ppn:332291049 Philipps-Universität Marburg analytical performance Bomert, Martin Bomert Martin monograph German multiplex assay system 2013-09-24 Publikationsserver der Universitätsbibliothek Marburg Universitätsbibliothek Marburg In the course of the prospective multicentre cohortstudy PASTURE/ EFRAIM, factors that influence the development of childhood asthma and allergy were investigated. One part has been the creation of an image of the different T-cell-profiles by measuring cytokines at different time points. The usually used ELISA for cytokine measurement is regarded as the gold standard, but is very time-, money-, personell- and sample-consuming. Therefore, cytokine measurement should be switched to a high-throughput system, the „cytometric bead array assay (CBA)“-system from BD Biosciences. The aim of this paper is to validate the CBA-method, to ínvestigate the quality of measurements in comparison to ELISA and the comparability of both methods. The comparison of methods based on 556 samples of fourty study members which were measured in parallel according to the manufacturer’s manual. The ELISA measurements were performed according to the PASTURE-/ EFRAIM- study protocol. The results concerning sample carryover, calibration stability, analytical and functional sensitivity, imprecision, recovery and linearity show, that CBA generates at least as stable and reliable results as the ELISA and is, in most cases, superiour with regard to sample consumption, cost effectiveness, reproducibility, and usability. With regard to the comparability of identical samples, the measurement results show correlations coefficients (Pearson) between 0,854 (TNF-a) and 0,930 (IL-10) for undiluted samples. The concordance corellation coefficient according to Lin, as well as the graphical illustration according to Bland and Altman shows a fair agreement of measurement results for low concentrations and bad agreement in high concentrations. The present results allow the comparison of the results with certain limitations between 5 pg/ml and 100 pg/ml for IL-5, for IL-10 between 20 pg/ml and 300 pg/ml, for IFN-g and TNF-a between 20 pg/ml and 1000 pg/ml. The reason can most likely be seen in the use of different antibody pairs, the existence of heterophilic antibodys, and dilution effects. A solution to the lack of agreement of the measured values between the two methods can be the division to categories. The membership to a certain category may then be compared between CBA and ELISA. 2013-10-07 Immunologie Im Rahmen der prospektiven multizentrischen Kohortenstudie PASTURE/ EFRAIM wurden Einflussfaktoren auf die Entstehung von kindlichem Asthma und Allergien untersucht. Ein Bestandteil war die Erstellung des T-Effektorzell-Profils der Teilnehmer mittels Zytokinmessung zu verschiedenen Zeitpunkten. Der für die Zytokinmessung verwendete ELISA wird derzeit als Goldstandard betrachtet, ist jedoch sehr Zeit-, Kosten-, Personal- und Probenintensiv. Die Messung der Zytokine sollte somit auf ein Hochdurchsatzverfahren, das „cytometric bead array assay (CBA)“-System der Firma BD Biosciences umgestellt werden. Ziel dieser Arbeit war es, die CBA-Methode zu validieren, sie auf ihre Messqualität im Vergleich zum ELISA und auf die Vergleichbarkeit von Messergebnissen beider Methoden zu untersuchen. Für den Vergleich der Messergebnisse beider Methoden wurden 556 Proben von vierzig Studienteilnehmern parallel per CBA und ELISA gemäß Herstelleranweisung gemessen. Die Messungen mittels ELISA wurden auf die gleiche Weise durchgeführt wie die Messungen der Studienproben im Rahmen der PASTURE-/ EFRAIM-Studie zuvor. Die Ergebnisse hinsichtlich Probenverschleppung, Kalibrationsstabilität, Analytischer und Funktioneller Sensitivität, Präzision, Wiederfindung und Linearität belegen, dass die CBA-Methode mindestens so stabile und verlässliche Messwerte generiert, wie die ELISA-Methode. In den meisten Fällen ist sie dabei jedoch bei Probenverbrauch, Kosteneffizienz, Reproduzierbarkeit und Benutzerfreundlichkeit überlegen. Die Vergleichbarkeit von Messergebnissen identischer Proben zeigt, dass die Ergebnisse zwischen ELISA und CBA mit Korrelationskoeffizienten (Pearson) zwischen 0,854 (TNF-a) und 0,930 (IL-10) für unverdünnt gemessene Proben korrelieren. Die Konkordanzkoeffizienten nach Lin, sowie die Darstellung der Messergebnisse nach Bland und Altman zeigen nur eine gute bis mäßige Übereinstimmung der Messergebnisse in niedrigen Konzentrationsbereichen und eine schlechte Übereinstimmung in hohen Konzentrationsbereichen. Die vorliegenden Ergebnisse lassen mit Einschränkungen einen Vergleich der Werte zwischen 5 pg/ml bis maximal 100 pg/ml für IL-5 zu, für IL-10 von 20 pg/ml bis 300 pg/ml, für IFN-g und TNF-a von 20 pg/ml bis 1000 pg/ml. Die Ursache liegt höchstwahrscheinlich in den unterschiedlichen Antikörpern der beiden Methoden, am Vorliegen von heterophilen Antikörpern, sowie an Verdünnungseffekten. Eine Lösung der dargestellten Probleme kann die Aufteilung der Messergebnisse in Kategorien sein. Die Zugehörigkeit zu einer Kategorie kann dann zwischen den Messmethoden verglichen werden. Medizin urn:nbn:de:hebis:04-z2013-05861 cytokine measurement Comparison of methods to measure cytokines in the context of large-scale epidemiologic studies Medical sciences Medicine Medizin https://archiv.ub.uni-marburg.de/diss/z2013/0586/cover.png 2013 https://doi.org/10.17192/z2013.0586 Methodenvergleich zur Messung von Zytokinen im Rahmen großer epidemiologischer Studien BLAND, J. M. und D. G. ALTMAN (2003). Applying the right statistics: analyses of measurement studies. Ultrasound in obstetrics gynecology : the official journal of the International Society of Ultrasound in Obstetrics and Gynecology, 22(1):85– 93. ALM, J. S., J. SWARTZ, G. LILJA, A. SCHEYNIUS und G. PERSHAGEN (1999). Atopy in children of families with an anthroposophic lifestyle. Lancet, 353(9163):1485–8. PFEFFERLE, PETRA INA, G. BUECHELE, N. BLUEMER, M. ROPONEN, M. J. EGE, S. KRAUSS-ETSCHMANN, J. GENUNEIT, A. HYVARINEN, M.-R. HIRVONEN, R. LAUENER, J. PEKKANEN, J. RIEDLER, J. C. DALPHIN, B. BRUNEKEEF, C. BRAUN-FAHRLANDER, E. VON MUTIUS und H. RENZ (2010). Cord blood cytokines are modulated by maternal farming activities and consumption of farm dairy products during pregnancy: the PASTURE Study. The Journal of allergy and clinical immunology, 125(1):108–15.e1–3. KHAN, SAMEENA S., M. S. SMITH, D. REDA, A. F. SUFFREDINI und J. P. J. MCCOY (2004). Multiplex bead array assays for detection of soluble cytokines: comparisons of sensitivity and quantitative values among kits from multiple manufacturers. Cytometry. Part B, Clinical cytometry, 61(1):35–9. Variable estimates of cytokine levels produced by commercial ELISA kits: results using international cytokine standards. Journal of immunological methods, 186(2):171–9. NICOLAI, T. und E. VON MUTIUS (1996). Respiratory hypersensitivity and environmen- tal factors: East and West Germany. Toxicol Lett, 86(2-3):105–13. PRABHAKAR, UMA, E. EIRIKIS, M. REDDY, E. SILVESTRO, S. SPITZ, C. N. PENDLEY, H. M. DAVIS und B. E. MILLER (2004). Validation and comparative analysis of a multiplexed assay for the simultaneous quantitative measurement of Th1/Th2 cytokines in human serum and human peripheral blood mononuclear cell culture supernatants. Journal of immunological methods, 291(1-2):27–38. YOUNG, SHIH-HOUNG, J. M. ANTONINI, J. R. ROBERTS, A. D. ERDELY und P. C. ZEIDLER-ERDELY (2008). Performance evaluation of cytometric bead assays for the measurement of lung cytokines in two rodent models. Journal of immunological methods, 331(1-2):59–68. TOEPLITZ, BARBARA K., A. I. COHEN, P. T. FUNKE, W. L. PARKER und J. Z. GOUGOUTAS (1979). Structure of ionomycin -a novel diacidic polyether antibiotic having high affinity for calcium ions. Journal of the American Chemical Society, 101(12):3344–53. VON EHRENSTEIN, O. S., E. VON MUTIUS, S. ILLI, L. BAUMANN, O. BOHM und R. VON KRIES (2000). Reduced risk of hay fever and asthma among children of farmers. Clin Exp Allergy, 30(2):187–93. LENG, SEAN X., J. E. MCELHANEY, J. D. WALSTON, D. XIE, N. S. FEDARKO und G. A. KUCHEL (2008). ELISA and multiplex technologies for cytokine measurement in inflammation and aging research. J Gerontol A Biol Sci Med Sci, 63(8):879–84. RICHENS, JOANNA L., R. A. URBANOWICZ, R. METCALF, J. CORNE, P. O'SHEA und L. FAIRCLOUGH (2010). Quantitative validation and comparison of multiplex cytokine kits. Journal of biomolecular screening, 15(5):562–8. MAECKER, HOLDEN T., J. HASSLER, J. K. PAYNE, A. SUMMERS, K. COMATAS, M. GHANAYEM, M. A. MORSE, T. M. CLAY, H. K. LYERLY, S. BHATIA, S. A. GHANEKAR, V. C. MAINO, C. DELAROSA und M. L. DISIS (2008). Precision and linearity targets for validation of an IFNgamma ELISPOT, cytokine flow cytometry, and tetramer assay using CMV peptides. BMC immunology, 9:9. Cord blood allergen-specific IgE is associated with reduced IFN-gamma production by cord blood cells: the Protection against Allergy-Study in Rural Environments (PA- STURE) Study. The Journal of allergy and clinical immunology, 122(4):711–6. MCINNES, IAIN B. und G. SCHETT (2011). The pathogenesis of rheumatoid arthritis. The New England journal of medicine, 365(23):2205–19. NOWAK, D., J. HEINRICH, R. JORRES, G. WASSMER, J. BERGER, E. BECK, S. BOCZOR, M. CLAUSSEN, H. E. WICHMANN und H. MAGNUSSEN (1996). Prevalence of respiratory symptoms, bronchial hyperresponsiveness and atopy among adults: west and east Germany. Eur Respir J, 9(12):2541–52. ASHER, M. INNES, S. MONTEFORT, B. BJORKSTEN, C. K. W. LAI, D. P. STRACHAN, S. K. WEILAND und H. WILLIAMS (2006). Worldwide time trends in the preva- lence of symptoms of asthma, allergic rhinoconjunctivitis, and eczema in childhood: ISAAC Phases One and Three repeat multicountry cross-sectional surveys. Lancet, 368(9537):733–43. LIN, L. I. (1989). A concordance correlation coefficient to evaluate reproducibility. Biome- trics, 45(1):255–68. KAY, A. B. (2001). Allergy and allergic diseases. First of two parts. N Engl J Med, 344(1):30–7. AGNELLO, DAVIDE, C. S. R. LANKFORD, J. BREAM, A. MORINOBU, M. GADINA, J. J. O'SHEA und D. M. FRUCHT (2003). Cytokines and transcription factors that regulate T helper cell differentiation: new players and new insights. J Clin Immunol, 23(3):147–61. MORGAN, EDWARD, R. VARRO, H. SEPULVEDA, J. A. EMBER, J. APGAR, J. WIL- SON, L. LOWE, R. CHEN, L. SHIVRAJ, A. AGADIR, R. CAMPOS, D. ERNST und A. GAUR (2004). Cytometric bead array: a multiplexed assay platform with applications in various areas of biology. Clinical immunology (Orlando, Fla.), 110(3):252–66. RAY, CHAD A., R. R. BOWSHER, W. C. SMITH, V. DEVANARAYAN, M. B. WILLEY, J. T. BRANDT und R. A. DEAN (2005). Development, validation, and implementation of a multiplex immunoassay for the simultaneous determination of five cytokines in human serum. Journal of pharmaceutical and biomedical analysis, 36(5):1037–44. BORG, LONE, J. KRISTIANSEN, J. M. CHRISTENSEN, K. F. JEPSEN und L. K. POUL- SEN (2002). Evaluation of accuracy and uncertainty of ELISA assays for the determina- tion of interleukin-4, interleukin-5, interferon-gamma and tumor necrosis factor-alpha. Clinical chemistry and laboratory medicine : CCLM / FESCC, 40(5):509–19. MUTIUS, ERIKA VON und D. VERCELLI (2010). Farm living: effects on childhood asthma and allergy. Nature reviews. Immunology, 10(12):861–8. MARTIN, R. F. (2000). General deming regression for estimating systematic bias and its confidence interval in method-comparison studies. Clinical chemistry, 46(1):100–4. STRACHAN, D. P. (1989). Hay fever, hygiene, and household size. BMJ, 299(6710):1259– 60. BAUER, STEFAN, D. HANGEL und P. YU (2007). Immunobiology of toll-like receptors in allergic disease. Immunobiology, 212(6):521–33. ROMAGNANI, SERGIO (2004). Immunologic influences on allergy and the TH1/TH2 balance. J Allergy Clin Immunol, 113(3):395–400. AVERBECK, MARCO, C. GEBHARDT, F. EMMRICH, R. TREUDLER und J. C. SIMON (2007). Immunologic principles of allergic disease. J Dtsch Dermatol Ges, 5(11):1015– 28. Is the prevalence of asthma declining? Systematic review of epidemiological studies. Allergy, 65(2):152–67. VI B LITERATURVERZEICHNIS MURPHY, KENNETH M., P. TRAVERS, M. WALPORT, C. A. JANEWAY, L. SEID- LER und M. EHRENSTEIN (2009). Janeway Immunologie. Spektrum Akad. Verl, Heidelberg, 7. Aufl. LANGE, S. und R. BENDER (2007). Lineare Regression und Korrelation. Deutsche medizinische Wochenschrift (1946), 132 Suppl 1:e9–11. BLAND, J. M. und D. G. ALTMAN (1999). Measuring agreement in method comparison studies. Statistical methods in medical research, 8(2):135–60. KUMAR, HIMANSHU, T. KAWAI und S. AKIRA (2011). Pathogen recognition by the innate immune system. International reviews of immunology, 30(1):16–34. WADHWA, M. und R. THORPE (1997). Standardization and calibration of cytokine immunoassays: meeting report and recommendations. Cytokine, 9(11):791–3. BLAND, J. M. und D. G. ALTMAN (1986). Statistical methods for assessing agreement between two methods of clinical measurement. Lancet, 1(8476):307–10. RENZ, HARALD (2004). The biology of T-cells in allergy and asthma: beyond the TH1/TH2 concept. Pediatr Pulmonol Suppl, 26:40–1. O'GARRA, A. und N. ARAI (2000). The molecular basis of T helper 1 and T helper 2 cell differentiation. Trends Cell Biol, 10(12):542–50. LIU, W. S. und C. A. HECKMAN (1998). The sevenfold way of PKC regulation. Cell Signal, 10(8):529–42. TIMMONS, BRIAN W., M. J. HAMADEH und M. A. TARNOPOLSKY (2009). Two me- thods for determining plasma IL-6 in humans at rest and following exercise. European journal of applied physiology, 105(1):13–8. BEASLEY, A, U. KEIL, E. VON MUTIUS und N. PEARCE (1998). Worldwide variation in prevalence of symptoms of asthma, allergic rhinoconjunctivitis, and atopic ecze- ma: ISAAC. The International Study of Asthma and Allergies in Childhood (ISAAC) Steering Committee. Lancet, 351(9111):1225–32. STRACHAN, D. P. (1997). Allergy and family size: a riddle worth solving. Clin Exp Allergy, 27(3):235–6. NICOLAI, T., B. BELLACH, E. V. MUTIUS, W. THEFELD und H. HOFFMEISTER (1997). Increased prevalence of sensitization against aeroallergens in adults in West compared with East Germany. Clin Exp Allergy, 27(8):886–92. PEARCE, N., J. SUNYER, S. CHENG, S. CHINN, B. BJORKSTEN, M. BURR, U. KEIL, H. R. ANDERSON und P. BURNEY (2000). Comparison of asthma prevalence in the ISAAC and the ECRHS. ISAAC Steering Committee and the European Community Respiratory Health Survey. International Study of Asthma and Allergies in Childhood. Eur Respir J, 16(3):420–6. J. SUNYER und M. WJST (2001). The European Community Respiratory Health Sur- vey: what are the main results so far? European Community Respiratory Health Survey II. Eur Respir J, 18(3):598–611. NANDAKUMAR, SUBHADRA, C. W. MILLER und U. KUMARAGURU (2009). T regula- tory cells: an overview and intervention techniques to modulate allergy outcome. Clin Mol Allergy, 7:5. MUTIUS, E. VON und S. SCHMID (2006). The PASTURE project: EU support for the improvement of knowledge about risk factors and preventive factors for atopy in Europe. Allergy, 61(4):407–13. MANICASSAMY, SANTHAKUMAR und B. PULENDRAN (2009). Modulation of adaptive immunity with Toll-like receptors. Semin Immunol, 21(4):185–93. PAPPU, BHANU P., P. ANGKASEKWINAI und C. DONG (2008). Regulatory mechanisms of helper T cell differentiation: new lessons learned from interleukin 17 family cytokines. Pharmacol Ther, 117(3):374–84. VIGNALI, DARIO A. A., L. W. COLLISON und C. J. WORKMAN (2008). How regula- tory T cells work. Nature reviews. Immunology, 8(7):523–32. LEMANSKE, ROBERT F. JR und W. W. BUSSE (2010). Asthma: clinical expression and molecular mechanisms. The Journal of allergy and clinical immunology, 125(2 Suppl 2):S95–102. Ellington AA, Kullo IJ, Bailey KR, Klee GG. Antibody-based protein multiplex platforms: technical and operational challeng- es. Clin Chem 2010;56:186–93. WAMBRE, ERIK, E. A. JAMES und W. W. KWOK (2012). Characterization of CD4+ T cell subsets in allergy. Current opinion in immunology, 24(6):700–6. LARCHE, MARK, C. A. AKDIS und R. VALENTA (2006). Immunological mechanisms of allergen-specific immunotherapy. Nature reviews. Immunology, 6(10):761–71. CBA Medizin 2013-10-07 opus:5138 doctoralThesis application/pdf Allergie ELISA ths Prof. Dr. Renz Harald Renz, Harald (Prof. Dr.)