Selective inhibition of the C5a chemotactic cofactor function of the vitamin D binding protein by 1,25(OH)2 vitamin D3.
The Vitamin D binding protein (DBP) is a multifunctional plasma protein that can significantly enhance the chemotactic response to complement fragment C5a. The chemotactic cofactor function of DBP requires cell surface binding in order to mediate this process. The goal of this study was to investigate the effect of ligating DBP with its two primary physiological ligands, Vitamin D and G-actin, on both binding to neutrophils and the ability to enhance chemotaxis to C5a. There was no difference in neutrophil binding between of the holo (bound) forms versus the apo (unbound) form of radioiodinated DBP, indicating that the cell binding region of DBP is likely distinct from the Vitamin D sterol and G-actin binding sites. Likewise, G-actin, 25(OH)D3, and G-actin plus 25(OH)D3 bound to DBP did not alter its capacity to enhance chemotaxis toward C5a. However, the active form of Vitamin D (1,25(OH)2D3) completely eliminated the chemotactic cofactor function of DBP. Dose-response curves demonstrated that as little as 1pM 1,25(OH)2D3 significantly inhibited chemotaxis enhancement. Moreover, at physiological concentrations 1,25(OH)2D3 needs to be bound to DBP to mediate the inhibitory effect. Neutrophil chemotaxis to optimal concentrations of C5a, formyl peptide, CXCL8 or leukotriene B4 was not altered by 1,25(OH)2D3, indicating that the active vitamin does not have a global inhibitory effect on neutrophil chemotaxis. Finally, inhibition of cell surface alkaline phosphatase (AP) with sodium orthovanadate completely reversed the inhibitory effect of 1,25(OH)2D3. These results indicate that the cell binding and co-chemotactic functions of DBP are not altered when the protein binds G-actin and/or Vitamin D. Furthermore, the co-chemotactic signal from DBP can be eliminated or counteracted by 1,25(OH)2D3.
Shah
Anisha B
AB
Department of Pathology, Health Sciences Center, Stony Brook University School of Medicine, Stony Brook, NY 11794-8691, USA.
DiMartino
Stephen J
SJ
Trujillo
Glenda
G
Kew
Richard R
RR
D000199:Actins / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology*; D002630:Chemotactic Factors / Q000037:antagonists & inhibitors* / Q000378:metabolism; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D015936:Complement C5a / Q000037:antagonists & inhibitors* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007457:Iodine Radioisotopes; D008024:Ligands; D009504:Neutrophils / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D014638:Vanadates / Q000494:pharmacology; D014809:Vitamin D-Binding Protein / Q000037:antagonists & inhibitors* / Q000378:metabolism
D000199:Actins; D002630:Chemotactic Factors; D007457:Iodine Radioisotopes; D008024:Ligands; D014809:Vitamin D-Binding Protein; D014638:Vanadates; D015936:Complement C5a; D002117:Calcitriol