Figure 7. Specific decrease in Vγ4/Vδ7 γδT1 cells in the lungs of SAP-deficient mice.
(A) Uniform manifold approximation and projection (UMAP) representation of B6 (left) and B6.Sh2d1a-/- (right) lung γδ T cells, pooled from 3 mice per strain. (B) SAP-dependent variation in lung γδ T cell cluster frequencies. The frequency of each B6 and B6.Sh2d1a-/- cluster as a percentage of all lung γδ T cells is shown. (C) UMAP representation of selected TRG and TRD chain V-segment usage (blue) among B6 and B6.Sh2d1a-/- lung γδ T cells. (D) SAP-dependent decrease in lung γδT1 TRGV4/TRDV7. Normalized TRDV counts in B6 and B6.Sh2d1a-/- lung TRGV4+ γδT1 clusters (C2/C3/C7/C9). (E) SAP-dependent decrease in SLAMF7+ lung Vγ4 T cells. Representative contour plots of CD44 and SLAMF7 expression in B6 (left) and B6.Sh2d1a-/- (right) lung Vγ4 γδ T cells are shown at left. Relative frequencies and counts of CD44+SLAMF7+ lung Vγ4 cells is shown at right. Data represent the cumulative data from five independent experiments, ****p < 0.0001 using unpaired t-test. (F) SAP-dependent skewing of the lung γδT17 TCR repertoire. Amino acid length distributions of B6 and B6.Sh2d1a-/- TRGV4+ CDR3γ (left) and TRDV2+ CDR3δ (right) sequences in the lung Vγ4 γδT17 C1 cluster. The top 10 clonotypes are color-coded and all other clonotypes are shown in gray. Bars represent the frequency of γδ T cells in the c1 cluster. (G) TCR clonotype bubble plots depicting the top 50 lung TRGV4 clonotypes in the C1 γδT17 cluster. Bubble size indicates clonotype frequency as a percentage of C1 Vγ4 (n = 62 B6, 104 B6.Sh2d1a-/-) T cells, and colors correspond to specific TRDV chains. Selected clonotypes are numbered and their corresponding CDR3γ and CDR3δ sequences are displayed below.