Repertoire characterization and validation of gB-specific human IgGs directly cloned from humanized mice vaccinated with dendritic cells and protected against HCMV

Abstract

Human cytomegalovirus (HCMV) causes serious complications to immune compromised hosts. Dendritic cells (iDCgB) expressing granulocyte-macrophage colony-stimulating factor, interferon-alpha and HCMV-gB were developed to promote de novo antiviral adaptive responses. Mice reconstituted with a human immune system (HIS) were immunized with iDCgB and challenged with HCMV, resulting into 93% protection. Immunization stimulated the expansion of functional effector memory CD8 + and CD4 + T cells recognizing gB. Machine learning analyses confirmed bone marrow T/CD4 + , liver B/IgA + and spleen B/IgG + cells as predictive biomarkers of immunization (�87% accuracy). CD8 + and CD4 + T cell responses against gB were validated. Splenic gB-binding IgM-/IgG + B cells were sorted and analyzed at a single cell level. iDCgB immunizations elicited human-like IgG responses with

CAR-T Cells Targeting Epstein-Barr Virus gp350 Validated in a Humanized Mouse Model of EBV Infection and Lymphoproliferative Disease

“…We have shown that HIS mice develop functional human memory CD4 + and CD8 + T cells and fully mature IgG + and IgA + B cells reactive against human herpes antigens. 32 , 37 , 39 We have also shown that challenge of HIS mice with EBV/B95-8 resulted in LPD (perivascular lesions with abundant tumor infiltrating lymphocyte [TILs]) and a pathology recapitulating DLBCL (large and necrotic lesions with fewer TILs). 32 A hallmark of EBV LPD progression toward DLBCL was emergence of highly activated/exhausted PD-1 + CD8 + T cells.…”

Building the Next Generation of Humanized Hemato-Lymphoid System Mice

“…Given that the germinal center is the primary site of B cell-T cell collaboration, antibody affinity maturation and class switching, multiple groups sought to improve LN formation and germinal center development ( 158 ). Adoptive transfer of autologous human DCs lentivirally transduced to express GM-CSF, IFNα, and human cytomegalovirus (HCMV) viral antigen enhances LN development in humanized NRG mice, and by extension promotes T cell-B cell collaboration and the production of neutralizing anti-HCMV antibodies ( 161 , 162 ). Human fetal organ co-transplantation also partially rescues secondary lymphoid organ development.…”

Effective high-throughput isolation of fully human antibodies targeting infectious pathogens

“…We have successfully used and validated the described approach for effective isolation of antigenspecific antibodies against human immunodeficiency virus 1 (HIV-1) 36 , human cytomegalovirus (HCMV) 45 , Ebola virus 34 and SARS-CoV-2 37 , as well as hepatitis C virus (HCV) and Middle East respiratory syndrome coronavirus (MERS-CoV, unpublished data). However, the pipeline is broadly applicable to a plethora of pathogens.…”

“…1, stage I, . We and others have successfully performed single B-cell sorting with numerous pathogen-derived surface proteins to identify pathogen-specific B cells 34,[36][37][38][43][44][45] .…”