Journal of Neurology

, Volume 255, Issue 5, pp 723–731 | Cite as

Gene expression in human peripheral blood mononuclear cells upon acute ischemic stroke

  • C.  Grond-GinsbachEmail author
  • M. Hummel
  • T. Wiest
  • S. Horstmann
  • K. Pfleger
  • M. Hergenhahn†
  • M. Hollstein
  • U. Mansmann
  • A. J. Grau
  • S. Wagner


Background and purpose

Ischemic stroke provokes a systemic inflammatory response. The purpose of this study was to characterize this response on the gene expression level in circulating mononuclear leukocytes from acute ischemic stroke (AIS) patients.


RNA from peripheral blood mononuclear cells (PBMCs) of AIS patients (24 + 2 hours after onset of symptoms) was analyzed with Affymetrix U133A GeneChips using a pooled design. We compared the gene expression signature from AIS patients (n = 20), stroke survivors (n = 15), patients with acute traumatic brain injury (ATBI, n = 15) and healthy control subjects without vascular risk factors (n = 15).


Expression levels of 9682 probe sets with present calls on each GeneChip were compared. Between AIS patients and stroke survivors or between AIS patients and ATBI patients there were no significant differences in expression values of single genes after correction for multiple testing. However, comparison of the PBMC expression profiles from AIS patients and healthy subjects revealed significantly different expression (p = 0.012) of a single probe set, specific for phosphodiesterase 4 D (PDE4D). In order to detect modest expression differences in multiple genes with a presumed cumulative effect we studied the gene expression of functional groups of genes by global statistical tests. Analysis of 11 gene groups revealed differential expression between AIS patients and healthy subjects for genes involved in the inflammatory response (GeneOntology GO:0006954). Genes encoding the N-formyl peptide receptor-like 1 (FPRL1), interleukin-1 receptor antagonist (IL1RN) and complement component 3a receptor 1 (C3AR1) contributed most to the observed difference.


This transcriptome analysis did not identify significant changes between circulating mononuclear cells from AIS patients 24 hours after stroke and closely matched stroke survivors. However, comparing AIS patients with healthy control subjects revealed measurable differences in PDE4D and in inflammatory response genes when considered as a set.

Key words

acute ischemic stroke acute traumatic brain injury gene expression blood 


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Supplementary material

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  1. 1.
    Baird AE (2007) Blood genomics in human stroke. Stroke 38:694–698PubMedCrossRefGoogle Scholar
  2. 2.
    Bakay M, Chen YW, Borup R, Zhao P, Nagaraju K, Hoffman EP (2002) Sources of variability and effect of experimental approach on expression profiling data interpretation. BMC Bioinformatics 3:4PubMedCrossRefGoogle Scholar
  3. 3.
    Barnum SR, Ames RS, Maycox PR, Hadingham SJ, Meakin J, Harrison D, Parsons AA (2002) Expression of the complement C3a and C5a receptors after permanent focal ischemia: An alternative interpretation. Glia 38:169–173PubMedCrossRefGoogle Scholar
  4. 4.
    Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. Journal of the Royal Statistical Society, Series, pp 289–300Google Scholar
  5. 5.
    Chiang N, Serhan CN, Dahlen SE, Drazen JM, Hay DWP, Rovati GE, Shimizu T, Yokomitz T, Brink C (2006) The lipoxin receptor ALX: Potent ligandspecific and steroselective actions in vivo. Pharmacol Rev 58:463–487PubMedCrossRefGoogle Scholar
  6. 6.
    Chtanova T, Newton R, Liu SM, Weininger L, Young TR, Silva DG, Bertoni F, Rinaldi A, Chappaz S, Sallusto F, Rolph MS, Mackay CR (2005) Identification of T cell-restricted genes and signatures for different T cell responses, using a comprehensive collection of microarray datasets. J Immunol 175:7837–7847PubMedGoogle Scholar
  7. 7.
    Connolly PH, Caiozzo VJ, Zaldivar F, Nemet D, Larson J, Hung SP, Heck JD, Hatfield GW, Cooper DM (2004) Effects of exercise on gene expression in human peripheral blood mononuclear cells. J Appl Physiol 97:1461–1469PubMedCrossRefGoogle Scholar
  8. 8.
    Cowell RM, Plane JM, Silverstein FS (2003) Complement activation contributes to hypoxic ischemic brain injury in neonatal rats. J. Neurosci 23:9459–9468PubMedGoogle Scholar
  9. 9.
    Emsley HC, Smith CJ, Georgiou RF, Vail A, Hopkins SJ, Rothwell NJ, Tyrell PJ, acute stroke investigators (2005) A randomised phase II study of interleukin-1 receptor antagonist in acute stroke patients. J Neurol Neurosurg Psychiatry 76:1366–1372PubMedCrossRefGoogle Scholar
  10. 10.
    Feezor RJ, Baker HV, Mindrinos M, Hayden D, Tannahill CL, Brownstein BH, Fay A, MacMillan S, Laramie J, Xiao W, Moldawer LL, Cobb JP, Laudanski K, Miller-Graziano CL, Maier RV, Schoenfeld D, Davis RW, Tompkins RG; and the Inflammation and Host Response to Injury, Large-Scale Collaborative Research Program (2004) Whole blood and leukocyte RNA isolation for gene expression analyses. Physiol Genomics 19:247– 254PubMedCrossRefGoogle Scholar
  11. 11.
    Gentleman RC, Carey VJ, Bates DM, Bolstad B, Dettling M, Dudoit S, Ellis B, Gautier L, Ge Y, Gentry J, Hornik K, Hothorn T, Huber W, Iacus S, Irizarry R, Leisch F, Li C, Maechler M, Rossini AJ, Sawitzki G, Smith C, Smyth G, Tierney L, Yang JY, Zhang J (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5:R80PubMedCrossRefGoogle Scholar
  12. 12.
    Gretarsdottir S, Sveinbjornsdottir S, Jonsson HH, Jakobsson F, Einarsdottir E, Agnarsson U, Shkolny D, Einarsson G, Gudjonsdottir HM, Valdimarsson EM, Einarsson OB, Thorgeirsson G, Hadzic R, Jonsdottir S, Reynisdottir ST, Bjarnadottir SM, Gudmundsdottir T, Gudlaugsdottir GJ, Gill R, Lindpaintner K, Sainz J, Hannesson HH, Sigurdsson GT, Frigge ML, Kong A, Gudnason V, Stefansson K, Gulcher JR (2002) Localization of a susceptibility gene for common forms of stroke to 5q12. Am J Hum Genet 70:593–603PubMedCrossRefGoogle Scholar
  13. 13.
    Gretarsdottir S, Thorleifsson G, Reynisdottir ST, Manolescu A, Jonsdottir S, Jonsdottir T, Gudmundsdottir T, Bjarnadottir SM, Einarsson OB, Gudjonsdottir HM, Hawkins M, Gudmundsson G, Gudmundsdottir H, Andrason H, Gudmundsdottir AS, Sigurdardottir M, Chou TT, Nahmias J, Goss S, Sveinbjornsdottir S, Valdimarsson EM, Jakobsson F, Agnarsson U, Gudnason V, Thorgeirsson G, Fingerle J, Gurney M, Gudbjartsson D, Frigge ML, Kong A, Stefansson K, Gulcher JR (2003) The gene encoding phosphodiesterase 4D confers risk of ischemic stroke. Nat Gen 353:131–138CrossRefGoogle Scholar
  14. 14.
    Grond-Ginsbach C, Horstmann S, Hummel M, Wiest T, Honold C, Pfleger K, Hergenhahn M, Hollstein M, Weninger A, Knyazev Y, Mansmann U, Grau AJ, Wagner S (2007) Gene expression in peripheral blood mononuclear cells (PBMCs) from stroke survivors. (Manuscript submitted)Google Scholar
  15. 15.
    Helgadottir A, Manolescu A, Thorleifsson G, Gretarsdottir S, Jonsdottir H, Thorsteinsdottir U, Samani NJ, Gudmundsson G, Grant SF, Thorgeirsson G, Sveinbjornsdottir S, Valdimarsson EM, Matthiasson SE, Johannsson H, Gudmundsdottir O, Gurney ME, Sainz J, Thorhallsdottir M, Andresdottir M, Frigge ML, Topol EJ, Kong A, Gudnason V, Hakonarson H, Gulcher JR, Stefansson K (2004) The gene encoding 5-lipoxygenase activating protein confers risk of myocardial infarction and stroke. Nat Genet 36:233–239PubMedCrossRefGoogle Scholar
  16. 16.
    Iaonnidis JPA (2005) Why most published research findings are false. PLoS Medicine 2:696–701Google Scholar
  17. 17.
    Jolly RA, Goldstein KM, Wei T, Gao H, Chen P, Huang S, Colet JM, Ryan TP, Thomas CE, Estrem ST (2005) Pooling samples within microarray studies: a comparative analysis of rat liver transcription response to prototypical toxicants. Physiol Genomics 22:346–355PubMedCrossRefGoogle Scholar
  18. 18.
    Lamb J, Ramaswamy S, Ford HL, Contreras B, Martinez MV, Kitrell FS, Zahnow CA, Patterson N, Golub TR, Ewen ME (2003) A mechanism of cyclin D1 action encoded in the patterns of gene expression in human cancer. Cell 114:323–334PubMedCrossRefGoogle Scholar
  19. 19.
    Liu J, Walter E, Stenger D, Thach D (2006) Effects of globin mRNA reduction methods on gene expression profiles from whole blood. J Mol Diagnos 8:551–558CrossRefGoogle Scholar
  20. 20.
    Malcolm KC, Arndt PG, Manos EJ, Jones DA, Worthen GS (2003) Microarray analysis of lipopolysaccharidetreated human neutrophils. Am J Physiol 284:L663–L670Google Scholar
  21. 21.
    Mansmann U, Meister R (2005) Testing differential gene expression in functional groups. Goeman’s global test versus an ANCOVA approach. Methods Inf Med 44:449–453PubMedGoogle Scholar
  22. 22.
    Mocco J, Mack WJ, Ducruet AF, Susonov SA, Sughrue ME, Hassid BG, Nair MN, Laufer I, Komotar RJ, Claire M, Holland H, Pinsky DJ, Connolly ES (2006) Complement factor. Complement component C3 mediates inflammatory injury following focal cerebral ischemia. Circulation Res 99:209–217PubMedCrossRefGoogle Scholar
  23. 23.
    Moore DF, Li H, Jeffries N, Wright V, Cooper RA, Elkahloun A, Gelderman MP, Zudaire E, Blevins G, Yu H, Goldin E, Baird AE (2005) Using peripheral blood mononuclear cells to determine a gene expression profile of acute ischemic stroke: a pilot investigation. Circulation 111:212–221PubMedCrossRefGoogle Scholar
  24. 24.
    Rosand J, Bayley N, Rost N, de Bakker PI (2006) Many hypotheses but no replication for the association between PDE4D and stroke. Nature Genetics 38:1091–1092PubMedCrossRefGoogle Scholar
  25. 25.
    Rothwell N, Allan S, Toulmond S (1997) The role of interleukin 1 in acute neurodegeneration and stroke: pathophysiological and therapeutic implications. J Clin Invest 100:2648–2652PubMedCrossRefGoogle Scholar
  26. 26.
    Sebastiani P, Ramoni MF, Nolan V, Baldwin CT, Steinberg MH (2005) Genetic dissection and prognostic modeling of overt stroke in sickle cell anemia. Nature Genetics 37:435–440PubMedCrossRefGoogle Scholar
  27. 27.
    Sharp FR, Lit L, Xu H, Apperson M, Walker W, Wong B, Gilbert DL, Hershey G, Glauser TA (2006) Genomics of brain and blood: progress and pitfalls. Epilepsia 47:1603–1607PubMedCrossRefGoogle Scholar
  28. 28.
    Sharp FR, XU H, Lit L, Walker W, Pinter J, Apperson M, Verro P (2007) Genomic profiles of stroke in blood. Stroke 38:691–693PubMedCrossRefGoogle Scholar
  29. 29.
    Subrahmanyam Y, Yamaga S, Prashar Y, Lee HH, Hoe NP, Kluger Y, Gerstein M, Goguen JD, Newburger PE, Weissman SM (2001) RNA expression patterns change dramatically in human neutrophils exposed to bacteria. Blood 97:2457–2468PubMedCrossRefGoogle Scholar
  30. 30.
    Tang Y, Xu H, Du X, Lit L, Walker W, Lu A, Ran R, Gregg JP, Reilly M, Pancioli A, Khoury JC, Sauerbeck LR, Carrozzella JA, Spilker J, Clark J, Wagner KR, Jauch EC, Chang DJ, Verro P, Broderick JP, Sharp FR (2006) Gene expression in blood changes rapidly in neutrophils and monocytes after ischemic stroke in humans: a microarray study. J Cereb Blood Flow Metab 26:1089–1102PubMedCrossRefGoogle Scholar
  31. 31.
    Wang Y, Chang CF, Morales M, Chou J, Chen HL, Chiang YH, Lin SZ, Cadet J, Deng X, Wang JY, Chen SY, Kaplan PL, Hoffer BJ (2001) Bone morphogenetic protein-6 reduces ischemia-induced brain damage in rats. Stroke 32:2170– 2178PubMedCrossRefGoogle Scholar
  32. 32.
    Worrall BB, Brott TG, Brown RD, Brown WM, Rich SS, Arepalli S, Wavrant-De Vrièze F, Duckworth J, Singleton AB, Hardy J, Meschia JF (2007) IL1RN VNTR polymorphism in ischemic stroke: analysis in 3 populations. Stroke 38:1189–1196PubMedCrossRefGoogle Scholar
  33. 33.
    Wu Z, Irizarry RA, Gentleman R, Martinez Murillo F, Spencer F (2004) A Model Based Background Adjustment for Oligonucleotide Expression Arrays. J Am Statist Assoc 99:909–917CrossRefGoogle Scholar
  34. 34.
    Yue L, Reisdorf WC (2005) Pathway and ontology analysis: emerging approaches connecting transcriptome data and clinical endpoints. Curr Mol Med 5:11–21PubMedCrossRefGoogle Scholar
  35. 35.
    Zakharkin OS, Kim K, Mehta T, Chen L, Barnes S, Scheirer KE, Parrish RS, Allison DB, Page GP (2005) Sources of variation in Affymetrix microarray experiments. BMC Bioinformatics 6:214PubMedCrossRefGoogle Scholar

Copyright information

© Steinkopff-Verlag 2008

Authors and Affiliations

  • C.  Grond-Ginsbach
    • 1
    Email author
  • M. Hummel
    • 2
  • T. Wiest
    • 1
  • S. Horstmann
    • 1
  • K. Pfleger
    • 1
  • M. Hergenhahn†
    • 3
  • M. Hollstein
    • 3
  • U. Mansmann
    • 2
  • A. J. Grau
    • 4
  • S. Wagner
    • 1
  1. 1.Dept. of NeurologyUniversity of HeidelbergHeidelbergGermany
  2. 2.IBE, Medical School LMUMünchenGermany
  3. 3.Dept. of Genetic Alterations in CarcinogenesisGerman Cancer Research CenterHeidelbergGermany
  4. 4.Dept. of NeurologyKlinikum der Stadt LudwigshafenLudwigshafen a. Rh.Germany

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