Aim:
To assess immune reconstitution dynamics in recipients of third-party VSTs early in the course of viral infection following aHSCT.
Method:
Epstein-Barr virus (EBV) and cytomegalovirus (CMV)-specific VSTs were administered to aHSCT recipients within 7 days of commencing antiviral treatment for these viruses. Multiparameter flow immunophenotyping was performed on post-infusion peripheral blood samples using a custom panel of 20 antibody markers covering 32 cell populations, identifying innate and adaptive subsets with a focus on T-subsets and activation markers. Viral antigen specificity in T-cell populations was assessed using virus-specific iTAg MHC class I tetramers where available based on VST and patient HLA-typing. Data was acquired using a BD FACSymphony flow cytometer. Immune reconstitution was assessed to 6 months post-last VST infusion.
Results:
28/30 treated patients achieved complete virological response (CR), defined as undetectable viral load by quantitative PCR by a median of 25 days post-infusion (range 10-161) without significant toxicity. Cytometry was performed on 162 post-infusion samples at multiple timepoints in 25 patients. CD8+ T-cell expansion (median pre-infusion: 0.31 x 109/L, 100-day post-infusion: 0.72 x109/L) was associated with control of viraemia. The predominant subset within both CD8+ and CD4+ populations was CD45RAnegCD62Lneg effector memory cells, with an increase over time in CD8+ CD45RAposCD62Lneg terminally differentiated effector memory cells. 12/19 patients tested with CMV/EBV-specific tetramers showed a rapid rise of virus-specific T-cells to between 1-13% of CD8+ T-cells, corresponding with virological CR, and persistence of these cells long term. The 2 patients who didn’t achieve CR showed poor recovery of adaptive immunity, with dominance of innate effectors.
Conclusion:
Early infusion of VSTs to treat infection post-aHSCT was associated with rapid immune reconstitution correlating with complete virological clearance. Immunophenotyping in this study will inform a randomised trial that is underway, comparing VSTs with antivirals to standard-of-care antivirals alone, to confirm clinical benefit.