Identification of inflammatory markers suitable for non-invasive, repeated measurement studies in biobehavioral research: A feasibility study

Autoři: H. M. Schenk aff001;  S. L. van Ockenburg aff001;  M. C. Nawijn aff002;  P. De Jonge aff001;  J. G. M. Rosmalen aff001
Působiště autorů: Interdisciplinary Center Psychopathology and Emotion regulation (ICPE), University Center for Psychiatry, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands aff001;  Department of Pathology and Medical Biology, GRIAC Research Institute, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands aff002
Vyšlo v časopise: PLoS ONE 14(9)
Kategorie: Research Article
doi: 10.1371/journal.pone.0221993



Studying the role of the immune system in the interaction between mental and physical health is challenging. To study individuals with an intensive, longitudinal study design that requires repetitive sampling in their daily life, non-invasive sampling techniques are a necessity. Urine can be collected in a non-invasive way, but this may be demanding for participants and little is known about fluctuation of inflammatory markers in urine over time. The aim of this study was to investigate the feasibility of non-invasive sampling, and to explore intra-individual differences in inflammatory markers in urine.

Materials & methods

Ten healthy individuals collected 24-hour urine for 63 consecutive days. In a pilot analysis, 39 inflammatory markers were examined for detectability in urine, stability over time and under storage conditions, and daily fluctuations. Multiplex analyses were used to quantify levels of eight selected markers: C-reactive protein (CRP), Fractalkine, Interleukin-1 receptor-antagonist (IL-1RA), interferon-α (IFNα), interferon-γ (IFNγ), Interferon gamma-induced protein 10 (IP10), Macrophage inflammatory protein-1β (MIP-1β), and Vascular Endothelial Growth Factor (VEGF). Cross-correlations were calculated between the overnight and 24-hour samples were calculated, to examine whether 24-hour urine could be replaced by the overnight portion for better feasibility. We examined intra- and interindividual differences in the levels of inflammatory markers in urine and the fluctuations thereof.


This study showed that levels of selected inflammatory markers can be detected in urine. Cross-correlation analyses showed that correlations between levels of inflammatory markers in the night portion and the 24-hour urine sample varied widely between individuals. In addition, analyses of time series revealed striking inter- and intra-individual variation in levels of inflammatory markers and their fluctuations.


We show that the assessment of urinary inflammatory markers is feasible in an intensive day-to-day study in healthy individuals. However, 24-hour urine cannot be replaced by an overnight portion to alleviate the protocol burden. Levels of inflammatory markers show substantial variation between and within persons.

Klíčová slova:

C-reactive proteins – Cytokines – Excretion – Inflammation – Interferons – Interleukins – Mental health and psychiatry – Urine


1. Gabbay V, Klein RG, Alonso CM, Babb JS, Nishawala M, De Jesus G, et al. Immune system dysregulation in adolescent major depressive disorder. J Affect Disord. 2009; doi: 10.1016/j.jad.2008.07.022 18790541

2. Prince M, Patel V, Saxena S, Maj M, Maselko J, Phillips MR, et al. No health without mental health. Lancet. 2007;370: 859–877. doi: 10.1016/S0140-6736(07)61238-0 17804063

3. Dahl J, Ormstad H, Aass HCD, Malt UF, Bendz LT, Sandvik L, et al. The plasma levels of various cytokines are increased during ongoing depression and are reduced to normal levels after recovery. Psychoneuroendocrinology. 2014; doi: 10.1016/j.psyneuen.2014.03.019 24845179

4. Wium-Andersen MK, Ørsted DD, Nielsen SF, Nordestgaard BG. Elevated C-reactive protein levels, psychological distress, and depression in 73, 131 individuals. JAMA Psychiatry. 2013;70: 176–84. doi: 10.1001/2013.jamapsychiatry.102 23266538

5. Rohleder N. Stimulation of Systemic Low-Grade Inflammation by Psychosocial Stress. Psychosom Med. 2014;76: 181–189. doi: 10.1097/PSY.0000000000000049 24608036

6. Sublette ME, Postolache TT. Neuroinflammation and depression: The role of indoleamine 2,3-dioxygenase (IDO) as a molecular pathway. Psychosomatic Medicine. 2012. doi: 10.1097/PSY.0b013e318268de9f 22923699

7. Reis HT. Why researchers should think “real-world”: A conceptual rationale. In: Mehl MR; Connor T, editor. Handbook of research methods for studying daily life. 2012. pp. 3–21.

8. Chiang JJ, Eisenberger NI, Seeman TE, Taylor SE. Negative and competitive social interactions are related to heightened proinflammatory cytokine activity. Proc Natl Acad Sci. 2012; doi: 10.1073/pnas.1120972109 22308464

9. Segerstrom SC, Miller GE. Psychological Stress and the Human Immune System: A Meta-Analytic Study of 30 Years of Inquiry NIH Public Access. Psychol Bull. 2004.

10. Hamaker EL. Why researchers should think “within-person”: A paradigmatic rationale. Handbook of research methods for studying daily life. New York: Guilford Press; 2012. pp. 43–61.

11. van Ockenburg SL, Booij SH, Riese H, Rosmalen JGM, Janssens KAM. How to assess stress biomarkers for idiographic research? Psychoneuroendocrinology. 2015; doi: 10.1016/j.psyneuen.2015.08.002 26318629

12. Shiffman S, Stone A a., Hufford MR. Ecological Momentary Assessment. Annu Rev Clin Psychol. 2008;4: 1–32. doi: 10.1146/annurev.clinpsy.3.022806.091415 18509902

13. Haberkorn J, Burbaum C, Fritzsche K, Geser W, Fuchs D, Ocaña-Peinado FM, et al. Day-to-day cause-effect relations between cellular immune activity, fatigue and mood in a patient with prior breast cancer and current cancer-related fatigue and depression. Psychoneuroendocrinology. 2013; doi: 10.1016/j.psyneuen.2013.03.001 23541233

14. Steptoe A, Hamer M, Chida Y. The effects of acute psychological stress on circulating inflammatory factors in humans: A review and meta-analysis. Brain, Behavior, and Immunity. 2007. doi: 10.1016/j.bbi.2007.03.011 17475444

15. Khan A. Detection and quantitation of forty eight cytokines, chemokines, growth factors and nine acute phase proteins in healthy human plasma, saliva and urine. J Proteomics. 2012; doi: 10.1016/j.jprot.2012.05.018 22634087

16. Prasad S, Tyagi AK, Aggarwal BB. Detection of inflammatory biomarkers in saliva and urine: Potential in diagnosis, prevention, and treatment for chronic diseases. Exp Biol Med. 2015;241: 783–799. doi: 10.1177/1535370216638770 27013544

17. Schubert C, Geser W, Noisternig B, Fuchs D, Welzenbach N, König P, et al. Stress system dynamics during “life as it is lived”: An integrative single-case study on a healthy woman. PLoS One. 2012; doi: 10.1371/journal.pone.0029415 22403606

18. Nayyar AS, Khan M, Vijayalakshmi KR, Suman B, Gayitri HC, Anitha M. Serum total protein, albumin and advanced oxidation protein products (AOPP)—implications in oral squamous cell carcinoma. Malays J Pathol. 2012;

19. Stiegel MA, Pleil JD, Sobus JR, Morgan MK, Madden MC. Analysis of inflammatory cytokines in human blood, breath condensate, and urine using a multiplex immunoassay platform. Biomarkers. 2015; doi: 10.3109/1354750X.2014.988646 25495125

20. Booij SH, Bos EH, Bouwmans MEJ, Van Faassen M, Kema IP, Oldehinkel AJ, et al. Cortisol and α-amylase secretion patterns between and within depressed and non-depressed individuals. PLoS One. 2015; doi: 10.1371/journal.pone.0131002 26148294

21. Slavish DC, Graham-Engeland JE, Smyth JM, Engeland CG. Salivary markers of inflammation in response to acute stress. Brain, Behavior, and Immunity. 2015. doi: 10.1016/j.bbi.2014.08.008 25205395

22. Hamaker EL, Dolan C V., Molenaar PCM. Statistical Modeling of the Individual: Rationale and Application of Multivariate Stationary Time Series Analysis. Multivariate Behav Res. 2005;40: 207–233. doi: 10.1207/s15327906mbr4002_3 26760107

23. van Ockenburg SL, Schenk HM, van der Veen A, van Rossum EFC, Kema IP, Rosmalen JGM. The relationship between 63 days of 24-h urinary free cortisol and hair cortisol levels in 10 healthy individuals. Psychoneuroendocrinology. 2016; doi: 10.1016/j.psyneuen.2016.07.220 27497154

24. Murakami K, Sasaki S, Takahashi Y, Uenishi K, Watanabe T, Kohri T, et al. Sensitivity and specificity of published strategies using urinary creatinine to identify incomplete 24-h urine collection. Nutrition. 2008;24: 16–22. doi: 10.1016/j.nut.2007.09.001 17996421

25. Honaker J, King G, Blackwell M. AMELIA II: A Program for Missing Data. J Stat Softw. 2011;45: 1–54. doi:

26. Peng W, Chen J, Jiang Y, Wu J, Shou Z, He Q, et al. Urinary fractalkine is a marker of acute rejection. Kidney Int. 2008; doi: 10.1038/ki.2008.459 18800027

27. Bhide AA, Cartwright R, Khullar V, Digesu GA. Biomarkers in overactive bladder. International Urogynecology Journal. 2013. doi: 10.1007/s00192-012-2027-1 23314226

28. Huan Y, Qin W, Gao Y. Factors to consider in the verification of urinary biomarkers. Science China Life Sciences. 2018. doi: 10.1007/s11427-017-9204-5 29524122

29. Scheiermann C, Kunisaki Y, Frenette PS. Circadian control of the immune system. Nat Rev Immunol. 2013;13: 190–198. doi: 10.1038/nri3386 23391992

30. McAlpine CS, Swirski FK. Circadian Influence on Metabolism and Inflammation in Atherosclerosis. Circ Res. 2016;119: 131–141. doi: 10.1161/CIRCRESAHA.116.308034 27340272

31. Patel SR, Zhu X, Storfer-Isser A, Mehra R, Jenny NS, Tracy R, et al. Sleep duration and biomarkers of inflammation. Sleep. 2009;32: 200–4. doi: 10.1093/sleep/32.2.200 19238807

32. Cermakian N, Lange T, Golombek D, Sarkar D, Nakao A, Shibata S, et al. Crosstalk between the circadian clock circuitry and the immune system. Chronobiol Int. 2013; doi: 10.3109/07420528.2013.782315 23697902

33. Wright KP, Drake AL, Frey DJ, Fleshner M, Desouza CA, Gronfier C, et al. Influence of sleep deprivation and circadian misalignment on cortisol, inflammatory markers, and cytokine balance. Brain Behav Immun. 2015; doi: 10.1016/j.bbi.2015.01.004 25640603

34. Alireza S, Forough S, Khosro S, Omid A. Night work and inflammatory markers. Indian J Occup Environ Med. 2011;15: 38. doi: 10.4103/0019-5278.82996 21808500

35. Dorn LD, Gayles JG, Engeland CG, Houts R, Cizza G, Denson LA. Cytokine Patterns in Healthy Adolescent Girls. Psychosom Med. 2016; 1. doi: 10.1097/PSY.0000000000000321 27187849

36. Kleiner G, Marcuzzi A, Zanin V, Monasta L, Zauli G. Cytokine levels in the serum of healthy subjects. Mediators Inflamm. 2013; doi: 10.1155/2013/434010 23533306

37. De Melo EN, Deda L, Har R, Reich HN, Scholey JW, Daneman D, et al. The urinary inflammatory profile in gluten free diet—Adherent adolescents with type 1 diabetes and celiac disease. J Diabetes Complications. 2016; doi: 10.1016/j.jdiacomp.2015.11.020 26790575

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