Supplementary Materials

Supplementary Material for:

Circulating TFH cells, serological memory, and tissue compartmentalization shape human influenza-specific B cell immunity

Marios Koutsakos, Adam K. Wheatley, Liyen Loh, E. Bridie Clemens, Sneha Sant, Simone Nüssing, Annette Fox, Amy W. Chung, Karen L. Laurie, Aeron C. Hurt, Steve Rockman, Martha Lappas, Thomas Loudovaris, Stuart I. Mannering, Glen P. Westall, Michael Elliot, Stuart G. Tangye, Linda M. Wakim, Stephen J. Kent, Thi H. O. Nguyen,* Katherine Kedzierska*

*Corresponding author. Email: kkedz{at}unimelb.edu.au (K.K.); tho.nguyen{at}unimelb.edu.au (T.H.O.N.)

Published 14 February 2018, Sci. Transl. Med. 10, eaan8405 (2018)
DOI: 10.1126/scitranslmed.aan8405

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Gating strategy for circulating ASCs and activated cTFH1 cells.
  • Fig. S2. Specific activation of cTFH1 cells after IIV.
  • Fig. S3. Gating strategy for influenza-specific B cells in PBMC.
  • Fig. S4. Validation of rHA probe staining.
  • Fig. S5. BCR analysis of single rHA+ B cells.
  • Fig. S6. Frequency of IgA+ cells in IgGIgDIgMrHA+ B cells in healthy adults.
  • Fig. S7. Numbers of isotype-specific rHA+ (H3N2/Swi) B cells.
  • Fig. S8. rHA+ B cell kinetics by ELISPOT.
  • Fig. S9. CD20 expression by CD21loCD27+ B cells and ASCs.
  • Fig. S10. Gating strategy for ex vivo live virus ICS.
  • Fig. S11. Vaccination does not induce CD8+ and innate T cell responses.
  • Fig. S12. Gating strategy for T cell phenotyping.
  • Fig. S13. Fold change in influenza-specific B cells during repeated vaccination.
  • Fig. S14. Validating of rHA probes in human tissues.
  • Fig. S15. B cell isotype distributions in paired tissue samples.
  • Table S1. Details of vaccination cohorts.
  • Table S2. Cohorts of human tissues.
  • Table S3. FACS panel for ASCs and cTFH cells.
  • Table S4. FACS panel for influenza-specific T cells after influenza virus infection.
  • Table S5. FACS panel for phenotyping T cells after vaccination.
  • Table S6. FACS panel for influenza-specific B cells in 2015 cohort.
  • Table S7. FACS panel for influenza-specific B cells in 2016 cohort.
  • Table S8. FACS panel for influenza-specific B cells in human tissues.

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Other Supplementary Material for this manuscript includes the following:

  • Table S9 (Microsoft Excel format). Primary data.