Molecular and functional characterization of antibodies from PfCSP-reactive memory B cells and plasmablasts. Molecular and functional characterization.

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Presentation transcript:

Molecular and functional characterization of antibodies from PfCSP-reactive memory B cells and plasmablasts. Molecular and functional characterization of antibodies from PfCSP-reactive memory B cells and plasmablasts. (A) PfSPZ-CVac immunization scheme (9). Blood samples were taken at day 7 (d7) after Pf exposure I, II, and III. (B to F) Ig gene sequence analysis of PfCSP-reactive memory B cells and plasmablasts from all (B to D) or individual (E and F) donors, as indicated. (B) Mean and SEM IGH isotype frequency. (C) IGHV absolute SHM counts and mean (red lines). (D) Mean and SEM ratio of replacement (R) to silent (S) SHM in CDRs compared with FWRs. (E) Clonal expansion. (F) Clonal expansion and diversity (day 7 after exposure III). Individual cell clusters are colored; unique sequences are white (top). Corresponding subsampling analysis (bottom). Dots represent independent analyses. Red lines show mean. Blue lines indicate observed values as in (E). (G to J) Recombinant monoclonal PfCSP memory B cell antibodies from eight donors at day 7 after Pf exposure I, II, and III. Representative optical density (OD) (G) and AUC values from PfCSP (H) and NANP (I) ELISA. Red and blue lines in (G) indicate positive (2A10) and negative (mGO53) controls. Antibody sporozoite inhibitory activity in traversal assays (J). (K to M) Affinity, frequencies (K), and kinetic on-rate (L) and off-rate (M) constants of NANP-reactive antibodies at day 7 after inoculation II and III. (H to J, L, and M) Red lines indicate mean. Data are representative of at least two independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, two tailed Mann-Whitney test (H to J, L, and M) and χ2 test (K). n numbers are provided in tables S1 and S2. ns, not significant. Rajagopal Murugan et al. Sci. Immunol. 2018;3:eaap8029 Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works