Abstract
Plasma cells and their secreted antibodies play a central role in the long‐term protection against chronic viral infection. However, due to experimental limitations, a comprehensive description of linked genotypic, phenotypic, and antibody repertoire features of plasma cells (gene expression, clonal frequency, virus specificity, and affinity) has been challenging to obtain. To address this, we performed single‐cell transcriptome and antibody repertoire sequencing of the murine BM plasma cell population following chronic lymphocytic choriomeningitis virus infection. Our single‐cell sequencing approach recovered full‐length and paired heavy‐ and light‐chain sequence information for thousands of plasma cells and enabled us to perform recombinant antibody expression and specificity screening. Antibody repertoire analysis revealed that, relative to protein immunization, chronic infection led to increased levels of clonal expansion, class‐switching, and somatic variants. Furthermore, antibodies from the highly expanded and class‐switched (IgG) plasma cells were found to be specific for multiple viral antigens and a subset of clones exhibited cross‐reactivity to nonviral and autoantigens. Integrating single‐cell transcriptome data with antibody specificity suggested that plasma cell transcriptional phenotype was correlated to viral antigen specificity. Our findings demonstrate that chronic viral infection can induce and sustain plasma cell clonal expansion, combined with significant somatic hypermutation, and can generate cross‐reactive antibodies.
Highlights
IntroductionThe humoral immune response plays a critical role in defending the host from a variety of pathogens, whereby the terminally www.eji-journal.eu
We performed FACS to isolate BM PCs based on CD138+, TACI+ CD19lo, and B220lo expression [21] and performed single-cell sequencing (scSeq) of transcriptomes and antibody repertoires using the 5’ GEX and V(D)J protocols from 10× Genomics
We were able to recover thousands of cells for each mouse (Supporting information Fig. S1C and S1D), with chronic Lymphocytic choriomeningitis virus (LCMV) infection resulting in the highest proportion of class-switched IgG producing BM PCs and clones (Fig. 1B and C, Supporting information Fig. S1E and S1F): we detected 5337 IgG-expressing BM PCs (Fig. 1B) corresponding to 1653 unique clones (Fig. 1C)
Summary
The humoral immune response plays a critical role in defending the host from a variety of pathogens, whereby the terminally www.eji-journal.eu. Lymphocytic choriomeningitis virus (LCMV) represents a wellstudied pathogen that can establish chronic (dose-dependent) infections in mice. Such infections can eventually be controlled and cleared due to the emergence of plasma cells secreting neutralizing antibodies. It has further been established that neutralizing antibodies of the class-switched IgG isotype are especially crucial for the resolution of chronic LCMV infection [5, 6]. These neutralizing IgG antibodies are directed against the LCMV surface glycoprotein complex (GPC) and only emerge several months after initial infection. LCMV infection results in plasma cells producing antibodies specific to unrelated antigens such as OVA and dinitrophenol (DNP) or autoantigens (i.e., ds and ss DNA, insulin, or thyroglobulin) [8, 9], whereby these autoantibody titers can even be comparable to the concomitantly induced antiviral antibody response [10]
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