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. 1994 Aug;97(2):293–298. doi: 10.1111/j.1365-2249.1994.tb06083.x

Cytokine mRNA expression in endomyocardial biopsies during acute rejection from human heart transplants.

C C Baan 1, N E van Emmerik 1, A H Balk 1, W G Quint 1, B Mochtar 1, N H Jutte 1, H G Niesters 1, W Weimar 1
PMCID: PMC1534709  PMID: 8050179

Abstract

The immune response to an allograft is regulated by cytokines produced by cells infiltrating the allograft. However, the immunopathogenesis of allograft rejection is not completely understood. To investigate the role of cytokines after clinical heart transplantation, we analysed the expression of cytokine genes in sequentially taken endomyocardial biopsies (EMB) by using the reverse transcriptase-polymerase chain reaction (RT-PCR). We analysed 44 EMB from 11 recipients: 21 EMB before or during rejection, and 23 EMB without histological evidence of acute rejection. A strong correlation was found between IL-2 gene expression and histologically proved rejection (16/21 versus 1/23 without rejection, P < 0.001; chi 2 test). Also, expression of IL-4 and IL-6 genes was more often found in EMB during rejection than in EMB without signs of rejection (IL-4, 62% versus 35%; and IL-6, 81% versus 39%, respectively). No relation with rejection or with immunological quiescence was observed for the presence of IL-10 gene transcripts. IL-10, but also IL-6 mRNA were detectable in donor heart tissue before transplantation (9/10). In contrast, IL-2 and IL-4 gene transcripts were absent in these samples. These differences could not be explained by the presence or absence of T cells, since the gene for the constant region of the beta-chain (C beta) of the T cell receptor (TCR) not only was expressed in post-transplant EMB but also in pre-transplant donor heart tissue. Our results provide strong evidence that the immunoregulatory cytokines IL-2, IL-4 and IL-6 are important local regulators in the graft during acute rejection. The role of IL-10 in the immunologic response to the transplanted organ needs further investigation.

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Selected References

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  1. Billingham M. E., Cary N. R., Hammond M. E., Kemnitz J., Marboe C., McCallister H. A., Snovar D. C., Winters G. L., Zerbe A. A working formulation for the standardization of nomenclature in the diagnosis of heart and lung rejection: Heart Rejection Study Group. The International Society for Heart Transplantation. J Heart Transplant. 1990 Nov-Dec;9(6):587–593. [PubMed] [Google Scholar]
  2. Brenner C. A., Tam A. W., Nelson P. A., Engleman E. G., Suzuki N., Fry K. E., Larrick J. W. Message amplification phenotyping (MAPPing): a technique to simultaneously measure multiple mRNAs from small numbers of cells. Biotechniques. 1989 Nov-Dec;7(10):1096–1103. [PubMed] [Google Scholar]
  3. Chen W. F., Zlotnik A. IL-10: a novel cytotoxic T cell differentiation factor. J Immunol. 1991 Jul 15;147(2):528–534. [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Daane C. R., van Besouw N. M., van Emmerik N. E., Baan C. C., Balk A. H., Jutte N. H., Niesters B., Vaessen L. M., Weimar W. Discrepancy between mRNA expression and production of IL-2 and IL-4 by cultured graft infiltrating cells propagated from endomyocardial biopsies. Transpl Int. 1994;7 (Suppl 1):S627–S628. doi: 10.1111/j.1432-2277.1994.tb01459.x. [DOI] [PubMed] [Google Scholar]
  6. Dallman M. J., Larsen C. P., Morris P. J. Cytokine gene transcription in vascularised organ grafts: analysis using semiquantitative polymerase chain reaction. J Exp Med. 1991 Aug 1;174(2):493–496. doi: 10.1084/jem.174.2.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dallman M. J., Montgomery R. A., Larsen C. P., Wanders A., Wells A. F. Cytokine gene expression: analysis using northern blotting, polymerase chain reaction and in situ hybridization. Immunol Rev. 1991 Feb;119:163–179. doi: 10.1111/j.1600-065x.1991.tb00583.x. [DOI] [PubMed] [Google Scholar]
  8. Dallman M. J., Porter A. C., Larsen C. P., Morris P. J. Lymphokine production in allografts--analysis of RNA by northern blotting. Transplant Proc. 1989 Feb;21(1 Pt 1):296–298. [PubMed] [Google Scholar]
  9. Dallman M. J., Roake J., Hughes D., Toogood G., Morris P. J. Sequential analysis of IL-2 gene transcription in renal transplants. Transplantation. 1992 Mar;53(3):683–685. doi: 10.1097/00007890-199203000-00038. [DOI] [PubMed] [Google Scholar]
  10. Fiorentino D. F., Zlotnik A., Vieira P., Mosmann T. R., Howard M., Moore K. W., O'Garra A. IL-10 acts on the antigen-presenting cell to inhibit cytokine production by Th1 cells. J Immunol. 1991 May 15;146(10):3444–3451. [PubMed] [Google Scholar]
  11. Gassel A. M., Hansmann M. L., Radzun H. J., Weyand M. Human cardiac allograft rejection. Correlation of grading with expression of different monocyte/macrophage markers. Am J Clin Pathol. 1990 Sep;94(3):274–279. doi: 10.1093/ajcp/94.3.274. [DOI] [PubMed] [Google Scholar]
  12. Hawes G. E., Struyk L., van den Elsen P. J. Differential usage of T cell receptor V gene segments in CD4+ and CD8+ subsets of T lymphocytes in monozygotic twins. J Immunol. 1993 Mar 1;150(5):2033–2045. [PubMed] [Google Scholar]
  13. Jordan S. C., Kondo T., Prehn J., Marchevsky A., Waters P. Cytokine gene activation in rat lung allografts: analysis by northern blotting. Transplant Proc. 1991 Feb;23(1 Pt 1):604–606. [PubMed] [Google Scholar]
  14. Kishimoto T., Akira S., Taga T. Interleukin-6 and its receptor: a paradigm for cytokines. Science. 1992 Oct 23;258(5082):593–597. doi: 10.1126/science.1411569. [DOI] [PubMed] [Google Scholar]
  15. Lansdorp P. M., Aarden L. A., Calafat J., Zeiljemaker W. P. A growth-factor dependent B-cell hybridoma. Curr Top Microbiol Immunol. 1986;132:105–113. doi: 10.1007/978-3-642-71562-4_14. [DOI] [PubMed] [Google Scholar]
  16. Lowry R. P., Wang K., Vernooij B., Harcus D. Lymphokine transcription in vascularized mouse heart grafts: effects of "tolerance" induction. Transplant Proc. 1989 Feb;21(1 Pt 1):72–73. [PubMed] [Google Scholar]
  17. Marchuk D., McCrohon S., Fuchs E. Complete sequence of a gene encoding a human type I keratin: sequences homologous to enhancer elements in the regulatory region of the gene. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1609–1613. doi: 10.1073/pnas.82.6.1609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Martinez O. M., Krams S. M., Sterneck M., Villanueva J. C., Falco D. A., Ferrell L. D., Lake J., Roberts J. P., Ascher N. L. Intragraft cytokine profile during human liver allograft rejection. Transplantation. 1992 Feb;53(2):449–456. doi: 10.1097/00007890-199202010-00035. [DOI] [PubMed] [Google Scholar]
  19. Martinez O. M., Villanueva J. C., Lake J., Roberts J. P., Ascher N. L., Krams S. M. IL-2 and IL-5 gene expression in response to alloantigen in liver allograft recipients and in vitro. Transplantation. 1993 May;55(5):1159–1166. doi: 10.1097/00007890-199305000-00042. [DOI] [PubMed] [Google Scholar]
  20. May L. T., Ghrayeb J., Santhanam U., Tatter S. B., Sthoeger Z., Helfgott D. C., Chiorazzi N., Grieninger G., Sehgal P. B. Synthesis and secretion of multiple forms of beta 2-interferon/B-cell differentiation factor 2/hepatocyte-stimulating factor by human fibroblasts and monocytes. J Biol Chem. 1988 Jun 5;263(16):7760–7766. [PubMed] [Google Scholar]
  21. Merville P., Pouteil-Noble C., Wijdenes J., Potaux L., Touraine J. L., Banchereau J. Detection of single cells secreting IFN-gamma, IL-6, and IL-10 in irreversibly rejected human kidney allografts, and their modulation by IL-2 and IL-4. Transplantation. 1993 Mar;55(3):639–646. doi: 10.1097/00007890-199303000-00032. [DOI] [PubMed] [Google Scholar]
  22. Morgan C. J., Pelletier R. P., Hernandez C. J., Teske D. L., Huang E., Ohye R., Orosz C. G., Ferguson R. M. Alloantigen-dependent endothelial phenotype and lymphokine mRNA expression in rejecting murine cardiac allografts. Transplantation. 1993 Apr;55(4):919–924. doi: 10.1097/00007890-199304000-00042. [DOI] [PubMed] [Google Scholar]
  23. Noronha I. L., Hartley B., Cameron J. S., Waldherr R. Detection of IL-1 beta and TNF-alpha message and protein in renal allograft biopsies. Transplantation. 1993 Oct;56(4):1026–1029. [PubMed] [Google Scholar]
  24. O'Garra A. Interleukins and the immune system 2. Lancet. 1989 May 6;1(8645):1003–1005. doi: 10.1016/s0140-6736(89)92640-8. [DOI] [PubMed] [Google Scholar]
  25. Pham S. M., Yoshida Y., Aeba R., Hattler B. G., Iwaki Y., Zeevi A., Hardesty R. L., Griffith B. P. Interleukin-6, a marker of preservation injury in clinical lung transplantation. J Heart Lung Transplant. 1992 Nov-Dec;11(6):1017–1024. [PubMed] [Google Scholar]
  26. Ruan X. M., Qiao J. H., Trento A., Czer L. S., Blanche C., Fishbein M. C. Cytokine expression and endothelial cell and lymphocyte activation in human cardiac allograft rejection: an immunohistochemical study of endomyocardial biopsy samples. J Heart Lung Transplant. 1992 Nov-Dec;11(6):1110–1115. [PubMed] [Google Scholar]
  27. Takeuchi T., Lowry R. P., Konieczny B. Heart allografts in murine systems. The differential activation of Th2-like effector cells in peripheral tolerance. Transplantation. 1992 Jun;53(6):1281–1294. [PubMed] [Google Scholar]
  28. Van Oers M. H., Van der Heyden A. A., Aarden L. A. Interleukin 6 (IL-6) in serum and urine of renal transplant recipients. Clin Exp Immunol. 1988 Feb;71(2):314–319. [PMC free article] [PubMed] [Google Scholar]
  29. Vandenbroecke C., Caillat-Zucman S., Legendre C., Noel L. H., Kreis H., Woodrow D., Bach J. F., Tovey M. G. Differential in situ expression of cytokines in renal allograft rejection. Transplantation. 1991 Mar;51(3):602–609. doi: 10.1097/00007890-199103000-00012. [DOI] [PubMed] [Google Scholar]
  30. Vieira P., de Waal-Malefyt R., Dang M. N., Johnson K. E., Kastelein R., Fiorentino D. F., deVries J. E., Roncarolo M. G., Mosmann T. R., Moore K. W. Isolation and expression of human cytokine synthesis inhibitory factor cDNA clones: homology to Epstein-Barr virus open reading frame BCRFI. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1172–1176. doi: 10.1073/pnas.88.4.1172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Whitehead B. F., Stoehr C., Wu C. J., Patterson G., Burchard E. G., Theodore J., Clayberger C., Starnes V. A. Cytokine gene expression in human lung transplant recipients. Transplantation. 1993 Oct;56(4):956–961. doi: 10.1097/00007890-199310000-00034. [DOI] [PubMed] [Google Scholar]
  32. Winslow S. G., Henkart P. A. Polyinosinic acid as a carrier in the microscale purification of total RNA. Nucleic Acids Res. 1991 Jun 25;19(12):3251–3253. doi: 10.1093/nar/19.12.3251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wu C. J., Lovett M., Wong-Lee J., Moeller F., Kitamura M., Goralski T. J., Billingham M. E., Starnes V. A., Clayberger C. Cytokine gene expression in rejecting cardiac allografts. Transplantation. 1992 Aug;54(2):326–332. doi: 10.1097/00007890-199208000-00024. [DOI] [PubMed] [Google Scholar]
  34. Yssel H., De Waal Malefyt R., Roncarolo M. G., Abrams J. S., Lahesmaa R., Spits H., de Vries J. E. IL-10 is produced by subsets of human CD4+ T cell clones and peripheral blood T cells. J Immunol. 1992 Oct 1;149(7):2378–2384. [PubMed] [Google Scholar]
  35. Zhao X. M., Frist W. H., Yeoh T. K., Miller G. G. Expression of cytokine genes in human cardiac allografts: correlation of IL-6 and transforming growth factor-beta (TGF-beta) with histological rejection. Clin Exp Immunol. 1993 Sep;93(3):448–451. doi: 10.1111/j.1365-2249.1993.tb08199.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zhao X. M., Yeoh T. K., Hiebert M., Frist W. H., Miller G. G. The expression of acidic fibroblast growth factor (heparin-binding growth factor-1) and cytokine genes in human cardiac allografts and T cells. Transplantation. 1993 Nov;56(5):1177–1182. doi: 10.1097/00007890-199311000-00025. [DOI] [PubMed] [Google Scholar]
  37. de Waal Malefyt R., Abrams J., Bennett B., Figdor C. G., de Vries J. E. Interleukin 10(IL-10) inhibits cytokine synthesis by human monocytes: an autoregulatory role of IL-10 produced by monocytes. J Exp Med. 1991 Nov 1;174(5):1209–1220. doi: 10.1084/jem.174.5.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]

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