Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself
<p>Abstract</p> <p>Background</p> <p>Toll-like receptor 4 (TLR4) is activated by bacterial endotoxin, a prototypical pathogen-associated molecular pattern (PAMP). It has been suggested that TLR4 can also be activated by damage-associated molecular pattern (DAMP) protein...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-03-01
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| Series: | Journal of Inflammation |
| Subjects: | |
| Online Access: | http://www.journal-inflammation.com/content/9/1/11 |
| _version_ | 1811250901781839872 |
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| author | Luong Michael Zhang Yanyu Chamberlain Tim Zhou Tianhui Wright Jill F Dower Ken Hall J Perry |
| author_facet | Luong Michael Zhang Yanyu Chamberlain Tim Zhou Tianhui Wright Jill F Dower Ken Hall J Perry |
| author_sort | Luong Michael |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Toll-like receptor 4 (TLR4) is activated by bacterial endotoxin, a prototypical pathogen-associated molecular pattern (PAMP). It has been suggested that TLR4 can also be activated by damage-associated molecular pattern (DAMP) proteins such as HSP70. It remains a challenge to provide unequivocal evidence that DAMP proteins themselves play a role in TLR4 activation, as the DAMP proteins used are often contaminated with endotoxin and other TLR ligands introduced during protein expression and/or purification.</p> <p>Results</p> <p>Here we report that the activation of TLR4 on primary human macrophage cultures by recombinant HSP70 is not solely due to contaminating endotoxin. Polymyxin B pretreatment of HSP70 preparations to neutralize contaminating endotoxin caused significant reductions in the amount of TNF-α induced by the recombinant protein as determined by ELISA. However, digestion of HSP70 with Proteinase K-agarose beads also dramatically reduced the TNF-α response of macrophages to HSP70, while leaving levels of contaminating endotoxin largely unchanged relative to controls.</p> <p>Conclusions</p> <p>These results indicate that the stimulatory effect of recombinant HSP70 requires both the presence of endotoxin and structural integrity of the heat shock protein itself.</p> |
| first_indexed | 2024-04-12T16:11:41Z |
| format | Article |
| id | doaj.art-4719907bb5d7423ab8493c67a80b9dab |
| institution | Directory Open Access Journal |
| issn | 1476-9255 |
| language | English |
| last_indexed | 2024-04-12T16:11:41Z |
| publishDate | 2012-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Inflammation |
| spelling | doaj.art-4719907bb5d7423ab8493c67a80b9dab2022-12-22T03:25:52ZengBMCJournal of Inflammation1476-92552012-03-01911110.1186/1476-9255-9-11Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itselfLuong MichaelZhang YanyuChamberlain TimZhou TianhuiWright Jill FDower KenHall J Perry<p>Abstract</p> <p>Background</p> <p>Toll-like receptor 4 (TLR4) is activated by bacterial endotoxin, a prototypical pathogen-associated molecular pattern (PAMP). It has been suggested that TLR4 can also be activated by damage-associated molecular pattern (DAMP) proteins such as HSP70. It remains a challenge to provide unequivocal evidence that DAMP proteins themselves play a role in TLR4 activation, as the DAMP proteins used are often contaminated with endotoxin and other TLR ligands introduced during protein expression and/or purification.</p> <p>Results</p> <p>Here we report that the activation of TLR4 on primary human macrophage cultures by recombinant HSP70 is not solely due to contaminating endotoxin. Polymyxin B pretreatment of HSP70 preparations to neutralize contaminating endotoxin caused significant reductions in the amount of TNF-α induced by the recombinant protein as determined by ELISA. However, digestion of HSP70 with Proteinase K-agarose beads also dramatically reduced the TNF-α response of macrophages to HSP70, while leaving levels of contaminating endotoxin largely unchanged relative to controls.</p> <p>Conclusions</p> <p>These results indicate that the stimulatory effect of recombinant HSP70 requires both the presence of endotoxin and structural integrity of the heat shock protein itself.</p>http://www.journal-inflammation.com/content/9/1/11EndotoxinHeat shock protein 70Polymyxin BProteinase KTLR4 |
| spellingShingle | Luong Michael Zhang Yanyu Chamberlain Tim Zhou Tianhui Wright Jill F Dower Ken Hall J Perry Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself Journal of Inflammation Endotoxin Heat shock protein 70 Polymyxin B Proteinase K TLR4 |
| title | Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself |
| title_full | Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself |
| title_fullStr | Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself |
| title_full_unstemmed | Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself |
| title_short | Stimulation of TLR4 by recombinant HSP70 requires structural integrity of the HSP70 protein itself |
| title_sort | stimulation of tlr4 by recombinant hsp70 requires structural integrity of the hsp70 protein itself |
| topic | Endotoxin Heat shock protein 70 Polymyxin B Proteinase K TLR4 |
| url | http://www.journal-inflammation.com/content/9/1/11 |
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