34 HBD3-enhanced Porphyromonas gingivalis rHagB Cytokine Responses in Dendritic Cells

Wednesday, March 21, 2012: 2:30 p.m. - 4 p.m.
Presentation Type: Oral Session
L.E. HARVEY1, A. PROGULSKE-FOX2, and K. BROGDEN1, 1Dows Institute for Dental Research, University of Iowa, Iowa City, IA, 2Dept. of Oral Biology, University of Florida, Gainesville, FL
Previously, we found that HBD3, co-administered with recombinant hemagglutinin B (rHagB) from Porphyromonas gingivalis attenuates a chemokine and pro-inflammatory cytokine response in human myeloid dendritic cells.  Objectives: The objective of this study was to determine if the chemokine and cytokine response of dendritic cells induced by rHagB is still attenuated when HBD3 is administered prior to exposure with rHagB; when administered simultaneously with rHagB; or when administered after exposure to rHagB.  Methods: 5 X 105 human myeloid dendritic cells in 25 cm2 flasks were i) first exposed to 0.2 uM HBD3 and then 0.02 uM rHagB, ii) given 0.2 uM HBD3 and0.02 uM rHagB simultaneously, but not together, and iii) first given 0.02 uM rHagB and then 0.2 uM HBD3.  At 0, 1, 2, 4, 8, and 16 hours post exposure, culture supernatants were removed for the determination of 22 chemokines and cytokines using a commercial multiplexed fluorescent bead-based immunoassay (Millipore, Billerica, MA) in the Luminex 100 IS Instrument (Luminex, Austin, TX).  Results:   By 16 hours, rHagB alone induced a robust chemokine and cytokine response.  The timing of HBD3 administration was significant ( P < 0.05).  HBD3 co-incubated with rHagB (for 30 min prior to exposure) attenuated a chemokine and pro-inflammatory cytokine response to rHagB.  However if HBD3 was administered 1 hour prior, rHagB induced an early, robust, and sustained pro-inflammatory cytokine response.  If HBD3 and rHagB were administered simultaneously, but not together or if HBD3 was administered 1 hour after, then rHagB induced a modestly robust and slightly sustained pro-inflammatory cytokine response.  Conclusions: The results show that the timing of HBD3 administration is important and HBD3 has the capacity to modulate the cytokine and chemokine response of dendritic cells to rHagB.  Supported by funds from NIH, NIDCR R01 DE014390.
This abstract is based on research that was funded entirely or partially by an outside source: Dental Reasearch Grant, University of Iowa, College of Dentistry NIH, NIDCR R01 DE014390

Keywords: Antigens-antibodies, Cytokine and Inflammatory mediators