title: Allergen specific responses in cord and adult blood are differentially modulated in the presence of endotoxins.
abstract: BACKGROUND
  Endotoxins are common contaminants in allergen preparations and affect antigen-specific cellular responses. Distinct effects of endotoxin on cells in human umbilical cord and adult blood are poorly defined.
  
  
  OBJECTIVE
  To examine the effect of endotoxins in allergen preparations on cellular responses in human cord and peripheral blood (PB).
  
  
  METHODS
  The endotoxin content in beta lactoglobulin (BLG), the peanut allergen Ara h 1 and the major birch pollen allergen Bet v 1 was assessed. Proliferation and cytokine response of mononuclear cells towards contaminated and lipopolysaccharide (LPS)-free allergens were evaluated at different time-points. Fractions of contaminated BLG were generated and assayed on their immuno-stimulatory capacity. The involvement of toll-like receptor (TLR) 2 and 4 was investigated by blocking antibodies and TLR-transfected human embryonic kidney cells.
  
  
  RESULTS
  The proliferative response of cord blood (CB)-derived mononuclear cells towards allergen-preparations at day 3 was related to the level of LPS contamination. At day 7, proliferation was also detected in the absence of endotoxin. Cytokine production in CB was strongly affected by the content of endotoxin, TLR-4 dependent and not related to the allergen content. Allergen- and endotoxin-induced proliferative responses were generally significantly higher in CB than in adult blood.
  
  
  CONCLUSIONS
  Endotoxins in allergen preparations confound allergen-specific cellular responses. The impact of these contaminations varies with the blood source (CB vs. PB), the type of allergen and is time- and dose-dependent.
authors.affiliation: Department of Pediatrics, Medical University of Vienna, Vienna, Austria.
authors.forename: T
authors.initials: T
authors.lastname: Eiwegger
authors.forename: E
authors.initials: E
authors.lastname: Mayer
authors.forename: S
authors.initials: S
authors.lastname: Brix
authors.forename: I
authors.initials: I
authors.lastname: Schabussova
authors.forename: E
authors.initials: E
authors.lastname: Dehlink
authors.forename: B
authors.initials: B
authors.lastname: Bohle
authors.forename: V
authors.initials: V
authors.lastname: Barkholt
authors.forename: Z
authors.initials: Z
authors.lastname: Szépfalusi
mesh_terms: D000328:Adult; D000485:Allergens / Q000276:immunology* / Q000494:pharmacology; D052179:Antigens, Plant / Q000187:drug effects / Q000276:immunology; D002460:Cell Line; D016207:Cytokines / Q000187:drug effects / Q000276:immunology; D004731:Endotoxins / Q000276:immunology* / Q000494:pharmacology; D005312:Fetal Blood / Q000187:drug effects / Q000276:immunology*; D006023:Glycoproteins / Q000276:immunology / Q000494:pharmacology; D006801:Humans; D007782:Lactoglobulins / Q000276:immunology* / Q000494:pharmacology; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology*; D008565:Membrane Proteins; D010940:Plant Proteins / Q000276:immunology / Q000494:pharmacology; D013745:Tetanus Toxoid / Q000276:immunology / Q000494:pharmacology; D051195:Toll-Like Receptor 2 / Q000276:immunology; D051197:Toll-Like Receptor 4 / Q000276:immunology / Q000378:metabolism
chemical_list: D000485:Allergens; D052179:Antigens, Plant; C500889:Ara h 1 protein, Arachis hypogaea; D016207:Cytokines; D004731:Endotoxins; D006023:Glycoproteins; D007782:Lactoglobulins; D008565:Membrane Proteins; D010940:Plant Proteins; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D013745:Tetanus Toxoid; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4
references.citation: Int Arch Allergy Immunol. 1994 Aug;104(4):358-65
references.pmid: 7518714
references.citation: J Immunol. 1995 Feb 15;154(4):1932-40
references.pmid: 7530747
references.citation: Exp Hematol. 2005 Dec;33(12):1508-20
references.pmid: 16338494
references.citation: Allergy. 2000 May;55(5):470-5
references.pmid: 10843428
references.citation: J Allergy Clin Immunol. 2003 Dec;112(6):1216-22
references.pmid: 14657886
references.citation: Clin Exp Allergy. 1998 Nov;28(11):1313-6
references.pmid: 9824400
references.citation: Clin Exp Allergy. 1996 Jul;26(7):780-8
references.pmid: 8842551
references.citation: Pediatr Allergy Immunol. 1996 Aug;7(3):109-16
references.pmid: 9116874
references.citation: Pediatr Res. 1997 Jan;41(1):128-31
references.pmid: 8979301
references.citation: J Biol Chem. 2003 Jan 3;278(1):174-9
references.pmid: 12403778
references.citation: J Immunol. 1998 May 15;160(10):4730-7
references.pmid: 9590218
references.citation: Arch Dis Child. 1992 Aug;67(8):1003-7
references.pmid: 1520001
references.citation: J Immunol. 2004 Sep 1;173(5):3084-92
references.pmid: 15322168
references.citation: J Clin Immunol. 2005 Jul;25(4):329-37
references.pmid: 16133989
references.citation: Int Arch Allergy Immunol. 2004 Nov;135(3):216-20
references.pmid: 15475660
references.citation: Pediatr Allergy Immunol. 1995 May;6(2):85-90
references.pmid: 7581725
references.citation: J Clin Invest. 2007 Jun;117(6):1637-46
references.pmid: 17549258
references.citation: Int Arch Allergy Immunol. 1997 May-Jul;113(1-3):75-9
references.pmid: 9130488
references.citation: Pediatr Res. 2000 Sep;48(3):404-7
references.pmid: 10960510
references.citation: Clin Exp Allergy. 2006 Sep;36(9):1130-7
references.pmid: 16961712
references.citation: Lancet. 2000 Dec 2;356(9245):1900-2
references.pmid: 11130390
references.citation: J Allergy Clin Immunol. 2000 Sep;106(3):530-6
references.pmid: 10984374
references.citation: Transplantation. 2007 Jul 15;84(1):23-30
references.pmid: 17627233
references.citation: J Allergy Clin Immunol. 1998 Apr;101(4 Pt 1):514-20
references.pmid: 9564805
references.citation: Toxicology. 2000 Nov 2;152(1-3):37-45
references.pmid: 11090938
references.citation: Clin Exp Allergy. 1997 Jan;27(1):28-35
references.pmid: 9117877
references.citation: Int Arch Allergy Immunol. 1993;102(3):301-3
references.pmid: 8219784
references.citation: Lancet. 1999 Jan 16;353(9148):196-200
references.pmid: 9923875