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The effect of extended freeze-dried or heat-killed BCG and TLR agonists on the human phagocyte NADPH oxidase

J Moreira L A P Talero; Bernardo Boris Jorge Vargaftig; Antonio Condino-Neto; Meeting of the Brazilian Society for Immunology (31. 2006 Búzios)

Abstracts São Paulo, SP: Brazilian Society for Immunology, 2006

São Paulo 2006

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  • Título:
    The effect of extended freeze-dried or heat-killed BCG and TLR agonists on the human phagocyte NADPH oxidase
  • Autor: J Moreira
  • L A P Talero; Bernardo Boris Jorge Vargaftig; Antonio Condino-Neto; Meeting of the Brazilian Society for Immunology (31. 2006 Búzios)
  • Assuntos: IMUNOLOGIA; FARMACOLOGIA
  • É parte de: Abstracts São Paulo, SP: Brazilian Society for Immunology, 2006
  • Notas: Disponível em CD-ROM
  • Descrição: Introduction and Objectives: Toll-like receptors (TLR) are important in the recognition of different microbial pathogens by the immune system, triggering inflammatory responses. They are involved in the activation of several transcription factors such as nuclear factor (NF)- B, which may be involved in the activation of the NADPH oxidase. Different Bacillus Calmette- Guérin (BCG) preparations have been tested on Th1/Th2 balance restoration in asthma. We studied the effects of two different preparations of inactivated BCG, extended freeze-dried (EFD) and heat-killed (HK), and different Toll-receptor agonists on the human NADPH oxidase, using either the U937 cell line and human peripheral blood mononuclear cells (PBMC). Significant differences were determined by ANOVA test (p=0.05). Methods and Results: PBMC were obtained from healthy individuals through density gradient. U937 cells were cultured in RPMI medium supplemented with fetal calf serum. NADPH oxidase activation was evaluated by respiratory burst, accessed by cytochrome c reduction, according to Condino-Neto (J. Immunol. 161:4960-7, 1998), and gp91phox and p47phox gene expression, by RT-PCR. Incubation of U937 cells (48 or 72h, both n=5) with EFD and HK BCG (MOI 20:1) did not increase the PMA-induced superoxide release or affected gp91phox gene expression by U937 cells. Synthetic bacterial lipoprotein-PAM-3CSK-4 (TLR-2/TLR-1 ligand) and syndiacylated-FSL- 1 (TLR-2/TLR-6) as well as
    LPS (TLR-4) and zymosan (TLR-5) did not alter the gp91phox expression in U937 cells when incubated for 24, 48 and 72h. Though there was a decrease of gp91phox expression in PBMC cells incubated for 24h with HK BCG (54.2%, p<0.001, n=4) in relation to control. EFD BCG also produced decrease (16.7%), but not significant. Futhermore, it was observed a reduction of p47phox expression in PBMC when incubated with EFD (22%) and HK BCG (32%), nevertheless not significant (n=3). Conclusion: The incubation of non differentiated U937 cells with EFD and HK BCG or TLR ligands do not alter NADPH oxidase activation. However, in differentiated cells as PBMC, these BCGs seem to inhibit this enzymatic complex.
  • Editor: São Paulo
  • Data de criação/publicação: 2006
  • Formato: res. IN.006.
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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