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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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mRNA expression of TLR-4 in the spleen and thymus: the results in this figure (a) represents the differential transcription of tlr-4 gene on LPS administration in thymus and spleen of Swiss albino mice from triplicate experiments; lane 1 represents TLR-4 mRNA in thymus from control mice (CT), lane 2 thymus from LPS treated mice at 6 hours after treatment (LT6), lane 3 thymus from LPS treated mice at 12 hours after treatment (LT12), lane 4 thymus from LPS treated mice at 24 hours after treatment (LT24), lane 5 represents spleen from control mice (CS), lane 6 spleen from LPS treated mice at 6 hours after treatment (LS6), lane 7 spleen from LPS treated mice at 12 hours after treatment (LS12), and lane 8 spleen from LPS treated mice at 24 hours post treatment (LS24). (b) Is the diagrammatic representation of the fold difference in their expression compare to control and among different time periods of treatment. Molecular weight marker (100 bp). The following symbols indicates the significant change in the expression of TLR-4 between: *control versus LPS treated tissue, #LPS treatment for 6 hours versus LPS treatment for 12 and 24 hours, %LPS treatment for 12 hours versus LPS treatment 24 hours. Significant change (P < 0.05).
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fig1: mRNA expression of TLR-4 in the spleen and thymus: the results in this figure (a) represents the differential transcription of tlr-4 gene on LPS administration in thymus and spleen of Swiss albino mice from triplicate experiments; lane 1 represents TLR-4 mRNA in thymus from control mice (CT), lane 2 thymus from LPS treated mice at 6 hours after treatment (LT6), lane 3 thymus from LPS treated mice at 12 hours after treatment (LT12), lane 4 thymus from LPS treated mice at 24 hours after treatment (LT24), lane 5 represents spleen from control mice (CS), lane 6 spleen from LPS treated mice at 6 hours after treatment (LS6), lane 7 spleen from LPS treated mice at 12 hours after treatment (LS12), and lane 8 spleen from LPS treated mice at 24 hours post treatment (LS24). (b) Is the diagrammatic representation of the fold difference in their expression compare to control and among different time periods of treatment. Molecular weight marker (100 bp). The following symbols indicates the significant change in the expression of TLR-4 between: *control versus LPS treated tissue, #LPS treatment for 6 hours versus LPS treatment for 12 and 24 hours, %LPS treatment for 12 hours versus LPS treatment 24 hours. Significant change (P < 0.05).

Mentions: Expression of TLR4 mRNA was examined in fresh tissue isolates from control and LPS challenged mice at 6 hours, 12 hours, and 24 hours after treatment. In case of thymus (Figure 1(a)) the expression of TLR-4 mRNA peaked significantly at 12 hours after treatment as compared to the untreated mice as well as to 6 hours and 24 hours of LPS treatment. On the other hand in spleen the expression of TLR-4 kept on increasing significantly with time after LPS administration at peaked at 24 hours. The change in their expression in represented in (Figure 1(b)). The relevant difference in the degree of expression between thymus and spleen is suggestive to their varied cell type as well as their function in inflammation.


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)

mRNA expression of TLR-4 in the spleen and thymus: the results in this figure (a) represents the differential transcription of tlr-4 gene on LPS administration in thymus and spleen of Swiss albino mice from triplicate experiments; lane 1 represents TLR-4 mRNA in thymus from control mice (CT), lane 2 thymus from LPS treated mice at 6 hours after treatment (LT6), lane 3 thymus from LPS treated mice at 12 hours after treatment (LT12), lane 4 thymus from LPS treated mice at 24 hours after treatment (LT24), lane 5 represents spleen from control mice (CS), lane 6 spleen from LPS treated mice at 6 hours after treatment (LS6), lane 7 spleen from LPS treated mice at 12 hours after treatment (LS12), and lane 8 spleen from LPS treated mice at 24 hours post treatment (LS24). (b) Is the diagrammatic representation of the fold difference in their expression compare to control and among different time periods of treatment. Molecular weight marker (100 bp). The following symbols indicates the significant change in the expression of TLR-4 between: *control versus LPS treated tissue, #LPS treatment for 6 hours versus LPS treatment for 12 and 24 hours, %LPS treatment for 12 hours versus LPS treatment 24 hours. Significant change (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: mRNA expression of TLR-4 in the spleen and thymus: the results in this figure (a) represents the differential transcription of tlr-4 gene on LPS administration in thymus and spleen of Swiss albino mice from triplicate experiments; lane 1 represents TLR-4 mRNA in thymus from control mice (CT), lane 2 thymus from LPS treated mice at 6 hours after treatment (LT6), lane 3 thymus from LPS treated mice at 12 hours after treatment (LT12), lane 4 thymus from LPS treated mice at 24 hours after treatment (LT24), lane 5 represents spleen from control mice (CS), lane 6 spleen from LPS treated mice at 6 hours after treatment (LS6), lane 7 spleen from LPS treated mice at 12 hours after treatment (LS12), and lane 8 spleen from LPS treated mice at 24 hours post treatment (LS24). (b) Is the diagrammatic representation of the fold difference in their expression compare to control and among different time periods of treatment. Molecular weight marker (100 bp). The following symbols indicates the significant change in the expression of TLR-4 between: *control versus LPS treated tissue, #LPS treatment for 6 hours versus LPS treatment for 12 and 24 hours, %LPS treatment for 12 hours versus LPS treatment 24 hours. Significant change (P < 0.05).
Mentions: Expression of TLR4 mRNA was examined in fresh tissue isolates from control and LPS challenged mice at 6 hours, 12 hours, and 24 hours after treatment. In case of thymus (Figure 1(a)) the expression of TLR-4 mRNA peaked significantly at 12 hours after treatment as compared to the untreated mice as well as to 6 hours and 24 hours of LPS treatment. On the other hand in spleen the expression of TLR-4 kept on increasing significantly with time after LPS administration at peaked at 24 hours. The change in their expression in represented in (Figure 1(b)). The relevant difference in the degree of expression between thymus and spleen is suggestive to their varied cell type as well as their function in inflammation.

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