TLR-Mediated Production of Reactive Oxygen Species and Tumor Necrosis Factor Alpha by Human Peripheral Blood Neutrophils


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This paper presents the study on TLR-mediated production of reactive oxygen species and tumor necrosis factor alpha by peripheral blood neutrophils in healthy donors stimulated with zymosan (TLR2/6 ligand), peptidoglycan (TLR2/1 ligand), and lipopolysaccharide (TLR4 ligand). Luminol- and lucigen-independent chemiluminescence was used to detect the production of reactive oxygen species. The concentration of tumor necrosis factor alpha was measured by enzyme immunoassay. The plots of dependence of the light sums of luminol- and lucigenin-dependent chemiluminescence on the concentration of each ligand were shaped as saturation curves. The comparison of the light sums of lucigenin-dependent chemiluminescence (the production of superoxide anion radical) and luminol-dependent chemiluminescence (the total production of reactive oxygen species) showed that the contribution of NADPH oxidase to the total TLR-mediated production of oxidants can reach 40–50%. Stimulation indices were calculated to compare the ability of TLR ligands to stimulate the production of reactive oxygen species and tumor necrosis factor alpha by neutrophils. It has been established that the activation of neutrophils with zymosan leads to higher (more than 8-fold) production of reactive oxygen species rather than production of tumor necrosis factor alpha. Unlike zymosan, lipopolysaccharide stimulated the production of tumor necrosis factor alpha to a greater extent (by more than 2 times) than the production of reactive oxygen species. Peptidoglycan takes an intermediate position between these ligands. Thus, the production of effector molecules (reactive oxygen species and tumor necrosis factor alpha) by human peripheral blood neutrophils depends on the nature of the TRL ligand.

Sobre autores

Yu. Teselkin

Pirogov Russian National Research Medical University

Autor responsável pela correspondência
Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

M. Khoreva

Pirogov Russian National Research Medical University

Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

A. Veselova

Pirogov Russian National Research Medical University

Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

I. Babenkova

Pirogov Russian National Research Medical University

Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

A. Osipov

Pirogov Russian National Research Medical University

Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

L. Gankovskaya

Pirogov Russian National Research Medical University

Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

Yu. Vladimirov

Pirogov Russian National Research Medical University

Email: teselkin-box@mail.ru
Rússia, Moscow, 117997

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