Transcriptional expression levels of chicken collectins are affected by avian influenza A virus inoculation

Mammalian collectins have been found to play an important role in the defense against influenza A virus H9N2 inoculation, but for chicken collectins this has not yet been clarified. The aim of this study was to determine the effect of avian influenza A virus (AIV) inoculation on collectin gene expression in the respiratory tract of chickens and whether this was affected by age. For this purpose 1- and 4-week-old chickens were inoculated intratracheally with PBS or H9N2 AIV. Chickens were killed at 0, 8, 16 and 24 h postinoculation and trachea and lung were harvested for analysis. Viral RNA expression and mRNA expression of chicken collectins 1 and 2 (cCL-1 and cCL-2), chicken lung lectin (cLL) and chicken surfactant protein A (cSP-A) were determined using real-time quantitative RT-PCR. In lung, a decrease in mRNA expression of cCL-2, cLL and cSP-A after inoculation with H9N2 was seen in both 1- and 4-week-old birds, although at different time points, while in trachea changes were only seen in 4-week-old birds and expression was increased. Moreover, collectin expression correlated with viral RNA expression in lung of 1-week-old birds. These results suggest that both age and location in the respiratory tract affect changes in collectin mRNA expression after inoculation with H9N2 and indicate a possible role for collectins in the host response to AIV in the respiratory tract of chickens

The serine protease domain of MASP-3: enzymatic properties and crystal structure in complex with ecotin.

International audienceMannan-binding lectin (MBL), ficolins and collectin-11 are known to associate with three homologous modular proteases, the MBL-Associated Serine Proteases (MASPs). The crystal structures of the catalytic domains of MASP-1 and MASP-2 have been solved, but the structure of the corresponding domain of MASP-3 remains unknown. A link between mutations in the MASP1/3 gene and the rare autosomal recessive 3MC (Mingarelli, Malpuech, Michels and Carnevale,) syndrome, characterized by various developmental disorders, was discovered recently, revealing an unexpected important role of MASP-3 in early developmental processes. To gain a first insight into the enzymatic and structural properties of MASP-3, a recombinant form of its serine protease (SP) domain was produced and characterized. The amidolytic activity of this domain on fluorescent peptidyl-aminomethylcoumarin substrates was shown to be considerably lower than that of other members of the C1r/C1s/MASP family. The E. coli protease inhibitor ecotin bound to the SP domains of MASP-3 and MASP-2, whereas no significant interaction was detected with MASP-1, C1r and C1s. A tetrameric complex comprising an ecotin dimer and two MASP-3 SP domains was isolated and its crystal structure was solved and refined to 3.2 Å. Analysis of the ecotin/MASP-3 interfaces allows a better understanding of the differential reactivity of the C1r/C1s/MASP protease family members towards ecotin, and comparison of the MASP-3 SP domain structure with those of other trypsin-like proteases yields novel hypotheses accounting for its zymogen-like properties in vitro

Alteration of the level of salivary cortisol under psychological stress and its relationship with rumination and personality traits

Background and Objective: Gender differences in biobehavioral responses to environmental stressors and experience of psychological stress should be identified. This study was done to evaluate the changes of the level of salivary cortisol under psychological stress and its relationship with rumination and personality traits. Methods: In this case-control study, for 45 medical students, The NEO Personality Inventory-Short Form and emotional control questionnaire (ECQ) were filed two months before the final examination. Saliva samples were taken from students in the non-stress (control) and examination stress conditions. Salivary cortisol levels were measured by ELISA method. Results: Gender differences were not observed in the level of salivary cortisol under psychological stress. Significant difference was observed between the mean of salivary cortisol in the non-stress and under examination stress conditions. Positive correlation was found between traits of neuroticism (P<0.05) and rumination (P<0.05) with salivary cortisol as well as negative correlation between the traits openness to experience (P<0.05) and angery control (P<0.05) with salivary cortisol. Neuroticism, rumination and angery control may predict a substantial variance (32%) of salivary cortisol under exam stress. Conclusion: Psychological stress leads to increase in the secretion of salivary cortisol unrelated to gender. Subjects with different personality traits are prone to cortisol responses to stress based on their particular character

The membrane-type collectin CL-P1 is a scavenger receptor on vascular endothelial cells

American Society for Biochemistry and Molecular Biology, Katsuki, Ohtani ; Yasuhiko, Suzuki ; Souji, Eda ; Takao, Kawai ; Tetsuo, Kase ; Hiroyuki, Keshi ; Yoshinori, Sakai ; Atsushi, Fukuoh ; Takashi, Sakamoto ; Hiroyuki, Itabe ; Tatsuo, Suzutani ; Masahiro, Ogasawara ; Itsuro, Yoshida ; Nobutaka, Wakamiya, Journal of Biological Chemistry, 276(47), 2001, 44222-44228. authorCollectins are a family of C-type lectins that have collagen-like sequences and carbohydrate recognition domains (CRD). They are involved in host defense through their ability to bind to carbohydrate antigens of microorganisms. The scavenger receptors type A and MARCO are classical type scavenger receptors that have internal collagen-like domains. Here we describe a new scavenger receptor that is a membrane-type collectin from placenta (collectin placenta 1 (CL-P1)), which has a typical collectin collagen-like domain and a CRD. The cDNA has an insert of about 2.2 kilobases coding for a protein containing 742 amino acid residues. The deduced amino acid sequence shows that CL-P1 is a type II membrane protein, has a coiled-coil region, a collagen-like domain, and a CRD. It resembles type A scavenger receptors because the scavenger receptor cysteine-rich domain is replaced by a CRD. Northern analyses, reverse transcription-polymerase chain reaction, and immunohistochemistry show that CL-P1 is expressed in vascular endothelial cells but not in macrophages. By immunoblotting and flow cytometry CL-P1 appears to be a membrane glycoprotein of about 140 kDa in human umbilical vein or arterial endothelial cells, placental membrane extracts, and CL-P1 transfected Chinese hamster ovary cells. We found that CL-P1 can bind and phagocytose not only bacteria (Escherichia coli and Staphylococcus aureus) but also yeast (Saccharomyces cerevisiae). Furthermore, it reacts with oxidized low density lipoprotein (OxLDL) but not with acetylated LDL (AcLDL). These binding activities are inhibited by polyanionic ligands (polyinosinic acid, polyguanylic acid, dextran sulfate) and OxLDL but not by polycationic ligands (polyadenylic acid or polycytidylic acid), LDL, or AcLDL. These results indicate that CL-P1 might play important roles in host defenses that are different from those of soluble collectins in innate immunity