The influence of intention in conflict on the interventions and apologies of children in virtual conflict situations

To examine the influence of intention in conflict on the interventions and apologies of children in virtual conflict situations, we conducted experiments with 3- and 5-year-old girls and boys. Our findings suggest that the conflict's intention reflected the interventions, apologies and reasons. An age difference was observed. Five-year-old children gave more effective answers to resolve conflict situations, using both interventions and apologies more than the 3 year-old children. When asked about such interventions and apologies, many 5-year-old children suggested reasons that were focused on the victim's feelings and accepted responsibility for the conflict or expressed a sense of guilt. In contrast, many 3-year-olds decided their behaviors based on roles or evaluations of others. Furthermore, gender differences were observed Most of the girls selected an impartial intervention between victim and attacker; in contrast, many of the boys selected an intervention that supported only the victim. As an attacker, most girls answered apologetically. When questioned about such interventions and apologies, many girls made judgements of right and wrong themselves and accepted responsibility for the conflict or expressed a sense of guilt. In contrast, many of the boys decided their behaviors from the evaluations of others

The diacylated lipopeptide FSL-1 induces TLR2-mediated Th2 responses

The diacylated lipopeptide FSL-1 enhanced the generation of IgG antibodies in TLR2+/+ mice, but not in TLR2-/- mice, when administered together with hen egg lysozyme as an antigen. Escherichia coli lipopolysaccharide enhanced the generation of antigen-specific antibodies in both TLR2-/- and TLR2+/+ mice. In TLR2+/+ mice, the enhancement level by FSL-1 was similar to that by lipopolysaccharide. Analysis of the IgG antibodies subclass demonstrated that level of Th2-type IgG1 antibodies was higher than that of Th1-type IgG2a antibodies. Both FSL-1 and lipopolysaccharide induced production of IL-10 and IL-6 by splenocytes from TLR2+/+ mice. Lipopolysaccharide also induced production of these cytokines by splenocytes from TLR2-/- mice, but FSL-1 did not. Neither FSL-1 nor lipopolysaccharide induced IL-12p70 production by splenocytes from both types of mice. FSL-1 upregulated B7.2 expression in B220+ cells from TLR2+/+ mice but not those from TLR2-/- mice, whereas lipopolysaccharide upregulated B7.2 expression in B220+ cells from both types of mice. FSL-1 and, to a lesser extent, lipopolysaccharide activated mitogen-activated protein kinases in splenocytes. FSL-1 and, to a lesser extent,lipopolysaccharide induced the expression of c-Fos, which is known to be involved in Th2-type responses, in splenocytes. Thus, this study demonstrated that FSL-1 possessed TLR2-mediated Th2-type responses in vivo

CD14 directly binds to triacylated lipopeptides and facilitates recognition of the lipopeptides by the receptor complex of Toll-like receptors 2 and 1 without binding to the complex

It has demonstrated that the recognition of triacylated lipopeptides by Toll-like receptor (TLR) 2 requires TLR1 as a coreceptor. In the NF-κB reporter assay system in which human embryonic kidney 293 cells were transfected with TLR2 and TLR1 together with an NF-κB luciferase reporter gene, S-(2,3-bispalmitoyloxypropyl)-N-palmitoyl-Cys-Lys-Lys-Lys-Lys (Pam3CSK4) and Pam3CSSNA were recognized by TLR2/TLR1, but the recognition level was unexpectedly very low. However, cotransfection of CD14 drastically enhanced the recognition of triacylated lipopeptides by TLR2/TLR1. The CD14-induced enhancement did not occur without cotransfection of TLR1. Both CD14dS39-A48, a mutant with deletion of the part of possible N-terminal ligand-binding pocket, and anti-CD14 monoclonal antibody reduced the CD14-induced enhancement. Transfection of a TIR domain-deficient mutant of TLR2 (TLR2dE772-S784) or TLR1 (TLR1dQ636-K779) completely abrogated the CD14-induced enhancement. Soluble recombinant CD14 added extracellularly enhanced the recognition of Pam3CSSNA by TLR2/TLR1. Immunoprecipitation analysis demonstrated that CD14 was not associated with TLR2 but that TLR1 was associated with TLR2. In addition, surface plasmon resonance-based assay demonstrated that CD14 binds to Pam3CSK4 at a dissociation constant of 5.7 µM. This study suggests that CD14 directly binds to triacylated lipopeptides and facilitates recognition of the lipopeptides by the TLR2/TLR1 complex without binding to the receptor complex

Roles of N-linked glycans in the recognition of microbial lipopeptides and lipoproteins by TLR2

Details of roles of carbohydrates attached to Toll-like receptors (TLRs) in the recognition of pathogen-associated molecular patterns and in the formation of the functional receptor complex still remain unknown. This study was designed to determine whether the glycans linked at Asn114, Asn199, Asn414 and Asn442 residues of TLR2 ectodomain were involved in the recognition of diacylated lipopeptide and lipoprotein. Single and multiple mutants were transfected into human embryonic kidney (HEK) 293 cells together with a NF-κB luciferase reporter plasmid. All of these mutants were expressed on the surface. SDS-PAGE of the transfectants demonstrated that these mutants migrated lower than wild-type TLR2 and their molecular masses decreased as the number of mutated Asn residues increased. TLR2N114A, TLR2N199A and TLR2N414A as well as wild-type TLR2 induced NF-κB activation when stimulated with these ligands, whereas TLR2N442A failed to induce NF-κB activation. All of triple and quadruple mutants failed to induce NF-κB activation, but were associated with both wild-type TLR2 and TLR6 in the transfectants. TLR2N114A,N199A, TLR2N114A,N414A and, to a lesser extent, TLR2N114A,N442A, in which two N-linked glycans are speculated to be exposed to the concave surface of TLR2 solenoid, not only induce NF-κB activation but also are associated with wild-type TLR2 and TLR6. These results suggest that the glycan at Asn442 and at least two N-linked glycans speculated to be exposed to the concave surface of TLR2 solenoid are involved in the recognition of ligands by TLR2 and/or in formation or maturation of a functional TLR2 receptor complex

The synthetic analogue of mycoplasmal lipoprotein FSL-1 induces dendritic cell maturation through

GM-CSF-differentiated bone marrow-derived DCs (BMDCs) were stimulated with FSL-1 or E.coli LPS. FSL-1 induced the production of TNF-α and IL-12 by C57BL/6-derived BMDCs but not by BMDCs from Toll-like receptor 2-deficient (TLR2-/-) mice, whereas LPS induced the production of TNF-α and IL-12 by BMDCs derived from either type of mice. FSL-1 did not induce production of IL-10 by BMDCs from either type of mice, whereas LPS induced small amounts of IL-10 by BMDCs from both types of mice. FSL-1 upregulated the expression of CD80, CD86 and the MHC class II molecule IAb in both dose- and time-dependent manners on the surfaces of C57BL/6-derived BMDCs but not on the surface of TLR2-/--derived BMDCs, whereas LPS upregulated the expression of them on the surfaces of BMDCs from both types of mice. The expression of CD11c on the surfaces of C57BL/6-derived BMDCs was upregulated by stimulation with both FSL-1 and LPS up to 12 h and then the expression was downregulated.The results suggest that FSL-1 has activity to accelerate maturation of BMDCs and that the activity of FSL-1 is mediated by TLR2