequently digested into Fab and Fc. Fab was purified from the digested mixture by SDS-PAGE. The Fab containing gel-bands were excised, tryptic digested and measured on a nano-LC-Orbitrap-Mass-spectrometry system. Multivariate analysis of the mass spectrometry data by linear canonical discriminant analysis combined with stepwise logistic regression resulted in a 12-antibody-peptide model which was able to distinguish lung cancer patients from controls in a high risk population with a sensitivity of 84% and specificity of 90%. With our Fab-purification combined Orbitrap-mass-spectrometry approach, we found peptides from the variable-parts of antibodies which are shared among lung cancer patients.
References
[1]
Ferlay J, Soerjomataram I, Ervik M, Dikshit R, Eser S, et al.. (2012) GLOBOCAN 2012 v1.0, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 11 [Internet]. Lyon, France: International Agency for Research on Cancer.
[2]
Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics, 2010. CA Cancer J Clin 60: 277–300. doi: 10.3322/caac.20073
[3]
Aberle DR, Adams AM, Berg CD, Black WC, Clapp JD, et al. (2011) Reduced lung-cancer mortality with low-dose computed tomographic screening. N Engl J Med 365: 395–409. doi: 10.1056/nejmoa1102873
[4]
Horeweg N, van der Aalst CM, Vliegenthart R, Zhao Y, Xie X, et al. (2013) Volumetric computed tomography screening for lung cancer: three rounds of the NELSON trial. Eur Respir J 42: 1659–1667. doi: 10.1183/09031936.00197712
[5]
van Klaveren RJ, Oudkerk M, Prokop M, Scholten ET, Nackaerts K, et al. (2009) Management of lung nodules detected by volume CT scanning. N Engl J Med 361: 2221–2229. doi: 10.1056/nejmoa0906085
[6]
Anderson KS, LaBaer J (2005) The sentinel within: exploiting the immune system for cancer biomarkers. J Proteome Res 4: 1123–1133. doi: 10.1021/pr0500814
[7]
Baldwin RW (1955) Immunity to transplanted tumour: the effect of tumour extracts on the growth of homologous tumours in rats. Br J Cancer 9: 646–651.
[8]
Caron M, Choquet-Kastylevsky G, Joubert-Caron R (2007) Cancer immunomics using autoantibody signatures for biomarker discovery. Mol Cell Proteomics 6: 1115–1122. doi: 10.1074/mcp.r600016-mcp200
[9]
Gure AO, Altorki NK, Stockert E, Scanlan MJ, Old LJ, et al. (1998) Human lung cancer antigens recognized by autologous antibodies: definition of a novel cDNA derived from the tumor suppressor gene locus on chromosome 3p21.3. Cancer Res 58: 1034–1041.
[10]
Hanash S (2003) Harnessing immunity for cancer marker discovery. Nat Biotechnol 21: 37–38. doi: 10.1038/nbt0103-37
[11]
Mintz PJ, Kim J, Do KA, Wang X, Zinner RG, et al. (2003) Fingerprinting the circulating repertoire of antibodies from cancer patients. Nat Biotechnol 21: 57–63. doi: 10.1038/nbt774
[12]
Stockert E, Jager E, Chen YT, Scanlan MJ, Gout I, et al. (1998) A survey of the humoral immune response of cancer patients to a panel of human tumor antigens. J Exp Med 187: 1349–1354. doi: 10.1084/jem.187.8.1349
[13]
Liu L, Liu N, Liu B, Yang Y, Zhang Q, et al.. (2012) Are circulating autoantibodies to ABCC3 transporter a potential biomarker for lung cancer? J Cancer Res Clin Oncol.
[14]
Murphy KTP, Walport M (2008) Janeway’s immunobiology. Garland Science.
[15]
Saada R, Weinberger M, Shahaf G, Mehr R (2007) Models for antigen receptor gene rearrangement: CDR3 length. Immunol Cell Biol 85: 323–332. doi: 10.1038/sj.icb.7100055
[16]
Foreman AL, Lemercier B, Lim A, Kourlisky P, Kenny T, et al. (2008) VH gene usage and CDR3 analysis of B cell receptor in the peripheral blood of patients with PBC. Autoimmunity 41: 80–86. doi: 10.1080/08916930701619656
[17]
Scheid JF, Mouquet H, Ueberheide B, Diskin R, Klein F, et al. (2011) Sequence and structural convergence of broad and potent HIV antibodies that mimic CD4 binding. Science 333: 1633–1637. doi: 10.1126/science.1207227
[18]
VanDuijn MM, Dekker LJ, Zeneyedpour L, Smitt PA, Luider TM (2010) Immune responses are characterized by specific shared immunoglobulin peptides that can be detected by proteomic techniques. J Biol Chem 285: 29247–29253. doi: 10.1074/jbc.m110.139071
[19]
Weinstein JA, Jiang N, White RA 3rd, Fisher DS, Quake SR (2009) High-throughput sequencing of the zebrafish antibody repertoire. Science 324: 807–810. doi: 10.1126/science.1170020
[20]
Maat P, VanDuijn M, Brouwer E, Dekker L, Zeneyedpour L, et al. (2012) Mass spectrometric detection of antigen-specific immunoglobulin peptides in paraneoplastic patient sera. J Autoimmun 38: 354–360. doi: 10.1016/j.jaut.2012.02.002
[21]
van Iersel CA, de Koning HJ, Draisma G, Mali WP, Scholten ET, et al. (2007) Risk-based selection from the general population in a screening trial: selection criteria, recruitment and power for the Dutch-Belgian randomised lung cancer multi-slice CT screening trial (NELSON). Int J Cancer 120: 868–874. doi: 10.1002/ijc.22134
[22]
de Costa D, Broodman I, Vanduijn MM, Stingl C, Dekker LJ, et al. (2010) Sequencing and quantifying IgG fragments and antigen-binding regions by mass spectrometry. J Proteome Res 9: 2937–2945. doi: 10.1021/pr901114w
[23]
Lefranc MP, Giudicelli V, Ginestoux C, Jabado-Michaloud J, Folch G, et al. (2009) IMGT, the international ImMunoGeneTics information system. Nucleic Acids Res 37: D1006–1012. doi: 10.1093/nar/gkn838
[24]
Snedecor GW (1980) Statistical Methods. Iowa: The Iowa State University Press.
[25]
Armitage P, Berry GS (2002) Statistical Methods in Medical Research. Oxford: Blackwell Scientific.
[26]
Afifi A, Clark V, May S (2004) Computer-aided multivariate analysis. Florida: Chapman & Hall.
[27]
Kleinbaum DG, Kupper LL, Muller KE, Nizam A (1998) Applied regression analysis and other multivariable methods. California: Duxbury Press.
[28]
Arentz G, Thurgood LA, Lindop R, Chataway TK, Gordon TP (2012) Secreted human Ro52 autoantibody proteomes express a restricted set of public clonotypes. J Autoimmun 39: 466–470. doi: 10.1016/j.jaut.2012.07.003
[29]
Andersen PS, Haahr-Hansen M, Coljee VW, Hinnerfeldt FR, Varming K, et al. (2007) Extensive restrictions in the VH sequence usage of the human antibody response against the Rhesus D antigen. Mol Immunol 44: 412–422. doi: 10.1016/j.molimm.2006.02.021
[30]
Baranzini SE, Jeong MC, Butunoi C, Murray RS, Bernard CC, et al. (1999) B cell repertoire diversity and clonal expansion in multiple sclerosis brain lesions. J Immunol 163: 5133–5144.
[31]
Brichory F, Beer D, Le Naour F, Giordano T, Hanash S (2001) Proteomics-based identification of protein gene product 9.5 as a tumor antigen that induces a humoral immune response in lung cancer. Cancer Res 61: 7908–7912.
[32]
Brichory FM, Misek DE, Yim AM, Krause MC, Giordano TJ, et al. (2001) An immune response manifested by the common occurrence of annexins I and II autoantibodies and high circulating levels of IL-6 in lung cancer. Proc Natl Acad Sci U S A 98: 9824–9829. doi: 10.1073/pnas.171320598
[33]
Chapman CJ, Healey GF, Murray A, Boyle P, Robertson C, et al.. (2012) EarlyCDT-Lung test: improved clinical utility through additional autoantibody assays. Tumour Biol.
[34]
Koziol JA, Zhang JY, Casiano CA, Peng XX, Shi FD, et al. (2003) Recursive partitioning as an approach to selection of immune markers for tumor diagnosis. Clin Cancer Res 9: 5120–5126.
[35]
Khattar NH, Coe-Atkinson SP, Stromberg AJ, Jett JR, Hirschowitz EA (2010) Lung cancer-associated auto-antibodies measured using seven amino acid peptides in a diagnostic blood test for lung cancer. Cancer Biol Ther 10: 267–272. doi: 10.4161/cbt.10.3.12395
[36]
Zhong L, Coe SP, Stromberg AJ, Khattar NH, Jett JR, et al. (2006) Profiling tumor-associated antibodies for early detection of non-small cell lung cancer. J Thorac Oncol 1: 513–519. doi: 10.1097/01243894-200607000-00003
[37]
Pass HI, Johnson DH, Minna JD, Scagliotti GV, Turrisi AT (2010) Principles and practice of lung cancer. The official reference text of the IASLC. Lippincott Williams & Wilkins.
[38]
Villeneuve PJ, Mao Y (1994) Lifetime probability of developing lung cancer, by smoking status, Canada. Can J Public Health 85: 385–388.
[39]
Pecot CV, Li M, Zhang XJ, Rajanbabu R, Calitri C, et al. (2012) Added value of a serum proteomic signature in the diagnostic evaluation of lung nodules. Cancer Epidemiol Biomarkers Prev 21: 786–792. doi: 10.1158/1055-9965.epi-11-0932
