Journal: bioRxiv

Article Title: Single-cell sequencing reveals clonally expanded plasma cells during chronic viral infection produce virus-specific and cross-reactive antibodies

doi: 10.1101/2021.01.29.428852

Figure Lengend Snippet: Experimental overview of immunization schemes and antibody repertoire statistics. A. Serial immunization scheme with ovalbumin (OVA) and subsequent bone marrow plasma cell (BM PC) isolation using CD138+ magnetic beads. Subsequent antibody repertoire libraries were prepared using the 5’ single-cell immune profiling kit from 10X Genomics. B. Serial immunization scheme with the extracellular region of human tumor necrosis factor receptor 2 (TNFR2) and subsequent bone marrow plasma cell (BM PC) isolation using flow cytometry. BM PCs were sorted based on CD138 hi , TACI hi , B220 lo , CD19 lo . Subsequent antibody repertoire and transcriptome libraries were prepared using the 5’ single-cell immune profiling kit from 10X Genomics. C. The total number of cells recovered in each infected or immunized mouse. Only cells containing exactly one variable heavy (V H ) and variable light (V L ) chain were considered. D. The total number of clones recovered in each infected or immunized mouse. Clone was determined by grouping those B cells containing identical CDRH3+CDRL3 amino acid sequences. Only cells containing exactly one variable heavy (V H ) and variable light (V L ) chain were considered. E-F. Fraction of clones (E) and cells (F) corresponding to a given isotype in each infected or immunized mouse. Clone was determined by grouping those B cells containing identical CDRH3+CDRL3 amino acid sequences. Only cells containing exactly one variable heavy (V H ) and variable light (V L ) chain were considered.

Article Snippet: For the OVA immunized mouse, BM PCs were isolated using a CD138+ plasma cell isolation kit (Miltenyi, 130-092-530) following the instructions of the manufacturer.

Techniques: Clinical Proteomics, Isolation, Magnetic Beads, Flow Cytometry, Infection, Clone Assay

Journal: bioRxiv

Article Title: Single-cell sequencing reveals clonally expanded plasma cells during chronic viral infection produce virus-specific and cross-reactive antibodies

doi: 10.1101/2021.01.29.428852

Figure Lengend Snippet: Differentially expressed genes between the bone marrow plasma cells from mice either chronically infected with LCMV or immunized with human TNFR2. (A) Normalized expression of plasma cell genes Sdc1 ( CD138 ), Tnfrsf13b ( TACI ), Slamf7 , and Ptprc ( B220 ) for either mice infected with LCMV or immunized with human TNFR2. (B) Uniform manifold approximation project (UMAP) plots showing normalized gene expression for selected genes.

Article Snippet: For the OVA immunized mouse, BM PCs were isolated using a CD138+ plasma cell isolation kit (Miltenyi, 130-092-530) following the instructions of the manufacturer.

Techniques: Clinical Proteomics, Infection, Expressing, Gene Expression