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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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Related in: MedlinePlus

Expression of TLR-4 receptor in purified splenic macrophages and lymphocytes after in vitro stimulation with LPS. The results in (a) and (c) show the differential translation of the TLR-4 mRNA to the protein in the splenic macrophages and lymphocytes, respectively, after in vitro stimulation with LPS from a set of triplicate experiments. Lane 1 (Mφ Con) represents TLR-4 on untreated macrophages, lane 2 (Mφ + L60 m) represents TLR-4 expression on the macrophages treated with LPS for 60 minutes, lane 3 (Mφ + L90 m) represents TLR-4 expression on macrophages treated with LPS for 90 minutes, and lane 4 (Mφ + 120 m) represents TLR-4 expression on macrophages treated with LPS for 120 minutes. Lane 1 (Lym Con) represents TLR-4 on untreated lymphocyte, lane 2 (Lym + L60 m) represents TLR-4 expression on the lymphocyte treated with LPS for 60 minutes, lane 3 (Lym + L90 m) represents TLR-4 expression on lymphocyte treated with LPS for 90 minutes, and lane 4 (Lym + L120 m) represents TLR-4 expression on lymphocyte treated with LPS for 120 minutes. The following symbols represent the significant fold change in TLR-4 expression between: *control versus LPS treated cells, #LPS treatment for 60 min versus LPS treatment for 90 and 120 min, %LPS treatment for 90 min versus LPS treatment for 120 min. Significant change (P < 0.05).
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fig3: Expression of TLR-4 receptor in purified splenic macrophages and lymphocytes after in vitro stimulation with LPS. The results in (a) and (c) show the differential translation of the TLR-4 mRNA to the protein in the splenic macrophages and lymphocytes, respectively, after in vitro stimulation with LPS from a set of triplicate experiments. Lane 1 (Mφ Con) represents TLR-4 on untreated macrophages, lane 2 (Mφ + L60 m) represents TLR-4 expression on the macrophages treated with LPS for 60 minutes, lane 3 (Mφ + L90 m) represents TLR-4 expression on macrophages treated with LPS for 90 minutes, and lane 4 (Mφ + 120 m) represents TLR-4 expression on macrophages treated with LPS for 120 minutes. Lane 1 (Lym Con) represents TLR-4 on untreated lymphocyte, lane 2 (Lym + L60 m) represents TLR-4 expression on the lymphocyte treated with LPS for 60 minutes, lane 3 (Lym + L90 m) represents TLR-4 expression on lymphocyte treated with LPS for 90 minutes, and lane 4 (Lym + L120 m) represents TLR-4 expression on lymphocyte treated with LPS for 120 minutes. The following symbols represent the significant fold change in TLR-4 expression between: *control versus LPS treated cells, #LPS treatment for 60 min versus LPS treatment for 90 and 120 min, %LPS treatment for 90 min versus LPS treatment for 120 min. Significant change (P < 0.05).

Mentions: Expression of TLR-4 on splenic macrophages and lymphocytes was observed after LPS treatment in vitro for a short time interval of 60, 90, and 120 minutes. The expression on splenic macrophages increased markedly after 60 minutes of LPS treatment and peaked at 90 minutes as compared to untreated cells, but then a decrease in expression was observed after 30 more minutes (Figure 3(a)). In case splenic lymphocytes the expression was only seen to rise noticeably at 120 minutes of LPS treatment (Figure 3(c)). The change in their expression is represented in Figures 3(b) and 3(d), respectively. Expression of TLR-4 on thymic lymphocytes could not be detected on LPS treatment as the total lymphocytes isolated from untreated thymic was quantitatively not sufficient to carry out our in vitro model of study.


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)

Expression of TLR-4 receptor in purified splenic macrophages and lymphocytes after in vitro stimulation with LPS. The results in (a) and (c) show the differential translation of the TLR-4 mRNA to the protein in the splenic macrophages and lymphocytes, respectively, after in vitro stimulation with LPS from a set of triplicate experiments. Lane 1 (Mφ Con) represents TLR-4 on untreated macrophages, lane 2 (Mφ + L60 m) represents TLR-4 expression on the macrophages treated with LPS for 60 minutes, lane 3 (Mφ + L90 m) represents TLR-4 expression on macrophages treated with LPS for 90 minutes, and lane 4 (Mφ + 120 m) represents TLR-4 expression on macrophages treated with LPS for 120 minutes. Lane 1 (Lym Con) represents TLR-4 on untreated lymphocyte, lane 2 (Lym + L60 m) represents TLR-4 expression on the lymphocyte treated with LPS for 60 minutes, lane 3 (Lym + L90 m) represents TLR-4 expression on lymphocyte treated with LPS for 90 minutes, and lane 4 (Lym + L120 m) represents TLR-4 expression on lymphocyte treated with LPS for 120 minutes. The following symbols represent the significant fold change in TLR-4 expression between: *control versus LPS treated cells, #LPS treatment for 60 min versus LPS treatment for 90 and 120 min, %LPS treatment for 90 min versus LPS treatment for 120 min. Significant change (P < 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: Expression of TLR-4 receptor in purified splenic macrophages and lymphocytes after in vitro stimulation with LPS. The results in (a) and (c) show the differential translation of the TLR-4 mRNA to the protein in the splenic macrophages and lymphocytes, respectively, after in vitro stimulation with LPS from a set of triplicate experiments. Lane 1 (Mφ Con) represents TLR-4 on untreated macrophages, lane 2 (Mφ + L60 m) represents TLR-4 expression on the macrophages treated with LPS for 60 minutes, lane 3 (Mφ + L90 m) represents TLR-4 expression on macrophages treated with LPS for 90 minutes, and lane 4 (Mφ + 120 m) represents TLR-4 expression on macrophages treated with LPS for 120 minutes. Lane 1 (Lym Con) represents TLR-4 on untreated lymphocyte, lane 2 (Lym + L60 m) represents TLR-4 expression on the lymphocyte treated with LPS for 60 minutes, lane 3 (Lym + L90 m) represents TLR-4 expression on lymphocyte treated with LPS for 90 minutes, and lane 4 (Lym + L120 m) represents TLR-4 expression on lymphocyte treated with LPS for 120 minutes. The following symbols represent the significant fold change in TLR-4 expression between: *control versus LPS treated cells, #LPS treatment for 60 min versus LPS treatment for 90 and 120 min, %LPS treatment for 90 min versus LPS treatment for 120 min. Significant change (P < 0.05).
Mentions: Expression of TLR-4 on splenic macrophages and lymphocytes was observed after LPS treatment in vitro for a short time interval of 60, 90, and 120 minutes. The expression on splenic macrophages increased markedly after 60 minutes of LPS treatment and peaked at 90 minutes as compared to untreated cells, but then a decrease in expression was observed after 30 more minutes (Figure 3(a)). In case splenic lymphocytes the expression was only seen to rise noticeably at 120 minutes of LPS treatment (Figure 3(c)). The change in their expression is represented in Figures 3(b) and 3(d), respectively. Expression of TLR-4 on thymic lymphocytes could not be detected on LPS treatment as the total lymphocytes isolated from untreated thymic was quantitatively not sufficient to carry out our in vitro model of study.

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