Figure 3
Figure 3. Contraction of Tem TCR diversity but not Tnaive TCR diversity in CMV-reactivating patients. (A) Representative flow cytometry analysis illustrates the purity of CD8+ Tnaive and CD8+ Tem cells before (left plot) and after (middle and right plots) cell sorting. (B) Shown are representative graphs of the CD8+ naive (left column) and CD8+ Tem (right column) TCR landscape (showing frequencies of V and J gene combinations detected through deep sequencing) from 1 −CMV (blue) and 1 +CMV (red) patient. (C) Clonality of PBMC, CD8+ Tnaive, and CD8+ Tem are shown for −CMV and +CMV patients as measured by the inverse of the normalized Shannon entropy number. All data are mean ± SEM. *P ≤ .05; Wilcoxon rank-sum test. (D) The clonality of Tnaive and Tem CD8+ cells for each patient is compared with the percentage of Tem CD8+ cells detected via flow cytometry in each patient. Linear regression (R2 = 0.506) shows a correlation between expansion of Tem and increased Tem clonality in +CMV patients (red). No such relationship could be detected in −CMV patients (blue). (E) TCR diversity of CD8+ Tnaive and CD8+ Tem are shown for −CMV and +CMV patients as measured by the Gini coefficient.44 All data are mean ± SEM. *P ≤ .05; Wilcoxon rank-sum test. #Sorting purity of Tnaive from patient 001-008 could not be confirmed due to low yield.

Contraction of Tem TCR diversity but not Tnaive TCR diversity in CMV-reactivating patients. (A) Representative flow cytometry analysis illustrates the purity of CD8+ Tnaive and CD8+ Tem cells before (left plot) and after (middle and right plots) cell sorting. (B) Shown are representative graphs of the CD8+ naive (left column) and CD8+ Tem (right column) TCR landscape (showing frequencies of V and J gene combinations detected through deep sequencing) from 1 −CMV (blue) and 1 +CMV (red) patient. (C) Clonality of PBMC, CD8+ Tnaive, and CD8+ Tem are shown for −CMV and +CMV patients as measured by the inverse of the normalized Shannon entropy number. All data are mean ± SEM. *P ≤ .05; Wilcoxon rank-sum test. (D) The clonality of Tnaive and Tem CD8+ cells for each patient is compared with the percentage of Tem CD8+ cells detected via flow cytometry in each patient. Linear regression (R2 = 0.506) shows a correlation between expansion of Tem and increased Tem clonality in +CMV patients (red). No such relationship could be detected in −CMV patients (blue). (E) TCR diversity of CD8+ Tnaive and CD8+ Tem are shown for −CMV and +CMV patients as measured by the Gini coefficient.44  All data are mean ± SEM. *P ≤ .05; Wilcoxon rank-sum test. #Sorting purity of Tnaive from patient 001-008 could not be confirmed due to low yield.

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