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Principal Investigator: Susan Moir
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2024
Award: $1,480,637
Funding agency: National Institute of Allergy and Infectious Diseases
In 2024, we pursued studies encompassing three major themes: 1) investigating B-cell function in lymphoid tissues of PLWH; 2) exploring the clinical benefits of HIV neutralizing antibodies; and 3) contributing to studies on B cells in non-HIV diseases.
1) In 2024, we began performing spatial transcriptomic and phenotypic profiling of lymphoid tissues of PLWH who develop bNAbs with goal of addressing how lymphoid architecture and cellular interactions affect this process. This project began with the screening for bNAbs in serum collected from PLWH who underwent lymph node biopsies under a clinical research protocol, ClinicalTrials.gov identifier NCT00001316. We identified over 100 pairs of lymph node tissue sections fixed in formalin and embedded in paraffin (FFPE) with matching serum samples that were collected over a 30-year period from PLWH who were not receiving antiretroviral therapy (ART), and we screened for the presence of bNAbs with a global panel of HIV envelope-pseudotyped viruses developed for this purpose. In partnership with investigators in NCI and NICHD, we successfully performed a pilot study on the 10X Genomics Visium platform, demonstrating both feasibility and potential for discerning differences in the spatial landscape of specimens collected from four PLWH, two with and two without bNAb. However, when this approach was expanded to additional pairs of specimens that differed by bNAb profile, we found that the RNA in our older specimens, which are most samples given that they predate the availability of and recommendations to rapidly initiate ART, was too degraded for transcriptomics. In lieu, we have turned to the co-detection by indexing (CODEX) imaging platform to perform multiplexed phenotyping with help from the NCI Collaborative Protein Technology Resource (CPTR). This is an approach that is more likely to succeed given that protein is more stable than RNA in FFPE specimens.
2) In follow-up studies with longtime LIR collaborator Dr. Tae-Wook Chun on the role of broad and potent anti-HIV antibodies (bNAbs) in suppressing HIV in the absence of ART, we have been exploring how B cells contribute to the HIV-neutralizing antibody repertoire and how the virus evolves to evade the antibody response. To pursue these studies, we have identified and screened plasma from three groups of PLWH enrolled in several of our LIR observational clinical research protocols: 1) a small group of PLWH whose plasma HIV viremia appears to be controlled by autologous antibodies; 2) a small group of PLWH who develop bNAbs but whose virus is resistant to them; and 3) a large group of PLWH with varying profiles of resistance to well characterized bNAbs. From the first group, we identified an early-treated PLWH who stopped ART after 6.5 years as part of a study that included an analytical teatment interruption phase and whose IgG serum antibodies likely contributed to the viral suppression he maintained in the absence of ART for almost four years (after which he experienced superinfection with virus that was resistant to his prior antibodies). We cultured his B cells from the mid-point of his ART-free period in conditions of limiting dilution that allowed for the identification and cloning of antibodies that suppressed his virus in vitro. The antibodies originated from two related B-cell lineages and accounted for all the neutralizing activity present in the supernatants of his B-cell cultures. The cloned antibodies carried few somatic mutations yet were extremely potent in neutralizing autologous virus, although with limited activity against other HIV strains. As such, these are not bNAbs. In partnership with researchers at the University of Maryland, we have begun to characterize the properties of these two families of antibodies through mutational and functional experiments as well as visualization by cryo-electron microscopy. These analyses have revealed that the cloned antibodies target the V3/C4 region of gp120 and that their binding to cell-surface expressed autologous viral envelope was enhanced in the presence of CD4. These properties may explain the potency of the antibodies and why they may have been able to prevent viral escape and rebound for an extended period.
3) In 2024, we contributed to several collaborative efforts on HIV and other diseases that affect B cells. In partnership with our LIR colleague, Dr. Richard Davey, we used a flow cytometry panel that we had designed for probing B-cell responses to SARS-CoV-2 mRNA vaccination to evaluate B-cell responses to a recombinant Vesicular Stomatitis Virus-based vaccine containing the Zaire ebolavirus (EBOV) glycoprotein. The participants were enrolled in PREPARE (ClinicalTrials.gov Identifier: NCT02788227), a study that evaluated the safety and immunogenicity of the EBOV vaccine with a placebo-controlled arm for evaluating immune responses to a booster dose. Our contribution has been to measure B-cell responses to primary and secondary doses of the EBOV vaccine and to evaluate how different vaccine platforms can modulate memory B-cell responses. We found that the first dose of the EBOV vaccine elicited a predominantly IgM B-cell response, consistent with the reported IgM dominant antibody response. The booster dose induced a short-lived B cell response driven by IgG, again similar in isotype to the antibody response, yet the antibody response was significantly stronger in the boosted than placebo group at end of study while the B-cell response was not. Thus, this vaccine platform may favor the establishment of long-live plasma cells (responsible for serologic antibodies) over memory B cells, which is the opposite to the outcome observed for SARS-CoV-2 mRNA vaccines where antibody titers wane more rapidly than B-cell responses. We also contributed to NIAID studies on viral reservoirs and therapeutic interventions in PLWH (LIR, with Dr. Tae-Wook Chun); the role of metalloproteases in the life cycle of HIV (Dr. Peter Sun); and we contributed clinical specimens and our B-cell expertise to a study on the immunologic signatures of human health (Drs. Rachel Sparks and John Tsang). The remainder of our collaborative efforts have focused on COVID-19, details of which can be found in the annual report on COVID-19 research.
Terms: <19S Gamma Globulin><2019 novel corona virus><2019 novel coronavirus><2019-nCoV><7S Gamma Globulin><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Address><Affect><Annual Reports><Antibodies><Antibody Repertoire><Antibody Response><Antibody Therapy><Antibody titer measurement><Antigens><Architecture><Autologous><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Binding><Blood Plasma><Blood Plasma Cell><Blood Serum><COVID-19><COVID-19 virus><COVID19 virus><CV-19><Cell Culture Techniques><Cell Function><Cell Lineage><Cell Physiology><Cell Process><Cell surface><Cellular Function><Cellular Immune Function><Cellular Physiology><Cellular Process><Clinical><Clinical Research Protocols><Cloning><CoV-2><CoV2><Coronavirus Infectious Disease 2019><Cryo-electron Microscopy><Cryoelectron Microscopy><Detection><Development><Disease><Disease Progression><Disorder><Dose><Early identification><Ebola><Ebola Vaccines><Ebola Virus Vaccines><Electron Cryomicroscopy><Emerging Communicable Diseases><Emerging Infectious Diseases><Engineering / Architecture><Enrollment><Family><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Follow-Up Studies><Followup Studies><Formalin><Genetic Alteration><Genetic Change><Genetic defect><Genomics><Glycoproteins><Goals><HIV><HIV Antibodies><HIV Envelope Glycoprotein gp120><HIV Envelope Protein gp120><HIV Infections><HIV env Protein gp120><HIV envelope><HIV envelope protein><HIV-Associated Antibodies><HTLV-III Antibodies><HTLV-III Infections><HTLV-III gp120><HTLV-III-LAV Antibodies><HTLV-III-LAV Infections><Health><Human><Human Immunodeficiency Viruses><Human T-Lymphotropic Virus Type III Antibodies><Human T-Lymphotropic Virus Type III Infections><IgG><IgM><Immune response><Immunochemical Immunologic><Immunoglobulin G><Immunoglobulin M><Immunologic><Immunological><Immunological response><Immunologically><Immunologics><Immunology><In Vitro><Interruption><Investigators><Knowledge><LAV Antibodies><LAV-HTLV-III><Life Cycle><Life Cycle Stages><Lymph Node Tissue><Lymphadenopathy-Associated Antibodies><Lymphadenopathy-Associated Virus><Lymphatic Tissue><Lymphoid><Lymphoid Tissue><Maryland><Measures><Memory B Cell><Memory B-Lymphocyte><Metallopeptidases><Metalloproteases><Metalloproteinases><Microbial Superinvasion><Modern Man><Molecular Interaction><Mutation><NIAID><NICHD><National Institute of Allergy and Infectious Disease><National Institute of Child Health and Human Development><National Institute of Children's Health and Human Development><Non-Polyadenylated RNA><Outcome><Paraffin><Paraffin waxes and Hydrocarbon waxes><Participant><Pathogenesis><Persons><Phase><Phenotype><Pilot Projects><Placebo Control><Plasma><Plasma Cells><Plasma Serum><Plasmacytes><Process><Property><Proteins><RNA><RNA Gene Products><RNA Stability><RNA immunization><RNA vaccination><RNA vaccine><RNA-based vaccine><Recombinants><Recommendation><Regulation><Reporting><Research><Research Personnel><Research Resources><Research Specimen><Researchers><Resistance><Resistance profile><Resistant profile><Resources><Reticuloendothelial System, Serum, Plasma><Ribonucleic Acid><Role><SARS><SARS corona virus 2><SARS coronavirus disease><SARS-CO-V2><SARS-COVID-2><SARS-CoV disease><SARS-CoV-2><SARS-CoV2><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Safety><Sampling><Serology><Serum><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome CoV disease><Severe Acute Respiratory Syndrome coronavirus disease><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome related corona virus 2><Somatic Mutation><Specimen><Subcellular Process><Technology><Therapeutic Intervention><Time><Universities><VSV><Vaccination><Vaccine against ebola><Vaccines><Vesicular Stomatitis Virus><Vesicular stomatitis Indiana virus><Viral><Viral reservoir><Viremia><Virus><Virus reservoir><Virus-HIV><Visualization><Wuhan coronavirus><ZEBOV><Zaire Ebola virus><Zaire ebolavirus><antibody based therapies><antibody titering><antibody treatment><antibody-based therapeutics><antibody-based treatment><antiretroviral therapy><antiretroviral treatment><arm><booster dose><booster shot><booster vaccine><cell culture><cell cultures><coronavirus disease 2019><coronavirus disease 2019 virus><coronavirus disease-19><coronavirus disease-19 virus><coronavirus infectious disease-19><cryo-EM><cryoEM><cryogenic electron microscopy><design><designing><developmental><enroll><experience><experiment><experimental research><experimental study><experiments><flow cytophotometry><genome mutation><gp120><gp120 ENV Glycoprotein><gp120(HIV)><hCoV19><host response><imaging platform><immune function><immune system response><immunogen><immunogenicity><immunoresponse><indexing><intervention therapy><life course><lymph node biopsy><mRNA immunization><mRNA vaccination><mRNA vaccine><mRNA-based vaccine><nCoV2><neutralizing antibody><peripheral blood><pilot study><placebo controlled><placebo group><plasmocyte><prevent><preventing><resistant><response><screening><screenings><sham group><social role><somatic variant><super infection><superinfection><transcriptomics><vaccine boost><vaccine platform><viraemia><viral sepsis><virusemia>