Abstract
Current methods are inadequate for the diagnosis of early chronic allograft rejection. The goal of this study was to determine whether ligation of anti‐HLA antibodies to endothelial cells is associated with a distinctive cytokine secretory pattern. Human iliac artery endothelial cells (HIAEC) cultured in vitro were incubated with w6/32, an anti‐HLA class I mAb. Culture supernatants collected daily for up to 4 days were tested for secretion of 13 cytokines using a multiplexed fluorescent microsphere immunoassay. Culture of HIAEC with medium containing mAb w6/32 supported the growth of HIAEC during the 4‐day study period. Levels of the pro‐inflammatory cytokines IL‐1β, IL‐6, IL‐8, and TNF‐α became significantly increased in supernatants of HIAEC incubated with the mAb w6/32. We conclude that ligation of anti‐HLA class I antibodies to HLA class I antigens in endothelial cells initiates an acute inflammatory process and detecting an inflammatory cytokine secretory pattern might be useful to diagnose sub‐clinical chronic allograft rejection. J. Clin. Lab. Anal. 23:157–160, 2009. © 2009 Wiley‐Liss, Inc.
Keywords: endothelial cells, HLA antibody, cytokines
| Abbreviations | |
|---|---|
| EGM | endothelial cell growth medium |
| HIAEC | human iliac artery endothelial cells. |
REFERENCES
- 1. Cornell LD, Smith RN, Colvin RB. Kidney transplantation: mechanisms of rejection and acceptance. Annu Rev Pathol 2008;3:189–220. [DOI] [PubMed] [Google Scholar]
- 2. Harris PE, Bian H, Reed EF. Induction of high affinity fibroblast growth factor receptor expression and proliferation in human endothelial cells by anti‐HLA antibodies: A possible mechanism for transplant atherosclerosis. J Immunol 1997;159:5697–5704. [PubMed] [Google Scholar]
- 3. Terasaki PI, Cai J. Human leukocyte antigen antibodies and chronic rejection: From association to causation. Transplantation 2008;86:377–383. [DOI] [PubMed] [Google Scholar]
- 4. Bian H, Harris PE, Reed EF. Ligation of HLA class I molecules on smooth muscle cells with anti‐HLA antibodies induces tyrosine phosphorylation, fibroblast growth factor receptor expression and cell proliferation. Int Immunol 1998;10:1315–1323. [DOI] [PubMed] [Google Scholar]
- 5. Mao Q, Terasaki PI, Cai J, et al. Extremely high association between appearance of HLA antibodies and failure of kidney grafts in a five‐year longitudinal study. Am J Transplant 2007;7:864–871. [DOI] [PubMed] [Google Scholar]
- 6. Mizutani K, Terasaki P, Rosen A, et al. Serial ten‐year follow‐up of HLA and MICA antibody production prior to kidney graft failure. Am J Transplant 2005;5:2265–2272. [DOI] [PubMed] [Google Scholar]
- 7. Worthington JE, Martin S, Al‐Husseini DM, Dyer PA, Johnson RW. Posttransplantation production of donor HLA‐specific antibodies as a predictor of renal transplant outcome. Transplantation 2003;75:1034–1040. [DOI] [PubMed] [Google Scholar]
- 8. Lee PC, Terasaki PI, Takemoto SK, et al. All chronic rejection failures of kidney transplants were preceded by the development of HLA antibodies. Transplantation 2002;74:1192–1194. [DOI] [PubMed] [Google Scholar]
- 9. Palmer SM, Davis RD, Hadjiliadis D, et al. Development of an antibody specific to major histocompatibility antigens detectable by flow cytometry after lung transplant is associated with bronchiolitis obliterans syndrome. Transplantation 2002;74:799–804. [DOI] [PubMed] [Google Scholar]
- 10. Jaramillo A, Smith MA, Phelan D, et al. Development of ELISA‐detected anti‐HLA antibodies precedes the development of bronchiolitis obliterans syndrome and correlates with progressive decline in pulmonary function after lung transplantation. Transplantation 1999;67:1155–1161. [DOI] [PubMed] [Google Scholar]
- 11. Stastny P, Lavingia B, Fixler DE, Yancy CW, Ring WS. Antibodies against donor human leukocyte antigens and the outcome of cardiac allografts in adults and children. Transplantation 2007;84:738–745. [DOI] [PubMed] [Google Scholar]
- 12. Tambur AR, Bray RA, Takemoto SK, et al. Flow cytometric detection of HLA‐specific antibodies as a predictor of heart allograft rejection. Transplantation 2000;70:1055–1059. [DOI] [PubMed] [Google Scholar]
- 13. Martins TB, Pasi BM, Pickering JW, Jaskowski TD, Litwin CM, Hill HR. Determination of cytokine responses using a multiplexed fluorescent microsphere immunoassay. Am J Clin Pathol 2002;118:346–353. [DOI] [PubMed] [Google Scholar]
- 14. Jaramillo A, Smith CR, Maruyama T, Zhang L, Patterson GA, Mohanakumar T. Anti‐HLA class I antibody binding to airway epithelial cells induces production of fibrogenic growth factors and apoptotic cell death: A possible mechanism for bronchiolitis obliterans syndrome. Hum Immunol 2003;64:521–529. [DOI] [PubMed] [Google Scholar]
- 15. Remick DG. Interleukin‐8. Crit Care Med 2005;33:S466–S467. [DOI] [PubMed] [Google Scholar]
- 16. Bradley JR. TNF‐mediated inflammatory disease. J Pathol 2008;214:149–160. [DOI] [PubMed] [Google Scholar]
- 17. Dinarello CA. Interleukin‐1beta. Crit Care Med 2005;33:S460–S462. [DOI] [PubMed] [Google Scholar]
- 18. Song M, Kellum JA. Interleukin‐6. Crit Care Med 2005;33:S463–S465. [DOI] [PubMed] [Google Scholar]