ZIKV-specific CD4⁺T cell TCR diversity.

(A) TCRβ CDR3 region diversity among CD4+T cells responding to ZIKV-specific epitopes. Wild type C57BL/6J mice (n = 4) were infected with 106 FFU ZIKV. At 10 DPI, splenocytes were harvested and stimulated with the indicated peptide or anti-CD3 in the presence of brefeldin A as described in Fig 4. CD19-, CD8-, CD4+, IFNγ+ cells were sorted separately for each peptide and DNA was extracted for multiplexed amplification of the TCRβ CDR3 region. Unique CDR3 sequences (clonotypes) are enumerated for cells responding to each peptide for each mouse. Each line represents the number of unique TCRβ clonotypes identified from individual peptide or αCD3 stimulated T cells from four mice. (B) Representation of unique TCRβV-J recombinations in cells responding to each stimulation. Circos graphs were generated to display the relative frequency of each specific TCRβV-J join detected in CD4+T cells responding to each epitope. The width of each ribbon is proportional to the number of unique clonotypes that share those specific TCRβV-J segments.