Detection of a Low Level and Heterogeneous B Cell Immune Response in Peripheral Blood of Acute Borreliosis Patients With High Throughput Sequencing

• Published in: Frontiers in immunology Vol. 10; p. 1105
• Main Authors: Kirpach, Josiane, Colone, Alessia, Bürckert, Jean-Philippe, Faison, William J, Dubois, Axel R S X, Sinner, Regina, Reye, Anna L, Muller, Claude P
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 16.05.2019
• Subjects: B cell repertoire; Borrelia; Borreliosis; Immunology; Lyme; tetramer; VlsE-C6; Lyme; in vitro stimulation; Borrelia; multicolor flow cytometry; Borreliosis; B cell repertoire; tetramer; VlsE-C6
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2019.01105

The molecular diagnosis of acute Borreliosis is complicated and better strategies to improve the diagnostic processes are warranted. High Throughput Sequencing (HTS) of human B cell repertoires after e.g., Dengue virus infection or influenza vaccination revealed antigen-associated "CDR3 signatures" which may have the potential to support diagnosis in infectious diseases. The human B cell immune response to sensu lato-the causative agent of Borreliosis-has mainly been studied at the antibody level, while less attention has been given to the cellular part of the humoral immune response. There are indications that actively influence the B cell immune response and that it is therefore not directly comparable to responses induced by other infections. The main goal of this study was to identify B cell features that could be used to support diagnosis of Borreliosis. Therefore, we characterized the B cell immune response in these patients by combining multicolor flow cytometry, single B cell receptor (BCR) sequencing, and B cell repertoire deep sequencing. Our phenotyping experiments showed, that there is no significant difference between B cell subpopulations of acute Borreliosis patients and controls. BCR sequences from individual epitope-reactive B cells had little in common between each other. HTS showed, however, a higher complementarity determining region 3 (CDR3) amino acid (aa) sequence overlap between samples from different timepoints in patients as compared to controls. This indicates, that HTS is sensitive enough to detect ongoing B cell immune responses in these patients. Although each individual's repertoire was dominated by rather unique clones, clustering of bulk BCR repertoire sequences revealed a higher overlap of IgG BCR repertoire sequences between acute patients than controls. Even if we have identified a few -associated CDR3aa sequences, they seem to be rather unique for each patient and therefore not suitable as biomarkers.