Fluorescence Adherence Inhibition Assay: A Novel Functional Assessment of Blocking Virus Attachment by Vaccine-Induced Antibodies.

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

Virus Attachment*; Antibodies, Neutralizing/*chemistry ; Antibodies, Neutralizing/*immunology ; Antibodies, Viral/*chemistry ; Antibodies, Viral/*immunology ; Fluorescent Dyes/*chemistry ; Viral Vaccines/*immunology; Animals ; Chlorocebus aethiops ; Erythrocytes/immunology ; Humans ; Models, Biological ; Poliovirus Vaccine, Inactivated/immunology ; Turkeys ; Vaccination ; Vero Cells

Neutralizing antibodies induced by vaccination or natural infection play a critically important role in protection against the viral diseases. In general, neutralization of the viral infection occurs via two major pathways: pre- and post-attachment modes, the first being the most important for such infections as influenza and polio, the latter being significant for filoviruses. Neutralizing capacity of antibodies is typically evaluated by virus neutralization assays that assess reduction of viral infectivity to the target cells in the presence of functional antibodies. Plaque reduction neutralization test, microneutralization and immunofluorescent assays are often used as gold standard virus neutralization assays. However, these methods are associated with several important prerequisites such as use of live virus requiring safety precautions, tedious evaluation procedure and long assessment time. Hence, there is a need for a robust, inexpensive high throughput functional assay that can be performed rapidly using inactivated virus, without extensive safety precautions. Herein, we report a novel high throughput Fluorescence Adherence Inhibition assay (fADI) using inactivated virus labeled with fluorescent secondary antibodies virus and Vero cells or erythrocytes as targets. It requires only few hours to assess pre-attachment neutralizing capacity of donor sera. fADI assay was tested successfully on donors immunized with polio, yellow fever and influenza vaccines. To further simplify and improve the throughput of the assay, we have developed a mathematical approach for calculating the 50% titers from a single sample dilution, without the need to analyze multi-point titration curves. Assessment of pre- and post-vaccination human sera from subjects immunized with IPOL®, YF-VAX® and 2013-2014 Fluzone® vaccines demonstrated high efficiency of the assay. The results correlated very well with microneutralization assay performed independently by the FDA Center of Biologics Evaluation and Research, with plaque reduction neutralization test performed by Focus Diagnostics, and with hemaglutination inhibition assay performed in-house at Sanofi Pasteur. Taken together, fADI assay appears to be a useful high throughput functional immunoassay for assessment of antibody-related neutralization of the viral infections for which pre-attachment neutralization pathway is predominant, such as polio, influenza, yellow fever and dengue.

Trop Med Int Health. 1999 Dec;4(12):867-71. (PMID: 10632996)
J Virol. 2003 Oct;77(19):10684-8. (PMID: 12970455)
Virol J. 2004 Dec 20;1:14. (PMID: 15610559)
Bull World Health Organ. 1962;27:699-707. (PMID: 13950710)
Int J Nanomedicine. 2006;1(1):59-64. (PMID: 17722262)
J Cell Biol. 1981 Dec;91(3 Pt 1):601-13. (PMID: 7328111)
Am J Trop Med Hyg. 1991 Jan;44(1):79-82. (PMID: 1996743)
J Virol Methods. 2004 Sep 15;120(2):207-15. (PMID: 15288964)
J Clin Microbiol. 2007 Nov;45(11):3777-80. (PMID: 17804661)
Vaccine. 2011 Aug 11;29(35):6008-16. (PMID: 21718741)
Arch Virol. 1993;130(3-4):441-55. (PMID: 8517795)
Anal Chem. 2004 Dec 15;76(24):7360-5. (PMID: 15595880)
J Virol. 2008 Jan;82(1):237-45. (PMID: 17928343)
Virology. 2010 Jun 5;401(2):228-35. (PMID: 20304456)
Retrovirology. 2011 Dec 06;8:99. (PMID: 22145853)
J Virol Methods. 2005 Sep;128(1-2):192-7. (PMID: 15955576)
Viral Immunol. 2008 Jun;21(2):123-32. (PMID: 18476771)
Clin Vaccine Immunol. 2008 Feb;15(2):177-81. (PMID: 18045884)
J Virol Methods. 2003 Jun 30;110(2):179-84. (PMID: 12798246)
PLoS One. 2008 Apr 23;3(4):e1993. (PMID: 18431480)
J Clin Microbiol. 1999 Apr;37(4):937-43. (PMID: 10074505)
J Virol Methods. 2003 Jun 9;110(1):67-71. (PMID: 12757922)
Am J Trop Med Hyg. 1983 May;32(3):565-8. (PMID: 6344672)

27305C0011 United States ES NIEHS NIH HHS; 27307C0011 United States ES NIEHS NIH HHS; 27398C0011 United States ES NIEHS NIH HHS

0 (Antibodies, Neutralizing)
0 (Antibodies, Viral)
0 (Fluorescent Dyes)
0 (Poliovirus Vaccine, Inactivated)
0 (Viral Vaccines)

Date Created: 20160211 Date Completed: 20160718 Latest Revision: 20240615

20240615

PMC4749260

10.1371/journal.pone.0144261

26863313