Lin Liu

Background: Donor–recipient sex mismatch, particularly female-to-male (F-to-M) transplantation, has been associated with an increased risk of chronic graft-vs.-host disease (cGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), but its relationship with early immune reconstitution remains incompletely characterized. Methods: In this single-center retrospective cohort of 294 patients transplanted between 2016 and 2025, patients were classified by donor–recipient sex combination (F-to-M, M-to-F, M-to-M, F-to-F). Multivariable regression was used for cGVHD and survival analyses, and lymphocyte subsets were measured by flow cytometry at months 1–3 in 170 patients. Results: The F-to-M group had the highest cGVHD incidence (51.1%) and was independently associated with cGVHD relative to M-to-M [odds ratio (OR) 2.62, 95% CI: 1.23–5.58; p = 0.013], which remained significant after adjustment for acute GVHD (OR: 2.48, 95% CI: 1.15–5.33; p = 0.020); in a sensitivity analysis restricted to patients with available pre-transplant measurable residual disease (MRD) data ( n = 199), the estimate was similar in magnitude but did not reach statistical significance after MRD adjustment (OR: 2.58, 95% CI: 0.95–6.99; p = 0.062). Overall survival differed marginally among the four groups (log-rank p = 0.049), with the lowest 3-year OS in the F-to-M group (58.0%, 95% CI: 35.4–77.1, vs. 83.3–89.5%); however, F-to-M transplantation was not independently associated with OS in the multivariable model [hazard ratio (HR) 2.01, 95% CI: 0.94–4.29; p = 0.071]. Cumulative non-relapse mortality was higher in F-to-M recipients (Gray's test p = 0.016), whereas relapse did not differ (Gray's test p = 0.536). F-to-M recipients showed a persistently higher CD4+/CD8+ ratio during months 1–3, mainly reflecting delayed CD8+ T-cell recovery. Within this subgroup, a lower 3-month NKT-like cell count was associated with cGVHD; an exploratory threshold of ≤14 cells/µL yielded an apparent area under the curve of 0.814 (95% CI: 0.618–0.965; bootstrap-corrected 0.810), and the F-to-M × log(NKT-like) interaction was significant ( p = 0.046). Conclusions: These findings suggest that F-to-M transplantation may be associated with a higher burden of cGVHD and with higher non-relapse mortality, accompanied by a characteristic early immune-reconstitution pattern, whereas its association with overall survival was not statistically significant after adjustment. Low 3-month NKT-like cell counts may serve as an exploratory candidate marker for cGVHD risk stratification, but require validation in larger prospective cohorts.

Background: Male testis is highly susceptible to irreversible damage from acute myeloid leukemia (AML) infiltration and subsequent chemotherapy. As a gene critical for sperm function, the role of SPAG6 in AML testicular infiltration and chemotherapy-induced injury remains poorly elucidated. Methods: SPAG6 expression was knocked down in C1498 leukemia cell line, and its impact on testicular infiltration was evaluated in vivo . RNA sequencing was performed to identify altered pathways. In parallel, SPAG6 was silenced in murine GC-1 spg spermatogonial cells, followed by assessment of reactive oxygen species (ROS) levels and cell proliferation. Results: Both AML infiltration and subsequent chemotherapy exerted profound damage on the testes. SPAG6 was highly expressed in normal mouse testes and was upregulated upon AML infiltration and chemotherapy. Knockdown of SPAG6 in leukemia cells was associated with suppressed proliferation and induced apoptosis in vitro , and reduced testicular infiltration in vivo ; chemotherapy further enhanced these effects. RNA sequencing and wound healing assay further indicated that SPAG6 depletion may impair cell adhesion and migration. Conversely, silencing SPAG6 in spermatogonial cells was linked to suppressed proliferation, elevated mitochondrial membrane potential and ROS levels, and these effects were further exacerbated under chemotherapeutic challenge. Conclusion: SPAG6 may exert cell-type-specific, opposing roles in AML-infiltrated testes: SPAG6 depletion is associated with suppressed leukemic cell infiltration, potentially through modulation of cell adhesion, while its deficiency correlates with increased chemotherapy-induced injury to spermatogonial cells.

The overall incidence of extramedullary disease (EMD) in acute myeloid leukemia (AML) ranges from 2% to 30.5%, and is potentially associated with short overall survival. However, the risk factors for EMD and the underlying molecular mechanisms remain poorly understood. Here, we analyzed a cohort of 118 adult patients with de novo AML, among whom 27 (22.88%) developed EMD in various body sites, with tissue involvement as the most common (16/118 [13.56%]). Mesothelin (MSLN) expression, detectable at both transcript and protein levels, can serve as an independent risk factor for EMD in patients with AML. Multivariate analyses revealed that MSLN positivity at transcript levels (hazard ratio [HR], 3.43; 95% confidence interval [CI], 1.44-8.20; P = .006) and MSLN positivity on leukemic blasts detected by flow cytometry (HR, 3.28; 95% CI, 1.37-7.86; P = .008) are independently correlated with EMD risk. Mechanistically, MSLN overexpression promotes cell proliferation, metastasis, and invasion in AML cells. MSLN greatly facilitates cell-cell adhesion by binding with mucin 16 (MUC16), resulting in extramedullary dissemination. MSLN also activates the phosphoinositide 3-kinase (PI3K) signaling pathway and upregulates neural cell adhesion molecules (CAMs) CD56 and neural cell adhesion molecule 2 (NCAM2) through interaction with MUC16. Our study highlights the importance of MSLN expression, particularly identified by flow cytometry, as an independent risk factor for EMD in patients with AML, and elucidates its role in modulating PI3K signaling and CAMs in AML cells.

B-cell reconstitution is closely associated with prognosis, infection risk, and graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the impact of different calcineurin inhibitors (CNIs) on B-cell recovery remains unclear. We retrospectively analyzed 312 allo-HSCT recipients receiving cyclosporine (CsA, n = 211) or tacrolimus (FK506, n = 101). B-cell proportions in bone marrow and peripheral lymphocyte subsets were compared, and propensity score matching was applied to balance confounders. Multivariate analysis demonstrated that FK506 was independently associated with superior B-cell reconstitution at 3 months (OR 2.93, 95% CI 1.49–5.76, p = 0.010) and 4 months post-transplant (OR 3.53, 95% CI 1.11–11.15, p = 0.024). Higher B-cell proportions at 3–5 months were associated with a significantly lower risk of viral infection. Mediation analysis further revealed that improved B-cell reconstitution at day + 90 partially mediated the protective effect of FK506 against viral infection. These findings suggest that FK506 promotes improved B-cell recovery and may reduce viral infection risk after allo-HSCT.

Purpose Achieving high-quality immune reconstitution (IR) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is directly associated with the patient’s ability to resist infections, prevent tumor relapses, and suppress graft-versus-host disease (GVHD). The status of CD4 + T cells is critical for reconstituting adaptive immunity after transplantation; however, the patterns of reconstitution and their influencing factors remain unclear. Methods This retrospective cohort study involved 164 patients who underwent myeloablative allo-HSCT from April 2016 to May 2023. Based on the early post-transplantation CD4 + T cell counts, the cohort was divided into two groups: 73 patients with high-quality IR (HIR) and 34 patients with low-quality IR (LIR). LIR was associated with an increased risk of Epstein–Barr virus (EBV) reactivation following transplantation. Plasma EBV viral load was monitored weekly using quantitative PCR for the first 100 days post-transplantation. Results The LIR group had a higher viral load at the time of the first EBV reactivation and a significantly earlier first reactivation compared to the HIR group. No significant differences were observed in overall survival, GVHD incidence, or relapse rates between the groups. A CD4 + T cell count of < 60 cells/μL within 30 days post-transplantation was associated with a higher incidence of EBV viremia compared to the control group. Conclusion These findings suggest that early CD4 + T cell counts may serve as a predictive marker for post-transplant EBV infection.

Background: Diffuse large B-cell lymphoma (DLBCL) is the most prevalent form of non-Hodgkin’s lymphoma globally. SPAG5, a mitotic spindle protein, plays a significant role in DLBCL, where its abnormal expression is often associated with tumor growth, chemotherapy resistance, local recurrence, and poor prognosis. Methods: A comprehensive analysis of SPAG5 expression across various cancer types was conducted using Timer 2.0 and Sanger Box 3.0. Subsequently, the expression levels of SPAG5 in DLBCL were investigated in comparison to normal samples. Receiver operating characteristic (ROC) curve was then generated to evaluate the diagnostic performance of SPAG5 for DLBCL. Furthermore, the functional role of SPAG5 was characterized, and its impact on the immune microenvironment of DLBCL patients was analyzed. Its potential in predicting immune checkpoint status and responses to immunotherapy was also evaluated. Results: SPAG5 expression demonstrated significant heterogeneity across various cancer types, with a marked upregulation in DLBCL. The diagnostic efficacy of SPAG5 was moderate, yielding an area under curve (AUC) of 0.75. SPAG5 exerted a multifaceted influence on DLBCL progression by regulating critical cellular processes, including cell cycle dynamics, chromosomal segregation, and DNA homeostasis. Notably, patients with elevated SPAG5 expression had poorer survival outcomes than those with low expression. Analysis of the tumor immune microenvironment revealed a distinct pattern: high SPAG5 expression correlated with increased infiltration of resting natural killer (NK) cells, while being associated with reduced presence of regulatory T cells (Tregs) and follicular helper T cells (Tfh). Conclusion: Our bioinformatics study elucidated the expression profile, diagnostic potential, and prognostic significance of SPAG5 in DLBCL, emphasizing the complex interplay between SPAG5 expression and the tumor immune landscape. Our findings suggested SPAG5 could be a candidate prognostic marker and potential therapeutic target for DLBCL.