Limulus Clotting Factor C is a multi-domain serine protease that triggers horseshoe crab hemolymph clotting in the presence of trace amounts of bacterial lipopolysaccharides. Here we describe and functionally characterize an homologous molecule, designated as IrFC, from the hard tick Ixodes ricinus. Tick Factor C consists of an N-terminal cysteine-rich domain, four complement control protein (sushi) modules, an LCCL domain, a truncated C-lectin domain and a C-terminal trypsin-type domain. Developmental expression profiling by quantitative real-time PCR revealed that the irfc mRNA is expressed in all stages including eggs. In tissues dissected from adult I. ricinus females, the irfc mRNA is present mainly in tick hemocytes and accordingly, indirect immunofluorescence microscopy localized IrFC intracellularly, in tick hemocytes. Irfc mRNA levels were markedly increased upon injection of sterile saline, or different microbes, demonstrating that the irfc gene transcription occurs in response to injury. This indicates a possible role of IrFC in hemolymph clotting and/or wound healing, although these defense mechanisms have not been yet definitely demonstrated in ticks. RNAi silencing of irfc expression resulted in a significant reduction in phagocytic activity of tick hemocytes against the Gram-negative bacteria Chryseobacterium indologenes and Escherichia coli, but not against the yeast, Candida albicans. This result suggests that IrFC plays a role in the tick primordial complement system and as such possibly mediates transmission of tick-borne pathogens.
- MeSH
- Borrelia imunologie MeSH
- Candida albicans imunologie MeSH
- Escherichia coli imunologie MeSH
- exprese genu MeSH
- fagocytóza MeSH
- klíště enzymologie genetika imunologie mikrobiologie MeSH
- komplement fyziologie MeSH
- messenger RNA biosyntéza genetika MeSH
- Micrococcus luteus imunologie MeSH
- molekulární sekvence - údaje MeSH
- prekurzory enzymů biosyntéza genetika MeSH
- přirozená imunita MeSH
- proteiny členovců biosyntéza genetika MeSH
- serinové endopeptidasy biosyntéza genetika MeSH
- upregulace imunologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
OBJECTIVE: Progelatinase B/proMMP-9 has recently been identified as an indicator of pleural inflammation, presumably originating from granulocytes. The aim of this study was to verify the origin of progelatinase B by simultaneous estimation of specific markers of neutrophil recruitment and activation in pleural effusions following induced pleurisy and pleural injury. MATERIAL AND METHODS: Sixty-three samples of pleural fluid from patients undergoing therapeutic talc pleurodesis (n = 8) and explorative thoracoscopy (n = 3) collected before and at different time intervals after the intervention were analyzed for progelatinase B and neutrophil gelatinase-associated lipocalin (NGAL)-gelatinase complex by substrate electrophoresis, for myeloperoxidase (MPO) and interleukin-8 (IL-8) by immunoadsorbent sandwich assay, as well as for leukocyte count, C-reactive protein (CRP) and total protein (TP). RESULTS: A significant increase in free and NGAL-complexed progelatinase B, MPO and IL-8 was recorded within 48 h following treatment in all subjects. Progelatinase B was strongly correlated with NGAL-gelatinase complex (r = 0.88, p = 0.001), MPO (r = 0.81, p = 0.001), neutrophil count (r = 0.75, p = 0.01) and IL-8 (r = 0.71, p = 0.001), but not with CRP and TP. CONCLUSIONS: The results support the neutrophil origin of the proenzyme, which confirms progelatinase B as an indicator of a local inflammatory reaction. Quantifying the inflammatory reaction may be helpful in the evaluation of both the technical variants of therapeutic pleurodesis and finer discrimination of paraneoplastic effusions.
- MeSH
- biologické markery metabolismus MeSH
- degranulace buněk fyziologie MeSH
- financování organizované MeSH
- interleukin-8 metabolismus MeSH
- kolagenasy biosyntéza MeSH
- lidé středního věku MeSH
- lidé MeSH
- lipokaliny MeSH
- matrixová metaloproteinasa 9 MeSH
- metaloendopeptidasy biosyntéza MeSH
- neutrofily enzymologie fyziologie patologie MeSH
- peroxidasa metabolismus MeSH
- pleura enzymologie patologie zranění MeSH
- pleurální výpotek enzymologie patologie MeSH
- pleuritida enzymologie patologie MeSH
- prekurzory enzymů biosyntéza MeSH
- proteiny akutní fáze metabolismus MeSH
- protoonkogenní proteiny metabolismus MeSH
- senioři MeSH
- želatinasy biosyntéza MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
