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Principal Investigator: Pamela Schwartzberg
Organization: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
Fiscal Year: 2023
Award: $38,953
Funding agency: National Institute of Allergy and Infectious Diseases
Peripheral blood, tonsil tissues, and adenoid tissues were obtained from 110 children who underwent tonsillectomy at Children's National Hospital from late 2020 to 2021 with the goal of: 1) evaluating the prevalence of prior COVID-19 and epidemiologic risk factors for infection; 2) evaluating the nature of SARS-CoV-2-specific immune responses in the oropharyngeal tissue; and 3)comparing the immune profile in the mucosal tissue and blood of COVID-19-convalescent children and children without prior COVID-19. All patients had to be PCR negative for SARS-CoV-2 prior to surgery. We identified 24 participants with evidence of prior COVID-19 through serologic testing and identification of B cells that recognize the RBD region of the SARS-CoV-2 spike protein by flow cytometry. Only 11 of these participants had a prior positive PCR test for SARS-CoV-2 (25-303 days prior to surgery) and were aware of their prior infection. Participants found to be COVID-19-convalescent during this early period of the pandemic were more likely to be Hispanic and to live in a zip code with a poverty level >10% (Mudd et al. Laryngoscope, 2022).
With high dimensional flow cytometry and CITE-seq, we identified SARS-CoV-2-specific memory B cells in the tissue derived from robust geminal center (GC) responses, with evidence of somatic hypermutation and class switching; we also noted overlapping clones in the tonsil and adenoid suggesting migration of cells between these two oropharyngeal tissues. We also found persistent expansion of B and T cell populations involved in the GC and anti-viral response including GC B cells, CD57+PD-1hi GC-Tfh, and CD57+PD1+ CD8 TRM cells, as well as CXCR3+ (IFN-gamma-associated) Tfh populations in COVID-19-convalescent subjects. These changes were most notable in the adenoids, with fewer changes observed in peripheral blood. With CITE-seq, we found more expanded CD8+ and CD4+ T cell clones in convalescent samples, some of which had CDR3 sequences that have been reported to be SARS-CoV-2 specific. Lastly, we found persistent viral RNA by ddPCR as long as 10 months after acute infection. In summary, our results indicate persistent SARS-CoV-2-specific immunity in the upper respiratory tract weeks to months after infection (Xu et al Nat. Immunol. 2023). However, whether intramuscular vaccination elicits similar tissue immunity in the oropharyngeal lymphoid tissue remained an important question.
In the last year, we characterized SARS-CoV-2-specific B cell responses in tonsils, adenoids, and peripheral blood of children who had been immunized with a SARS-CoV-2 mRNA vaccine and compared these samples with those from children infected with SARS-CoV-2 or both infected and immunized. Notably, we found SARS-CoV-2-specific B cells in the tonsils and adenoids of children both post-infection and post-vaccination. These were primarily memory B cells but also a small portion of GC B cells, indicating that immune memory can be found and maintained in the upper respiratory tract after intramuscular vaccination as well as after infection. Nonetheless, vaccination and infection generated different quantities and qualities of responses in both the mucosal tissues and blood, with memory B cells expressing different markers associated with distinct functional responses. Our work provides evidence for tissue-specific immunity post-vaccination, but also highlights differences between SARS-CoV-2 specific B cells generated post-infection and post-vaccination, which will provide important insight for evaluating mucosal vaccines (Xu et al, under revision).
Terms: <0-11 years old><2019 novel corona virus><2019 novel coronavirus><2019-nCoV><2019-nCoV S protein><2019-nCoV spike glycoprotein><2019-nCoV spike protein><Adenoidal structure><Adenoids><Anti-Viral Response><Antiviral Response><Awareness><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Blood><Blood Reticuloendothelial System><Body Tissues><CD183><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CD8><CD8B><CD8B1><CD8B1 gene><CITE sequencing><CITE-seq><CITEseq><CKR-L2><CMKAR3><COVID-19><COVID-19 S protein><COVID-19 infection><COVID-19 negative><COVID-19 spike glycoprotein><COVID-19 spike protein><COVID-19 test><COVID-19 tests><COVID-19 virus><COVID-19 virus infection><COVID19><COVID19 S protein><COVID19 infection><COVID19 negative><COVID19 spike glycoprotein><COVID19 spike protein><COVID19 test><COVID19 tests><COVID19 virus><CV-19><CV19><CXCR3><CXCR3 gene><Cell Body><Cell Locomotion><Cell Migration><Cell Movement><Cells><Cellular Indexing of Transcriptomes and Epitopes by Sequencing><Cellular Migration><Cellular Motility><Chemokine (C-X-C Motif) Receptor 3><Child><Child Youth><Children (0-21)><Class Switching><Class Switchings><Clone Cells><CoV-2><CoV2><Epidemiology><Evaluation><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><G Protein-Coupled Receptor 9><GPR9><Germinal Center><Goals><Helper Cells><Helper T-Cells><Helper T-Lymphocytes><Helper-Inducer T-Cells><Helper-Inducer T-Lymphocyte><Hispanic><Hospitals><IFN-Gamma><IFN-g><IFN-γ><IFNG><IFNγ><IP10><IP10 Receptor><IP10-Mig receptor><IP10-R><Ig Somatic Hypermutation><Immune><Immune Interferon><Immune memory><Immune response><Immunes><Immunity><Immunize><Immunoglobulin Class Switching><Immunoglobulin Class Switchings><Immunoglobulin Somatic Hypermutation><Immunologic Memory><Immunological Memory><Immunological response><Impoverished><In Situ><Inducer Cells><Inducer T-Lymphocytes><Infection><Interferon Gamma><Interferon Type II><Intramuscular><Isotype Switching><Isotype Switchings><LYT3><Laryngoscopes><Lymphatic Tissue><Lymphoid Tissue><Memory B Cell><Memory B-Lymphocyte><Mig Receptor><Mig-R><MigR><Motility><Mucosa><Mucosal Tissue><Mucous Membrane><Nature><Operative Procedures><Operative Surgical Procedures><Oropharyngeal><Oropharynx><Oropharynxs><PD 1><PD-1><PD1><Palatine Tonsil><Participant><Patients><Pharyngeal Tonsil><Population><Poverty><Prevalence><RNA vaccine><RNA-based vaccine><Reporting><Risk Factors><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 S protein><SARS-CoV-2 infection><SARS-CoV-2 negative><SARS-CoV-2 spike glycoprotein><SARS-CoV-2 spike protein><SARS-CoV-2 test><SARS-CoV-2 tests><SARS-CoV2><SARS-CoV2 S protein><SARS-CoV2 infection><SARS-CoV2 spike glycoprotein><SARS-CoV2 spike protein><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Sampling><Serology test><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 CoV 2><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 coronavirus 2 S protein><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome coronavirus 2 spike glycoprotein><Severe acute respiratory syndrome coronavirus 2 spike protein><Severe acute respiratory syndrome related corona virus 2><Site><Structure of germinal center of lymph node><Surgical><Surgical Interventions><Surgical Procedure><T-Cells><T-Lymphocyte><T4 Cells><T4 Lymphocytes><Tissues><Tonsil><Tonsillectomy><Upper respiratory tract><Vaccination><Work><Wuhan coronavirus><acute infection><adaptive immune response><anamnestic reaction><cell motility><cellular indexing of transcriptomes and epitopes by single cell sequencing><corona virus disease 2019><coronavirus disease 2019><coronavirus disease 2019 S protein><coronavirus disease 2019 infection><coronavirus disease 2019 negative><coronavirus disease 2019 spike glycoprotein><coronavirus disease 2019 spike protein><coronavirus disease 2019 test><coronavirus disease 2019 tests><coronavirus disease 2019 virus><coronavirus disease-19><coronavirus disease-19 virus><coronavirus infectious disease-19><epidemiologic><epidemiological><flow cytophotometry><hCoV19><high dimensionality><host response><immune system response><immunoresponse><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus 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