FIGURE 5. The TREC-enriched CD8⁺CD103⁺ naive CD8⁺ T cell subset, but not the TREC-enriched CD4⁺CD31⁺ naive CD4⁺ T cell subset, increased at week 24 after treatment in both Leuprolide treated and untreated patients.

Multiparameter flow cytometric analysis of CD4⁺CD45RA⁺RO⁻CD31⁺ and CD8⁺CD45RA⁺RO⁻CD27⁺CD103⁺ subsets was performed using Frozen/thawed PBMC from 32 patients at different time points. Cells were stained with anti-CD4-AmCyan, CD8-Pacific Blue, CD45RA-FITC, CD103-PE, CD127-PerCP-Cy5.5, CD45RO-PE-Cy7, CD27-APC-H7 and CD31-Alexa Fluor 647. The gating strategy to study CD4⁺CD45RA⁺RO⁻CD31⁺ (A) and CD8⁺CD45RA⁺RO⁻CD27⁺CD103⁺ (B) subsets is shown. A comparison of CD4⁺CD45RA⁺RO⁻CD31⁺ (C) or CD8⁺CD45RA⁺RO⁻CD27⁺CD103⁺ (D) subpopulations between baseline and week 24 post-treatment of all patients shows that only an increase in the CD8⁺CD45RA⁺RO⁻CD27⁺CD103⁺ subpopulation correlates with the increase of TREC levels in PBMC.