Surface-bound bovine serum albumin carrier protein as present in recombinant cytokine preparations amplifies T helper 17 cell polarization.
Understanding of T helper 17 lineage (TH17) polarization has been significantly promoted by cell culture experiments that reduce the complexity of the in vivo environment. We here investigated TH17amplification by coating of cytokine preparations. Cytokine preparations coated to the surface compared to the same amount given in solution significantly enhanced TH17polarization assessed by flow cytometry and interleukin (IL)-17A, IL-17F and RORγt mRNA expression. T cell proliferation and TH1polarization were similarly enhanced while TREGpolarization was impeded. TH17amplification was replicated by coating the plate with low amounts of FCS or albumin as used as carrier protein for cytokines (0.5 μl 0.1%). It was unaltered by filtration, protein digestion and arylhydrocarbon receptor blockade, not replicated by LPS and independent of integrin stimulation. TH17amplification required anti-CD3 stimulation and was T cell intrinsic. Supernatants of CD4+cells polarized on coated cytokine preparations with carrier albumin conferred amplification to fresh splenocytes. Coating markedly elevated CD4+IL-22 mRNA expression and IL-22 blockade significantly reduced TH17amplification. Our data show TH17amplification by coated albumin in the low amounts present in recombinant cytokine preparations. This unexpected adjuvant like effect underscores the need for controls also for temporal and spatial factors in cell culture.
Department of Internal Medicine, Division of Nephrology and Hypertension, Hannover Medical School, Hannover, Germany.|Department of Nephrology, Tongji Hospital, Huazhong University of Science and Technology, China.
Lei
L
Dong
Department of Internal Medicine, Division of Nephrology and Hypertension, Hannover Medical School, Hannover, Germany.
Alexandra
A
Helmke
Institute for Molecular Medicine, University of Mainz, Germany.
Ari
A
Waisman
Department of Internal Medicine, Division of Nephrology and Hypertension, Hannover Medical School, Hannover, Germany.
Hermann
H
Haller
Institute for Toxicology, Proteomics Unit, Hannover Medical School, Hannover, Germany.
Andreas
A
Pich
Department of Internal Medicine, Division of Nephrology and Hypertension, Hannover Medical School, Hannover, Germany.
Sibylle
S
von Vietinghoff
D000818:Animals; D002417:Cattle; D016207:Cytokines / Q000494:pharmacology*; D004337:Drug Carriers / Q000493:pharmacokinetics*; D051379:Mice; D018345:Mice, Knockout; D011994:Recombinant Proteins / Q000494:pharmacology; D012710:Serum Albumin, Bovine; D058504:Th17 Cells / Q000166:cytology / Q000276:immunology*
D016207:Cytokines; D004337:Drug Carriers; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine