Allogeneic hematopoietic stem cell transplant (alloSCT) recipients often experience late-onset human cytomegalovirus (HCMV) reactivations following termination of letermovir prophylaxis. Letermovir prophylaxis extends the window for protective B- and T-cell reconstitution; however, our understanding of humoral responses and their contribution to HCMV immune control remains limited.

Combining serological and flow cytometric analyses in 42 HCMV-seropositive alloSCT recipients, we herein provide the first comprehensive longitudinal (days 90-270 post-transplant) characterization of HCMV-specific humoral responses, natural killer (NK)-cell phenotypes, and γδ T cells in the letermovir era.

HCMV controllers showed predominantly IgG-driven responses, higher pre-reactivation Vδ1+ γδ T-cell frequencies, and stronger expansion of “memory-like” NK cells than patients with clinically significant CMV infection (csCMVi). In contrast, csCMVi patients showed delayed HCMV-specific IgG production, IgM-skewed humoral responses, and stronger post-reactivation expansion of memory B cells and Vδ1+ γδ T cells. Early γδ T-cell reconstitution by day 90 was predictive of future HCMV control. HCMV-specific IgG levels correlated only weakly with γδ T cells but showed distinct associations with “memory-like” NK-cell reconstitution in HCMV controllers, suggesting synergisms between humoral and cellular immunity.

Collectively, these findings highlight a need to study anti-HCMV immune protection beyond type-1 T cells and refine risk stratification models in alloSCT patients by inclusion of novel immune markers such as γδ T-cell frequencies and phenotypes. Leveraging the extended B-cell reconstitution window created by letermovir, novel immunotherapies (e.g., therapeutic antibodies) and future vaccines might boost humoral anti-HCMV immunity and benefit from synergisms with γδ T cells and “memory-like” NK cells in improving HCMV control.

The authors have declared no competing interest.

This research was funded by a grant from the German Research Council (DFG) within the DFG Research Unit FOR 2830 (KR 5761/1-2, project number 398367752 (to S.K.); EI 269/10-2, project number 398367752 (to H.E.); DO 1275/7-2 (to L.D.); HE2526/9-2 (to H.H.)). S.K. was further supported by a fellowship of the Interdisciplinary Center for Clinical Research (IZKF) and the trust Forschung Hilft.

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This study was approved by the Ethics Committees of the University of Wuerzburg, Germany (protocol code 17/19-sc). Written informed consent was obtained from all patients.

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Data sharing statement: The datasets generated and analysed in this study are available from the corresponding author upon reasonable request. Please contact Sabrina Kraus (Kraus_S3ukw.de). Individual patient data will not be shared.

The datasets generated and analysed in this study are available from the corresponding author upon reasonable request.