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1.
Infect Immun ; 75(4): 1926-32, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17283098

RESUMO

Lipoteichoic acid (LTA) is a cell surface glycoconjugate of gram-positive bacteria and is reported to activate the innate immune system. We previously reported that purified LTA obtained from Enterococcus hirae has no immunostimulating activity, but a subfraction (Eh-AF) in an LTA fraction possesses activity. In this study, we established a mouse monoclonal antibody neutralizing the activity of Eh-AF and investigated its inhibitory effects. Monoclonal antibody (MAbEh1) was established by the immunization of BALB/c mice with Eh-AF, followed by hybridoma screening based on its inhibitory effect for the production of interleukin-6 (IL-6) induced by Eh-AF. MAbEh1 neutralized the production of IL-6 by LTA fraction from not only E. hirae but also Staphylococcus aureus, while it failed to block that of lipopolysaccharide, suggesting that the antibody recognized a common active structure(s) in LTA fractions. Synthetic glycolipids in these LTAs did not induce cytokine production, at least in our system. Interestingly, the antibody was found to inhibit the activity of immunostimulating synthetic lipopeptides, Pam(3)CSK(4) and FSL-1. These results suggest that MAbEh1 neutralizes the activity of lipoprotein-like compounds which is responsible for the activity of the LTA fraction of E. hirae and S. aureus.


Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Enterococcus/imunologia , Lipopolissacarídeos/imunologia , Lipoproteínas/imunologia , Ácidos Teicoicos/imunologia , Animais , Linhagem Celular Tumoral , Interleucina-6/metabolismo , Lipopolissacarídeos/isolamento & purificação , Lipoproteínas/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Modelos Animais , Testes de Neutralização , Staphylococcus aureus/imunologia , Ácidos Teicoicos/isolamento & purificação
2.
J Bone Miner Metab ; 24(5): 373-9, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16937269

RESUMO

Multinucleated cell formation is crucial for osteoclastogenesis, and the expression of nuclear factor of activated T cells (NFAT)2 (NFATc1) is essential for this process. We previously found, using mouse RAW264 cells, that culture at high cell density blocked progression to the multinucleated cell stage induced by stimulation with receptor activator of nuclear factor kappaB ligand (RANKL). Here, we have confirmed this finding in a bone marrow cell system and extended the analysis further. A high cell density appeared to cause a change in the composition of the culture medium accompanying downregulation of NFAT2 expression, and we identified L-serine (LSer) as essential for the expression of NFAT2 induced by RANKL. Namely, culture at high cell density caused a depletion of LSer in the medium. Consequently, L-Ser appeared to exert its effect at an early stage under the regular conditions used for inducing the expression of c-Fos, an upstream regulator of NFAT2. D-Ser, an enantiomer of L-Ser, showed no NFAT2-inducing activity. The expression of NFAT2, using a retrovirus vector, could compensate for the depletion of L-Ser and resume the progression to the multinucleated cell stage. These results demonstrate a novel role for L-Ser in RANKL-induced osteoclastogenesis in vitro.


Assuntos
Proteínas de Transporte/fisiologia , Diferenciação Celular/fisiologia , Glicoproteínas de Membrana/fisiologia , Fatores de Transcrição NFATC/genética , Osteoclastos/citologia , Serina/fisiologia , Animais , Linhagem Celular , Macrófagos/fisiologia , Camundongos , Fatores de Transcrição NFATC/biossíntese , Ligante RANK , Receptor Ativador de Fator Nuclear kappa-B
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