Cytokine signatures in chronic fatigue syndrome patients: a Case Control Study and the effect of anakinra treatment

Megan E Roerink, Hans Knoop, Ewald M Bronkhorst, Henk A Mouthaan, Luuk J A C Hawinkels, Leo A B Joosten, Jos W M van der Meer, Megan E Roerink, Hans Knoop, Ewald M Bronkhorst, Henk A Mouthaan, Luuk J A C Hawinkels, Leo A B Joosten, Jos W M van der Meer

Abstract

Background: Cytokine disturbances have been suggested to be associated with the Chronic Fatigue Syndrome/Myalgic encephalomyelitis (CFS/ME) for decades.

Methods: Fifty female CFS patients were included in a study on the effect of the interleukin-1-receptor antagonist anakinra or placebo during 4 weeks. EDTA plasma was collected from patients before and directly after treatment. At baseline, plasma samples were collected at the same time from 48 healthy, age-matched female neighborhood controls. A panel of 92 inflammatory markers was determined in parallel in 1 μL samples using a 'proximity extension assay' (PEA) based immunoassay. Since Transforming growth factor beta (TGF-β) and interleukin-1 receptor antagonist (IL-1Ra) were not included in this platform, these cytokines were measured with ELISA.

Results: In CFS/ME patients, the 'normalized protein expression' value of IL-12p40 and CSF-1 was significantly higher (p value 0.0042 and 0.049, respectively). Furthermore, using LASSO regression, a combination of 47 markers yielded a prediction model with a corrected AUC of 0.73. After correction for multiple testing, anakinra had no effect on circulating cytokines. TGF-β did not differ between patients and controls.

Conclusions: In conclusion, this study demonstrated increased IL-12p40 and CSF-1 concentrations in CFS/ME patients in addition to a set of predictive biomarkers. There was no effect of anakinra on circulating cytokines other than IL-1Ra.

Trial registration: ClinicalTrials.gov Identifier: NCT02108210 , Registered April 2014.

Keywords: Anakinra; CSF-1; Chronic fatigue syndrome; IL-12p40; Proximity extension assay.

Figures

Fig. 1
Fig. 1
Normalized Protein Expression Units (NPX) values for patients with chronic fatigue syndrome (CFS/ME, n = 50) compared to healthy controls (HC, n = 48). Displays linear NPX values for patients and controls. P values were derived by analysis of covariance
Fig. 2
Fig. 2
Concentration of IL-1Ra for patients treated with either anakinra or placebo. Displays IL-1Ra concentrations ± SEM in patients treated with either anakinra or placebo before and after treatment
Fig. 3
Fig. 3
Treatment effect of the 4 week intervention period on circulating cytokines. Displays the difference and 95% confidence intervals between study arms for protein values after treatment (anakinra:controls)

References

    1. Fukuda K, et al. The chronic fatigue syndrome: a comprehensive approach to its definition and study. International Chronic Fatigue Syndrome Study Group. Ann Intern Med. 1994;121(12):953–959. doi: 10.7326/0003-4819-121-12-199412150-00009.
    1. Reeves WC, et al. Identification of ambiguities in the 1994 chronic fatigue syndrome research case definition and recommendations for resolution. BMC Health Serv Res. 2003;3(1):25. doi: 10.1186/1472-6963-3-25.
    1. Solomon L, Reeves WC. Factors influencing the diagnosis of chronic fatigue syndrome. Arch Intern Med. 2004;164(20):2241–2245. doi: 10.1001/archinte.164.20.2241.
    1. National Collaborating Centre for Primary Care UK. Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (or Encephalopathy) Diagnosis and Management of Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (or Encephalopathy) in Adults and Children. London: National Collaborating Centre for Primary Care; 2007.
    1. Collin SM, et al. Chronic fatigue syndrome (CFS) symptom-based phenotypes in two clinical cohorts of adult patients in the UK and The Netherlands. J Psychosom Res. 2016;81:14–23. doi: 10.1016/j.jpsychores.2015.12.006.
    1. Klimas NG, Broderick G, Fletcher MA. Biomarkers for chronic fatigue. Brain Behav Immun. 2012;26(8):1202–1210. doi: 10.1016/j.bbi.2012.06.006.
    1. Loebel M, et al. Deficient EBV-specific B- and T-cell response in patients with chronic fatigue syndrome. PLoS ONE. 2014;9(1):e85387. doi: 10.1371/journal.pone.0085387.
    1. Lunde S, et al. Serum BAFF and APRIL Levels, T-lymphocyte subsets, and immunoglobulins after B-cell depletion using the monoclonal anti-CD20 antibody rituximab in myalgic encephalopathy/chronic fatigue syndrome. PLoS ONE. 2016;11(8):e0161226. doi: 10.1371/journal.pone.0161226.
    1. Hornig M, et al. Distinct plasma immune signatures in ME/CFS are present early in the course of illness. Sci Adv. 2015;1(1):e1400121. doi: 10.1126/sciadv.1400121.
    1. Broderick G, et al. Cytokine expression profiles of immune imbalance in post-mononucleosis chronic fatigue. J Transl Med. 2012;10:191. doi: 10.1186/1479-5876-10-191.
    1. Brenu EW, et al. Longitudinal investigation of natural killer cells and cytokines in chronic fatigue syndrome/myalgic encephalomyelitis. J Transl Med. 2012;10:88. doi: 10.1186/1479-5876-10-88.
    1. Rajeevan MS, et al. Pathway-focused genetic evaluation of immune and inflammation related genes with chronic fatigue syndrome. Hum Immunol. 2015;76(8):553–560. doi: 10.1016/j.humimm.2015.06.014.
    1. Schlauch KA, et al. Genome-wide association analysis identifies genetic variations in subjects with myalgic encephalomyelitis/chronic fatigue syndrome. Transl Psychiatry. 2016;6:e730. doi: 10.1038/tp.2015.208.
    1. Blundell S, et al. Chronic fatigue syndrome and circulating cytokines: a systematic review. Brain Behav Immun. 2015;50:186–195. doi: 10.1016/j.bbi.2015.07.004.
    1. Montoya JG, et al. Cytokine signature associated with disease severity in chronic fatigue syndrome patients. Proc Natl Acad Sci U S A. 2017;114(34):E7150–E7158. doi: 10.1073/pnas.1710519114.
    1. Russell L, et al. Illness progression in chronic fatigue syndrome: a shifting immune baseline. BMC Immunol. 2016;17:3. doi: 10.1186/s12865-016-0142-3.
    1. Fletcher MA, et al. Plasma cytokines in women with chronic fatigue syndrome. J Transl Med. 2009;7:96. doi: 10.1186/1479-5876-7-96.
    1. de Jager W, et al. Prerequisites for cytokine measurements in clinical trials with multiplex immunoassays. BMC Immunol. 2009;10:52. doi: 10.1186/1471-2172-10-52.
    1. Aguilar-Mahecha A, et al. The effect of pre-analytical variability on the measurement of MRM-MS-based mid- to high-abundance plasma protein biomarkers and a panel of cytokines. PLoS ONE. 2012;7(6):e38290. doi: 10.1371/journal.pone.0038290.
    1. Solier C, Langen H. Antibody-based proteomics and biomarker research—current status and limitations. Proteomics. 2014;14(6):774–783. doi: 10.1002/pmic.201300334.
    1. Assarsson E, et al. Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability. PLoS ONE. 2014;9(4):e95192. doi: 10.1371/journal.pone.0095192.
    1. Roerink ME, et al. Cytokine inhibition in chronic fatigue syndrome patients: study protocol for a randomized controlled trial. Trials. 2015;16:439. doi: 10.1186/s13063-015-0971-z.
    1. Roerink ME, et al. Interleukin-1 as a mediator of fatigue in disease: a narrative review. J Neuroinflamm. 2017;14(1):16. doi: 10.1186/s12974-017-0796-7.
    1. Roerink ME, et al. Cytokine inhibition in patients with chronic fatigue syndrome: a randomized trial. Cytokine inhibition in patients with chronic fatigue syndrome. Ann Intern Med. 2017;166(8):557–564. doi: 10.7326/M16-2391.
    1. Kim J, Shin W. How to do random allocation (randomization) Clin Orthop Surg. 2014;6(1):103–109. doi: 10.4055/cios.2014.6.1.103.
    1. Vercoulen JH, Alberst M, Bleijenberg G. De Checklist Individual Strength (CIS) Gedragstherapie. 1999;32:131–136.
    1. Vercoulen JH, et al. Dimensional assessment of chronic fatigue syndrome. J Psychosom Res. 1994;38(5):383–392. doi: 10.1016/0022-3999(94)90099-X.
    1. Wiborg JF, et al. Randomised controlled trial of cognitive behaviour therapy delivered in groups of patients with chronic fatigue syndrome. Psychother Psychosom. 2015;84(6):368–376. doi: 10.1159/000438867.
    1. Derogatis LR, Savitz KL. Handbook of psychological assessment in primary care settings. Vol. 236. New Jersey: Lawrence Erlbaum Associates; 2000. pp. 297–334.
    1. Jo C, et al. Random-graft polymer-directed synthesis of inorganic mesostructures with ultrathin frameworks. Angew Chem Int Ed Engl. 2014;53(20):5117–5121.
    1. Landi A, et al. Reductions in circulating levels of IL-16, IL-7 and VEGF-A in myalgic encephalomyelitis/chronic fatigue syndrome. Cytokine. 2016;78:27–36. doi: 10.1016/j.cyto.2015.11.018.
    1. Hawinkels LJ, et al. Tissue level, activation and cellular localisation of TGF-beta1 and association with survival in gastric cancer patients. Br J Cancer. 2007;97(3):398–404. doi: 10.1038/sj.bjc.6603877.
    1. RC Team. R: A language and environment for statistical computing. R Foundation for Statistical Computing. 2016. . Accessed on 1 Nov 2016.
    1. Tibshirani R. Regression shrinkage and selection via the Lasso. J Roy Stat Soc. 1996;58(1):267–288.
    1. Hanley JA, McNeil BJ. The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology. 1982;143(1):29–36. doi: 10.1148/radiology.143.1.7063747.
    1. Steyerberg EW. Clinical prediction models: a practical approach to development, validation, and updating. New York: Springer; 2009.
    1. Croxford AL, Kulig P, Becher B. IL-12-and IL-23 in health and disease. Cytokine Growth Factor Rev. 2014;25(4):415–421. doi: 10.1016/j.cytogfr.2014.07.017.
    1. Zundler S, Neurath MF. Interleukin-12: functional activities and implications for disease. Cytokine Growth Factor Rev. 2015;26(5):559–568. doi: 10.1016/j.cytogfr.2015.07.003.
    1. Cua DJ, et al. Interleukin-23 rather than interleukin-12 is the critical cytokine for autoimmune inflammation of the brain. Nature. 2003;421(6924):744–748. doi: 10.1038/nature01355.
    1. White AT, et al. Severity of symptom flare after moderate exercise is linked to cytokine activity in chronic fatigue syndrome. Psychophysiology. 2010;47(4):615–624.
    1. Khaiboullina SF, et al. Cytokine expression provides clues to the pathophysiology of Gulf War illness and myalgic encephalomyelitis. Cytokine. 2015;72(1):1–8. doi: 10.1016/j.cyto.2014.11.019.
    1. Smylie AL, et al. A comparison of sex-specific immune signatures in Gulf War illness and chronic fatigue syndrome. BMC Immunol. 2013;14:29. doi: 10.1186/1471-2172-14-29.
    1. Hurteau JA, et al. Evaluation of recombinant human interleukin-12 in patients with recurrent or refractory ovarian cancer: a Gynecologic Oncology Group Study. Gynecol Oncol. 2001;82(1):7–10. doi: 10.1006/gyno.2001.6255.
    1. van Herpen CM, et al. Intratumoral administration of recombinant human interleukin 12 in head and neck squamous cell carcinoma patients elicits a T-helper 1 profile in the locoregional lymph nodes. Clin Cancer Res. 2004;10(8):2626–2635. doi: 10.1158/1078-0432.CCR-03-0304.
    1. Ries CH, et al. CSF-1/CSF-1R targeting agents in clinical development for cancer therapy. Curr Opin Pharmacol. 2015;23:45–51. doi: 10.1016/j.coph.2015.05.008.
    1. Muller AF, et al. Neutralization of colony-stimulating factor 1 receptor prevents sickness behavior syndrome by reprogramming inflammatory monocytes to produce IL-10. Brain Behav Immun. 2015;48:78–85. doi: 10.1016/j.bbi.2015.02.027.
    1. Hawinkels LJ, Ten Dijke P. Exploring anti-TGF-beta therapies in cancer and fibrosis. Growth Factors. 2011;29(4):140–152. doi: 10.3109/08977194.2011.595411.
    1. Blakytny R, et al. Latent TGF-beta1 activation by platelets. J Cell Physiol. 2004;199(1):67–76. doi: 10.1002/jcp.10454.
    1. Rondina MT, et al. Whole blood flow cytometry measurements of in vivo platelet activation in critically-Ill patients are influenced by variability in blood sampling techniques. Thromb Res. 2012;129(6):729–735. doi: 10.1016/j.thromres.2011.11.031.
    1. Clark LV, et al. Cytokine responses to exercise and activity in patients with chronic fatigue syndrome: Case control study. Clin Exp Immunol. 2017;190:360–371. doi: 10.1111/cei.13023.
    1. Norris LA, Bonnar J. Effect of oestrogen dose on whole blood platelet activation in women taking new low dose oral contraceptives. Thromb Haemost. 1994;72(6):926–930.
    1. Fernandez-Martinez E, et al. Immunomodulatory effects by oral contraceptives in normal and cholestatic female rats: role of cytokines. Int Immunopharmacol. 2014;21(1):10–19. doi: 10.1016/j.intimp.2014.03.022.
    1. van Kuppeveld FJ, van der Meer JW. XMRV and CFS–the sad end of a story. Lancet. 2012;379(9814):e27–e28. doi: 10.1016/S0140-6736(11)60899-4.
    1. Page TH, et al. Nonsteroidal anti-inflammatory drugs increase TNF production in rheumatoid synovial membrane cultures and whole blood. J Immunol. 2010;185(6):3694–3701. doi: 10.4049/jimmunol.1000906.
    1. Endres S, et al. In vitro production of IL 1 beta, IL 1 alpha, TNF and IL2 in healthy subjects: distribution, effect of cyclooxygenase inhibition and evidence of independent gene regulation. Eur J Immunol. 1989;19(12):2327–2333. doi: 10.1002/eji.1830191222.
    1. Tosato G, Jones KD. Interleukin-1 induces interleukin-6 production in peripheral blood monocytes. Blood. 1990;75(6):1305–1310.
    1. Emsley HC, et al. A randomised phase II study of interleukin-1 receptor antagonist in acute stroke patients. J Neurol Neurosurg Psychiatry. 2005;76(10):1366–1372. doi: 10.1136/jnnp.2004.054882.
    1. Van Tassell BW, et al. Enhanced interleukin-1 activity contributes to exercise intolerance in patients with systolic heart failure. PLoS ONE. 2012;7(3):e33438. doi: 10.1371/journal.pone.0033438.
    1. Pusch G, et al. Early Dynamics of P-selectin and interleukin 6 predicts outcomes in ischemic stroke. J Stroke Cerebrovasc Dis. 2015;24(8):1938–1947. doi: 10.1016/j.jstrokecerebrovasdis.2015.05.005.
    1. Lewith G, et al. Complementary and alternative healthcare use by participants in the PACE trial of treatments for chronic fatigue syndrome. J Psychosom Res. 2016;87:37–42. doi: 10.1016/j.jpsychores.2016.06.005.
    1. Boneva RS, et al. Use of medications by people with chronic fatigue syndrome and healthy persons: a population-based study of fatiguing illness in Georgia. Health Qual Life Outcomes. 2009;7:67. doi: 10.1186/1477-7525-7-67.
    1. Mueller-Using S, et al. Factors associated with performing tuberculosis screening of HIV-positive patients in Ghana: LASSO-based predictor selection in a large public health data set. BMC Public Health. 2016;16:563. doi: 10.1186/s12889-016-3239-y.

Source: PubMed

Спонсоры и соавторы

Заболевания

Медикаментозные вмешательства

Подписаться