Abstract
Previous studies demonstrated that no significant relationships exist between salivary and serum IL-6 in resting conditions and following exercise and that appropriate saliva collection procedures allow to avoid analytical drawbacks. This investigation aimed to: (a) compare the effects of two methods of saliva collection on IL-6 assay; (b) search for correlation between salivary and serum IL-6 in resting and post-exercise conditions; (c) evaluate the IL-6 response to isometric contractions. Seventeen sedentary subjects and fifteen athletes underwent one blood and two salivary draws: saliva was collected chewing on cotton salivettes and using a plastic straw (SA method and ST method, respectively). Afterwards, the athletes only completed a fatiguing isometric exercise of the knee extensors and blood and saliva were sampled after the exercise. In the entire group (n = 32), ST method produced higher IL-6 levels than SA method and serum sampling. The exercise elicited significant responses of lactate, serum IL-6, salivary IL-6 (by ST method): salivary IL-6 values using the ST collection method were higher at each sampling point than with the SA method. The correlation analyses applied to both resting levels in the entire group and absolute changes above baseline in the athlete group showed that: (1) no significant relationships exist between serum and salivary IL-6 levels; (2) the greater the salivary IL-6 measurement, the higher the resultant inaccuracy of the SA method; (3) significant correlations exist between isometric force and mechanical fatigue during exercise and peaks of lactate and serum IL-6. These data provided demonstration of a cotton-interference effect for the results of salivary IL-6 assay and confirmed the lack of significant correlation between salivary and serum IL-6 in resting and post-exercise conditions.
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References
Aleksandra Nielsen A, Nederby Nielsen J, Schmedes A, Brandslund I, Hey H (2005) Saliva Interleukin-6 in patients with inflammatory bowel disease. Scand J Gastroenterol 40:1444–1448
Anderson DJ, Hector MP, Linden RW (1996) The effects of unilateral and bilateral chewing, empty clenching and simulated bruxism, on the masticatory-parotid salivary reflex in man. Exp Physiol 81:305–312
Bland JM, Altman DG (1986) Statistical methods for assessing agreement between two methods of clinical measurement. Lancet 1:307–310
Boumba D, Skopouli FN, Moutsopoulos HM (1995) Cytokine mRNA expression in the labial salivary gland tissues from patients with primary Sjogren’s syndrome. Br J Rheumatol 34:326–333
Croisier JL, Camus G, Venneman I, Deby-Dupont G, Juchmes-Ferir A, Lamy M, Crielaard JM, Deby C, Duchateau J (1999) Effects of training on exercise-induced muscle damage and interleukin 6 production. Muscle Nerve 22:208–212
Dennis RA, Trappe TA, Simpson P, Carroll C, Huang BE, Nagarajan R, Bearden E, Gurley C, Duff GW, Evans WJ, Kornman K, Peterson CA (2004) Interleukin-1 polymorphisms are associated with the inflammatory response in human muscle to acute resistance exercise. J Physiol 560:617–626
Edwards KM, Burns VE, Ring C, Carroll D (2006) Individual differences in the interleukin-6 response to maximal and submaximal exercise tasks. J Sports Sci 24:855–862
Fischer CP (2006) Interleukin-6 in acute exercise and training: what is the biological relevance? Exerc Immunol Rev 12:6–33
Fischer CP, Hiscock NJ, Penkowa M, Basu S, Vessby B, Kallner A, Sjoberg LB, Pedersen BK (2004) Supplementation with vitamins C and E inhibits the release of interleukin-6 from contracting human skeletal muscle. J Physiol 558:633–645
Formanek M, Knerer B, Temmel A, Thurnher D, Millesi W, Kornfehl J (1998) Oral keratinocytes derived from the peritonsillar mucosa express the proinflammatory cytokine IL-6 without prior stimulation. J Oral Pathol Med 27:202–206
Formanek M, Knerer B, Kornfehl J (1999) Cytokine expression of human oral keratinocytes. ORL J Otorhinolaryngol Relat Spec 61:103–107
Fox RI, Kang HI, Ando D, Abrams J, Pisa E (1994) Cytokine mRNA expression in salivary gland biopsies of Sjogren’s syndrome. J Immunol 152:5532–5539
Helge JW, Stallknecht B, Pedersen BK, Galbo H, Kiens B, Richter EA (2003) The effect of graded exercise on IL-6 release and glucose uptake in human skeletal muscle. J Physiol 546:299–305
Hiscock N, Chan MH, Bisucci T, Darby IA, Febbraio MA (2004) Skeletal myocytes are a source of interleukin-6 mRNA expression and protein release during contraction: evidence of fiber type specificity. FASEB J 18:992–994
Jensen Kjeilen JC, Brodin P, Aars H, Berg T (1987) Parotid salivary flow in response to mechanical and gustatory stimulation in man. Acta Physiol Scand 131:169–175
Kirschbaum C, Hellhammer DH (1989) Salivary cortisol in psychobiological research: an overview. Neuropsychobiology 22:150–169
Margeli A, Skenderi K, Tsironi M, Hantzi E, Matalas AL, Vrettou C, Kanavakis E, Chrousos G, Papassotiriou I (2005) Dramatic elevations of interleukin-6 and acute-phase reactants in athletes participating in the ultradistance foot race spartathlon: severe systemic inflammation and lipid and lipoprotein changes in protracted exercise. J Clin Endocrinol Metab 90:3914–3918
Minetto M, Rainoldi A, Gazzoni M, Terzolo M, Borrione P, Termine A, Saba L, Dovio A, Angeli A, Paccotti P (2005) Differential responses of serum and salivary interleukin-6 to acute strenuous exercise. Eur J Appl Physiol 93:679–686
Minetto MA, Rainoldi A, Gazzoni M, Ganzit GP, Saba L, Paccotti P (2006) Interleukin-6 response to isokinetic exercise in elite athletes: relationships to adrenocortical function and to mechanical and myoelectric fatigue. Eur J Appl Physiol 98:373–382
Okada N, Kobayashi M, Mugikura K, Okamatsu Y, Hanazawa S, Kitano S, Hasegawa K (1997) Interleukin-6 production in human fibroblasts derived from periodontal tissues is differentially regulated by cytokines and a glucocorticoid. J Periodontal Res 32:559–569
Ostrowski K, Hermann C, Bangash A, Schjerling P, Nielsen JN, Pedersen BK (1998) A trauma-like elevation of plasma cytokines in humans in response to treadmill running. J Physiol 15:889–894
Parkar M, Tabona P, Newman H, Olsen I (1998) IL-6 expression by oral fibroblasts is regulated by androgen. Cytokine 10:613–619
Pedersen BK, Hoffman-Goetz L (2000) Exercise and the immune system: regulation, integration, and adaptation. Physiol Rev 80:1055–1081
Pedersen BK, Febbraio M (2005) Muscle-derived interleukin-6 – a possible link between skeletal muscle, adipose tissue, liver, and brain. Brain Behav Immun 19:371–376
Pedersen BK, Steensberg A, Fischer C, Keller C, Ostrowski K, Schjerling P (2001) Exercise and cytokines with particular focus on muscle-derived IL-6. Exerc Immunol Rev 7:18–31
Penkowa M, Keller C, Keller P, Jauffred S, Pedersen BK (2003) Immunohistochemical detection of interleukin-6 in human skeletal muscle fibers following exercise. FASEB J 17:2166–2168
Petersen AM, Pedersen BK (2005) The anti-inflammatory effect of exercise. J Appl Physiol 98:1154–1162
Plomgaard P, Penkowa M, Pedersen BK (2005) Fiber type specific expression of TNF-alpha, IL-6 and IL-18 in human skeletal muscles. Exerc Immunol Rev 11:53–63
Pyne DB, Boston T, Martin DT, Logan A (2000) Evaluation of the Lactate Pro blood lactate analyser. Eur J Appl Physiol 82:112–116
Shirtcliff EA, Granger DA, Schwartz E, Curran MJ (2001) Use of salivary biomarkers in biobehavioral research: cotton-based sample collection methods can interfere with salivary immunoassay results. Psychoneuroendocrinology 26:165–173
Steensberg A, van Hall G, Osada T, Sacchetti M, Saltin B, Klarlund Pedersen B (2000) Production of interleukin-6 in contracting human skeletal muscles can account for the exercise-induced increase in plasma interleukin-6. J Physiol 529:237–242
Steensberg A, Febbraio MA, Osada T, Schjerling P, van Hall G, Saltin B, Pedersen BK (2001) Interleukin-6 production in contracting human skeletal muscle is influenced by pre-exercise muscle glycogen content. J Physiol 537:633–639
Vgontzas AN, Papanicolaou DA, Bixler EO, Lotsikas A, Zachman K, Kales A, Prolo P, Wong ML, Licinio J, Gold PW, Hermida RC, Mastorakos G, Chrousos GP (1999) Circadian interleukin-6 secretion and quantity and depth of sleep. J Clin Endocrinol Metab 84:2603–2607
Yamamoto T, Kita M, Kimura I, Oseko F, Terauchi R, Takahashi K, Kubo T, Kanamura N (2006) Mechanical stress induces expression of cytokines in human periodontal ligament cells. Oral Dis 12:171–175
Yao C, Wei W, Li X, Hosoi K (2005) Acute phase protein induction by experimental inflammation in the salivary gland. J Oral Pathol Med 34:364–367
Acknowledgments
We wish to thank Prof. E. Ghigo (Division of Endocrinology and Metabolism, Department of Internal Medicine, University of Turin, Turin, Italy) and Prof. R. Merletti (LISiN, Department of Electronics, Polytechnic of Turin, Turin, Italy) for their constructive criticisms in the manuscript editing and M. Gollin and P. Bertano (SUISM, University of Turin, Turin, Italy) for valuable assistance in the sample collection.
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Minetto, M.A., Gazzoni, M., Lanfranco, F. et al. Influence of the sample collection method on salivary interleukin–6 levels in resting and post-exercise conditions. Eur J Appl Physiol 101, 249–256 (2007). https://doi.org/10.1007/s00421-007-0484-x
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DOI: https://doi.org/10.1007/s00421-007-0484-x