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The release of MMP-3 was not detected in untreated control (Figure 2(c), lane 1) or cells treated with either 10 nM HMGB1 (Figure 2(c), lane 11) or MTDs composed of 10 nM fMLF and 10 [micro]g/mL CpG DNA (Figure 2(c), lane 13).
In a study by Zhang et al., mtDNA or fMLF alone could upregulate MMP-8 expression by PMNs.
Same lack of response was observed when chondrocytes were challenged with combinations of CpG DNA and fMLF that showed effectiveness in upregulating MMP-8 in PMNs.
Caption: FIGURE 1: In the initial two experiments with bovine chondrocytes, HMGB1 synergized with IL-1[beta] on upregulating metalloproteinase production while MTDs (fMLF and CpG DNA) or HMGB1 alone showed little effect.
The fragment recognized by NFPRb demonstrated the characteristic NFPRb sensitivity to prior exposure of PMN to fMLF, suggesting it was derived from FPR1 as well as being something we had not observed previously in suspensions of PMN.
Each pair of neighboring lanes corresponds to the solubilized well content after a 5 min exposure of the adherent cells to vehicle (-) or 1 [micro]M fMLF (+).
In these lanes, the 60 kDa FPR1 and 25 kDa bands both show indistinguishable sensitivity to fMLF exposure of PMN ([EC.sub.50] of approximately 20-40 nM), suggesting that the 25K species carries the C-terminal epitope [17, 23, 37] recognized by NFPRb and is therefore a C-terminal proteolytic fragment of FPR1.
This region contains no serine/threonine residues and shows no posttranslational modification after exposure of cells to fMLF .
PAF is generally considered as a nonactivating or very poor/weak ROS inducer , contrasting the activity induced by many other chemoattractants such as fMLF which is regarded as a strong inducer.
Non-activating concentrations of PAF primed neutrophils to enhanced ROS production with fMLF as the second agonist [31, 32], which is a well known phenomenon and it was to be expected since PAF is a potent secretagogue (supplementary Figure 2, ).
Accordingly, when binding of the chemoattractant fMLF to FPR1 takes place at low temperature (<15[degrees]C), the activating signalling state of the receptor is bypassed, and it is directly deactivated/desensitized, but the receptor can be reactivated/resensitized by cytoskeleton-disrupting drugs (Figure 6).
A rise in [[[Ca.sup.2+]].sub.i] depending on an emptying of the stores is by definition not inhibited through addition of a [Ca.sup.2+] chelator (i.e., EGTA) (; shown for fMLF in supplementary Figure 4).
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