Llama nanoantibodies with therapeutic potential against human norovirus diarrhea.

Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE

Original Publication: San Francisco, CA : Public Library of Science

Antibodies, Monoclonal/*immunology ; Capsid Proteins/*immunology ; Diarrhea/*prevention & control ; Gastroenteritis/*prevention & control ; Immunization, Passive/*methods ; Single-Chain Antibodies/*immunology; Animals ; Antibodies, Monoclonal/administration & dosage ; Antibodies, Monoclonal/genetics ; Antibodies, Monoclonal/isolation & purification ; Antibody Specificity ; Camelids, New World ; Capsid Proteins/chemistry ; Capsid Proteins/genetics ; Cell Surface Display Techniques ; Diarrhea/immunology ; Diarrhea/virology ; Epitope Mapping ; Epitopes/chemistry ; Epitopes/immunology ; Gastroenteritis/immunology ; Gastroenteritis/virology ; Gene Library ; Hemagglutination Inhibition Tests ; Humans ; Immune Sera/chemistry ; Immunization ; Male ; Norovirus/drug effects ; Norovirus/immunology ; Norovirus/pathogenicity ; Protein Binding ; Recombinant Proteins/administration & dosage ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/immunology ; Single-Chain Antibodies/administration & dosage ; Single-Chain Antibodies/genetics ; Single-Chain Antibodies/isolation & purification ; Swine

Noroviruses are a major cause of acute gastroenteritis, but no vaccines or therapeutic drugs are available. Llama-derived single chain antibody fragments (also called VHH) are small, recombinant monoclonal antibodies of 15 kDa with several advantages over conventional antibodies. The aim of this study was to generate recombinant monoclonal VHH specific for the two major norovirus (NoV) genogroups (GI and GII) in order to investigate their potential as immunotherapy for the treatment of NoV diarrhea. To accomplish this objective, two llamas were immunized with either GI.1 (Norwalk-1968) or GII.4 (MD2004) VLPs. After immunization, peripheral blood lymphocytes were collected and used to generate two VHH libraries. Using phage display technology, 10 VHH clones specific for GI.1, and 8 specific for GII.4 were selected for further characterization. All VHH recognized conformational epitopes in the P domain of the immunizing VP1 capsid protein, with the exception of one GII.4 VHH that recognized a linear P domain epitope. The GI.1 VHHs were highly specific for the immunizing GI.1 genotype, with only one VHH cross-reacting with GI.3 genotype. The GII.4 VHHs reacted with the immunizing GII.4 strain and showed a varying reactivity profile among different GII genotypes. One VHH specific for GI.1 and three specific for GII.4 could block the binding of homologous VLPs to synthetic HBGA carbohydrates, saliva, and pig gastric mucin, and in addition, could inhibit the hemagglutination of red blood cells by homologous VLPs. The ability of Nov-specific VHHs to perform well in these surrogate neutralization assays supports their further development as immunotherapy for NoV treatment and immunoprophylaxis.

J Biol Chem. 2009 Jan 30;284(5):3273-3284. (PMID: 19010777)
J Clin Microbiol. 2008 Dec;46(12):3971-9. (PMID: 18842943)
Vaccine. 2006 May 8;24(19):4130-7. (PMID: 16616802)
J Med Virol. 2008 Aug;80(8):1461-7. (PMID: 18551595)
Vaccine. 2012 May 21;30(24):3580-6. (PMID: 22469864)
Arch Virol. 2013 Oct;158(10):2059-68. (PMID: 23615870)
N Engl J Med. 2013 Mar 21;368(12):1121-30. (PMID: 23514289)
Antiviral Res. 2014 May;105:80-91. (PMID: 24583027)
J Clin Invest. 2013 Sep;123(9):3829-38. (PMID: 23925294)
J Infect Dis. 2015 Mar 15;211(6):870-8. (PMID: 25210140)
J Gen Virol. 1998 Mar;79 ( Pt 3):479-87. (PMID: 9519826)
Emerg Infect Dis. 2008 Aug;14(8):1224-31. (PMID: 18680645)
Pediatr Transplant. 2011 Nov;15(7):718-21. (PMID: 21883746)
J Agric Food Chem. 2010 Jan 27;58(2):918-26. (PMID: 20039674)
J Virol. 2007 Jun;81(11):5949-57. (PMID: 17392366)
J Clin Virol. 2009 Jan;44(1):1-8. (PMID: 19084472)
J Mol Recognit. 2005 Jul-Aug;18(4):307-17. (PMID: 15952219)
PLoS Pathog. 2012;8(5):e1002705. (PMID: 22615565)
Vaccine. 1993;11(2):273-81. (PMID: 8382422)
J Virol. 2009 Nov;83(22):11890-901. (PMID: 19759138)
J Clin Microbiol. 1993 Aug;31(8):2185-91. (PMID: 8396590)
J Virol. 2012 Jul;86(13):7414-26. (PMID: 22532688)
Nat Med. 2006 May;12(5):580-4. (PMID: 16604085)
Virology. 2014 Apr;454-455:139-44. (PMID: 24725940)
Appl Environ Microbiol. 2012 Feb;78(3):786-94. (PMID: 22138991)
J Virol. 2007 Nov;81(22):12298-306. (PMID: 17855545)
Virology. 1994 Apr;200(1):319-25. (PMID: 8128635)
Adv Protein Chem. 2002;61:119-60. (PMID: 12461823)
BMC Microbiol. 2001;1:24. (PMID: 11710959)
J Virol. 2011 Jan;85(1):231-42. (PMID: 20980508)
Cancer Res. 2004 Apr 15;64(8):2853-7. (PMID: 15087403)
N Engl J Med. 2011 Dec 8;365(23):2178-87. (PMID: 22150036)
Cancer Lett. 2013 Jul 1;334(2):237-44. (PMID: 22902507)
J Clin Microbiol. 2003 Jun;41(6):2367-71. (PMID: 12791850)
Planta. 2012 Aug;236(2):703-14. (PMID: 22526499)
PLoS Pathog. 2013;9(5):e1003334. (PMID: 23658521)
Biotechniques. 1991 Nov;11(5):620-7. (PMID: 1804254)
Science. 1999 Oct 8;286(5438):287-90. (PMID: 10514371)
Arch Virol. 1996;141(5):865-75. (PMID: 8678832)
J Virol. 2002 Dec;76(23):12335-43. (PMID: 12414974)
J Virol. 2013 Mar;87(5):2803-13. (PMID: 23269783)
J Clin Microbiol. 2006 Mar;44(3):992-8. (PMID: 16517888)
J Mol Recognit. 1999 Sep-Oct;12(5):279-84. (PMID: 10556875)
Virol J. 2009 Sep 25;6:150. (PMID: 19781066)
J Gen Virol. 2005 Oct;86(Pt 10):2799-2806. (PMID: 16186235)
J Gen Virol. 2010 Oct;91(Pt 10):2449-56. (PMID: 20592107)
Clin Infect Dis. 2014 Feb;58(4):483-91. (PMID: 24300042)
Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1254-6. (PMID: 7877964)
PLoS One. 2010 May 05;5(5):e10482. (PMID: 20463957)
J Mol Biol. 1991 Dec 5;222(3):581-97. (PMID: 1748994)
J Virol. 2012 Jan;86(2):873-83. (PMID: 22090098)
J Virol. 2014 Jul;88(13):7244-55. (PMID: 24648459)
Transplant Proc. 2014 Dec;46(10):3603-5. (PMID: 25498097)
Anal Biochem. 2004 Feb 15;325(2):301-7. (PMID: 14751265)
BMC Biotechnol. 2012 Sep 07;12:59. (PMID: 22953695)
Proc Natl Acad Sci U S A. 2008 Jul 8;105(27):9175-80. (PMID: 18599458)
Virology. 2006 Mar 15;346(2):312-23. (PMID: 16343580)
Antiviral Res. 2013 Mar;97(3):293-300. (PMID: 23267830)
J Virol. 2015 Mar;89(5):2718-30. (PMID: 25520510)
Annu Rev Biochem. 2013;82:775-97. (PMID: 23495938)
Science. 1988 May 20;240(4855):1038-41. (PMID: 3285470)
Mol Biotechnol. 2013 Jun;54(2):590-601. (PMID: 23015323)
PLoS One. 2012;7(8):e42788. (PMID: 22880110)
Anal Biochem. 2006 Feb 1;349(1):136-47. (PMID: 16337141)
J Virol. 1992 Nov;66(11):6527-32. (PMID: 1328679)
PLoS One. 2013 Jul 19;8(7):e69379. (PMID: 23894462)
Anal Biochem. 1991 Nov 1;198(2):268-77. (PMID: 1724720)
Emerg Infect Dis. 2014 Jun;20(6):1016-8. (PMID: 24857806)
J Infect Dis. 2006 Dec 1;194(11):1580-8. (PMID: 17083044)
J Clin Microbiol. 2009 Sep;47(9):2855-62. (PMID: 19625473)
Antimicrob Agents Chemother. 2001 Oct;45(10):2807-12. (PMID: 11557473)
PLoS One. 2013 Jun 21;8(6):e67592. (PMID: 23805319)
J Biol Chem. 2006 Jul 14;281(28):19478-88. (PMID: 16675449)
J Med Virol. 2014 Dec;86(12):2055-64. (PMID: 24531909)
J Virol. 2011 May;85(9):4057-70. (PMID: 21345963)
Protein Eng Des Sel. 2014 Oct;27(10):339-49. (PMID: 24946948)
Arthritis Rheum. 2006 Jun;54(6):1856-66. (PMID: 16736523)
Emerg Infect Dis. 2008 Oct;14(10):1546-52. (PMID: 18826817)
Virology. 2006 Dec 5-20;356(1-2):179-87. (PMID: 16934306)
J Infect Dis. 2010 Oct 15;202(8):1212-8. (PMID: 20815703)
Vaccine. 2006 Jun 12;24(24):5220-34. (PMID: 16650512)
PLoS One. 2012;7(7):e41625. (PMID: 22844506)
J Virol Methods. 2012 Dec;186(1-2):126-31. (PMID: 22867844)
N Engl J Med. 2012 Nov 29;367(22):2126-32. (PMID: 23190223)
Virus Res. 2010 Aug;151(2):142-7. (PMID: 20417671)
J Virol. 2005 Jun;79(12):7402-9. (PMID: 15919896)
J Virol. 2014 Dec;88(24):14184-96. (PMID: 25275120)
J Clin Microbiol. 1997 Jul;35(7):1909-14. (PMID: 9196224)
J Immunol. 2011 Oct 15;187(8):4229-35. (PMID: 21930965)
J Virol. 2013 Sep;87(17):9547-57. (PMID: 23785216)
Clin Vaccine Immunol. 2012 Feb;19(2):284-7. (PMID: 22190401)
J Virol. 2008 Oct;82(19):9753-64. (PMID: 18632867)
J Infect Dis. 2002 May 1;185(9):1335-7. (PMID: 12001052)
Trends Microbiol. 2011 Aug;19(8):382-8. (PMID: 21705222)

United States Intramural NIH HHS

0 (Antibodies, Monoclonal)
0 (Capsid Proteins)
0 (Epitopes)
0 (Immune Sera)
0 (Recombinant Proteins)
0 (Single-Chain Antibodies)

Date Created: 20150813 Date Completed: 20160506 Latest Revision: 20231111

20250114

PMC4534396

10.1371/journal.pone.0133665

26267898