Document text
Principal Investigator: Jiang Zhu
Organization: INTERNATIONAL AIDS VACCINE INITIATIVE
Fiscal Year: 2019
Award: $280,575
Funding agency: National Institute of Allergy and Infectious Diseases
Project Summary
Broadly neutralizing antibodies (bnAbs) recognize conserved epitopes on the envelope glycoprotein (Env) of
the human immunodeficiency virus type-1 (HIV-1). Atomic structures of the BG505 SOSIP.664 gp140 trimer
have provided a rational framework for Env-based HIV-1 vaccine design. The Wyatt group recently developed
a cleavage-independent, native flexibly linked (NFL) trimer, which offers a promising alternative for Env-based
vaccine design. Vaccine delivery is a critical issue yet to be addressed in developing well-folded trimers toward
vaccine products. In a recent review, Schiller et al. promoted a virus-like particle (VLP) approach for HIV-1
vaccine delivery. Other delivery systems include mRNA and virus vectors. For the NFL trimer, it remains
unclear which delivery system will produce the most robust immune response to enable protection against HIV-
1 infection. We hypothesize that a quantitative readout of B-cell response will facilitate rational evaluation of
vaccine candidates since a robust cross-neutralizing antibody response is expected for an effective HIV-1
vaccine. Next-generation sequencing (NGS) has been widely used to study the diversity and maturation of
bnAbs. We have developed a series of novel NGS technologies for the analysis of dynamic B-cell responses in
natural infection, animal immunization, and human vaccine trials. In Project 3 of this HIVRAD application, we
will investigate the Env-specific B-cell response to four vaccines based on the NFL trimer and different delivery
systems in various animal models – inbred mice, VelocImmune mice, rabbits and non-human primates (NHPs).
In Aim 1, we will deep sequence the B-cell response of inbred mice, VelocImmune mice, and rabbits immunized
with four vaccines. We will characterize the basic repertoire properties such as germline gene usage, degree of
SHM, and CDR3 length and identify distinctive repertoire patterns associated with each vaccine. For each
animal model, we will compare the B-cell responses to four vaccines. We will compare the B-cell responses in
hu-mice and human vaccine trials. In Aim 2, we will deep sequence the B-cell response in NHPs immunized
with four vaccines. Similarly, we will characterize the basic repertoire properties and identify distinctive patterns
associated with four vaccines in the NHP model. We will compare the B-cell responses to four vaccines and
compare the B-cell responses in NHPs and human vaccine trials. In Aim 3, we will study the developmental
pathways of Env-specific B-cell lineages elicited by four vaccines. We will trace the lineages of monoclonal
antibodies (mAbs) identified by single B-cell sorting (Project 2, Wyatt) and microfluidics-based single-cell
analysis (Project 3, Zhu) in NGS repertoires. We will investigate whether the vaccine-elicited neutralizing mAbs
resemble known bnAbs and their developmental pathways. Project 3, together with Project 1 (RNA and VSV
vector), Project 2 (NFL trimer, VLP, and serum Ab analysis), and two Cores on in-vivo study and gene
expression profiling, will constitute a comprehensive HIVRAD program towards an effective HIV-1 vaccine.
Terms: <AIDS Virus><ATGN><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Address><Affinity Chromatography><Animal Model><Animal Models and Related Studies><Animals><Antibodies><Antibody Response><Antibody titer measurement><Antigenic Determinants><Antigens><B blood cells><B cell><B cell repertoire><B cells><B-Cells><B-Lymphocytes><B-cell><Binding Determinants><Blood><Blood Reticuloendothelial System><Blood Serum><Cell Body><Cell Isolation><Cell Lineage><Cell Segregation><Cell Separation><Cell Separation Technology><Cells><Clinical Treatment Moab><Data><Development><Domestic Rabbit><Epitopes><Evolution><Expression Signature><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profile><Gene Expression Profiling><General Viruses><Genes><Glycoproteins><HIV><HIV-1><HIV-I><HIV1><Human><Human Immunodeficiency Virus Type 1><Human Immunodeficiency Viruses><Human immunodeficiency virus 1><Immune response><Immunization><Immunize><Immunologic Sensitization><Immunologic Stimulation><Immunological Sensitization><Immunological Stimulation><Immunological response><Immunostimulation><Inbred Mouse><Individual><Infection><Investigators><LAV-HTLV-III><Length><Link><Lymphadenopathy-Associated Virus><Macaca><Macaca mulatta><Macaque><Messenger RNA><Mice><Mice Mammals><Microfluidics><Modeling><Modern Man><Monoclonal Antibodies><Murine><Mus><NGS Method><NGS system><Non-Polyadenylated RNA><Oryctolagus cuniculus><Outcome><Pathway interactions><Pattern><Phase><Property><RNA><RNA Gene Products><Rabbits><Rabbits Mammals><Reporting><Research Personnel><Researchers><Rhesus><Rhesus Macaque><Rhesus Monkey><Ribonucleic Acid><Series><Serum><Structure><System><Technology><Transcript Expression Analyses><Transcript Expression Analysis><VSV><Vaccination><Vaccine Design><Vaccines><Vesicular Stomatitis Virus><Vesicular stomatitis Indiana virus><Virus><Virus-HIV><Virus-like particle><affinity purification><antibody titering><base><cell sorting><design><designing><develop a vaccine><development of a vaccine><developmental><flexibility><flexible><gene expression analysis><gene expression assay><gene expression pattern><gene expression signature><host response><humanized mice><humanized mouse><immunogen><immunoresponse><in vivo><longitudinal analysis><mAbs><mRNA><model of animal><model organism><mouse model><murine model><neutralizing antibody><neutralizing mAb><neutralizing monoclonal antibodies><new technology><next gen sequencing><next generation sequencing><nextgen sequencing><non-human primate><nonhuman primate><novel><novel technologies><pathway><programs><response><single cell analysis><transcriptional profiling><transcriptional signature><vaccination study><vaccination trial><vaccine candidate><vaccine delivery><vaccine development><vaccine evaluation><vaccine formulation><vaccine screening><vaccine study><vaccine testing><vaccine trial><vector><virus-like nanoparticles><viruslike particle><µfluidic>