Establishing a positive correlation between CD5 expression and self-reactivity in BCR WT B-1 cells. Establishing a positive correlation between CD5 expression.

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Establishing a positive correlation between CD5 expression and self-reactivity in BCR WT B-1 cells. Establishing a positive correlation between CD5 expression and self-reactivity in BCR WT B-1 cells. (A) Flow cytometric analysis of peritoneal cavity (PerC) B cells 16 weeks after transplantation of E14.5 FL or ABM Lin−Sca1+cKit+ (LSK) HSPCs into lethally irradiated CD45.1+CD45.2+ congenic recipients. Lineage panel for E14.5 FL LSK: Ter119−B220−Gr1−CD3e−; ABM LSK: Ter119−B220−Gr1−CD11b−CD3e−; PerC B cells: Ter119−Gr1−CD3e−. (B) CD5neg, CD5low, CD5int, and CD5hi gating strategy for WT adult PerC B-1 cells (Lin−CD19+CD43+CD23−). (C) Histogram overlay showing post sort analysis of the populations defined in (B). (D) Relative distribution of IgHM CDR3 sequences as determined by high-throughput VDJseq of the indicated populations. (E) Gini index of IgHM CDR3 sequence reads in sorted CD5neg, CD5low, CD5int, and CD5hi populations (left to right). (F) Stacked bar graph indicates the combined frequency of IGHV-11 (white) and IGHV-12 (gray) containing IgHM CDR3 sequence reads. (G) Frequency of PtC liposome–reactive cells in the indicated B-1 populations as assessed by FACS. *P ≤ 0.05, ***P ≤ 0.001, ****P ≤ 0.0001 (n = 9 from three experiments). (H) Frequency of CDR3 containing ≥1 N-nucleotide additions at the N1 and N2 junctions combined. VDJseq data (B to F and H) are representative of two biological and technical replicates. Stijn Vanhee et al. Sci. Immunol. 2019;4:eaax4453 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works