Multiple sclerosis (MS) is a complex autoimmune disease with strong genetic components, but its genetic mechanisms remain largely underexplored. We aimed to pinpoint causal genes and evaluate their utility for MS risk prediction. We integrated MS genome-wide association study summaries with brain-derived splicing quantitative trait loci (sQTLs) and expression quantitative trait loci (eQTLs) via summary-data-based Mendelian randomization (SMR) and colocalization analyses to identify potential causal genes. Weighted gene coexpression network analysis (WGCNA) of the E-MTAB-5151 dataset identified MS-associated gene modules. LASSO regression determined the core gene signature. GO and KEGG enrichment analyses, immune infiltration, and gene set enrichment analysis (GSEA) explored the biological relevance. Using an independent protein quantitative trait loci (pQTL) dataset, key genes were further validated for pQTL-MS associations. SMR identified 28 sQTL genes and 66 eQTL genes for MS, 23 and 51 of which passed the colocalization tests, respectively. WGCNA identified three MS-associated modules, and their intersection with SMR genes prioritized 23 key genes. Functional enrichment analysis of the module genes and SMR genes highlighted the consistent involvement of immune-related pathways in MS, including lymphocyte activation and NF-κB signalling. LASSO regression established a 10–gene signature (ACP2, IL7, MYNN, RGS1, SAE1, SP140, TRAF3, TSPAN31, TYMP, and ZC2HC1A) with high predictive accuracy (AUC = 0.983 in internal validation; AUC > 0.70 across three external datasets). Immune infiltration analysis revealed a consistent immune cell expression pattern, in which the expression of MS risk genes was positively associated with naive CD4+ T cells and resting mast cells, but negatively associated with activated mast cells. In contrast, MS protective genes exhibited the opposite pattern. Furthermore, the integration of the MS genome-wide association study statistics validated ZC2HC1A and TRAF3 at the protein level. GSEA further linked both genes to the Hedgehog signalling pathway. Integrating genomic, transcriptomic, and proteomic data, we identified candidate causal genes for MS with robust evidence. ZC2HC1A and TRAF3 have emerged as promising biomarkers and mechanistic candidates for MS. Future follow-up functional studies are warranted to elucidate their molecular roles in MS pathogenesis.

Adult-onset Still’s disease (AOSD) is a rare, systemic autoinflammatory disorder predominantly affecting young adults. It is characterized by polyarthritis, systemic inflammation with high spiking fevers, a transient salmon-pink rash, hepatosplenomegaly, and elevated ferritin levels. Its clinical presentation is highly heterogeneous and lacks specificity, and there are currently no unified diagnostic criteria. Differential diagnosis requires the exclusion of infections, malignancies, other rheumatic diseases, and adverse drug reactions. We report the case of a woman in her 80s in whom the diagnosis of adult-onset Still’s disease was delayed owing to her advanced age, atypical rash, and positive antinuclear antibody status, despite significantly elevated serum ferritin levels. This case highlights that AOSD can affect older individuals, including those aged ≥ 80 years, and should be considered in the differential diagnosis of these patients when presenting with unexplained fever, especially if accompanied by rashes and hyperferritinemia. Moreover, a positive antinuclear antibody test does not rule out the diagnosis of AOSD. Timely and appropriate treatment can improve the prognosis of older patients with adult-onset Still’s disease.

BACKGROUND:Hypoplastic myelodysplastic syndrome (hMDS) is an uncommon MDS subtype, accounting for approximately 10-15% of cases, characterized by bone marrow hypocellularity, severe cytopenias, and features of immune dysregulation. Its clinical course overlaps with aplastic anemia, yet optimal management for lower-risk patients remains undefined, particularly in Asian populations. METHODS:We retrospectively analyzed 48 patients with lower-risk hMDS (IPSS-R very low/low/intermediate) treated at Tianjin Medical University General Hospital from 2013 to 2021. Patients received cyclosporine A (CsA, n = 16), hypomethylating agents (HMAs, n = 10), low-dose HMAs (LD-HMAs, n = 11), or best supportive care (BSC, n = 11). Responses were assessed by modified IWG criteria, and survival was evaluated by Kaplan-Meier and Cox regression analyses. RESULTS:Median age was 61 years (range 16-84), with 60.4% male. Somatic mutations were present in 64.6%, and PNH clones in 29.2%. Six-month overall response rates were 75.0% for CsA, 70.0% for HMAs, 54.5% for LD-HMAs, and 36.4% for BSC (p = 0.220). CsA achieved significantly longer complete remission (median 75 months) and hematologic improvement durations (p < 0.05). Median overall survival (OS)/progression-free survival (PFS) for CsA were 78.0/76.0 months, compared with 17.0/6.8 for HMAs, 31.0/15.0 for LD-HMAs, and 36.0/15.0 for BSC (p < 0.01). HMAs were associated with increased grade 3-4 infections. CONCLUSION:CsA provides superior response durability, survival outcomes, and safety compared with other approaches, supporting its use as first-line therapy in lower-risk hMDS. Integration of immune and molecular profiling may refine individualized treatment strategies.

Objectives Myelodysplastic neoplasm (MDS) patients are at a high risk of infections, contributing significantly to morbidity and mortality. While neutrophil dysfunction is considered a primary factor, specific functional defects remain elusive.Methods We conducted a comprehensive study involving 90 participants, including controls and de novo MDS patients. We utilized the TAXIScan-FL system to evaluate neutrophil chemotaxis towards leukotriene B4 (LTB4). The global reactive oxygen species (ROS) production by neutrophils were measured by chemiluminescence assay, neutrophil alkaline phosphatase (NAP) was evaluated by enzymatic staining.Results MDS patients, irrespective of absolute neutrophil count (ANC) levels, exhibited elevated empirical antimicrobial therapy (EAT) rate compared to controls. Neutrophil migration towards LTB4 was notably impaired, demonstrating reduced velocity and directionality. Interestingly, MDS patients with high ANC still displayed poor directionality and slower migration. MDS patients also had compromised ROS and NAP activity. A noteworthy correlation was observed between EAT rate and chemotactic directionality parameters.Conclusion MDS patients face a heightened risk of infection, potentially attributed to impaired neutrophil chemotactic speed and directionality, alongside compromised ROS and NAP activity. Notably, chemotactic directionality emerged as a pivotal factor correlated with antimicrobial therapy. These insights hold significant clinical implications for managing infections in MDS patients, underscoring the importance of targeting specific neutrophil defects for more effective therapeutic strategies.

Background: As one of the top ten malignancies in China with high morbidity and mortality rates, hematologic malignancies expose patients to a significant risk of physical and mental health deterioration during diagnosis and treatment. Although mindfulness-based interventions have been widely promoted and utilized in clinical settings as an effective approach to improving emotional and physical well-being in cancer patients, the disease-specific nature and the generalized intervention framework hinder the practical application of mindfulness-based cancer rehabilitation in patients with hematologic malignancies。 Objective: To develop a mindfulness-based cancer rehabilitation program for patients with hematologic malignancies, providing practical guidance to improve their physical and psychological symptoms. Method: A systematic search was conducted for domestic and international literature on mindfulness-based interventions for patients with hematologic malignancies, covering publications from January 1, 1982, to December 26, 2023. Seventeen eligible studies were analyzed and synthesized to provide evidence for key aspects of the mindfulness-based cancer rehabilitation program, including themes, facilitators, intervention duration, frequency, and session length. Using the Liverpool mindfulness model as the theoretical framework, the research team held group discussions to determine the facilitators, intervention duration, frequency, and session length based on the follow-up cycles of hematologic malignancy patients and individual psychological assessment differences. Key stages where the model's mindfulness operational phases could integrate with practice were identified, and rehabilitation themes were formulated. Considering practical feasibility, the preliminary framework of the mindfulness-based cancer rehabilitation program was developed by incorporating existing hematology clinical environments and nursing workflows. Qualitative interviews were conducted to refine implementation details, resulting in the initial draft of the program. From July to September 2024, two rounds of Delphi expert consultations involving 19 specialists from eight provinces and cities were conducted to finalize the intervention protocol. Results: The effective response rate for both rounds of the expert consultation questionnaires was 100%, with an expert authority coefficient of 0.88. After the second round of consultation, the Kendall's W coefficient for item rationality was 0.220 (P < 0.001). The final version of the mindfulness-based cancer rehabilitation program for patients with hematologic malignancies was established, encompassing the following components: intervention targets (eligible patient criteria)，intervention facilitators (qualifications and roles), intervention setting and format (e.g., individual/group sessions, in-person/remote delivery), intervention timing (disease phase-specific windows), intervention intensity (duration, frequency, and session length)，intervention procedures (step-by-step protocols), training themes and content (mindfulness modules tailored to hematologic malignancies). Conclusion: The mindfulness-based cancer rehabilitation program for patients with hematologic malignancies, constructed based on the Liverpool Mindfulness Model, demonstrates necessity, logical coherence, scientific rigor, and clinical applicability, thereby providing an evidence-based foundation for clinical psychological nursing practice in this patient population.

Objective This experiment will explore the role of TIGIT/PVR signaling pathway in the pathogenesis of MDS immune tolerance through in vitro co-culture of NK cells and MDSC cells. Methods Flow cytometry was used to detect the expression percentage of MDSCs and CD155 on MDSCs in the bone marrow of MDS patients and controls. The expression of NK cell surface receptors (NKG2D, NKp30, NKp46), secreted cytokines (perforin, granzyme B, CD107a, IFN-gamma) and NK cell apoptosis rate were detected by flow cytometry to evaluate the effect of MDSCs on NK cell function. Results The number of MDSCs in bone marrow of MDS patients was notably higher than that of the control group (8.39 +/- 7.01 vs 2.31 +/- 1.65, P = 0.0001). Compared with the control group, the expression of CD155 on MDSCs in MDS group was increased (31.81 +/- 21.33 vs. 10.49 +/- 6.53, P < 0.0001). After NK cells were co-cultured with MDSCs, NKG2D, NKp30, NKp46, CD107a, IFN-gamma, perforin and granzyme B were decreased, and the NK function partially recovered after the addition of inhibitors. Conclusion Compared with the normal control, MDSCs and CD155 on MDSCs were highly expressed in MDS patients. After co-culture with MDSCs, the expression of NK cells' surface receptors decreased, the secretion of cytokines decreased and the apoptosis rate increased. After blocking TIGIT/CD155 pathway, NK cell function was reversed, but NK cell apoptosis was not reduced.

Objectives: Myelodysplastic syndromes (MDS) are a group of myeloid malignancies characterized by peripheral blood cytopenia and hematopoietic dysplasia that often progress to acute myeloid leukemia (AML). Increased apoptosis of normal hematopoietic cells and decreased apoptosis of malignant clonal hematopoietic cells in patients with MDS is some of the mechanisms leading to ineffective hematopoietic cells in the bone marrow. S100 calcium-binding protein A6 (S100A6) is upregulated in many malignancies. The overexpression of S100A6 in these malignancies has been associated with proliferation, migration, and invasion phenotypes in cancer cells, and we aimed to investigate the expression of S100A6 in CD34(+) cells and the relationship between S100A6 expression and apoptosis of CD34(+) cells in high-risk patients with MDS. Methods: We measured S100A6 mRNA expression in bone marrow (BM) CD34(+) cells from highrisk patients with MDS using RT-PCR. Next, we examined S100A6 expression in CD34(+) cells using flow cytometry. We also analyzed the correlation between CD34(+) cell apoptosis and S100A6 expression in high-risk patients with MDS. Results: Our data showed increased S100A6 mRNA expression in CD34(+) cells in patients with MDS (1.05 +/- 0.69 vs. 0.17 +/- 0.12; P<0.01). The expression of S100A6 in BM CD34(+) cells also increased (58.40 +/- 13.18 vs. 45.83 +/- 15.01). The expression of S100A6 in CD34(+) cells and apoptosis of CD34(+) cells were negatively correlated in patients (r = 0.75; P < 0.01). Conclusions: Collectively, S100A6 may be a potential marker of CD34(+) cells in high-risk patients with MDS and may participate in the pathological behaviors of CD34(+) cells, such as evasion of apoptosis. Thus, S100A6 may be a potential target for eliminating minimal residual disease.

Myeloid-derived suppressor cells (MDSC) induced cellular immune deficiency and bone marrow inflammatory microenvironment play an important role in the pathogenesis and progression of myelodysplastic syndrome (MDS), but the underlying mechanism remains unclear. Here, we revealed that immune checkpoint protein TIM3 and CEACAM1 were highly demonstrated on MDSC and CD8+ T cells in MDS patients. CD8+ T cells were reduced in number and function and presented a exhaustion state. The levels of pro-inflammatory cytokines (IL-1β, IL-18) and CEACAM1 were raised in bone marrow supernatants and MDSC culture supernatants. Blocking or neutralizing TIM3/CEACAM1 and IL-1β/IL-18 partially reversed exhaustion of CD8+ T cells. Moreover, TIM3 correlated with NF-κB /NLRP3 inflammatory pathway. The levels of NF-κB/NLRP3/Caspase-1/IL-1β and IL-18 were all increased in MDSC of MDS. Co-culturing MDSC from MDS patients with rhCEACAM1 enhanced NF-κB/NLRP3/Caspase-1/IL-1β and IL-18 levels, whereas blocking TIM3 could partially reverse the above manifestations. These results indicated that TIM3/CEACAM1 pathway involved in CD8+ T cells exhaustion and might activate the NF-κB/NLRP3/Caspase-1 pathway in MDSC, increasing pro-inflammatory cytokines secretion in MDS bone marrow microenvironment. This study provided a basis for applying immune checkpoint inhibitors that could simultaneously modulate pro-inflammatory cytokine secretion and enhance anti-tumour immune function in the treatment of MDS.

BackgroundIntracerebral hemorrhage (ICH) is the devastating subtype of stroke with cardiovascular complications, resulting in high rates of mortality and morbidity with the release of inflammatory factors. Previous studies have demonstrated that activation of α7nAChR can reduce immune and inflammation-related diseases by triggering the cholinergic anti-inflammatory pathway (CAIP). α7nAChR mediates protection from nervous system inflammation through AMPK-mTOR-p70S6K-associated autophagy. Therefore, the purpose of this study is to explore whether the activation of α7nAChR improves cerebral and cardiac dysfunction after ICH through autophagy.MethodsMale C57BL/6 mice were randomly divided into five groups (1): Control + saline (2), ICH+ saline (3), ICH + PNU-282987 (4), ICH+ PNU-282987 + MLA (5), ICH + PNU-282987 + 3-MA. The neurological function was evaluated at multiple time points. Brain water content was measured at 3 days after ICH to assess the severity of brain edema. PCR, immunofluorescence staining, and Western Blot were performed at 7 days after ICH to detect inflammation and autophagy. Picro-Sirius Red staining was measured at 30 days after ICH to evaluate myocardial fibrosis, echocardiography was performed at 3 and 30 days to measure cardiac function.ResultsOur results indicated that the PNU-282987 reduced inflammatory factors (MCP-1, IL-1β, MMP-9, TNF-α, HMGB1, TLR2), promoted the polarization of macrophage/microglia into anti-inflammatory subtypes(CD206), repaired blood-brain barrier injury (ZO-1, Claudin-5, Occludin), alleviated acute brain edema and then recovered neurological dysfunction. Echocardiography and PSR indicated that activation of α7nAChR ameliorated cardiac dysfunction. Western Blot showed that activation of α7nAChR increased autophagy protein (LC3, Beclin) and decreased P62. It demonstrated that the activation of α7nAChR promotes autophagy and then recovers brain and heart function after ICH.ConclusionsIn conclusion, PNU-282987 promoted the cerebral and cardiac functional outcomes after ICH in mice through activated α7nAChR, which may be attributable to promoting autophagy and then reducing inflammatory reactions after ICH.

The lysosomal thiol reductase GILT catalyzes the reduction of disulfide bonds of protein antigens, facilitating antigen-presenting cells (APCs) to present antigen to T cells. However, whether GILT expression in tumor cells can be associated with improved T cell-mediated anti-tumor responses remains unknown. Here, we identify that GILT is able to facilitate anti-tumor immune surveillance via promoting MHC class I mediated-antigen presentation in colon carcinoma. By using mice model bearing colon tumors, we find that GILT inhibites tumor growth in vivo with more leucocytes infiltration but has no effect on tumor cell development in vitro in terms of proliferation, cell cycle and migration. Furthermore, by using transgenic OT-I mice, we recognize the tumor-expressing OVA peptide, a surrogate tumor antigen, we find that GILT is capable of enhancing MHC class I mediated antigen presentation and improving specific CD8+ T cell anti-tumor responses in murine colon carcinoma. These findings propose the boost of GILT-MHC-I axis in tumors as a viable option for immune system against cancer.

INTRODUCTION:Myelodysplastic syndromes (MDS) are a heterogeneous group of diseases which are prone to progress into acute myeloid leukemia (AML). Iron overload (IOL) caused by transfusion occurred in most MDS patients. But how IOL influences MDS progression has not been clarified yet.METHODS:Herein, we collected clinical data from 143 MDS patients to investigate the impacts of IOL on patients survival and AML transformation.RESULTS:We found that median survival time, 3-year survival rate, leukemia-free survival (LFS) time were significantly shorter in patients with IOL than those with non-iron overload (NIOL) (P = 0.040; P = 0.044; P = 0.037). Besides, IOL was more likely to be found in higher-risk subgroups (assessed by IPSS and WPSS) of MDS patients which also promoted 2-year AML transformation. Furthermore, the serum ferritin (SF) was significantly correlated with the overall survival (OS) of MDS patients (r = -0.311, P < 0.05). The concentrations of both intracellular iron and reactive oxygen species (ROS) in CD34+ cells of bone marrow were higher in the IOL group than the NIOL group, respectively (P = 0.0426; P = 0.0185). Moreover, ROS level was closely correlated with the percentage of bone marrow blasts (r = 0.7200, P = 0.0370). Collectively, IOL threatened the survival of MDS patients and promoted AML transformation.CONCLUSION:Elevated intracellular iron and ROS in CD34+ cells of bone marrow could accelerate the abnormal proliferation of blasts.

ObjectiveTargeting immune checkpoints, such as PD-1, represents a promising approach for cancer immunotherapy, achieving long-term disease remission rates in numerous types of cancer. T cell immunoglobulin and ITIM domain (TIGIT) is a checkpoint receptor associated with the antitumor roles of NK and T cells. Notably, the blockade of TIGIT has been revealed as a potential promising approach in cancer immunotherapy. However, the therapeutic potential of blocking TIGIT in myelodysplastic syndrome (MDS) remains unclear and further research is required to reveal their role.MethodsFresh peripheral blood (PB) and bone marrow (BM) were obtained from patients with MDS and healthy donors (HDs) at the Tianjin Medical University General Hospital between January 21 2018 and March 22 2019. The present study investigated the expression levels of TIGIT on NK and T cells using flow cytometry (FCM) and PCR. In addition, other checkpoint receptors, such as CD226 and PD-1, were also investigated. To determine the mechanisms of antitumor immunity, the functions of NK and T cells expressing TIGIT were determined.ResultsTIGIT was found to be highly expressed on NK and T cells of the PB, where it was involved in disease progression and the immune escape of MDS. The high expression levels of TIGIT were associated with decreased NK and T cell function, and significantly lower secretions of activation factors, such as CD107a, IFN-γ and TNF-α. Notably, blocking TIGIT enhanced the antitumor effects of NK and T cells.ConclusionThe results of the present study suggested that targeting TIGIT alone or in combination with PD-1 may be a promising anticancer therapeutic strategy in MDS.

Background: Chromosomal abnormalities play an important role in the diagnosis and prognosis of patients with myelodysplastic syndromes (MDSs). The single-nucleotide polymorphism array (SNP-A) technique has gained popularity due to its improved resolution compared to that of metaphase cytogenetic (MC) analysis. Methods: A total of 376 individuals were recruited from two medical centers in China, including 350 patients and 26 healthy individuals. Among these patients, 200 were diagnosed with de novo MDS, 25 with myeloproliferative neoplasm (MPN), 63 with primary acute myeloid leukemia (AML), and 62 with idiopathic cytopenia of undetermined significance (ICUS). We evaluated the significance of abnormal chromosomes detected by SNP-A in the diagnosis and prognosis of MDS-related disorders. Results: (1) When certain chromosomal abnormalities could not be detected by conventional MC methods, these abnormalities could be detected more efficiently by the SNP-A method. With SNP-A, the detection rates of submicroscopic or cryptic aberrations in the MDS, MPN, and AML patients with normal MC findings were 32.8, 30.8, and 30%, respectively. (2) The chromosomal abnormalities detected by SNP-A had a very important value for the prognosis of patients with MDSs, especially in the low-risk group. The survival of patients with abnormal chromosomes detected by SNP-A was significantly lower than that of patients with no detected chromosomal abnormalities; this difference was observed in overall survival (OS) (P = 0.001) and progression-free survival (PFS) [24 months vs. not reach (NR); P = 0.008]. The patients with multiple chromosomal abnormalities detected by SNP-A had an inferior prognosis, and SNP-A abnormalities (≥3 per patient) were found to be an independent predictor of poor prognosis in patients with MDSs [hazard ratio (HR) = 2.40, P = 0.002]. (3) Patients with ICUS may progress to myeloid malignancies, but most patients often maintain a stable ICUS status for many years without progression. An ICUS patient found to have an MDS-related karyotype would be rediagnosed with MDS. SNP-A can efficiently detect chromosomal abnormalities, which would be important for assessing the evolution of ICUS. In our study, 17 ICUS patients with SNP-A-detected abnormalities developed typical MDSs. Conclusions: SNP-A can help evaluate the prognosis of patients with MDSs and better assess the risk of disease progression for patients with ICUS.

CD8(+) T cells play a central role in antitumour immunity, which often exhibit 'exhaustion' in the setting of malignancy and chronic viral infection due to T cell immunoglobulin and mucin domain 3 (TIM3) and myeloid-derived suppressor cells (MDSCs). Our team previously found that overactive MDSCs and exhausted TIM3(+)CD8(+) T cells were observed in myelodysplastic syndromes (MDS) patients. However, it is not obvious whether MDSCs suppress CD8(+) T cells through TIM3/Gal-9 pathway. Here, Gal-9, as the ligand of TIM3, was overexpressed in MDSCs. The levels of Gal-9 in bone marrow supernatants, serum and culture supernatants of MDSCs from MDS patients were elevated. CD8(+) T cells from MDS or normal controls produced less perforin and granzyme B and exhibited increased early apoptosis after co-culture with MDSCs from MDS. Meanwhile, the cytokines produced by CD8(+) T cells could be partially restored by TIM3/Gal-9 pathway inhibitors. Furthermore, CD8(+) T cells produced less perforin and granzyme B after co-culture with excess exogenous Gal-9, and the function of CD8(+) T cells was similarly restored by TIM3/Gal-9 pathway inhibitors. Expression of Notch1, EOMES (associated with perforin and granzyme B secretion), p-mTOR and p-AKT (associated with cell proliferation) was decreased in CD8(+) T cells from MDS after co-culture with excess exogenous Gal-9. These suggested that MDSCs might be the donor of Gal-9, and TIM3/Gal-9 pathway might be involved in CD8(+) T cells exhaustion in MDS, and that TIM3/Gal-9 pathway inhibitor might be the promising candidate for target therapy of MDS in the future.

T cell immunoglobulin and mucin domain 3 (TIM3) expression is associated with immunosuppression and clinical outcomes in many diseases. However, the specific mechanism of TIM3 in immune system has not been clarified. In order to illustrate the mechanism of TIM3 in immune system, we analyzed the expression, function and regulation of TIM3 in T helper (Th)1 cells, Th2 cells, Th17 cells and regulatory T cells (Treg) through flow cytometry in patients with myelodysplastic syndrom (MDS). Our data showed elevated proportion of Th2 and Treg cells, while the proportion of Th1 and Th17 cells decreased in patients with MDS (p<0.05) and the expression of TIM3 increased in Th1, Th17 and Treg cells in patients with MDS when compared with expression in control patients (p<0.05). The secretion of transforming growth factor-β in TIM3+Treg cells decreased in patients with MDS. These findings suggested that TIM3 might affect immune helper systems by regulating Treg cells and related immune cells. Therefore, studying the role of the TIM3 pathway in MDS is necessary and may help to provide a new way to explore the pathogenesis and treatments of MDS.

Myelodysplastic syndromes (MDS) are clonal stem cell disorders characterized by ineffective hematopoiesis that lead to leukemia. Disorders of the immune system serve important functions in the pathophysiology and progression of this disease. Different levels or mechanisms of natural killer (NK) cells in patients with MDS have been measured in previous studies, making it challenging to understand the pathogenesis of NK cytotoxicity. The present study investigated the frequency of NK cell-mediated antibody-dependent cellular cytotoxicity and explored the function of NK cells by their activating receptors, inhibition signals, degranulation and cytotoxicity factors. In the present study, levels of cluster of differentiation (CD)3-CD56+ NK cells, CD16+-expressing NK cells and subset CD56dim NK cells were decreased in the peripheral blood of patients with MDS. Altered expression of NK protein 44, NK group 2 member D, killer cell immunoglobulin-like receptor 2DL1 (KIR2DL1) and KIR2DL3 on NK cell effector signaling pathways may trigger tumor cell lysis in patients with MDS. The weak cellular adhesion and decreased cytotoxicity of NK cells may lead to ineffective antitumor activity in MDS. These observations suggested that NK cells may serve as immunological determinants in MDS and may permit the development of NK cell-based immunotherapy for the treatment of patients with MDS.

OBJECTIVE:To analyze the subtype and functional biomarker expression changes of natural kill cells(NK) in peripheral blood of patients with myelodysplastic syndrome(MDS) and normal people, so as to evaluate the relationships between these changes and hematopoietic functions and to explore the role of NK cells in the pathogenesis of MDS.METHODS:The quantity of NK cells and the expression of biomarkers(NKp30,NKp46,NKG2A) on NK cells were detected by flow cytometry in 35 MDS patients from 2015 to 2016 in our hospital and 34 normal controls. The correlation between these changes and hematopoietic functions, including the percentages of neutrophil(ANC), hemoglobin in peripheral blood and the hematopoietic function in bone marrow(CD34+%) were evaluated.RESULTS:The percentage and quantity of NK cells and CD56dim NK cells in MDS patients were significantly lower than those in normal controls(P<0.05); the percentage of CD56bright NK cells was higher than that in controls. The percentage of CD56dim NK cells in NK cells of MDS patients was significantly lower than that of controls; the percentage of CD56bright NK cells in NK cells of MDS patients was significantly higher than that of controls. The expression of NKp30 and NKp46 of MDS patients was significantly lower than that of controls. In MDS group, the percentage of NK cells and CD56dim NK cells of peripheral blood lymphocytes in high risk MDS group was significantly lower than that in low risk MDS group. The percentage of NK and CD56dim NK cells negatively correlated with that of CD34+% in bone marrow, but positively correlated with ANC and Hb. The CD34+% in bone marrow negatively correlated with expression of NKp46, but positively correlated with expression of NKG2A.CONCLUSION:The decrease of NK number and function may cause the immune surveillance and lead to disease progression.

Objective: To explore characteristic and function of peripheral blood mononuclear cells (PBMNC) -induced macrophages in patients with myelodysplastic syndrome (MDS) to couple with its progression. Methods: A total of 24 MDS patients (11 low-risk patients and 13 high-risk group patients) referred to Department of Hematology of Tianjin Medical University General Hospital and normal controls were enrolled from September 2014 to December 2015. PBMNC was stimulated with GM-CSF to transform to macrophages. The morphology of macrophages was observed by microscope. The quantity of macrophages, CD206 and SIRPα on surface of macrophages were detected by flow cytometry. The phagocytic function of macrophages was analyzed by fluorescence microscopy and flow cytometry. Results: The morphology of macrophages from MDS patients was abnormal. The percentage of transformed macrophages was (5.17±3.47) % in patients with MDS, which was lower than that in controls significantly[ (66.18±13.43) %, t=3.529, P=0.001]. The expression of CD206 on macrophages from MDS patients was significantly lower than that of controls[ (9.73±2.59) % vs (51.15±10.82) %, t=4.551, P<0.001]. The SIRPα level of macrophages from MDS patients was significantly lower than that of controls [ (0.51±0.09) % vs (0.77±0.06) %, t=2.102, P=0.043]. The phagocytic index and the percentage of phagocytic of macrophages from MDS patients were significantly lower than those of macrophages from normal controls[0.45±0.08 vs 0.92±0.07, t=-6.253, P=0.008; (23.69±3.22) % vs (42.75±2.13) %, t=-6.982, P=0.006 respectively]by flow cytometry. The phagocytic index of MDS patients was significantly lower than that of controls (0.24±0.04 vs 0.48±0.96, t=3.464, P=0.001) by fluorescence microscopy. Conclusion: The quantity, recognization receptors and phagocytosis of PBMNC-induced macrophages decreased in MDS patients.

Objective: To investigate the frequencies and biological characteristics of CD25 positive hematopoietic stem cells (HSC) in myelodysplastic syndromes. Methods: The expression of CD25 on HSC in bone marrow derived from patients with untreated MDS patients, untreated AML patients and normal controls were accessed by flow cytometry (FCM). The correlation analysis was done between CD25(+) HSC and clinical parameters in MDS patients. Results: The expression of CD25 on HSC (CD34(+) CD38-cells) in MDS patients (28.81%) was significantly higher than that in normal controls (9.41%, P=0.020), which similar to that in AML patients (32.54%, P=0.410). The CD25 expression on HSC was positively correlated with the CD123 expression on HSC (r=0.602, P=0.008). The expression of CD25 on HSC in high-risk MDS group (53.27%) based on IPSS score was significantly higher than that in low-risk MDS group (18.66%, P=0.003). In MDS patients, CD25(+) HSC were negatively correlated with the counts of neutrophils (r=-0.684, P=0.002) and platelets (r=-0.561, P=0.015), while positively correlated with the percentage of blasts in bone marrow (r=0.596, P=0.009). The CD25 expression on erythroblasts had a significant positive correlation with red blood cell counts in MDS patients (r=0.536, P=0.012). Conclusions: CD25 was over-expressed on HSC in MDS patients, especially in high-risk MDS patients. Increased CD25(+) HSC was correlated with progression of MDS. Low-expression of CD25 on erythroblasts might correlate with anemia in MDS patients. CD25 could be a specific marker of LSC in MDS, and could involve in the mechanisms of development and progression of MDS.

Objective: To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) . Methods: Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA(+) nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA(+) nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA(+) nucleated RBC was detected by Western blot. Results: Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA(+) nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, P<0.001) , and lower than that in low-risk MDS patients (0.40±0.16, P=0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA(+) nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , P<0.017) . The expression of ULK1 mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , P<0.017]. The level of mTOR mRNA in GlycoA(+) nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , P<0.017]. The level of LC3B in GlycoA(+) nucleated RBC was negatively correlated with the HGB (r=0.529, P=0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42. Conclusion: Autophagy is impaired in nucleated RBC of MDS patients.