To the Editor
Allogeneic hematopoietic stem cell transplantation (HSCT) is an effective therapy for hematologic malignancies. During hematopoiesis reconstitution, the dynamic interactions between the graft’s immune cells and the recipient’s tissues lead to alloreactivity, which can be associated with the positive graft-versus-tumor effect but can also lead to a severe complication like graft-versus-host disease (GVHD).
Early after HSCT, T-cell homeostasis in the recipient is controlled by the peripheral expansion of alloreactive and pathogen-specific donor memory T cells or residual recipient cells [1]. Later, the reconstitution proceeds by de novo production of naïve T cells from donor precursors migrating into the thymus from the recipient’s bone marrow [2, 3]. CD8+ T cells require up to 1 year to reach physiological values [4, 5], while CD4+ T cells take 2 years or even longer, depending on patients’ conditions, such as GVHD, to achieve standard counts and full function [6].
Using high throughput sequencing of the TCR beta complementarity-determining region 3 (CDR3), we reported the TCR repertoire in a cohort of 116 patients 1-year post-HSCT. Compared to the donor’s repertoire before transplantation, we observed a more oligoclonal profile that depends on the patient’s and donor’s age [7]. Moreover, we demonstrated that the cytomegalovirus (CMV) serologic status and CMV infection/reactivation post-HSCT are the most significant clinical features influencing the reconstitution of the repertoire. In this study, we had the opportunity to analyze 26 patients of the previous cohort 5 to 6 years after HSCT. The first study was performed on the bulk of T cells; therefore, we aimed to explore the diversity of TCR repertoire according to a subset of CD4+ and CD8+ T-cell populations. For this part, T-cell differentiation was assessed in 25 other patients who received HSCT from related and unrelated donors (Supplementary Table 1). To evaluate CD4+ and CD8+ T-cell differentiation, we determined the frequency of naïve (Tnaive), stem-memory (Tscm), central memory (Tcm), effector memory (Tem), and RA-expressing memory (Temra) T cells in pre-Tx and post-Tx.1 (i.e., at 1 year) samples. In post-Tx samples, we observed a reduced proportion (<5%) of naïve T cells in both populations (Fig. 1A). In contrast, Tcm and, more noticeably, Tem and CD8+ Temra cells reached significantly higher frequencies than before HSCT. In the CD8+ compartment (Fig. 1B), we observed a drastic decrease in Tnaive cells and an increased proportion of Tem and Temra cells in the D+/R- and D+/R+ groups. A less marked but still noticeable decrease of Tnaive cells was also observed in the D-/R- and D-/R+ groups. Regarding the CD4+ compartment, Temra cells only represented a small portion of the repertoire at both time points. Globally, the most notable change post-Tx was in the D-/R+ group with an increased frequency of Tcm cells (Fig. 1B).
The repertoire diversity (Fig. 1C) switched from polyclonal values in donors to more oligoclonal profile in recipients at 1 year, with the CD8+ repertoire showing the most extreme reduction of diversity. Furthermore, the change in clonality was more evident in CMV-positive samples, particularly in the donor/recipient positive pairs. We observed an increased cumulative frequency and number of CMV-specific clonotypes in the CD8+ compartment post-HSCT (Fig. 1D), better highlighted in the D+/R+ pairs.
In the second part, we evaluated the long-term evolution of the TCR repertoire in 26 patients from our original cohort of 116 patients [7] at 5–6 years post-HSCT. The diversity was significantly reduced in 1-year post-HSCT samples (post-Tx.1) (p = 1.3e-6). However, the repertoire’s diversity remained quite stable when comparing 5–6 years post-HSCT samples (post-Tx.5) to post-Tx.1 samples (p = 0.18, Fig. 2A, left). This was confirmed in the D-/R-, D+/R- and D-/R+ groups as the clonality did not change over time (Fig. 2B left). Conversely, in D+/R+ pairs exhibiting the most drastic shift of clonality at 1 year, the repertoire showed an opposite trend with the recovery of slightly more diverse profiles. While the overlap of clonotypes remained very low in recipients at 5–6 years compared to their donor’s profile, a greater overlap was observed between the two post-HSCT time points (Fig. 2A, right), especially in D+/R+ pairs (Fig. 2B, right). The degree of clonal expansion increased in the post-Tx.1 group and, to a lesser extent, in post-Tx.5 compared to pre-Tx, mainly when the recipient was positive for CMV (Fig. 2C). Most expanded clonotypes post-HSCT in D+/R+ pairs were public (Fig. 2D). Interestingly, a decrease in the cumulative frequency of expanded clonotypes occurred between post-Tx.1 and post-Tx.5 in D+/R+, especially when there was no documented CMV infection/reactivation (Fig. 2D).
Although the TCR repertoire changed drastically regarding clonal diversity within the first year post-HCST, it evolved to a more stable profile throughout the follow-up of 5–6 years. Interestingly, a large TCR overlap was observed between 1 and 5 years in patients with a documented CMV infection/reactivation within the first year. CMV has been shown to drive the expansion of virus-specific memory T cells that accumulate over time, a process called T-cell memory inflation [8]. Remarkably, in the D+/R+ group, CMV infection/reactivation seemed to induce a T cell response that involves highly expanded public TCR clones, notably at 1-year post-HSCT.
Our previous publication did not assess the TCR diversity according to CD4+ or CD8+ subpopulations. These data were generated in an independent cohort, indicating that the TCR diversity is more pronounced in the CD4+ T cell compartment. A peripheral expansion of memory CD4+ and CD8+ T cells at the expense of naïve T cells during the first year post-HSCT was observed. It could be due to a reduced naïve pool linked to a reduced thymic output or the lymphopenic environment at the periphery [9]. Patients at risk for CMV reactivation were almost deprived of CD8+ Tnaive cells, similar to a previous report [10]. The CD8+ T cells exhibited the most skewed TCR profile, which might be driven by Tem or Temra subsets [10].
In summary, the diversity of the TCR repertoire post-HSCT established at 1 year remains stable over time. Furthermore, the TCR repertoire diversity is reduced in the CD8+ compartment, compared to the CD4+, mainly influenced by the CMV status.
Data availability
The datasets generated during and/or analyzed during the current study are available in the Yareta repository, https://doi.org/10.26037/yareta:ue7dlyulbzelzoqbek34g47ovm.
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Acknowledgements
This study was supported by the Swiss National Science Foundation (grant #310030_173237/1), the Academic Society of the University of Geneva, IRGHET (International Research Group on unrelated Hematopoietic stem cell Transplantation), the Dr. Henri Dubois-Ferrière Dinu Lippatti foundation and the Philanthropy Settlement. The authors are grateful to the technicians of the LNRH for their most efficient support for HLA typing and collection of samples.
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ZCS and SB were responsible for designing and writing the protocol, conducting the search, extracting and analyzing the data, interpreting results, and writing the manuscript; AS provided feedback and reviewed the manuscript; SFL: designed the protocol, interpreted results, reviewed and approved the final version of the manuscript; SML, ACM, AP, FS, and YC provided data, reviewed and approved the final version of the manuscript; JV designed and wrote the protocol, supervised the search, interpreted results, wrote the manuscript.
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YC: consulting fees from MSD, Novartis, Incyte, BMS, Pfizer, Abbvie, Roche, Jazz, Gilead, Amgen, Astra-Zeneca, Servier; Travel support from MSD, Roche, Gilead, Amgen, Incyte, Abbvie, Janssen, Astra-Zeneca, Jazz. The other authors declare no competing interests.
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Calderin Sollet, Z., Schäfer, A., Ferrari-Lacraz, S. et al. CMV serostatus and T-cell repertoire diversity 5 years after allogeneic hematopoietic stem cell transplantation. Leukemia 37, 948–951 (2023). https://doi.org/10.1038/s41375-023-01836-w
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DOI: https://doi.org/10.1038/s41375-023-01836-w