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Characterization of Toll-like receptor-4 (TLR-4) in the spleen and thymus of Swiss albino mice and its modulation in experimental endotoxemia.

Ghosh C, Bishayi B - J Immunol Res (2015)

Bottom Line: The diverse pattern of TLR-4 expression on different cell population indicated their distinct functional activity in LPS-endotoxemia.It may be hypothesized that the expression patterns of TLR-4 could be different based on the anatomical localization and the varying bacterial milieu or bacterial endotoxin encountered in each anatomical location.Thus, blocking TLR-4 or administering IL-6 or IL-10 might impart protection against endotoxemia in the clinical field.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Immunology Laboratory, University of Calcutta, University Colleges of Science and Technology, 92 APC Road, Kolkata, West Bengal 700009, India.

ABSTRACT
Expression of innate immune receptors varies among organs and species and within different strains among the same species; thus, periodic classification of different pattern recognition receptors in the available strains is necessary to initiate different therapeutic approaches to combat inflammation. On characterization of TLR-4 in spleen and thymus of Swiss albino mice--with no reports of TLR-4 expression--induced with endotoxemia, it was found that the mode of expression varied among the organs at both mRNA and protein level in a time-dependent manner. Their functionality was verified by measuring proinflammatory and anti-inflammatory cytokines. In the in vitro study using isolated macrophages and lymphocytes from the same organs, the expression of TLR-4 after a shorter period of LPS stimulation was verified. The results substantiated the potent role of macrophage on LPS challenge compared to lymphocytes. The diverse pattern of TLR-4 expression on different cell population indicated their distinct functional activity in LPS-endotoxemia. It may be hypothesized that the expression patterns of TLR-4 could be different based on the anatomical localization and the varying bacterial milieu or bacterial endotoxin encountered in each anatomical location. Thus, blocking TLR-4 or administering IL-6 or IL-10 might impart protection against endotoxemia in the clinical field.

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Serum cytokine levels in Swiss albino mice after experimental endotoxemia. Serum levels of (a) TNF-α; (b) IL-6; (c) IFN-γ; and (d) IL-10 in control and LPS treated mice at 3, 6, 12, 24, 48, and 72 hours after treatment showed time-dependent variation with the onset of endotoxemia (values are expressed as Mean ± SD from triplicate experiments and are significant (P < 0.05) from 6 mice in each group). The following symbols represent the significant difference between: *control versus LPS treated serum, #LPS treatment for 3 hours versus LPS treatment for 6, 12, 24, 48, and 72 hours, %LPS treatment for 6 hours versus LPS treatment for 12, 24, 48, and 72 hours, $LPS treatment for 12 hours versus LPS treatment for 24, 48, and 72 hours.
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fig5: Serum cytokine levels in Swiss albino mice after experimental endotoxemia. Serum levels of (a) TNF-α; (b) IL-6; (c) IFN-γ; and (d) IL-10 in control and LPS treated mice at 3, 6, 12, 24, 48, and 72 hours after treatment showed time-dependent variation with the onset of endotoxemia (values are expressed as Mean ± SD from triplicate experiments and are significant (P < 0.05) from 6 mice in each group). The following symbols represent the significant difference between: *control versus LPS treated serum, #LPS treatment for 3 hours versus LPS treatment for 6, 12, 24, 48, and 72 hours, %LPS treatment for 6 hours versus LPS treatment for 12, 24, 48, and 72 hours, $LPS treatment for 12 hours versus LPS treatment for 24, 48, and 72 hours.

Mentions: Quantitative analysis of some proinflammatory and anti-inflammatory serum cytokine was done to attest the functionality of TLR-4 and to examine its regulatory role in inflammation in our in vivo model of endotoxin induced inflammation. We examined proinflammatory cytokines like TNF-α, IFN-γ, and IL-6. Serum IL-10 level was measured, being the most important and widely studied anti-inflammatory cytokine so far. The analysis was done at 3, 6, 12, 24, 48, and 72 hours after LPS challenge. The serum level of TNF-α increased with time from the earlier hours of LPS treatment and peaked at 24 hours and then gradually started decreased in the next period of study (Figure 5(a)). The serum level of IL-6 was found to rise slowly after treatment and it peaked at 24 hours but sustained at that level in the later hours of the study (Figure 5(b)). The secretion of IFN-γ was irregular initially, raised at 3 hours post treatment and then returned to the basal level at 6 and 12 hours. Again increased significantly at 24 hours compared to normal and the earlier hours of treatment but started to return to normal in the 48th hour of our study and returned to normal at 72nd hour (Figure 5(c)). The serum level of anti-inflammatory cytokine IL-10 started rising in the late hours of LPS treatment and kept on increasing even at 72 hours of treatment (Figure 5(d)).


Characterization of Toll-like receptor-4 (TLR-4) in the spleen and thymus of Swiss albino mice and its modulation in experimental endotoxemia.

Ghosh C, Bishayi B - J Immunol Res (2015)

Serum cytokine levels in Swiss albino mice after experimental endotoxemia. Serum levels of (a) TNF-α; (b) IL-6; (c) IFN-γ; and (d) IL-10 in control and LPS treated mice at 3, 6, 12, 24, 48, and 72 hours after treatment showed time-dependent variation with the onset of endotoxemia (values are expressed as Mean ± SD from triplicate experiments and are significant (P < 0.05) from 6 mice in each group). The following symbols represent the significant difference between: *control versus LPS treated serum, #LPS treatment for 3 hours versus LPS treatment for 6, 12, 24, 48, and 72 hours, %LPS treatment for 6 hours versus LPS treatment for 12, 24, 48, and 72 hours, $LPS treatment for 12 hours versus LPS treatment for 24, 48, and 72 hours.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4352500&req=5

fig5: Serum cytokine levels in Swiss albino mice after experimental endotoxemia. Serum levels of (a) TNF-α; (b) IL-6; (c) IFN-γ; and (d) IL-10 in control and LPS treated mice at 3, 6, 12, 24, 48, and 72 hours after treatment showed time-dependent variation with the onset of endotoxemia (values are expressed as Mean ± SD from triplicate experiments and are significant (P < 0.05) from 6 mice in each group). The following symbols represent the significant difference between: *control versus LPS treated serum, #LPS treatment for 3 hours versus LPS treatment for 6, 12, 24, 48, and 72 hours, %LPS treatment for 6 hours versus LPS treatment for 12, 24, 48, and 72 hours, $LPS treatment for 12 hours versus LPS treatment for 24, 48, and 72 hours.
Mentions: Quantitative analysis of some proinflammatory and anti-inflammatory serum cytokine was done to attest the functionality of TLR-4 and to examine its regulatory role in inflammation in our in vivo model of endotoxin induced inflammation. We examined proinflammatory cytokines like TNF-α, IFN-γ, and IL-6. Serum IL-10 level was measured, being the most important and widely studied anti-inflammatory cytokine so far. The analysis was done at 3, 6, 12, 24, 48, and 72 hours after LPS challenge. The serum level of TNF-α increased with time from the earlier hours of LPS treatment and peaked at 24 hours and then gradually started decreased in the next period of study (Figure 5(a)). The serum level of IL-6 was found to rise slowly after treatment and it peaked at 24 hours but sustained at that level in the later hours of the study (Figure 5(b)). The secretion of IFN-γ was irregular initially, raised at 3 hours post treatment and then returned to the basal level at 6 and 12 hours. Again increased significantly at 24 hours compared to normal and the earlier hours of treatment but started to return to normal in the 48th hour of our study and returned to normal at 72nd hour (Figure 5(c)). The serum level of anti-inflammatory cytokine IL-10 started rising in the late hours of LPS treatment and kept on increasing even at 72 hours of treatment (Figure 5(d)).

Bottom Line: The diverse pattern of TLR-4 expression on different cell population indicated their distinct functional activity in LPS-endotoxemia.It may be hypothesized that the expression patterns of TLR-4 could be different based on the anatomical localization and the varying bacterial milieu or bacterial endotoxin encountered in each anatomical location.Thus, blocking TLR-4 or administering IL-6 or IL-10 might impart protection against endotoxemia in the clinical field.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Immunology Laboratory, University of Calcutta, University Colleges of Science and Technology, 92 APC Road, Kolkata, West Bengal 700009, India.

ABSTRACT
Expression of innate immune receptors varies among organs and species and within different strains among the same species; thus, periodic classification of different pattern recognition receptors in the available strains is necessary to initiate different therapeutic approaches to combat inflammation. On characterization of TLR-4 in spleen and thymus of Swiss albino mice--with no reports of TLR-4 expression--induced with endotoxemia, it was found that the mode of expression varied among the organs at both mRNA and protein level in a time-dependent manner. Their functionality was verified by measuring proinflammatory and anti-inflammatory cytokines. In the in vitro study using isolated macrophages and lymphocytes from the same organs, the expression of TLR-4 after a shorter period of LPS stimulation was verified. The results substantiated the potent role of macrophage on LPS challenge compared to lymphocytes. The diverse pattern of TLR-4 expression on different cell population indicated their distinct functional activity in LPS-endotoxemia. It may be hypothesized that the expression patterns of TLR-4 could be different based on the anatomical localization and the varying bacterial milieu or bacterial endotoxin encountered in each anatomical location. Thus, blocking TLR-4 or administering IL-6 or IL-10 might impart protection against endotoxemia in the clinical field.

Show MeSH
Related in: MedlinePlus