Supplementary Materials

Supplementary Material for:

Direct measurement of T cell receptor affinity and sequence from naïve antiviral T cells

Shu-Qi Zhang, Patricia Parker, Ke-Yue Ma, Chenfeng He, Qian Shi, Zhonghao Cui, Chad M. Williams, Ben S. Wendel, Amanda I. Meriwether, Mary Alice Salazar, Ning Jiang*

*Corresponding author. Email: jiang{at}austin.utexas.edu

Published 1 June 2016, Sci. Transl. Med. 8, 341ra77 (2016)
DOI: 10.1126/scitranslmed.aaf1278

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Nonspecific adhesion on primary CD8+ T cells is negligible and not correlated with pMHC site density.
  • Fig. S2. Multivalency of pMHC does not affect 2D affinity measurement.
  • Fig. S3. 2D affinity kinetic curves for three additional CD8+ T cell clones with varying affinities.
  • Fig. S4. Streptamer staining is fully reversible, and streptamers stain with the same intensity as conventional tetramers.
  • Fig. S5. Estimating the error of single-cell 2D affinity measurements.
  • Fig. S6. Peptide titration curves of 15 HCV-specific CD8+ T cell clones.
  • Fig. S7. Conventional 2D affinity measurement on HCV-specific CD8+ T cell clones shows similar correlations as single-cell 2D affinity.
  • Fig. S8. Gating strategy for sorting HCV-streptamer+ T cells.
  • Fig. S9. Ratio of high-affinity/low-affinity HCV-specific CD8+ T cells based on functional potential.
  • Fig. S10. Amplification efficiency of affinity-tested cells.
  • Fig. S11. TRAV and TRBV usage of affinity-tested primary HCV-specific CD8+ T cells by donor.
  • Table S1. Comparison of single-cell 2D affinity derived from a primary T cell and conventional 2D affinity derived from the T cell’s in vitro expanded clone.
  • Table S2. Single-cell 2D affinity and SPR 3D affinity for the native and peptide variants of NY-ESO-1 against 1G4 TCR.
  • Table S3. Median 2D affinity and ratio of high/low 2D affinity from primary HCV-specific CD8+ T cells.
  • Table S4. Single-cell 2D affinity and correlated TCRα and TCRβ CDR3 sequences.
  • Reference (29)

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