Development of New Immunological Assays and Animal Models to Evaluate Vaccine Safety and Efficacy Against Emerging Diseases
Hana Golding, PhD
Office of Vaccines Research and Review
Division of Viral Products
Laboratory of Retroviruses
Hana Golding is currently the Chief of the Laboratory of Retrovirus at the Division of Viral Products, Center for Biologics Evaluation and Research (CBER), FDA. Dr. Golding received her Ph.D. degree from Oregon Health Sciences University, Portland, Oregon, and her postdoctoral training at the Experimental Immunology Branch, NCI, NIH. Dr. Golding Joined the Division of Viral Products, CBER in 1987 and was appointed the Chief of the Laboratory of Retrovirus Research in 1993. To date, Dr. Golding has authored more than 200 research papers and book chapters on immunology, virology, and infectious diseases topics.
The main areas of research projects in the Golding lab include:
- Vaccines against viral pathogens, including pandemic influenza, RSV, Ebola, Zika, HIV, and SARS-CoV-2
- Evaluation of vaccine safety (in vivo/in vitro) and immune responses, including new methods for increasing antibody avidity and epitope diversity
- Vaccine adjuvants, including their mode of action and impact on immune responses, as well as identifying new biomarkers predictive of adjuvant safety and efficacy in people.
Industry and academic researchers are developing new vaccines against viruses, including pandemic influenza strains of avian origin (H5N1, H7N7, H7N9, H9N9), respiratory syncytial virus (RSV), Ebola, Zika virus, and now SARS-CoV-2. Novel "universal influenza vaccines" are also under development. Many of these vaccines are being developed and tested together with novel adjuvants (agents that stimulate or increase immune response to vaccines).
Therefore, in addition to evaluating the safety and efficacy of the vaccine itself, FDA must also evaluate the combination of vaccine and its adjuvant. Moreover, evaluation of each viral vaccine presents unique challenges that agency regulators must address individually.
The goal of our program is to develop new and improved tools for monitoring vaccine safety and efficacy. FDA will then share these new tools with the regulated industry to enable use of these tools in their research and development efforts.
The availability of new tools could reduce the time it takes to develop new vaccines against influenza, RSV, Ebola, Zika, and SARS-CoV-2, and help FDA to quickly recognize potential safety problems with vaccines that use new adjuvants. Thus, our work will contribute significantly to public health in the U.S. and globally by facilitating development and approval of new vaccines that prevent serious diseases.
Our laboratory is working to provide new insights that will facilitate the development of effective vaccines against highly pathogenic viruses, including vaccines against avian influenza (H5, H7, H9, H2) based on the globular head of the influenza hemagglutinin (HA) from the corresponding viruses. We are also exploring new protective targets in RSV attachment protein (G), Ebola glycoprotein GP, and the new SARS-CoV-2 virus.
Future vaccines for influenza strains with pandemic potential will likely be combined with adjuvants in order to improve their immunogenicity, to generate heterosubtypic neutralizing activity, and to reduce the amount of antigen required for vaccination. Reducing amounts needed would be an important advantage for global vaccination campaigns.
In parallel with vaccine development, we are working to improve the analytical tools available for comparing immune responses generated by different vaccine candidates. Our laboratory initiated the use of whole Genome Phage Display Libraries (GFPDL) for comprehensive analyses of antibody repertoires. Additionally, we use surface plasmon resonance (SPR) to quantitate antibody binding to properly folded viral surface proteins and to measure antibody affinity maturation. These tools are used in the evaluation of convalescent sera and monoclonal antibodies, and to compare immune sera from recipients of adjuvanted vs. unadjuvanted influenza vaccines.
Both GFPDL and SPR technologies have been developed for RSV, which causes high morbidity in newborn children and in frail elderly individuals. Detailed knowledge of the changes in anti-RSV reactivity in different age groups is key to successful vaccination. New RSV vaccines are in advanced development and will be evaluated in the very old, the very young (newborn infants), and pregnant mothers.
In response to the Ebola epidemic in West Africa and the outbreaks of Zika virus infections in the Americas, multiple vaccine initiatives are ongoing. We developed novel methods to better explore the antibody repertoires following Ebola and Zika infection and vaccination.
In response to the current COVID-19 pandemic, our lab developed several analytical tools for in-depth analyses of humoral immune responses to SARS-CoV-2 infection and vaccination. These include Whole-Genome-Fragment-Phage-Display-Libraries (GFPDL) covering both the spike protein and internal genes; pseudovirus neutralization assays against the original Wuhan strain and against multiple variants of concern; ACE2/RBD blocking ELISAs;, and surface plasmon resonance (SPR)-based assays to measure antibody affinity against spike protein subdomains and receptor binding domain (RBD) with escape mutations. Human monocytes are also being used in a project to understand the pathogenesis of COVID-19 that involves cytokine storm and high levels of fever-inducing molecules. A role for antibody complexes is also under investigation.
The new analytical tools developed in our lab will help to find correlates of protection against influenza, RSV, Ebola, Zika, and SARS-CoV-2. New tools could also contribute to the design of improved vaccines with broad cross-protection potential.
Some new adjuvants contain components that can cause unacceptable reactogenicity and systemic adverse reactions. Often these adverse reactions are undetectable in pre-clinical toxicity studies in small animals, due to species differences in the cellular receptor targeted by the novel adjuvants or because of their lower sensitivity to drug-associated toxicities. Therefore, we are developing rapid in vitro screening assays based on human cells to evaluate the activities of new adjuvants and to identify parameters that will predict unacceptable toxicities, including fever, in humans.
Our program is also continuing in the application of whole-body bioimaging of animals challenged with recombinant RSV-luciferase. This approach, in addition to the use of advanced statistic tools, provides a practical, quantitative, and humane approach to studying virus dissemination in animal models and to evaluating the efficacy of novel vaccines and therapies against smallpox and RSV.
Innovation and Regulatory Science-Research Summary: Study of antibody response to SARS-CoV-2 spike proteins could help inform vaccine design | FDA
- Nature 2021 May 10 [Epub ahead of print]
Neutralizing antibody vaccine for pandemic and pre-emergent coronaviruses.
Saunders KO, Lee E, Parks R, Martinez DR, Li D, Chen H, Edwards RJ, Gobeil S, Barr M, Mansouri K, Alam SM, Sutherland LL, Cai F, Sanzone AM, Berry M, Manne K, Bock KW, Minai M, Nagata BM, Kapingidza AB, Azoitei M, Tse LV, Scobey TD, Spreng RL, Rountree RW, DeMarco CT, Denny TN, Woods CW, Petzold EW, Tang J, Oguin TH 3rd, Sempowski GD, Gagne M, Douek DC, Tomai MA, Fox CB, Seder R, Wiehe K, Weissman D, Pardi N, Golding H, Khurana S, Acharya P, Andersen H, Lewis MG, Moore IN, Montefiori DC, Baric RS, Haynes BF
- Clin Infect Dis 2021 Apr 16 [Epub ahead of print]
Impact of convalescent plasma therapy on SARS CoV-2 antibody profile in COVID-19 patients.
Tang J, Grubbs G, Lee Y, Golding H, Khurana S
- Sci Adv 2021 Mar 5;7(10):eabf2467
Longitudinal antibody repertoire in "mild" versus "severe" COVID-19 patients reveals immune markers associated with disease severity and resolution.
Ravichandran S, Lee Y, Grubbs G, Coyle EM, Klenow L, Akasaka O, Koga M, Adachi E, Saito M, Nakachi I, Ogura T, Baba R, Ito M, Kiso M, Yasuhara A, Yamada S, Sakai-Tagawa Y, Iwatsuki-Horimoto K, Imai M, Yamayoshi S, Yotsuyanagi H, Kawaoka Y, Khurana S
- Nat Commun 2021 Feb 22;12(1):1221
Antibody affinity maturation and plasma IgA associate with clinical outcome in hospitalized COVID-19 patients.
Tang J, Ravichandran S, Lee Y, Grubbs G, Coyle EM, Klenow L, Genser H, Golding H, Khurana S
- Clin Transl Med 2021 Feb;11(2):e281
Bromelain inhibits SARS-CoV-2 infection via targeting ACE-2, TMPRSS2, and spike protein.
Sagar S, Rathinavel AK, Lutz WE, Struble LR, Khurana S, Schnaubelt AT, Mishra NK, Guda C, Palermo NY, Broadhurst MJ, Hoffmann T, Bayles KW, Reid SPM, Borgstahl GEO, Radhakrishnan P
- Mol Immunol 2020 Dec;128:139-49
Production of fever mediator PGE(2) in human monocytes activated with MDP adjuvant is controlled by signaling from MAPK and p300 HAT: key role of T cell derived factor.
Liu F, Romantseva T, Park YJ, Golding H, Zaitseva M
- Int J Mol Sci 2020 Nov 28;21(23):E9055
Secretome analysis of inductive signals for BM-MSC transdifferentiation into salivary gland progenitors.
Mona M, Kobeissy F, Park YJ, Miller R, Saleh W, Koh J, Yoo MJ, Chen S, Cha S
- Sci Transl Med 2020 Oct 28;12(567):eaaz4997
Nonhuman primates exposed to Zika virus in utero are not protected against reinfection at 1 year postpartum.
Vannella KM, Stein S, Connelly M, Swerczek J, Amaro-Carambot E, Coyle EM, Babyak A, Winkler CW, Saturday G, Gai ND, Hammoud DA, Dowd KA, Valencia LP, Ramos-Benitez MJ, Kindrachuk J, Pierson TC, Peterson KE, Brenchley JM, Whitehead SS, Khurana S, Herbert R, Chertow DS
- Viruses 2020 Oct 8;12(10):E1140
Autoreactivity of broadly neutralizing influenza human antibodies to human tissues and human proteins.
Khurana S, Hahn M, Klenow L, Golding H
- Sci Transl Med 2020 Jul 1;12(550):eabc3539
Antibody signature induced by SARS-CoV-2 spike protein immunogens in rabbits.
Ravichandran S, Coyle EM, Klenow L, Tang J, Grubbs G, Liu S, Wang T, Golding H, Khurana S
- iScience 2020 Mar 27;23(3):100920
Human antibody repertoire following Ebola virus infection and vaccination.
Fuentes S, Ravichandran S, Coyle EM, Klenow L, Khurana S
- Cell Host Microbe 2020 Feb 12;27(2):262-76
Longitudinal human antibody repertoire against complete Ebola virus proteome following natural infection reveals immune markers of protection.
Khurana S, Ravichandran S, Hahn M, Coyle EM, Spencer SW, Zak SE, Kindrachuk J, Davey Jr. RT, Dye JM, Chertow DS
- J Infect Dis 2020 Feb 15;221(4):636-46
Antigenic fingerprinting of RSV-A infected hematopoietic cell transplant recipients reveals importance of mucosal anti-RSV-G antibodies in control of RSV infection in humans.
Fuentes S, Hahn M, Chilcote K, Chemaly RF, Shah DP, Ye X, Avadhanula V, Piedra PA, Golding H, Khurana S
- Vaccine 2020 Jan 22;38(4):800-7
Expansion of the 1st WHO international standard for antiserum to respiratory syncytial virus to include neutralisation titres against RSV subtype B: an international collaborative study.
McDonald JU, Rigsby P, Atkinson E, Engelhardt OG, Study Participants
- Lancet Respir Med 2019 Nov;7(11):951-63
Anti-influenza hyperimmune intravenous immunoglobulin for adults with influenza A or B infection (FLU-IVIG): a double-blind, randomised, placebo-controlled trial.
Davey RT Jr, Fernandez-Cruz E, Markowitz N, Pett S, Babiker AG, Wentworth D, Khurana S, Engen N, Gordin F, Jain MK, Kan V, Polizzotto MN, Riska P, Ruxrungtham K, Temesgen Z, Lundgren J, Beigel JH, Lane HC, Neaton JD
- Sci Signal 2019 Oct 8;12(602):eaat6023
T cell-derived soluble glycoprotein GPIbalpha mediates PGE2 production in human monocytes activated with the vaccine adjuvant MDP.
Liu F, Endo Y, Romantseva T, Wu WW, Akue A, Shen RF, Golding H, Zaitseva M
- NPJ Vaccines 2019 Oct 14;4:42
Harmonization of Zika neutralization assays by using the WHO International Standard for anti-Zika virus antibody.
Mattiuzzo G, Knezevic I, Hassall M, Ashall J, Myhill S, Faulkner V, Hockley J, Rigsby P, Wilkinson DE, Page M, collaborative study participants
- Cell 2019 Sep 5;178(6):1313-28
Antibiotics-driven gut microbiome perturbation alters immunity to vaccines in humans.
Hagan T, Cortese M, Rouphael N, Boudreau C, Linde C, Maddur MS, Das J, Wang H, Guthmiller J, Zheng NY, Huang M, Uphadhyay AA, Gardinassi L, Petitdemange C, McCullough MP, Johnson SJ, Gill K, Cervasi B, Zou J, Bretin A, Hahn M, Gewirtz AT, Bosinger SE, Wilson PC, Li S, Alter G, Khurana S, Golding H, Pulendran B
- Proc Natl Acad Sci U S A 2019 Jul 23;116(30):15194-9
Antibody-dependent enhancement of influenza disease promoted by increase in hemagglutinin stem flexibility and virus fusion kinetics.
Winarski KL, Tang J, Klenow L, Lee J, Coyle EM, Manischewitz J, Turner HL, Takeda K, Ward AB, Golding H, Khurana S
- Nat Commun 2019 Jul 26;10(1):3338
Repeat vaccination reduces antibody affinity maturation across different influenza vaccine platforms in humans.
Khurana S, Hahn M, Coyle EM, King LR, Lin TL, Treanor J, Sant A, Golding H
- Nat Commun 2019 Apr 26;10(1):1943
Differential human antibody repertoires following Zika infection and the implications for serodiagnostics and disease outcome.
Ravichandran S, Hahn M, Belaunzaran-Zamudio PF, Ramos-Castaneda J, Najera-Cancino G, Caballero-Sosa S, Navarro-Fuentes KR, Ruiz-Palacios G, Golding H, Beigel JH, Khurana S
- J Virol 2019 Apr;93(8):e00169-19
Broad hemagglutinin-specific memory B cell expansion by seasonal influenza virus infection reflects early-life imprinting and adaptation to the infecting virus.
Tesini BL, Kanagaiah P, Wang J, Hahn M, Halliley JL, Chaves FA, Nguyen PQT, Nogales A, DeDiego ML, Anderson CS, Ellebedy AH, Strohmeier S, Krammer F, Yang H, Bandyopadhyay S, Ahmed R, Treanor JJ, Martinez-Sobrido L, Golding H, Khurana S, Zand MS, Topham DJ, Sangster MY
- J Infect Dis 2018 Dec 15;218(Suppl. 5):S636-48
Fully human immunoglobulin G from transchromosomic bovines treats nonhuman primates infected with Ebola virus Makona isolate.
Luke T, Bennett RS, Gerhardt DM, Burdette T, Postnikova E, Mazur S, Honko AN, Oberlander N, Byrum R, Ragland D, St Claire M, Janosko KB, Smith G, Glenn G, Hooper J, Dye J, Pal S, Bishop-Lilly KA, Hamilton T, Frey K, Bollinger L, Wada J, Wu H, Jiao JA, Olinger GG, Gunn B, Alter G, Khurana S, Hensley LE, Sullivan E, Jahrling PB
- J Infect Dis 2018 Dec 15;218(Suppl. 5):S597-602
Antibody repertoire of human polyclonal antibodies against Ebola virus glycoprotein generated after deoxyribonucleic acid and protein vaccination of transchromosomal bovines.
Fuentes S, Ravichandran S, Khurana S
- NPJ Vaccines 2018 Oct 1;3:40
AS03-adjuvanted H5N1 vaccine promotes antibody diversity and affinity maturation, NAI titers, cross-clade H5N1 neutralization, but not H1N1 cross-subtype neutralization.
Khurana S, Coyle EM, Manischewitz J, King LR, Gao J, Germain RN, Schwartzberg PL, Tsang JS, Golding H, CHI Consortium
- PLoS Pathog 2018 Aug 24;14(8):e1007262
Protective antigenic sites in respiratory syncytial virus G attachment protein outside the central conserved and cysteine noose domains.
Lee J, Klenow L, Coyle EM, Golding H, Khurana S
- Vaccine 2018 Jul 25;36(31):4657-62
Improving ability of RSV microneutralization assay to detect G-specific and cross-reactive neutralizing antibodies through immortalized cell line selection.
Boukhvalova MS, Mbaye A, Kovtun S, Yim KC, Konstantinova T, Getachew T, Khurana S, Falsey AR, Blanco JCG
- Cold Spring Harb Perspect Biol 2018 Apr 2;10(4):a029132
What is the predictive value of animal models for vaccine efficacy in humans? The importance of bridging studies and species-independent correlates of protection.
Golding H, Khurana S, Zaitseva M
- Vaccines 2018 Apr 27;6(2):24
Development and regulation of novel influenza virus vaccines: a United States young scientist perspective.
- J Virol 2018 Feb;92(4):e01588-17
Comparison of the efficacy of N9 neuraminidase-specific monoclonal antibodies against influenza A(H7N9) virus infection.
Wan H, Qi L, Gao J, Couzens LK, Jiang L, Gao Y, Sheng ZM, Fong S, Hahn M, Khurana S, Taubenberger JK, Eichelberger MC
- Cell Host Microbe 2017 Oct 11;22(4):471-83.e5
A potent germline-like human monoclonal antibody targets a pH-sensitive epitope on H7N9 influenza hemagglutinin.
Yu F, Song H, Wu Y, Chang SY, Wang L, Li W, Hong B, Xia S, Wang C, Khurana S, Feng Y, Wang Y, Sun Z, He B, Hou D, Manischewitz J, King LR, Song Y, Min JY, Golding H, Ji X, Lu L, Jiang S, Dimitrov DS, Ying T
- Antiviral Res 2017 Aug;144:8-20
Development of an animal model of progressive vaccinia in nu/nu mice and the use of bioluminescence imaging for assessment of the efficacy of monoclonal antibodies against vaccinial B5 and L1 proteins.
Zaitseva M, Thomas A, Meseda CA, Cheung CYK, Diaz CG, Xiang Y, Crotty S, Golding H
- J Transl Med 2017 Jul 10;15(1):155
Impaired B cell immunity in acute myeloid leukemia patients after chemotherapy.
Goswami M, Prince G, Biancotto A, Moir S, Kardava L, Santich BH, Cheung F, Kotliarov Y, Chen J, Shi R, Zhou H, Golding H, Manischewitz J, King L, Kunz LM, Noonan K, Borrello IM, Smith BD, Hourigan CS
- Clin Vaccine Immunol 2017 Mar 6;24(3):e00498-16
Preexisting immunity not frailty phenotype predicts influenza post vaccination titers among older veterans.
Van Epps P, Tumpey T, Pearce MB, Golding H, Higgins P, Hornick T, Burant C, Wilson BM, Banks R, Gravenstein S, Canaday DH
- Sci Rep 2017 Feb 10;7:42428
Preclinical evaluation of bacterially produced RSV-G protein vaccine: Strong protection against RSV challenge in cotton rat model.
Fuentes S, Klenow L, Golding H, Khurana S
- Vaccine 2017 Jan 23;35(4):694-702
Development of bioluminescence imaging of respiratory syncytial virus (RSV) in virus-infected live mice and its use for evaluation of therapeutics and vaccines.
Fuentes S, Arenas D, Moore MM, Golding H, Khurana S
- Nat Med 2016 Dec;22(12):1439-47
Human antibody repertoire after VSV-Ebola vaccination identifies novel targets and virus-neutralizing IgM antibodies.
Khurana S, Fuentes S, Coyle EM, Ravichandran S, Davey RT Jr, Beigel JH
- J Virol 2016 Oct 15;90(20):9383-93
Antigenic fingerprinting of antibody response following highly pathogenic H7N7 avian influenza virus exposure in humans: Evidence for anti-PA-X antibodies.
Khurana S, Chung KY, Coyle EM, Meijer A, Golding H
- Sci Rep 2016 May 27;6:26494
ICOS(+)PD-1(+)CXCR3(+) T follicular helper cells contribute to the generation of high-avidity antibodies following influenza vaccination.
Bentebibel SE, Khurana S, Schmitt N, Kurup P, Mueller C, Obermoser G, Palucka AK, Albrecht RA, Garcia-Sastre A, Golding H, Ueno H
- Sci Rep 2016 Apr 25;6:24897
Production of potent fully human polyclonal antibodies against Ebola Zaire virus in transchromosomal cattle.
Dye JM, Wu H, Hooper JW, Khurana S, Kuehne AI, Coyle EM, Ortiz RA, Fuentes S, Herbert AS, Golding H, Bakken RA, Brannan JM, Kwilas SA, Sullivan EJ, Luke TC, Smith G, Glenn G, Li W, Ye L, Yang C, Compans RW, Tripp RA, Jiao JA
- PLoS Pathog 2016 Apr 21;12(4):e1005554
Antigenic fingerprinting following primary RSV infection in young children identifies novel antigenic sites and reveals unlinked evolution of human antibody repertoires to fusion and attachment glycoproteins.
Fuentes S, Coyle EM, Beeler J, Golding H, Khurana S
- Neurol Neuroimmunol Neuroinflamm 2016 Jan 27;3(1):e196
Patients with MS under daclizumab therapy mount normal immune responses to influenza vaccination.
Lin YC, Winokur P, Blake A, Wu T, Manischewitz J, King LR, Romm E, Golding H, Bielekova B
- J Infect Dis 2015 Oct 15;212(8):1270-8
High affinity H7 head and stalk domain-specific antibody responses to an inactivated influenza H7N7 vaccine after priming with live attenuated influenza vaccine.
Halliley JL, Khurana S, Krammer F, Fitzgerald T, Coyle EM, Chung KY, Baker SF, Yang H, Martínez-Sobrido L, Treanor JJ, Subbarao K, Golding H, Topham DJ, Sangster MY
- MBio 2015 Aug 4;6(4):e01156-15
A simple flow-cytometric method measuring B cell surface immunoglobulin avidity enables characterization of affinity maturation to influenza A virus.
Frank GM, Angeletti D, Ince WL, Gibbs JS, Khurana S, Wheatley AK, Max EE, McDermott AB, Golding H, Stevens J, Bennink JR, Yewdell JW
- J Virol 2015 Mar 15;89(6):3295-307
Post-challenge administration of Brincidofovir protects normal and immune-deficient mice reconstituted with limited numbers of T cells from lethal challenge with IHD-J-Luc vaccinia virus.
Zaitseva M, McCullough KT, Cruz S, Thomas A, Diaz CG, Keilholz L, Grossi IM, Trost LC, Golding H
- PLoS One 2015 Jan 28;10(1):e0115476
Oral priming with replicating adenovirus serotype 4 followed by subunit H5N1 vaccine boost promotes antibody affinity maturation and expands H5N1 cross-clade neutralization.
Khurana S, Coyle EM, Manischewitz J, King LR, Ishioka G, Alexander J, Smith J, Gurwith M, Golding H