Upregulation of co-stimulatory cell-surface proteins by PorB is partially TLR2 dependent.

<p>WT and TLR2 KO BMDMs were stimulated in culture with PorB, Pam3CSK4 or LOS, then stained for markers of activation and examined by flow cytometry. In WT cells, PorB increased surface expression of CD14, CD40, CD54, CD69 and CD86 as compared to unstimulated cells. Knockout cells had decreased expression of CD14, CD40 and CD86, but still had higher expression of CD54 and CD69 when compared to basal levels. In the latter two cases, the TLR2 ligand Pam3CSK4 had no effect on TLR2 -/- cells. Histograms are from one representative sample of at least 3. Data represents one of two experiments.</p

PorB induction of pro-inflammatory cytokines in macrophages is MAPK dependent.

<p>(a,b) PorB potently induces IL-6 and TNFα. Supernatants were collected from WT BMDM stimulated with PorB, Pam3CSK4 or <i>N. meningitidis</i> LOS at multiple time points. At 12, 24 and 48 hours post-stimulation levels IL-6 (a) of TNFα (b) were significantly higher than unstimulated cells. (c,d) Induction of inflammatory cytokines by PorB is inhibited by blocking MAPK. PorB, Pam3CSK4 or LOS were added to cultures of WT mouse BMDM. Cells were inhibited with the carrier DMSO, or MAPK pathway inhibitors, and production of inflammatory cytokines was measured. All inhibitors tested showed significant reductions in the stimulatory activity of PorB on the production of IL-6 (c) and/or TNFα (d). U: UO126, an inhibitor of Erk1/2 by inhibiting MEK1/2. SB: SB203580, a direct inhibitor of p38. SP: SP600125, a direct inhibitor of jnk. Data represents one of three experiments.</p

Increases in antigen-specific IgG by PorB is dependent on TLR2 and MyD88.

<p>(a) Concentration of IgG antibody to Ova in C57Bl/6 mice as measured by ELISA in serum on from day 42. WT C57Bl/6, TLR2 ^-/- and MyD88 ^-/- mice were vaccinated on days 0, 14 and 28 with Ova, Ova + PorB or sham (PBS). Vaccines that included PorB showed significantly less elevation in IgG over vaccines that did not include PorB in TLR2 ^-/- mice, and no detectable effect in MyD88 ^-/- mice. None of the mice had detectable antibodies to Ova prior to vaccination (data not shown). (b) OD of specific subtypes of anti-Ova IgG at a 1:50 dilution of serum as measured by ELISA. In WT mice, IgG1 and IgG2b were the dominant subtypes indicative of a Th2-type response. These in turn decreased in the TLR2 ^-/- mice, consistent with the total IgG data. Data represents one of two experiments. * p<0.05.</p

IL-1β induction by PorB requires ATP to activate the inflammasome.

<p>WT BMDMs were stimulated with PorB, LPS, TNFα, or Pam₃CSK₄ at the indicated concentrations for 5 hours. After 5 hours, ATP was added to half the cells for 30 minutes. Supernatants were collected and analyzed by IL-1β ELISA. IL-1β release was not observed for PorB, LPS or Pam₃CSK₄ in the absence of ATP. In the presence of ATP, significantly increased IL-1β release was observed for all three TLR ligands (p<0.05). </p

Supplemental Material - Decompression Alone in the Setting of Adult Degenerative Lumbar Scoliosis and Stenosis: A Systematic Review and Meta-Analysis

Supplemental Material for Decompression Alone in the Setting of Adult Degenerative Lumbar Scoliosis and Stenosis: A Systematic Review and Meta-Analysis by Murray Echt, Rafael De la Garza Ramos, Eric Geng, Ula Isleem, Julia Schwarz, Steven Girdler, Andrew Platt, Adewale A Bakare, Richard G. Fessler, and Samuel K Cho in Global Spine Journal</p