Emerging infectious diseases

We use human monoclonal antibodies (MAbs) in functional and structural studies to understand mechanisms of viral entry, identify target epitopes that provide broad-spectrum protection and develop strategies that prevent viruses from undergoing neutralization escape. Our discoveries of highly conserved structural elements on the envelope glycoproteins of several virus families that serve as cross-neutralizing targets for Mab therapy has allowed us to make therapeutic inroads into the prevention and treatment of a wide range of human pathogenic viruses.

The Marasco Laboratory has also contributed to major advances in three areas of therapeutic Mab development through NIH-NIAID product development programs in Influenza A, Severe Acquired Respiratory Syndrome (SARS), Middle East Respiratory Syndrome coronavirus (MERS-CoV), West Nile virus (WNV) and HIV-1/AIDS infections. We are also actively involved in Biodefense programs and in the discovery of emerging human pathogens.

Influenza

We have identified a highly conserved epitope in the hemagglutinin (HA) stem of Group 1 and 2 human Influenza A viruses and a panel of broad-spectrum neutralizing human antibodies (BnAbs) against this epitope. Our finding of a “universal” vaccine target that can provide broad-spectrum protection against a wide range of influenza A viruses has lead to a paradigm shift in the field of influenza immunotherapy and vaccine development. We have several active discovery programs in this area of investigation.

We have also made important discoveries related to biased VH germline gene usage.  For instance anti-influenza HA stem-directed sBnAb are often encoded by rearranged IGHV1-69 germline genes that alone make contact with HA and prevent virus entry and emergence of escape mutants. In our most recent study, we investigated what structural requirements enable a rearranged IGHV1-69 Ab to become a potent cross-neutralizing antibody. We found that in addition to a critical amino acid triad consisting of a pair of anchor residues in CDR-H2 and a properly positioned CDR-H3 Tyr, distinctive VH-segment substitutions that arise in positions that are distinct from phase I activation-induced cytosine deaminase (AID) somatic hypermutation (SHM) hotspot motifs are often required.  As few as two V-segment SHM can fulfill this role which appears to facilitate the optimal binding of CDR-H2 Phe54 and CHR-H3-Tyr into adjacent hydrophobic pockets in the HA stem.  These studies provide new information on the SHM requirements for IGHV1-69-encoded B cells to produce IGHV1-69 encoded sBnAbs and suggest that there may exist alternative routes to their elicitation by vaccination.

Ying Fu, Zhen Zhang, Jared Sheehan, Yuval Avnir, Callie Ridenour, Thomas Sachnik, Jiusong Sun, M. Jaber Hossain, Li-Mei Chen, Quan Zhu, Ruben O. Donis, Wayne A. Marasco. A broadly neutralizing anti-influenza antibody reveals ongoing capacity of haemagglutinin-specific memory B cells to evolve. Nature Communications, 2016; 7: 12780 DOI: 10.1038/ncomms12780 

Avnir Y, Watson CT, Glanville J, Peterson EC, Tallarico AS, Bennett AS, Qin K, Fu Y, Huang CY, Beigel JH, Breden F, Zhu Q, Marasco WA.  IGHV1-69 polymorphism modulates anti-influenza antibody repertoires, correlates with IGHV utilization shifts and varies by ethnicity. Sci Rep. 2016 Feb 16;6:20842. PMID: 26880249

Avnir Y, Tallarico AS, Zhu Q, Bennett AS, Connelly G, Sheehan J, Sui J, Fahmy A, Huang C-Y, Cadwell G, Bankston LA, McGuire AT, Stamatatos L, Wagner G, Liddington RC, Marasco WA. Molecular Signatures of Hemagglutinin Stem-Directed Heterosubtypic Human Neutralizing Antibodies Against Influenza A Viruses. PLOS Pathogens. 2014 May 1;10(5):e1004103. PMID: 24788925  pdf

In the Media Jan 22, 2013, Boston Globe: Local scientists searching for flu fighters  pdf

Han T, Sui J, Bennett AS, Liddington RC, Donis RO, Zhu Q and Marasco WA. Fine epitope mapping of monoclonal antibodies against hemagglutinin of a highly pathogenic H5N1 influenza virus using yeast surface display. Biochem Biophys Res Commun. 2011 Jun 3;409(2):253-9. PMID: 21569761

Sui J, Sheehan J, Hwang WC, Bankston, LA, Burchett SK, Huang CY, Liddington R, Beigel JH, and Marasco WA. Wide Prevalence of Heterosubtypic Broadly Neutralizing Human Anti-Influenza A Antibodies. Clinical Infectious Diseases. 2011; PMID: 21460314.

Han T and Marasco, WA. Structural basis of influenza virus neutralization. Ann N Y Acad Sci. The Year in Immunology. 2011 Jan;1217(1):178-90. PMID: 21251008.

Wrammert J, Koutsonanos D, Li GM, Edupuganti S, Sui J, Morrissey M, McCausland M, Skountzou I, Hornig M, Lipkin WI, Mehta A, Razavi B, Del Rio C, Zheng NY, Lee JH, Huang M, Ali Z, Kaur K, Andrews S, Amara R, Wang Y, Das S, Yewdell J, Subbarao K, Marasco WA, Mulligan MJ, Compans R, Ahmed R, Wilson PC. Broadly cross-reactive antibodies dominate the acute B cell response against novel 2009 H1N1 influenza virus infection. J Exp Med. 2011 Jan 17;208(1):181-93. PMID: 21220454

Hashem AM, Van Domselaar G, Li C, Wang J, She YM, Cyr TD, Sui J, He R, Marasco WA, Li X. Universal Antibodies against the Highly Conserved Influenza Fusion Peptide Cross-Neutralize Several Subtypes of Influenza A Virus. Biochem Biophys Res Commun. 2010 Dec 10; 403(2):247-51. PMID: 21078301.

Sui J, Hwang WC, Perez S, Wei G, Aird D, Chen L, Santelli E, Stec B, Cadwell G, Ali M, Wan H, Murakami A, Yammanuru A, Han T, Cox N, Bankston LA, Donis RO, Liddington RC and Marasco WA. Structural and Functional Bases for Broad-Spectrum Neutralization of Avian and Human Influenza A Viruses. Nat Struct Mol Biol. 2009 Feb 22. PMID: 19234466 PMCID: PMC2692245.

Huang H, Li W, Sui J, Marasco WA, Choe H, Farzan M. Influenza A virus neuraminidase limits viral superinfection. J. Virol. 2008 May:82(10): 4834-43. Epub 2008 March 5. PMCID: PMC2346733

YouTube videos Attacking InfluenzaNIAID Influenza's Drifting Ways and How Influenza Pandemics Occur

DFCI Spotlight News: Immunology Research:  Developing a Common Influenza Vaccine

Middle East Respiratory Syndrome coronavirus (MERS-CoV)

MERS-CoV was first isolated in the Arabian Peninsula in 2012. Similar to the SARS-CoV, MERS-CoV causes severe respiratory tract infection, often in the lower respiratory tract, occasionally accompanied by renal disease and has shown a death rate of around 42%. We are the first laboratory to report on human neutralizing antibodies (nAbs) against the MERS-CoV. Using a well-characterized nonimmune human Ab-phage library and an unique panning strategy with proteoliposomes and cells immobilized Spike protein, we identify seven human nAbs with subnanomolar/nanomolar binding affinities that block the binding of MERS-CoV Spike protein with its hDPP4 receptor. Binding competition and escape mutant assays indicated that these seven nAbs recognize at least three different epitopes at the receptor binding domain (RBD)-hDPP4 interface. The study of viral evolution under Ab mediated immune pressure indicated that these nAbs can restrict MERS-CoV evolution by driving virus down an escape pathway that predominantly results in a significant cost in viral fitness. These human nAbs might serve as a potent ingredient in a therapeutic antibody cocktail that could be used for the prophylaxis and treatment of MERS. In addition, our studies suggest that the vulnerable spike protein-cellular receptor interface is a preferred target for protective vaccines.

Tang XC, Marasco WA.  Human neutralizing antibodies against MERS coronavirus: implications for future immunotherapy. Immunotherapy. 2015 Jul;7(6):591-4. PMID: 26098703  

Johnson RF, Bagci U, Keith L, Tang X, Mollura DJ, Zeitlin L, Qin J, Huzella L, Bartos CJ, Bohorova N, Bohorov O, Goodman C, Kim do H, Paulty MH, Velasco J, Whaley KJ, Johnson JC, Pettitt J, Ork BL, Solomon J, Oberlander N, Zhu Q, Sun J, Holbrook MR, Olinger GG, Baric RS, Hensley LE, Jahrling PB, Marasco WA. 3B11-N, a monoclonal antibody against MERS-CoV, reduces lung pathology in rhesus monkeys following intratracheal inoculation of MERS-CoV Jordan-n3/2012. Virology. 2016 Mar;490:49-58. PMID: 26828465.

Tang XC, Agnihothram SS, Jiao Y, Stanhope J, Graham RL, Peterson EC, Avnir Y, Tallarico AS, Sheehan J, Zhu Q, Baric RS, Marasco WA. Identification of human neutralizing antibodies against MERS-CoV and their role in virus adaptive evolution. PNAS April 28, 2014 pdf

SARS and other emerging coronaviruses (CoVs)

We were the first laboratory to report on broadly neutralizing human antibodies against the SARS-CoVs that caused a worldwide outbreak in 2003-2004. While we have not seen a repeat of this zoonotic transfer from animals to humans, we have discovered that a large reservoir of CoVs are circulating in bats globally. We have several active discovery programs in this area, including identification of BnAbs against a wide range of human and bat CoVs, the discovery of novel bat-CoV receptors and characterization of the bat antibodyome.

Menachery VD, Yount BL Jr, Sims AC, Debbink K, Agnihothram SS, Gralinski LE, Graham RL, Scobey T, Plante JA, Royal SR, Swanstrom J, Sheahan TP, Pickles RJ, Corti D, Randell SH, Lanzavecchia A, Marasco WA, Baric RS. SARS-like WIV1-CoV poised for human emergence. Proc Natl Acad Sci U S A. 2016 Mar 14.. PMID: 26976607

Menachery V, Yount BL Jr, Debbink K, Agnihothram S, Gralinski L, Plante JA, Graham R, Scobey T, Donadlson EF, Ge XY, Randell SH, Lanzavecchia A, Marasco WA, Shi ZL, Baric R. A SARS-like cluster of circulating bat coronaviruses pose threat for human emergence. Nat Med. 2015 Dec;21(12):1508-13. PMID: 26552008 

Freund N, Roitburd-Berman A, Sui J, Marasco WA, Gershoni JM. Reconstitution of the receptor binding motif of the SARS coronavirus. Protein Eng Des Sel. 2015 Dec;28(12):567-75. PMID: 26487711. 

Huang X, Dong W, Milewska A, Golda A, Qi Y, Zhu Q, Marasco W, Baric R, Sims A, Pyrc K, Li W, Sui J. HCoV-HKU1 Spike protein uses O-acetylated sialic acid as an attachment receptor determinant and employs HE protein as a receptor-destroying enzyme. J Virol. 2015 Jul 15;89(14):7202-13. PMID: 25926653

Sui J, Deming M, Rockx B, Liddington RC , Zhu QK, Baric RS, Marasco WA. Effects of human anti-spike protein receptor binding domain antibodies on severe acute respiratory syndrome coronavirus neutralization escape and fitness. J Virol. 2014 Dec;88(23):13769-80. PMID: 25231316

Sui, J., D. R. Aird, A. Tamin, A. Murakami, M. Yan, A. Yammanuru, H. Jing, B. Kan, X. Liu, Q. Zhu, Q. Yuan, G. P. Adams, W. J. Bellini, J. Xu, L. J. Anderson, and Marasco WA. Broadening of Neutralization Activity to Directly Block a Dominant Antibody-Driven SARS-Coronavirus Evolution Pathway. PLoS Pathogens 2008. 4:e1000197. PMID:18989460 PMCID: PMC2572002

Li W, Sui J, Huang IC, Kuhn JH, Radoshitzky SR, Marasco WA, Choe H, Farzan M. The S proteins of human coronavirus NL63 and severe acute respiratory syndrome coronavirus bind overlapping regions of ACE2. Virology. 2007 Oct 25;367(2):367-74. Epub 2007 Jul 12. PMID: 17631932

Hwang W, Lin Y, Santelli E, Sui J, Jaroszewski L, Stec B, Farzan M, Marasco WA and Liddington RC. Structural basis of neutralization by a human anti-severe SARS Spike Protein Antibody, 80R. J. Biol. Chem. 2006 Nov 10;281(45):34610-34616. PMID: 16954221

Tarnovitski N, Matthews LJ, Sui J, Gershoni JM, Marasco WA. Mapping a Neutralizing Epitope on the SARS Coronavirus Spike Protein: Computational Prediction Based on Affinity-selected Peptides. J Mol Biol 2006;359:190-201. PMID: 16630634

Li W, Zhang C, Moore MJ, Kuhn JH, Wing S-K, Vasilieva N, Sui J, Marasco WA, Guan Y, Choe H and Farzan M. Receptor and Viral Determinants of SARS-Coronavirus Adaptation to Human Antigotension-Converting Enzyme 2. EMBO J 2005;24(8):1634-43. PMCID: PMC1142572

Qin C, Wang J, Wei Q, She M, Marasco WA, Jiang H, Tu X, Zhu H, Ren L, Gao H, Guo L, Huang L, Yang R, Cong Z, Guo L, Wang Y, Liu Y, Sun Y, Duan S, Qu J, Chen L, Tong W, Ruan L, Peimao L, Zhang H, Zhang J, Zhang H, Liu D, Liu Q, He W, and Hong T. SARS Coronavirus-Infected Animal Model in Macaca mulata. J. Pathol. 2005. Jul;206(3):251-9. PMID: 15892035

Sui J, Li W, Roberts A, Matthews LJ, Murakami A, Vogel L, Wong SK, Subbarao K, Farzan M, and Marasco WA. Evaluation of Human mAb 80R for Immunoprophylaxis of SARS by an Animal Study, Epitope Mapping and Analysis of Spike Variants. J.Virol. 2005 May;79(10):5900-6. PMCID: PMC1091676

Sui J, Li W, Murakami A, Tamin A, Matthews LJ, Wong SK, Moore MJ, Tallarico AS, Olurinde M, Choe H, Anderson LJ, Bellini WJ, Farzan M, and Marasco WA. Potent Neutralization of (SARS) Coronavirus by a Human mAb to S1 protein that blocks receptor association. Proc Natl Acad Sci U S A. 2004 Feb 24;101(8):2536-41. PMCID: PMC356985

West Nile Virus (WNV), Zika and other flaviviruses

WNV is the causative agent of West Nile Virus encephalitis, an infection of the central nervous system in man that can result is severe debilitation or even death. There is currently no FDA approved WNV vaccine for man. We have isolated human Mabs that protect against WNV encephalitis in animal models. These Mabs are directed against a highly conserved neutralizing epitope on WNV E-protein. Importantly, these Mabs cross-neutralize a wide range of other human pathogenic flavirivuses including Denge virus serotypes 1-4. We are involved in US army (WRAIR) studies aimed at preventing flavivirus (Denge 1-4 viruses) infection of our special forces and as potentiating agents for Denge virus vaccines.

Sultana H, Foellmer HG, Neelakanta G, Oliphant T, Engle M, Ledozet M, Krishnan MJ, Bonafe N, Anthony K, Marasco WA, Kaplan P, Montgomey RR, Diamond MS, Koski RA and Fikrig E. The fusion loop peptide of the West Nile Virus Envelope Protein is Essential for Pathogenesis and Recognized by a Therapeutic Cross-Reactive Human Monoclonal Antibody. J Immunol. 2009 Jul 1;183(1):650-60. Epub 2009 Jun 17. PMID: 19535627.

Kanai R, Kar K, Anthony K, Gould LH, Ledizet M, Fikrig E, Marasco WA, Koski RA and Modis Y. Crystal structure of West Nile virus envelope glycoprotein reveals viral surface epitopes. J Virol 2006;80:11000-11008. PMCID: PMC1642136

Gould LH, Sui J, Foellmer H, Oliphant T, Wang T, Ledizet M, Murakami A, Noonan K, Lambeth C, Kar K, Anderson JF, de Silva AM, Diamond MS, Koski R, Marasco WA, and Fikrig E. Protection and therapeutic capacity of human single-chain-Fc fusion proteins against West Nile virus. J. Virol. 2005 Dec;79(23):14606-14613. PMCID: PMC1287547

HIV-1/AIDS

The Marasco Lab is developing genetic microbicides that can result in the prevention of mucosal transmission of HIV-1 with emphasis on blocking cervicovaginal transmission. We are also testing new human Mab-based immunotherapies against HIV-1 infection that are designed to reverse T-and B-cell exhaustion and restore a functional immune system so that these immune cells can kill HIV-1-infected cells and eliminate HIV-1 reservoirs. We have developed new “humanized mice” models that are being used to evaluate these strategies.

Kang W, Marasco WA, Tong HI, Byron M, Wu C, Shi Y, Sun S, Sun Y, Lu Y.  Anti-tat hutat2: Fc mediated protection against tat-induced neurotoxicity and HIV-1 replication in human monocyte-derived macrophages. J Neuroinflammation. 2014 Nov 22;11(1):195. PMID: 25416164

Abdel-Motal UM,  Harbison C, Han T, Pudney J, Anderson DJ, Zhu Q, Westmoreland S, Marasco WA.  Prolonged expression of an anti-HIV-1 gp120 minibody to the female rhesus macaque lower genital tract by AAV gene transfer. Gene Therapy. 2014 Sep;21(9):802-10. PMID: 24965083

Braun SE, Taube R, Zhu Q, Wong FE, Murakami A, Kamau E, Dweyer M, QIU G, Daigle J, Carville A, Johnson RP and Marasco WA.  In Vivo Selection of CD4+ T Cells Transduced with a Gamma-Retroviral Vector Expressing a Single-Chain Intrabody Targeting HIV-1 Tat.  Hum Gene Ther.  2012 Jun 26.  PMID: 22734618

Abdel-Motal U, Sarkis PTN, Han T, Pudney J, Anderson DJ, Zhu Q, Marasco WA. Anti-gp120 minibody gene transfer to female genital epithelial cells protects against HIV-1 virus challenge in vitro. PLoS One. 2011, 2011:6(10):e26473.  PMID: 22031835

Herschhorn A, Marasco WA, Hizi A. Antibodies and Lentiviruses that Specifically Recognize a T-cell Epitope Derived from HIV-1 Nef Protein and Presented by HLA-C. J Immunol. 2010 Dec 15; 185(12):7623-32. PMID: 21076072.

Si Z, Vandegraaff N, O'huigin C, Song B, Yuan W, Xu C, Perron M, Li X, Marasco WA, Engelman A, Dean M, Sodroski J. Evolution of a cytoplasmic tripartite motif (TRIM) protein in cows that restricts retroviral infection. Proc Natl Acad Sci U S A 2006 May 9;103(19):7454-59. PMCID: PMC1464360

Masiero S, Del Vecchio C, Gavioli R, Mattiuzzo G, Cusi MG, Micheli L, Gennari F, Siccardi A, Marasco WA, Palu G, Parolin C. T-cell engineering by a chimeric T-cell receptor with antibody-type specificity for the HIV-1 gp120. Gene Ther 2005 Feb;12:299-310 PMID: 15496956

Zhang D, Murakami A, Johnson RP, Sui J, Cheng J, and Marasco WA. Optimization of ex vivo activation and expansion of macaque primary CD4-enriched peripheral blood mononuclear cells for use in anti-HIV immunotherapy and gene therapy strategies. J Acquir Immune Defic Syndr., 2003 Mar:1;32(3):245-54. PMID: 12626883

Bai J, Sui J, Zhu R-Y, Tallarico AStC, Gennari F, Zhang D, and Marasco WA. Inhibition of Tat-Mediated Transactivation and HIV-1 Replication by Human Anti-hCyclin T1 Intrabodies. J. Biol. Chem., 2003;278:1433-1442. PMID: 12401780