title: In Vitro Effects of Thymoglobulin in Human Embryonic Kidney Cell Line (HEK293) in Culture.
abstract: BACKGROUND
  Anti-thymocyte globulins are polyclonal T-cell-depleting immunoglobulins used in induction of immunosuppression in kidney transplant recipients. Thymoglobulin is purified rabbit immunoglobulin (Ig)G, obtained by immunization of rabbits with fetal human thymus, which depletes T lymphocytes by complement-dependent lysis and apoptosis, reduces production of cytokines, and decreases expression of adhesion molecules in endothelial cells.
  
  
  METHODS
  To determine possible direct effects of Thymoglobulin on kidney cells during transplantation, we used the Human Embryonic Kidney cell line (HEK293) in culture. We measured membrane potential of the cells by use of the slow whole patch-clamp technique. We determined effects of Thymoglobulin on cell death and proliferation during hypoxia/re-oxygenation injury, using a hypoxic chamber.
  
  
  RESULTS
  Depolarizations of HEK293 cells caused by Thymoglobulin were concentration-dependent and membrane potential-dependent, showing direct effects of Thymoglobulin on the HEK293 cells, whereas rabbit anti-thymocyte globulin produced against Jurkat cells (ATG-F) and normal rabbit IgG had no effects. To determine the effects of Thymoglobulin in hypoxia/re-oxygenation conditions, cells were incubated for 24 hours with Thymoglobulin in an atmosphere with 5% CO2-95% N2at 37°C followed by 1 hour in atmosphere with 5% CO2-95% air at 37°C. The effects of hypoxia/re-oxygenation were detected by calculating cell death and determining the cell growth, using scratch test.
  
  
  CONCLUSIONS
  Thymoglobulin prevented the cell death induced by hypoxia and re-oxygenation conditions. In addition, it accelerated the cell growth (improved scratch wound-healing). This is the first study to show the direct effects of Thymoglobulin on kidney-derived epithelial cells, which may lead to better understanding of its effects in kidney transplantation.
authors: Department of Urology, University Hospital Merkur, Zagreb, Croatia.
authors: F
authors: F
authors: Jurenec
authors: Department of Medicine, University Hospital Merkur, Zagreb, Croatia.
authors: K
authors: K
authors: Mihovilović
authors: Department of Anatomy, Histology, and Embryology, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
authors: K
authors: K
authors: Špiranec
authors: Croatian Institute for Brain Research, Department of Physiology and Immunology, School of Medicine, University of Zagreb, Zagreb, Croatia.
authors: A
authors: A
authors: Sinđić
authors: Renal Division, Department of Medicine, University Hospital Merkur and School of Medicine, Zagreb, Croatia. Electronic address: mladen.knotek@kb-merkur.hr.
authors: M
authors: M
authors: Knotek
mesh_terms: D000818:Animals; D000961:Antilymphocyte Serum / Q000494:pharmacology*; D016923:Cell Death / Q000187:drug effects; D015687:Cell Hypoxia; D049109:Cell Proliferation; D006084:Graft Rejection / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization / Q000379:methods; D007074:Immunoglobulin G / Q000276:immunology; D007155:Immunologic Factors / Q000494:pharmacology*; D007165:Immunosuppression Therapy / Q000379:methods; D019169:Jurkat Cells / Q000276:immunology; D007668:Kidney / Q000187:drug effects*; D016030:Kidney Transplantation; D008212:Lymphocyte Depletion; D008564:Membrane Potentials / Q000187:drug effects; D011817:Rabbits
chemical_list: D000961:Antilymphocyte Serum; D007074:Immunoglobulin G; D007155:Immunologic Factors; C512542:thymoglobulin