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Tumorigenesis by Adenovirus Type 12 E1A

  • Hancheng Guan
  • Robert P. Ricciardi
Chapter
Part of the Current Cancer Research book series (CUCR)

Abstract

All human adenovirus serotypes are capable of transforming mammalian cells in vitro. Transformation requires only two viral oncogenes, E1A and E1B, which cause unconstrained cell proliferation by deregulating the cell cycle and blocking apoptosis, respectively. In contrast to the transformation omnipotence inherent in all 51 serotypes, only a few serotypes including adenovirus type 12 (Ad12) have the additional capability of generating tumors in immunocompetent rodents. The Ad12 E1A protein (E1A-12) is the key determinant of viral tumorigenesis. Specifically, E1A-12 downregulates Major Histocompatibility Complex (MHC) class I antigens on the cell surface by shutting off the class I gene transcription. This enables Ad12-transformed cells to evade lysis by cytotoxic T lymphocytes (CTLs). E1A-12 mediates MHC class I shutoff via regulating DNA binding and transcriptional activities of both activator NF-κB (p50/p65) and repressor COUP-TFII. On the one hand, E1A-12 prevents DNA binding of NF-κB to the class I enhancer R1 site to disable this Master Regulator of immune genes from stimulating transcription. By physically binding NF-κB, E1A-12 blocks protein kinase A from phosphorylating the critical residues on p50 and p65 that are required for NF-κB DNA binding and transactivation, respectively. On the other hand, E1A-12 induces binding of COUP-TFII repression complex to the class I enhancer R2 site. E1A-12 not only upregulates COUP-TFII expression, but also acts as a physical component of the repression complex to recruit or stabilize the histone deacetylases HDAC1 and HDAC8. Histone deacetylation by HDAC in the class I enhancer region causes chromatin condensation and transcription repression of MHC class I. These two E1A-12-mediated complementary mechanisms that involve DNA binding regulation of both NF-κB and COUP-TFII/HDAC, provide a “FAIL-SAFE” strategy to ensure persistent diminution of surface MHC class I antigens under variable physiological conditions. In addition, E1A-12 mediates another compelling mechanism that involves evasion of natural killer (NK) cells, which normally lyse cells upon loss of surface MHC class I antigens. Importantly, E1A-12 downregulates surface expression of all NKG2D activating ligands including RAE1-α, -β, -γ, -ε, MULT1, and H60. Notably, the mRNA levels of these NKG2D ligands are also downregulated, perhaps through a mechanism similar to MHC class I shutoff, since promoters of these ligands also contain binding sites for NF-κB and COUP-TFII. Finally, E1A-12-mediated tumorigenesis requires the function encoded by the unique 20-amino acid “Spacer” between the conserved regions CR2 and CR3. This Spacer plays an important role in inducing neuronal and tumor-related genes. Significantly, neuronal gene induction by E1A-12 requires not only the relief of transcription repression by the neuron-restrictive silencer factor (NRSF), but also the stimulation of neuronal promoters by certain transactivators. The loss of NRSF repression is mediated by E1A-12 via proteasomal degradation of the repressor in the nucleus. The E1A-12 mediated induction of neuronal and tumor-related genes may be essential for cell outgrowth and invasion in Ad12 tumorigenesis.

Keywords

Natural Killer Natural Killer Cell Major Histocompatibility Complex Class Neuronal Gene Human Adenovirus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgment

We wish to acknowledge Grant CA29797 from the National Institutes of Health (to R. P. R.).

References

  1. Ackrill AM, Blair GE (1988) Regulation of major histocompatibility class I gene expression at the level of transcription in highly oncogenic adenovirus transformed rat cells. Oncogene 3:483–487PubMedGoogle Scholar
  2. Albert ML, Darnell RB (2004) Paraneoplastic neurological degenerations: keys to tumour immunity. Nat Rev Cancer 4:36–44PubMedCrossRefGoogle Scholar
  3. Baldwin AS Jr (1996) The NF-kappa B and I kappa B proteins: new discoveries and insights. Annu Rev Immunol 14:649–683, ReviewPubMedCrossRefGoogle Scholar
  4. Bernards R, Houweling A, Schrier PI, Bos JL, van der Eb AJ (1982) Characterization of cells transformed by Ad5/Ad12 hybrid early region I plasmids. Virology 120:422–432PubMedCrossRefGoogle Scholar
  5. Bernards R, Schrier PI, Houweling A, Bos JL, van der Eb AJ, Zylstra M, Melief CJM (1983) Tumorigenicity of cells transformed by adenovirus type 12 by evasion of T-cell immunity. Nature 350:776–779CrossRefGoogle Scholar
  6. Brose K, Bland KS, Wang KH, Arnott D, Henzel W, Goodman CS, Tessier-Lavigne M, Kidd T (1999) Slit proteins bind Robo receptors and have an evolutionarily conserved role in repulsive axon guidance. Cell 96:795–806PubMedCrossRefGoogle Scholar
  7. Chen W, Böcker W, Brosius J, Tiedge H (1997) Expression of neural BC200 RNA in human tumours. J Pathol 183:345–351PubMedCrossRefGoogle Scholar
  8. Chinnadurai G (2004) Modulation of oncogenic transformation by the human adenovirus E1A C-terminal region. Curr Top Microbiol Immunol 273:139–161, ReviewPubMedGoogle Scholar
  9. Cook JL, Krantz CK, Routes BA (1996) Role of p300-family proteins in E1A oncogene ­induction of cytolytic susceptibility and tumor cell rejection. Proc Natl Acad Sci USA 93:13985–13990PubMedCrossRefGoogle Scholar
  10. Coulson JM (2005) Transcriptional regulation: cancer, neurons and the REST. Curr Biol 15:665–668, ReviewCrossRefGoogle Scholar
  11. Eager KB, Williams J, Breiding D, Pan S, Knowles B, Appela E, Ricciardi RP (1985) Expression of histocompatibility antigens H-2K, D, and L is reduced in adenovirus-12-transformed mouse cells and is restored by interferon γ. Proc Natl Acad Sci USA 82:5525–5529PubMedCrossRefGoogle Scholar
  12. Endter C, Dobner T (2004) Cell transformation by human adenoviruses. Curr Top Microbiol Immunol 273:163–214, ReviewPubMedGoogle Scholar
  13. Finley D (2009) Recognition and processing of ubiquitin-protein conjugates by the proteasome. Annu Rev Biochem 78:477–513PubMedCrossRefGoogle Scholar
  14. Flint J, Shenk T (1997) Viral transactivating proteins. Annu Rev Genet 31:177–212, ReviewPubMedCrossRefGoogle Scholar
  15. Friedman DJ, Ricciardi RP (1988) Adenovirus type 12 E1A gene represses accumulation of MHC class I mRNA at the level of transcription. Virology 165:303–305PubMedCrossRefGoogle Scholar
  16. Gallimore PH (1972) Tumor production in immunosuppressed rats with cells transformed in vitro by adenovirus type 2. J Gen Virol 16:99–102PubMedCrossRefGoogle Scholar
  17. Gallimore PH, Turnell AS (2001) Adenovirus E1A: remodelling the host cell, a life or death experience. Oncogene 20:7824–7835, ReviewPubMedCrossRefGoogle Scholar
  18. Garber ME, Troyanskaya OG, Schluens K, Petersen S, Thaesler Z, Pacyna-Gengelbach M, van de Rijn M, Rosen G, Perou C, Whyte RI, Altman RB, Brown PO, Botstein D, Petersen I (2001) Diversity of gene expression in adenocarcinoma of the lung. Proc Natl Acad Sci USA 98:13784–13789PubMedCrossRefGoogle Scholar
  19. Garcia-Lora A, Algarra I, Garrido F (2003) MHC class I antigens, immune surveillance and tumor immune escape. J Cell Physiol 195:346–355, ReviewPubMedCrossRefGoogle Scholar
  20. Ge R, Kralli A, Weinmann R, Ricciardi RP (1992) Down-regulation of the major histocompatibility complex class I enhancer in adenovirus type 12-transformed cells is accompanied by an increase in factor binding. J Virol 66:6969–6978PubMedGoogle Scholar
  21. Ge R, Liu X, Ricciardi RP (1994) E1A oncogene of adenovirus-12 mediates trans-repression of MHC class I transcription in Ad5/Ad12 somatic hybrid transformed cells. Virology 203:389–392PubMedCrossRefGoogle Scholar
  22. Grone J, Doebler O, Loddenkemper C, Hotz B, Buhr HJ, Bhargava S (2006) Robo1/Robo4: differential expression of angiogenic markers in colorectal cancer. Oncol Rep 15:1437–1443PubMedGoogle Scholar
  23. Gu H, Roizman B (2007) Herpes simplex virus-infected cell protein 0 blocks the silencing of viral DNA by dissociating histone deacetylases from the CoREST-REST complex. Proc Natl Acad Sci USA 104:17134–17139PubMedCrossRefGoogle Scholar
  24. Gu H, Liang Y, Mandel G, Roizman B (2005) Components of the REST/CoREST/histone deacetylase repressor complex are disrupted, modified, and translocated in HSV-1-infected cells. Proc Natl Acad Sci USA 102:7571–7576PubMedCrossRefGoogle Scholar
  25. Guan H, Smirnov DA, Ricciardi RP (2003) Identification of genes associated with adenovirus 12 tumorigenesis by microarray. Virology 309:114–124PubMedCrossRefGoogle Scholar
  26. Guan H, Hou S, Ricciardi RP (2005) DNA binding of repressor nuclear factor-kappaB p50/p50 depends on phosphorylation of Ser337 by the protein kinase A catalytic subunit. J Biol Chem 280:9957–9962PubMedCrossRefGoogle Scholar
  27. Guan H, Jiao J, Ricciardi RP (2008) Tumorigenic adenovirus type 12 E1A inhibits phosphorylation of NF-kappaB by PKAc, causing loss of DNA binding and transactivation. J Virol 82:40–48PubMedCrossRefGoogle Scholar
  28. Guan H, Williams JF, Ricciardi RP (2009) Induction of neuronal and tumor-related genes by adenovirus type 12 E1A. J Virol 83:651–661PubMedCrossRefGoogle Scholar
  29. Guardavaccaro D, Frescas D, Dorrello NV, Peschiaroli A, Multani AS, Cardozo T, Lasorella A, Iavarone A, Chang S, Hernando E, Pagano M (2008) Control of chromosome stability by the beta-TrCP-REST-Mad2 axis. Nature 452:365–369PubMedCrossRefGoogle Scholar
  30. Hayashi H, Tanaka K, Jay F, Khoury G, Jay G (1985) Modulation of the tumorigenicity of human adenovirus-12-transformed cells by interferon. Cell 43:263–267PubMedCrossRefGoogle Scholar
  31. Hohlweg U, Dorn A, Hosel M, Webb D, Buettner R, Doerfler W (2004) Tumorigenesis by adenovirus type 12 in newborn Syrian hamsters. Curr Top Microbiol Immunol 273:215–244, ReviewPubMedGoogle Scholar
  32. Hou S, Guan H, Ricciardi RP (2002) In adenovirus type 12 tumorigenic cells, major histocompatibility complex class I transcription shutoff is overcome by induction of NF-kappaB and relief of COUP-TFII repression. J Virol 76:3212–3220PubMedCrossRefGoogle Scholar
  33. Hou S, Guan H, Ricciardi RP (2003) Phosphorylation of serine 337 of NF-kappaB p50 is critical for DNA binding. J Biol Chem 278:45994–45998PubMedCrossRefGoogle Scholar
  34. Huebner RJ, Rowe WP, Lane WT (1962) Oncogenic effects in hamsters of human adenovirus types 12 and 18. Proc Natl Acad Sci USA 48:2051–2058PubMedCrossRefGoogle Scholar
  35. Ingvarsson S (1990) The myc gene family proteins and their role in transformation and differentiation. Semin Cancer Biol 1:359–369PubMedGoogle Scholar
  36. Ito H, Funahashi S, Yamauchi N, Shibahara J, Midorikawa Y, Kawai S, Kinoshita Y, Watanabe A, Hippo Y, Ohtomo T, Iwanari H, Nakajima A, Makuuchi M, Fukayama M, Hirata Y, Hamakubo T, Kodama T, Tsuchiya M, Aburatani H (2006) Identification of ROBO1 as a novel hepatocellular carcinoma antigen and a potential therapeutic and diagnostic target. Clin Cancer Res 12:3257–3264PubMedCrossRefGoogle Scholar
  37. Jiao J, Guan H, Lippa AM, Ricciardi RP (2010) The N terminus of adenovirus 12 E1A inhibits major histocompatibility complex class I expression by preventing phosphorylation of NF-κB p65 Ser276 through direct binding. J Virol 84:7668–7674PubMedCrossRefGoogle Scholar
  38. Kawagoe H, Kandilci A, Kranenburg TA, Grosveld GC (2007) Overexpression of N-Myc rapidly causes acute myeloid leukemia in mice. Cancer Res 67:10677–10685PubMedCrossRefGoogle Scholar
  39. Kenyon DJ, Raska K Jr (1986) Region E1a of highly oncogenic adenovirus 12 in transformed cells protects against NK but not LAK cytolysis. Virology 155:644–654PubMedCrossRefGoogle Scholar
  40. Kidd T, Brose K, Mitchell KJ, Fetter RD, Tessier-Lavigne M, Goodman CS, Tear G (1998) Roundabout controls axon crossing of the CNS midline and defines a novel subfamily of evolutionarily conserved guidance receptors. Cell 92:205–215PubMedCrossRefGoogle Scholar
  41. Kimura A, Israel A, Le Bail O, Kourilsky P (1986) Detailed analysis of the mouse H-2Kb promoter: enhancer-like sequences and their role in the regulation of class I gene expression. Cell 44:261–272PubMedCrossRefGoogle Scholar
  42. Kralli A, Ge R, Graeven U, Ricciardi RP, Weinmann R (1992) Negative regulation of the major histocompatibility complex class I enhancer in adenovirus type 12-transformed cells via a retinoic acid response element. J Virol 66:6979–6988PubMedGoogle Scholar
  43. Kushner DB, Ricciardi RP (1999) Reduced phosphorylation of p50 is responsible for diminished NF-κB binding to the major histocompatibility complex class I enhancer in adenovirus 12 transformed cells. Mol Cell Biol 99:2169–2179Google Scholar
  44. Kushner DB, Pereira DS, Liu X, Graham FL, Ricciardi RP (1996) The first exon of Ad12 E1A excluding the transactivation domain mediates differential binding of COUP-TF and NF-κB to the MHC class I enhancer in transformed cells. Oncogene 12:143–151PubMedGoogle Scholar
  45. Lanier LL (2005) NK cell recognition. Annu Rev Immunol 23:225–274, ReviewPubMedCrossRefGoogle Scholar
  46. Li CC, Dai RM, Chen E, Longo DL (1994) Phosphorylation of NF-KB1-p50 is involved in NF-kappa B activation and stable DNA binding. J Biol Chem 269:30089–30092PubMedGoogle Scholar
  47. Liu X, Ge R, Westmoreland S, Cooney AJ, Tsai SY, Tsai MJ, Ricciardi RP (1994) Negative regulation by the R2 site of the MHC class I enhancer in adenovirus-12 transformed cells correlates with high levels of COUP-TF binding. Oncogene 9:2183–2190PubMedGoogle Scholar
  48. Liu X, Ge R, Ricciardi RP (1996) Evidence for the involvement of a nuclear NF-κB inhibitor in global down-regulation of the major histocompatibility complex class I enhancer in adenovirus type12-transformed cells. Mol Cell Biol 16:398–404PubMedGoogle Scholar
  49. Lodoen MB, Lanier LL (2005) Viral modulation of NK cell immunity. Nat Rev Microbiol 3:59–69, ReviewPubMedCrossRefGoogle Scholar
  50. Majumder S (2006) REST in good times and bad: roles in tumor suppressor and oncogenic activities. Cell Cycle 5:1929–1935, ReviewPubMedCrossRefGoogle Scholar
  51. Makrigiannis AP, Anderson SK (2003) Regulation of natural killer cell function. Cancer Biol Ther 2:610–616, ReviewPubMedGoogle Scholar
  52. Mertsch S, Schmitz N, Jeibmann A, Geng JG, Paulus W, Senner V (2008) Slit2 involvement in glioma cell migration is mediated by Robo1 receptor. J Neurooncol 87:1–7PubMedCrossRefGoogle Scholar
  53. Nausch N, Florin L, Hartenstein B, Angel P, Schorpp-Kistner M, Cerwenka A (2006) Cutting edge: the AP-1 subunit JunB determines NK cell-mediated target cell killing by regulation of the NKG2D-ligand RAE-1epsilon. J Immunol 176:7–11PubMedGoogle Scholar
  54. Nomura M, Zou Z, Joh T, Takihara Y, Matsuda Y, Shimada K (1996) Genomic structures and characterization of Rae1 family members encoding GPI-anchored cell surface proteins and expressed predominantly in embryonic mouse brain. J Biochem 120:987–995PubMedGoogle Scholar
  55. Pahl HL (1999) Activators and target genes of Rel/NF-kappaB transcription factors. Oncogene 18:6853–6866, ReviewPubMedCrossRefGoogle Scholar
  56. Qiu Y, Cooney AJ, Kuratani S, DeMayo FJ, Tsai SY, Tsai MJ (1994) Spatiotemporal expression patterns of chicken ovalbumin upstream promoter-transcription factors in the developing mouse central nervous system: evidence for a role in segmental patterning of the diencephalon. Proc Natl Acad Sci USA 91:4451–4455PubMedCrossRefGoogle Scholar
  57. Ricciardi RP (1999) Adenovirus transformation and tumorigenicity. In: Seth P (ed) Adenoviruses: basic biology to gene therapy. RG Landes Co., Austin, TX, pp 217–227Google Scholar
  58. Ricciardi RP, Zhao B, Guan H (2006) Mechanism of tumorigenesis mediated by adenovirus-12 E1A. In: Tognon M (ed) Viral oncogenesis. Research Signpost, Kerala, India, pp 107–124Google Scholar
  59. Routes JM, Ryan S, Morris K, Takaki R, Cerwenka A, Lanier LL (2005) Adenovirus serotype 5 E1A sensitizes tumor cells to NKG2D-dependent NK cell lysis and tumor rejection. J Exp Med 202:1477–1482PubMedCrossRefGoogle Scholar
  60. Sawada Y, Föhring B, Shenk T, Raska K Jr (1985) Tumorigenicity of adenovirus transformed cells: region E1A of adenovirus 12 confers resistance to natural killer cells. Virology 147:413–421PubMedCrossRefGoogle Scholar
  61. Sawada Y, Raska K Jr, Shenk T (1988) Adenovirus type 5 and type 12 recombinant viruses containing heterologous E1 genes are viable, transform rat cells, but are not tumorigenic in rats. Virology 166:281–284PubMedCrossRefGoogle Scholar
  62. Scheidereit C (2006) IkappaB kinase complexes: gateways to NF-kappaB activation and transcription. Oncogene 25:6685–6705, ReviewPubMedCrossRefGoogle Scholar
  63. Schrader EK, Harstad KG, Matouschek A (2009) Targeting proteins for degradation. Nat Chem Biol 5:815–822PubMedCrossRefGoogle Scholar
  64. Schrier PI, Bernards R, Vaessen RTMJ, Houweling A, van der Eb AJ (1983) Expression of class I major histocompatibility antigens switched off by highly oncogenic adenovirus 12 in transformed rat cells. Nature 305:771–775PubMedCrossRefGoogle Scholar
  65. Smirnov DA, Hou S, Ricciardi RP (2000) Association of histone deacetylase with COUP-TF in tumorigenic Ad12 transformed cells and its potential role in shut-off of MHC class I transcription. Virology 268:319–328PubMedCrossRefGoogle Scholar
  66. Smirnov DA, Hou S, Liu X, Claudio E, Siebenlist UK, Ricciardi RP (2001) COUP-TFII is up-regulated in adenovirus type 12 tumorigenic cells and is a repressor of MHC class I transcription. Virology 284:13–19PubMedCrossRefGoogle Scholar
  67. Tanaka K, Isselbacher KJ, Khoury G, Jay G (1985) Reversal of oncogenesis by the expression of a histocompatibility complex class I gene. Science 228:26–30PubMedCrossRefGoogle Scholar
  68. Trentin JJ, Yabe Y, Taylor G (1962) The quest for human cancer viruses. Science 137:835–841PubMedCrossRefGoogle Scholar
  69. Vasavada R, Eager KB, Barbanti-Brodano G, Caputo A, Ricciardi RP (1986) Adenovirus type 12 early region 1A proteins repress class I HLA expression in transformed human cells. Proc Natl Acad Sci USA 83:5257–5261PubMedCrossRefGoogle Scholar
  70. Westbrook TF, Martin ES, Schlabach MR, Leng Y, Liang AC, Feng B, Zhao JJ, Roberts TM, Mandel G, Hannon GJ, Depinho RA, Chin L, Elledge SJ (2005) A genetic screen for candidate tumor suppressors identifies REST. Cell 121:837–848PubMedCrossRefGoogle Scholar
  71. Westbrook TF, Hu G, Ang XL, Mulligan P, Pavlova NN, Liang AC, Leng Y, Maehr R, Shi Y, Harper JW, Elledge SJ (2008) SCFbeta-TRCP controls oncogenic transformation and neural differentiation through REST degradation. Nature 452:370–374PubMedCrossRefGoogle Scholar
  72. Williams JF, Zhang Y, Williams MA, Hou S, Kushner D, Ricciardi RP (2004) E1A-based determinants of oncogenicity in human adenovirus groups A and C. Curr Top Microbiol Immunol 273:245–288, ReviewPubMedGoogle Scholar
  73. Xiao C, Ghosh S (2005) NF-kappaB, an evolutionarily conserved mediator of immune and inflammatory responses. Adv Exp Med Biol 560:41–45, ReviewPubMedCrossRefGoogle Scholar
  74. Yewdell JW, Bennink JR, Eager KB, Ricciardi RP (1988) CTL recognition of adenovirus-transformed cells infected with influenza virus: lysis by anti-influenza CTL parallels adenovirus-12-induced suppression of class I MHC molecules. Virology 162:236–238PubMedCrossRefGoogle Scholar
  75. Zallen JA, Yi BA, Bargmann CI (1998) The conserved immunoglobulin superfamily member SAX-3/Robo directs multiple aspects of axon guidance in C. elegans. Cell 92:217–227PubMedCrossRefGoogle Scholar
  76. Zhang Y, Dang C, Ma Q, Shimahara Y (2005) Expression of nerve growth factor receptors and their prognostic value in human pancreatic cancer. Oncol Rep 14:161–171PubMedGoogle Scholar
  77. Zhao B, Riciardi RP (2006) E1A is the component of the MHC class I enhancer complex that ­mediates HDAC1, 8 chromatin repression in adenovirus-12 tumorigenic cells. Virology 352:338–344PubMedCrossRefGoogle Scholar
  78. Zhao B, Hou S, Ricciardi RP (2003) Chromatin repression by COUP-TFII and HDAC dominates activation by NF-kappaB in regulating major histocompatibility complex class I transcription in adenovirus tumorigenic cells. Virology 306:68–76PubMedCrossRefGoogle Scholar
  79. Zhong H, SuYang H, Erdjument-Bromage H, Tempst P, Ghosh S (1997) The transcriptional activity of NF-kappaB is regulated by the IkappaB-associated PKAc subunit through a cyclic AMP-independent mechanism. Cell 89:413–424PubMedCrossRefGoogle Scholar
  80. Zhong H, Voll RE, Ghosh S (1998) Phosphorylation of NF-kappa B p65 by PKA stimulates transcriptional activity by promoting a novel bivalent interaction with the coactivator CBP/p300. Mol Cell 1:661–671PubMedCrossRefGoogle Scholar
  81. Zhong H, May MJ, Jimi E, Ghosh S (2002) The phosphorylation status of nuclear NF-kappa B determines its association with CBP/p300 or HDAC-1. Mol Cell 9:625–636PubMedCrossRefGoogle Scholar
  82. Zhou C, Tsai SY, Tsai M (2000) From apoptosis to angiogenesis: new insights into the roles of nuclear orphan receptors, chicken ovalbumin upstreampromoter-transcription factors, during development. Biochim Biophys Acta 1470:M63–M68, ReviewPubMedGoogle Scholar
  83. Zinkernagel RM, Doherty PC (1979) MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function and responsiveness. Adv Immunol 27:51–177PubMedCrossRefGoogle Scholar

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© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Microbiology, School of Dental MedicineUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Abramson Cancer CenterUniversity of Pennsylvania School of MedicinePhiladelphiaUSA

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