Lipoteichoic acid (LTA) is a Gram-positive cell surface molecule that is found in both a cell-bound form and cell-free form in the host during an infection. Hemoglobin (Hb) can synergize with LTA, a TLR2 ligand, to potently activate macrophage innate immune responses in a TLR2- and TLR4-dependent way. At low levels of LTA, the presence of Hb can result in a 200-fold increase in the secretion of IL-6 following macrophage activation. Six hours after activation, the macrophage genes that are most highly up-regulated by LTA plus Hb activation compared to LTA alone are cytokines, chemokines, receptors and interferon-regulated genes. Several of these genes exhibit a unique TLR4-dependent increase in mRNA levels that continued to rise more than eight hours after stimulation. This prolonged increase in mRNA levels could be the result of an extended period of NF-κB nuclear localization and the concurrent absence of the NF-κB inhibitor, IκBα, after stimulation with LTA plus Hb. Dynasore inhibition experiments indicate that an endocytosis-dependent pathway is required for the TLR4-dependent up-regulation of IL-6 secretion following activation with LTA plus Hb. In addition, interferon-β mRNA is present after activation with LTA plus Hb, suggesting that the TRIF/TRAM-dependent pathway may be involved. Hb alone can elicit the TLR4-dependent secretion of TNF-α from macrophages, so it may be the TLR4 ligand. Hb also led to secretion of high mobility group box 1 protein (HMGB1), which synergized with LTA to increase secretion of IL-6. The activation of both the TLR2 and TLR4 pathways by LTA plus Hb leads to an enhanced innate immune response.
References
[1]
Akira S, Takeda K, Kaisho T (2001) Toll-like receptors: critical proteins linking innate and acquired immunity. Nat Immunol 2: 675–680.
[2]
Kawai T, Akira S (2010) The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol 11: 373–384.
[3]
Underhill DM, Ozinsky A (2002) Toll-like receptors: key mediators of microbe detection. Curr Opin Immunol 14: 103–110.
[4]
Poltorak A, Smirnova I, He X, Liu MY, Van Huffel C, et al. (1998) Genetic and physical mapping of the Lps locus: identification of the toll-4 receptor as a candidate gene in the critical region. Blood Cells Mol Dis 24: 340–355.
[5]
Wright SD, Ramos RA, Tobias PS, Ulevitch RJ, Mathison JC (1990) CD14, a receptor for complexes of lipopolysaccharide (LPS) and LPS binding protein. Science 249: 1431–1433.
[6]
Nagai Y, Akashi S, Nagafuku M, Ogata M, Iwakura Y, et al. (2002) Essential role of MD-2 in LPS responsiveness and TLR4 distribution. Nat Immunol 3: 667–672.
[7]
Shimazu R, Akashi S, Ogata H, Nagai Y, Fukudome K, et al. (1999) MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med 189: 1777–1782.
[8]
Schumann RR, Leong SR, Flaggs GW, Gray PW, Wright SD, et al. (1990) Structure and function of lipopolysaccharide binding protein. Science 249: 1429–1431.
[9]
Jack RS, Fan X, Bernheiden M, Rune G, Ehlers M, et al. (1997) Lipopolysaccharide-binding protein is required to combat a murine gram-negative bacterial infection. Nature 389: 742–745.
[10]
Kirschning CJ, Schumann RR (2002) TLR2: cellular sensor for microbial and endogenous molecular patterns. Curr Top Microbiol Immunol 270: 121–144.
[11]
Hoebe K, Georgel P, Rutschmann S, Du X, Mudd S, et al. (2005) CD36 is a sensor of diacylglycerides. Nature 433: 523–527.
[12]
Husebye H, Halaas ？, Stenmark H, Tunheim G, Sandanger ？, et al. (2006) Endocytic pathways regulate Toll-like receptor 4 signaling and link innate and adaptive immunity. EMBO J 25: 683–692.
[13]
Kagan JC, Su T, Horng T, Chow A, Akira S, et al. (2008) TRAM couples endocytosis of Toll-like receptor 4 to the induction of interferon-beta. Nat Immunol 9: 361–368.
[14]
Dietrich N, Lienenklaus S, Weiss S, Gekara NO (2010) Murine toll-like receptor 2 activation induces type I interferon responses from endolysosomal compartments. PLoS One 5: e10250.
[15]
Fischer W (1988) Physiology of lipoteichoic acids in bacteria. Adv Microb Physiol 29: 233–302.
[16]
Neuhaus FC, Baddiley J (2003) A continuum of anionic charge: structures and functions of D-alanyl-teichoic acids in gram-positive bacteria. Microbiol Mol Biol Rev 67: 686–723.
[17]
Xia G, Kohler T, Peschel A (2010) The wall teichoic acid and lipoteichoic acid polymers of Staphylococcus aureus. Int J Med Microbiol 300: 148–154.
[18]
Joseph R, Shockman GD (1975) Synthesis and excretion of glycerol teichoic acid during growth of two streptococcal species. Infect Immun 12: 333–338.
[19]
Schneider O, Michel U, Zysk G, Dubuis O, Nau R (1999) Clinical outcome in pneumococcal meningitis correlates with CSF lipoteichoic acid concentrations. Neurology 53: 1584–1587.
[20]
Alkan ML, Beachey EH (1978) Excretion of lipoteichoic acid by group A streptococci. Influence of penicillin on excretion and loss of ability to adhere to human oral mucosal cells. J Clin Invest 61: 671–677.
[21]
Gerber J, Pohl K, Sander V, Bunkowski S, Nau R (2003) Rifampin followed by ceftriaxone for experimental meningitis decreases lipoteichoic acid concentrations in cerebrospinal fluid and reduces neuronal damage in comparison to ceftriaxone alone. Antimicrob Agents Chemother 47: 1313–1317.
[22]
Jin MS, Kim SE, Heo JY, Lee ME, Kim HM, et al. (2007) Crystal structure of the TLR1-TLR2 heterodimer induced by binding of a tri-acylated lipopeptide. Cell 130: 1071–1082.
[23]
Morath S, Stadelmaier A, Geyer A, Schmidt RR, Hartung T (2002) Synthetic lipoteichoic acid from Staphylococcus aureus is a potent stimulus of cytokine release. J Exp Med 195: 1635–1640.
[24]
Deininger S, Stadelmaier A, von Aulock S, Morath S, Schmidt RR, et al. (2003) Definition of structural prerequisites for lipoteichoic acid-inducible cytokine induction by synthetic derivatives. J Immunol 170: 4134–4138.
[25]
Hasty DL, Meron-Sudai S, Cox KH, Nagorna T, Ruiz-Bustos E, et al. (2006) Monocyte and macrophage activation by lipoteichoic Acid is independent of alanine and is potentiated by hemoglobin. J Immunol 176: 5567–5576.
[26]
Cox KH, Ofek I, Hasty DL (2007) Enhancement of macrophage stimulation by lipoteichoic acid and the costimulant hemoglobin is dependent on Toll-like receptors 2 and 4. Infect Immun 75: 2638–2641.
[27]
Buttari B, Profumo E, Petrone L, Pietraforte D, Siracusano A, et al. (2007) Free hemoglobin: a dangerous signal for the immune system in patients with carotid atherosclerosis? Ann N Y Acad Sci 1107: 42–50.
[28]
Mahtout H, Chandad F, Rojo JM, Grenier D (2011) Fusobacterium nucleatum binding to complement regulatory protein CD46 modulates the expression and secretion of cytokines and matrix metalloproteinases by oral epithelial cells. J Periodontol 82: 311–319.
[29]
Molloy EM, Cotter PD, Hill C, Mitchell DA, Ross RP (2011) Streptolysin S-like virulence factors: the continuing sagA. Nat Rev Microbiol 9: 670–681.
[30]
Chatellier S, Ihendyane N, Kansal RG, Khambaty F, Basma H, et al. (2000) Genetic relatedness and superantigen expression in group A streptococcus serotype M1 isolates from patients with severe and nonsevere invasive diseases. Infect Immun 68: 3523–3534.
[31]
Morath S, Geyer A, Hartung T (2001) Structure-function relationship of cytokine induction by lipoteichoic acid from Staphylococcus aureus. J Exp Med 193: 393–397.
[32]
Gao JJ, Xue Q, Zuvanich EG, Haghi KR, Morrison DC (2001) Commercial preparations of lipoteichoic acid contain endotoxin that contributes to activation of mouse macrophages in vitro. Infect Immun 69: 751–757.
[33]
Blasi E, Radzioch D, Durum SK, Varesio L (1987) A murine macrophage cell line, immortalized by v-raf and v-myc oncogenes, exhibits normal macrophage functions. Eur J Immunol 17: 1491–1498.
[34]
Zhang X, Goncalves R, Mosser DM (2008) The isolation and characterization of murine macrophages. Curr Protoc Immunol Chapter 14: Unit 14.11.
[35]
Nau GJ, Richmond JF, Schlesinger A, Jennings EG, Lander ES, et al. (2002) Human macrophage activation programs induced by bacterial pathogens. Proc Natl Acad Sci U S A 99: 1503–1508.
[36]
Korb M, Rust AG, Thorsson V, Battail C, Li B, et al. (2008) The Innate Immune Database (IIDB). BMC Immunol 9: 7.
[37]
Huang dW, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4: 44–57.
[38]
Huang dW, Sherman BT, Tan Q, Kir J, Liu D, et al. (2007) DAVID Bioinformatics Resources: expanded annotation database and novel algorithms to better extract biology from large gene lists. Nucleic Acids Res 35: W169–175.
[39]
Baeuerle PA, Baltimore D (1988) I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science 242: 540–546.
[40]
Brown K, Gerstberger S, Carlson L, Franzoso G, Siebenlist U (1995) Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation. Science 267: 1485–1488.
[41]
Traenckner EB, Pahl HL, Henkel T, Schmidt KN, Wilk S, et al. (1995) Phosphorylation of human I kappa B-alpha on serines 32 and 36 controls I kappa B-alpha proteolysis and NF-kappa B activation in response to diverse stimuli. EMBO J 14: 2876–2883.
[42]
Arenzana-Seisdedos F, Turpin P, Rodriguez M, Thomas D, Hay RT, et al. (1997) Nuclear localization of I kappa B alpha promotes active transport of NF-kappa B from the nucleus to the cytoplasm. J Cell Sci 110 (Pt 3): 369–378.
[43]
Long EM, Klimowicz AC, Paula-Neto HA, Millen B, McCafferty DM, et al. (2011) A subclass of acylated anti-inflammatory mediators usurp Toll-like receptor 2 to inhibit neutrophil recruitment through peroxisome proliferator-activated receptor gamma. Proc Natl Acad Sci U S A 108: 16357–16362.
[44]
Long EM, Millen B, Kubes P, Robbins SM (2009) Lipoteichoic acid induces unique inflammatory responses when compared to other toll-like receptor 2 ligands. PLoS One 4: e5601.
[45]
Macia E, Ehrlich M, Massol R, Boucrot E, Brunner C, et al. (2006) Dynasore, a cell-permeable inhibitor of dynamin. Dev Cell 10: 839–850.
[46]
Hirotani T, Yamamoto M, Kumagai Y, Uematsu S, Kawase I, et al. (2005) Regulation of lipopolysaccharide-inducible genes by MyD88 and Toll/IL-1 domain containing adaptor inducing IFN-beta. Biochem Biophys Res Commun 328: 383–392.
[47]
Kawai T, Adachi O, Ogawa T, Takeda K, Akira S (1999) Unresponsiveness of MyD88-deficient mice to endotoxin. Immunity 11: 115–122.
[48]
Yamamoto M, Sato S, Hemmi H, Hoshino K, Kaisho T, et al. (2003) Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301: 640–643.
[49]
Yu M, Wang H, Ding A, Golenbock DT, Latz E, et al. (2006) HMGB1 signals through toll-like receptor (TLR) 4 and TLR2. Shock 26: 174–179.
[50]
Park JS, Gamboni-Robertson F, He Q, Svetkauskaite D, Kim JY, et al. (2006) High mobility group box 1 protein interacts with multiple Toll-like receptors. Am J Physiol Cell Physiol 290: C917–924.
[51]
Kim JH, Kim SJ, Lee IS, Lee MS, Uematsu S, et al. (2009) Bacterial endotoxin induces the release of high mobility group box 1 via the IFN-beta signaling pathway. J Immunol 182: 2458–2466.
[52]
Lin T, Kwak YH, Sammy F, He P, Thundivalappil S, et al. (2010) Synergistic inflammation is induced by blood degradation products with microbial Toll-like receptor agonists and is blocked by hemopexin. J Infect Dis 202: 624–632.
[53]
Howe J, Garidel P, Roessle M, Richter W, Alexander C, et al. (2008) Structural investigations into the interaction of hemoglobin and part structures with bacterial endotoxins. Innate Immun 14: 39–49.
[54]
Fernandez PL, Dutra FF, Alves L, Figueiredo RT, Mour？o-Sa D, et al. (2010) Heme amplifies the innate immune response to microbial molecules through spleen tyrosine kinase (Syk)-dependent reactive oxygen species generation. J Biol Chem 285: 32844–32851.
[55]
Howe J, Hammer M, Alexander C, R？ssle M, Fournier K, et al. (2007) Biophysical characterization of the interaction of endotoxins with hemoglobins. Med Chem 3: 13–20.
[56]
Howe J, Richter W, Hawkins L, R？ssle M, Alexander C, et al. (2008) Hemoglobin enhances the biological activity of synthetic and natural bacterial (endotoxic) virulence factors: a general principle. Med Chem 4: 520–525.
[57]
Garg R, Qadri A (2010) Hemoglobin transforms anti-inflammatory Salmonella typhi virulence polysaccharide into a TLR-2 agonist. J Immunol 184: 5980–5987.
[58]
Van Gorp H, Delputte PL, Nauwynck HJ (2010) Scavenger receptor CD163, a Jack-of-all-trades and potential target for cell-directed therapy. Mol Immunol 47: 1650–1660.
[59]
Nielsen MJ, M？ller HJ, Moestrup SK (2010) Hemoglobin and heme scavenger receptors. Antioxid Redox Signal 12: 261–273.
[60]
Levy AP, Asleh R, Blum S, Levy NS, Miller-Lotan R, et al. (2010) Haptoglobin: basic and clinical aspects. Antioxid Redox Signal 12: 293–304.
[61]
Kaca W, Roth RI, Levin J (1994) Hemoglobin, a newly recognized lipopolysaccharide (LPS)-binding protein that enhances LPS biological activity. J Biol Chem 269: 25078–25084.
[62]
Roth RI, Levin J, Chapman KW, Schmeizl M, Rickles FR (1993) Production of modified crosslinked cell-free hemoglobin for human use: the role of quantitative determination of endotoxin contamination. Transfusion 33: 919–924.
[63]
Su D, Roth RI, Levin J (1999) Hemoglobin infusion augments the tumor necrosis factor response to bacterial endotoxin (lipopolysaccharide) in mice. Crit Care Med 27: 771–778.
[64]
Bodet C, Chandad F, Grenier D (2007) Hemoglobin and LPS act in synergy to amplify the inflammatory response. J Dent Res 86: 878–882.
[65]
Carrillo EH, Gordon LE, Richardson JD, Polk HC (2002) Free hemoglobin enhances tumor necrosis factor-alpha production in isolated human monocytes. J Trauma 52: 449–452.
[66]
Aukrust P, Halvorsen B, Yndestad A, Ueland T, ？ie E, et al. (2008) Chemokines and cardiovascular risk. Arterioscler Thromb Vasc Biol 28: 1909–1919.
[67]
Biber K, Zuurman MW, Homan H, Boddeke HW (2003) Expression of L-CCR in HEK 293 cells reveals functional responses to CCL2, CCL5, CCL7, and CCL8. J Leukoc Biol 74: 243–251.
[68]
O'Neill LA, Sheedy FJ, McCoy CE (2011) MicroRNAs: the fine-tuners of Toll-like receptor signalling. Nat Rev Immunol 11: 163–175.
[69]
Jennewein C, von Knethen A, Schmid T, Brüne B (2010) MicroRNA-27b contributes to lipopolysaccharide-mediated peroxisome proliferator-activated receptor gamma (PPARgamma) mRNA destabilization. J Biol Chem 285: 11846–11853.
[70]
Figueiredo RT, Fernandez PL, Mourao-Sa DS, Porto BN, Dutra FF, et al. (2007) Characterization of heme as activator of Toll-like receptor 4. J Biol Chem 282: 20221–20229.
[71]
Maitra U, Gan L, Chang S, Li L (2011) Low-dose endotoxin induces inflammation by selectively removing nuclear receptors and activating CCAAT/enhancer-binding protein δ. J Immunol 186: 4467–4473.
[72]
Park JS, Svetkauskaite D, He Q, Kim JY, Strassheim D, et al. (2004) Involvement of toll-like receptors 2 and 4 in cellular activation by high mobility group box 1 protein. J Biol Chem 279: 7370–7377.
[73]
Kokkola R, Andersson A, Mullins G, Ostberg T, Treutiger CJ, et al. (2005) RAGE is the major receptor for the proinflammatory activity of HMGB1 in rodent macrophages. Scand J Immunol 61: 1–9.
[74]
Hori O, Brett J, Slattery T, Cao R, Zhang J, et al. (1995) The receptor for advanced glycation end products (RAGE) is a cellular binding site for amphoterin. Mediation of neurite outgrowth and co-expression of rage and amphoterin in the developing nervous system. J Biol Chem 270: 25752–25761.
[75]
Agresti A, Lupo R, Bianchi ME, Müller S (2003) HMGB1 interacts differentially with members of the Rel family of transcription factors. Biochem Biophys Res Commun 302: 421–426.
[76]
Lotze MT, Tracey KJ (2005) High-mobility group box 1 protein (HMGB1): nuclear weapon in the immune arsenal. Nat Rev Immunol 5: 331–342.
[77]
Tsan MF (2011) Heat shock proteins and high mobility group box 1 protein lack cytokine function. J Leukoc Biol 89: 847–853.
[78]
Bonaldi T, Talamo F, Scaffidi P, Ferrera D, Porto A, et al. (2003) Monocytic cells hyperacetylate chromatin protein HMGB1 to redirect it towards secretion. EMBO J 22: 5551–5560.
[79]
Youn JH, Shin JS (2006) Nucleocytoplasmic shuttling of HMGB1 is regulated by phosphorylation that redirects it toward secretion. J Immunol 177: 7889–7897.
[80]
Youn JH, Oh YJ, Kim ES, Choi JE, Shin JS (2008) High mobility group box 1 protein binding to lipopolysaccharide facilitates transfer of lipopolysaccharide to CD14 and enhances lipopolysaccharide-mediated TNF-alpha production in human monocytes. J Immunol 180: 5067–5074.
[81]
Qin YH, Dai SM, Tang GS, Zhang J, Ren D, et al. (2009) HMGB1 enhances the proinflammatory activity of lipopolysaccharide by promoting the phosphorylation of MAPK p38 through receptor for advanced glycation end products. J Immunol 183: 6244–6250.
