技術(shù)天地
LPS那么多，你選對了嗎？
LPS是革蘭氏陰性細菌的主要成分，可被Toll樣受體4（TLR4）識別以激活先天免疫系統。通過(guò)信號級聯(lián)反應，導致NF-κB的活化和促炎性細胞因子的產(chǎn)生，因此LPS可以應用到免疫細胞的刺激、激活、分化等實(shí)驗中。但是LPS種類(lèi)好多，而且并不是所有的LPS都能夠激活TLR4受體，LPS-RS作為TLR4的拮抗劑，可以競爭性抑制LPS的活性。
在選訂LPS時(shí)，您是否有不知道選擇哪一種的困惑？在實(shí)驗效果不好的時(shí)候，您是否有懷疑過(guò)是不是使用的LPS不正確？希望以下的內容，能夠在您選擇產(chǎn)品時(shí)為您提供幫助。
圖1. 不同TLR4受體激動(dòng)劑的結構
不同的細菌種類(lèi)表達不同的LPS，其生物活性也有所不同。在某些細菌中，可能不存在O-抗原，表達粗糙型LPS。與光滑型LPS的典型表型相反，O-抗原的缺乏可調節免疫系統細胞識別LPS的過(guò)程。
|
Smooth (S)-form LPS |
Rough (R)-form LPS |
|
TLR4 agonist LPS-B5 from E. coli 055:B5 LPS-EB from E. coli 0111:B4 LPS-PG from P. gingivalis |
TLR4 agonist LPS-EK from E. coli K12 LPS-SM from Salmonella minnesota R595 TLR4 antagonist à LPS-RS from Rhodobacter sphaeroides |
LPS選擇指南
LPS-EB from E. coli 0111:B4
特點(diǎn)：
- E. coli 0111:B4菌株是已知可引起顯著(zhù)胃部疾病的致病性大腸埃希菌血清型
-文獻中引用最多的LPS菌株
-在體外和體內都可以發(fā)現廣泛的應用，包括巨噬細胞分化
應用實(shí)例：
-在體外誘導人巨噬細胞(M1)極化1
-刺激小鼠巨噬細胞表達siglec2
-誘導大西洋鱈魚(yú)和鮭魚(yú)單核細胞的炎癥反應3
-在小鼠模型中誘導膿毒性休克和急性肝衰竭4
-腹腔注射LPS治療小鼠低鐵血癥研究5
-誘導諾如病毒感染后人腸上皮細胞的形態(tài)學(xué)變化6
相關(guān)應用文獻
1.Vogel, D. Y. et al., 2014. Human macrophage polarization in vitro: maturation and activation methods compared. Immunobiology, 219(9), 695-703.
2. Nagala, M. et al., 2018. Expression of Siglec-E alters the proteome of lipopolysaccharide (LPS)-activated macrophages but does not affect LPS-driven cytokine production or toll-like receptor 4 endocytosis. Frontiers in immunology, 8, 1926.
3. Seppola, M. et al., 2015. Ultrapure LPS induces inflammatory and antibacterial responses attenuated in vitro by exogenous sera in Atlantic cod and Atlantic salmon. Fish & shellfish immunology, 44(1), 66-78.
4. Rialdi, A. et al., 2016. Topoisomerase 1 inhibition suppresses inflammatory genes and protects from death by inflammation. Science, 352(6289).
5. Guida, C. et al., 2015. A novel inflammatory pathway mediating rapid hepcidin-independent hypoferremia. Blood, The Journal of the American Society of Hematology, 125(14), 2265-2275.
6. Herbst-Kralovetz, M. M. et al., 2013. Lack of norovirus replication and histo-blood group antigen expression in 3-dimensional intestinal epithelial cells. Emerging infectious diseases, 19(3), 431.
LPS-EK from E. coli K12
特點(diǎn)：
-E. coli K12菌株為典型實(shí)驗室菌株
-具有結構異質(zhì)性
-一種主要用于體外試驗的強效TLR4激動(dòng)劑
應用實(shí)例：
-刺激骨髓源性巨噬細胞和人外周血單核細胞產(chǎn)生促炎細胞因子7
-誘導小鼠脾細胞的B細胞增殖8
-刺激小鼠造血和氣道上皮細胞用于哮喘研究9
-用于HIV感染后刺激樹(shù)突狀細胞和CD4+T細胞10
-誘導COVID-19患者PBMC產(chǎn)生IL-6、TNF-α和CCL211
-用于啟動(dòng)THP-1細胞以促進(jìn)炎癥小體活化12
相關(guān)應用文獻：
7. Saz-Leal, P. et al., 2018. Targeting SHIP-1 in myeloid cells enhances trained immunity and boosts response to infection. Cell reports, 25(5), 1118-1126.
8. Kang, S. S. et al., 2018. Staphylococcal LTA antagonizes the B cell-mitogenic potential of LPS. Scientific reports, 8(1), 1-12.
9. McAlees, J. W. et al., 2015. Distinct Tlr4-expressing cell compartments control neutrophilic and eosinophilic airway inflammation. Mucosal immunology, 8(4), 863-873.
10. Rosa, A. et al., 2015. HIV-1 Nef promotes infection by excluding SERINC5 from virion incorporation. Nature, 526(7572), 212-217.
11. Carvelli, J. et al., 2020. Association of COVID-19 inflammation with activation of the C5a–C5aR1 axis. Nature, 1-9.
12. Kroes, M. M. et al., 2019. Activation of human NK cells by Bordetella pertussis requires inflammasome activation in macrophages. Frontiers in Immunology, 10, 2030.
LPS-B5 from E. coli 055:B5
特點(diǎn)：
- E. coli 055：B5菌株是LAL試驗中經(jīng)常用作內毒素標準品的原型內毒素
-高致熱原性
-常用于體外細胞活化
應用實(shí)例：
-在小鼠巨噬細胞中產(chǎn)生促炎性細胞因子分泌13
-誘導小鼠脾臟B細胞分泌IL-614
-誘導大鼠骨髓源性MSCs4中IL-6和生長(cháng)因子的表達15
-誘導硬骨魚(yú)巨噬細胞中TNFα和IL-1β的表達16
-研究豬瘟病毒感染后豬組織中TLR4的mRNA表達17
相關(guān)應用文獻
13.Krukenberg, K. et al., 2015. Extracellular poly (ADP-ribose) is a pro-inflammatory signal for macrophages. Chemistry & biology, 22(4), 446-452.
14. Kiripolsky, J. et al., 2020. Activation of Myd88-Dependent TLRs Mediates Local and Systemic Inflammation in a Mouse Model of Primary Sj?gren's Syndrome. Frontiers in Immunology, 10, 2963.
15. Evaristo-Mendon?a, F. et al., 2019. Preconditioning of Rat Bone Marrow-Derived Mesenchymal Stromal Cells with Toll-Like Receptor Agonists. Stem cells international, 2019.
16. Bi, D. et al., 2018. Recognition of lipopolysaccharide and activation of NF-κB by cytosolic sensor NOD1 in teleost fish. Frontiers in immunology, 9, 1413.
17. Cao, Z. et al., 2018. Tissue expression of Toll-like receptors 2, 3, 4 and 7 in swine in response to the Shimen strain of classical swine fever virus. Molecular Medicine Reports, 17(5), 7122-7130
LPS-PG from Porphyromonas gingivalis
特點(diǎn)：
-牙周病機制中的一個(gè)重要毒力因子
-具有獨特的異質(zhì)化學(xué)結構，不同于傳統公認的腸道細菌來(lái)源的LPS
-通常與牙周炎相關(guān)研究相關(guān)的應用
應用實(shí)例：
-在小鼠和大鼠牙周炎模型中引起炎癥反應18.19
-研究牙齦卟啉單胞菌-誘導類(lèi)風(fēng)濕性關(guān)節炎患者樹(shù)突狀細胞的細胞因子應答20
-誘導人牙周膜(PDL)細胞中NF-κB活化21
-用于刺激人牙髓細胞(HDPCs)以研究mRNA表達22
-用于刺激破骨細胞前體細胞以研究破骨細胞基因調控23
相關(guān)應用文獻：
18. Park, J. et al., 2017. Fucoidan inhibits LPS-induced inflammation in vitro and during the acute response in vivo. International immunopharmacology, 43, 91-98.
19. Chang, P. C. et al., 2013. Irradiation by light‐emitting diode light as an adjunct to facilitate healing of experimental periodontitis in vivo. Journal of periodontal research, 48(2), 135-143.
20. Santegoets, K. C. et al., 2016. Impaired Porphyromonas gingivalis–induced tumor necrosis factor production by dendritic cells typifies patients with rheumatoid arthritis. Arthritis & Rheumatology, 68(4), 795-804.
21. G?lz, L. et al., 2015. Hypoxia and P. gingivalis synergistically induce HIF-1 and NF-κB activation in PDL cells and periodontal diseases. Mediators of inflammation, 2015.
22. Feng, Z. et al., 2018. METTL 3 regulates alternative splicing of MyD88 upon the lipopolysaccharide‐induced inflammatory response in human dental pulp cells. Journal of cellular and molecular medicine, 22(5), 2558-2568.
23. Chen, Z. et al., 2015. IL-1R/TLR2 through MyD88 divergently modulates osteoclastogenesis through regulation of nuclear factor of activated T cells c1 (NFATc1) and B lymphocyte-induced maturation protein-1 (Blimp1). Journal of Biological Chemistry, 290(50), 30163-30174.
LPS-SM from Salmonella minnesota R595
特點(diǎn)：
-僅由脂質(zhì)A和3-脫氧-D-甘露-辛酮糖酸組成
-通常用于體外細胞活化
應用實(shí)例：
-刺激肉雞血小板中促炎性細胞因子的產(chǎn)生24
-誘導PBMC分泌IL-6、TNF-α和IL-825
-刺激腹腔巨噬細胞產(chǎn)生IL-1β26
-啟動(dòng)小鼠骨髓來(lái)源的樹(shù)突狀細胞產(chǎn)生IL-1β前體和Caspase1前體27
-誘導大西洋鱈魚(yú)和鮭魚(yú)單核細胞的炎癥反應3
相關(guān)應用文獻
24. Ferdous, F. et al., 2008. Broiler chick thrombocyte response to lipopolysaccharide. Poultry science, 87(1), 61-63.
25. Berthet, J. et al., 2012. Human platelets can discriminate between various bacterial LPS isoforms via TLR4 signaling and differential cytokine secretion. Clinical Immunology, 145(3), 189-200.
26. Brykczynska, U. et al., 2020. Distinct Transcriptional Responses across Tissue-Resident Macrophages to Short-Term and Long-Term Metabolic Challenge. Cell Reports, 30(5), 1627-1643.
27. Ravindran, R. et al., 2016. The amino acid sensor GCN2 controls gut inflammation by inhibiting inflammasome activation. Nature, 531(7595), 523-527.
LPS-RS from Rhodobacter sphaeroides (TLR4 拮抗劑)
特點(diǎn)：
-LPS-RS Standard 是TLR2的強效激動(dòng)劑，而LPS-RS Ultrapure特異性拮抗TLR4
-在拮抗劑過(guò)量100倍時(shí)，可能會(huì )完全競爭性抑制LPS活性
-可通過(guò)LAL測定法檢測
應用實(shí)例：
-抑制人子宮內膜內皮細胞(HEECs)中TLR-4依賴(lài)性細胞因子分泌28
-抑制中耳膽脂瘤干細胞(ME-CSCs)中TLR4依賴(lài)性炎癥29
-研究LPS誘導的大鼠急性肺損傷的減輕30
-研究小鼠血管痙攣和神經(jīng)功能損傷的減輕31
-抑制TLR4誘導的CD14內吞作用32
相關(guān)應用文獻
28. Krikun, G. et al., 2012. Lipopolysaccharide appears to activate human endometrial endothelial cells through TLR‐4‐dependent and TLR‐4‐independent mechanisms. American journal of reproductive immunology, 68(3), 233-237.
29. Schürmann, M. et al., 2020. Stem Cell-Induced Inflammation in Cholesteatoma Is Inhibited by the TLR4 Antagonist LPS-RS. Cells, 9(1), 199.
30. Abdelmageed, M. E. et al., 2016. LPS-RS attenuation of lipopolysaccharide-induced acute lung injury involves NF-κB inhibition. Canadian Journal of Physiology and Pharmacology, 94(2), 140-146.
31. Kawakita, F. et al., 2017. Effects of Toll-like receptor 4 antagonists against cerebral vasospasm after experimental subarachnoid hemorrhage in mice. Molecular neurobiology, 54(8), 6624-6633.
32. Tan, Y. et al., 2015. Mechanisms of Toll-like receptor 4 endocytosis reveal a common immune-evasion strategy used by pathogenic and commensal bacteria. Immunity, 43(5), 909-922.
InvivoGen提供兩種純度的高品質(zhì)LPS
市售脂多糖(LPS)的純度可能存在顯著(zhù)差異。一些混合物中含有額外的細菌成分，例如肽聚糖或脂肽，能夠激活TLR2和TLR4受體。InvivoGen提供兩種等級的LPS：通過(guò)苯酚提取制備的Standard 級別LPS，能夠激活TLR2和TLR4，而連續酶處理的Ultrapure 級別的LPS不含脂蛋白，僅激活TLR4。InvivoGen是特異性激活TLR4的Ultrapure LPS的唯一提供者。
圖2.激活TLR2受體活性對比。使用不同濃度的其他品牌的LPS-EB和LPS-B5，InvivoGen的Ultrapure的LPS-EB和LPS-B5以及Pam3CSK4刺激HEK-Blue?hTLR2細胞。孵育后，通過(guò)讀取655 nm的OD值來(lái)確定NF-κB誘導的SEAP活性。
圖3.激活TLR4受體活性對比。用其他品牌的LPS-EB和InvivoGen的LPS-EB Ultrapure以不同濃度刺激HEK-Blue?hTLR4細胞。孵育后，通過(guò)讀取655 nm的OD值來(lái)確定NF-κB誘導的SEAP活性。
其實(shí)TLR4的激動(dòng)劑不只有LPS呦，InvivoGen還提供天然的和合成的LPS類(lèi)似物。后續小編還會(huì )繼續分享其他TLR4受體的選擇指南，歡迎關(guān)注。
InvivoGen 1977年成立于法國，是法國第一家生物科技公司。公司成立的初期階段主要提供細胞培養相關(guān)試劑，如抗支原體、細菌、真菌污染的抗生素以及基因篩選抗生素（Blasticidin, Puromycin, Hygromycin B, G418, Zeocin）。隨著(zhù)天然免疫研究熱點(diǎn)的興起，InvivoGen于2000年開(kāi)始致力于天然免疫相關(guān)產(chǎn)品的研發(fā)和生產(chǎn)，并成為全世界最優(yōu)質(zhì)、最專(zhuān)業(yè)的天然免疫相關(guān)產(chǎn)品生產(chǎn)商。InvivoGen為天然免疫研究提供一整套研究方案，包括不斷更新的模式識別受體（PRRs）激動(dòng)劑和拮抗劑，PRRs報告基因細胞以及天然免疫信號通路的誘導劑和抑制劑等。
全國免費熱線(xiàn)：400 819 7199 （工作日：8:30 - 17:30） Email：bj_info@dakewe.net
聯(lián)系我們
