Figure 6.
Re-immunization of individuals with persistent GCs promotes the development of affinity-matured PBs. (A) Schematic of study design. Three participants (04, 05, 11) received the 2019 QIV intramuscularly. Blood, FNAs of the ipsilateral axillary lymph nodes (LN), and bone marrow (BM) were collected prior to vaccination (week 0) and at the indicated weeks after vaccination. (B) ELISpot quantification of 2019 QIV–binding IgG-secreting PBs in blood at weeks 0, 1, and 2 after vaccination for three participants. (C) Median IGHV gene mutation frequency of QIV-binding PB clonal groups found after vaccination with 2018 and 2019 QIV. Lines between points indicate the clone was identified in the PB compartment after both vaccinations. Median IGHV nucleotide mutation frequency of all clonal groups is indicated above each column and the number of clones (n) is indicated in the bottom right of each panel (04, n = 73; 05, n = 122; 11, n = 65). P values were determined by paired t test. (D) Clustering was visualized via UMAP of B cells from blood, LN, and BM scRNA-seq samples in participant 05. Each dot represents a cell, colored by phenotype as defined by transcriptomic profiles. Naïve B cells (gold), PBs (red), ABCs (green), GC B cells (blue), LNPCs (red), RMBs (lavender), and plasma cells (PC, red) populations are pooled from all time points (first panel). QIV-specific cells at each week after vaccination are colored as described. (E) Median IGHV gene mutation frequency of QIV-binding GC B cell clonal groups found after vaccination with 2018 and 2019 QIV. Lines between points indicate the clone was identified in the GC B cell compartment after both vaccinations. Median IGHV nucleotide mutation frequency of all clonal groups is indicated above each column and the number of clones (n = 135) is indicated in the bottom right. P values were determined by paired t test. N.D., no data. See also Fig. S5 and Tables S3, S4, S5, S6, and S7.

Re-immunization of individuals with persistent GCs promotes the development of affinity-matured PBs. (A) Schematic of study design. Three participants (04, 05, 11) received the 2019 QIV intramuscularly. Blood, FNAs of the ipsilateral axillary lymph nodes (LN), and bone marrow (BM) were collected prior to vaccination (week 0) and at the indicated weeks after vaccination. (B) ELISpot quantification of 2019 QIV–binding IgG-secreting PBs in blood at weeks 0, 1, and 2 after vaccination for three participants. (C) Median IGHV gene mutation frequency of QIV-binding PB clonal groups found after vaccination with 2018 and 2019 QIV. Lines between points indicate the clone was identified in the PB compartment after both vaccinations. Median IGHV nucleotide mutation frequency of all clonal groups is indicated above each column and the number of clones (n) is indicated in the bottom right of each panel (04, n = 73; 05, n = 122; 11, n = 65). P values were determined by paired t test. (D) Clustering was visualized via UMAP of B cells from blood, LN, and BM scRNA-seq samples in participant 05. Each dot represents a cell, colored by phenotype as defined by transcriptomic profiles. Naïve B cells (gold), PBs (red), ABCs (green), GC B cells (blue), LNPCs (red), RMBs (lavender), and plasma cells (PC, red) populations are pooled from all time points (first panel). QIV-specific cells at each week after vaccination are colored as described. (E) Median IGHV gene mutation frequency of QIV-binding GC B cell clonal groups found after vaccination with 2018 and 2019 QIV. Lines between points indicate the clone was identified in the GC B cell compartment after both vaccinations. Median IGHV nucleotide mutation frequency of all clonal groups is indicated above each column and the number of clones (n = 135) is indicated in the bottom right. P values were determined by paired t test. N.D., no data. See also Fig. S5 and Tables S3, S4, S5, S6, and S7.

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