Transplant immunology最新文献_第4页

CXCL11 as potential predictive biomarker and therapeutic target of acute rejection after kidney transplantation CXCL11作为肾移植后急性排斥反应的潜在预测生物标志物和治疗靶点。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-05 DOI: 10.1016/j.trim.2025.102306

Jie Zhang, Chengjun Yu, Huyu Wang, Hanyu Xiao, Sheng Wen, Yi Hua, Guanghui Wei

Kidney transplantation is the optimal treatment method for chronic kidney disease. Although the short-term and long-term survival rates of transplanted kidneys have been significantly improved with the development of immunosuppressive agents, acute rejection remains the main risk factor threatening the survival of the allografts and patient. We utilized bioinformatics analysis to identify the predictive and therapeutic target of acute rejection after kidney transplantation. In the results, cytokines were considered as critical role in allografts acute rejection, and most cytokines were increased in the process of AR. According to the machine learning algorithm analysis and receiver operating characteristic curve results, CXCL11 was identified as the most valued cytokine in prediction of AR. Single-gene GSEA results showed CXCL11 was strongly associated with AR-related biological behavior. Subsequent analysis results showed the gene RELA regulate the expression of CXCL11 and mainly distribute in renal tubular epithelial cells. In cell experiments, LPS as the activator of NF-κB signaling pathway induced the expression of CXCL11. In animal experiments, compared to syn group, severe acute rejection occurs in allo group, and companied with severe inflammatory reaction and the expression of CXCL11, as the activation of NF-κB signaling pathway. CXCR3 specifically recognizes CXCL11 as one of its ligands, single cell analysis demonstrated CXCR3 and CD8 were co-expression on the T cells in the microenvironment of allografts. Finally, we demonstrated in allo group of rat kidney transplantation, there were a large number of CXCR3 + CD8+ T cells infiltrated the allografts. Conclusion, we utilized bioinformatics analysis tools, finally identified CXCL11 as the potential target for prediction and treatment in acute rejection after kidney transplantation.

肾移植是慢性肾脏疾病的最佳治疗方法。尽管随着免疫抑制剂的发展，移植肾脏的短期和长期存活率显著提高，但急性排斥反应仍然是威胁同种异体移植物和患者生存的主要危险因素。我们利用生物信息学分析来确定肾移植后急性排斥反应的预测和治疗靶点。结果表明，细胞因子在同种异体移植物急性排斥反应中起着至关重要的作用，大多数细胞因子在AR过程中升高。根据机器学习算法分析和受体工作特征曲线结果，CXCL11被确定为预测AR最有价值的细胞因子。单基因GSEA结果显示CXCL11与AR相关的生物学行为密切相关。随后的分析结果显示RELA基因调控CXCL11的表达，主要分布在肾小管上皮细胞中。在细胞实验中，LPS作为NF-κB信号通路的激活剂诱导CXCL11的表达。在动物实验中，与syn组相比，同种异体组出现严重的急性排斥反应，并伴有严重的炎症反应和CXCL11的表达，这是由于NF-κB信号通路的激活。CXCR3特异性识别CXCL11作为其配体之一，单细胞分析表明CXCR3和CD8在同种异体移植物微环境下的T细胞上共表达。最后，我们证明在大鼠肾移植同种异体组中，有大量的CXCR3 + CD8+ T细胞浸润同种异体移植物。结论利用生物信息学分析工具，最终确定了CXCL11作为预测和治疗肾移植急性排斥反应的潜在靶点。

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引用次数: 0

HLA-specific platelet trasfusion as part of desensitization strategy in patients with donor-specific anti-HLA antibodies before stem cell transplantation hla特异性血小板输注作为干细胞移植前供体特异性抗hla抗体患者脱敏策略的一部分。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-09 DOI: 10.1016/j.trim.2025.102310

Giuliana Lando , Roberto Crocchiolo , Giorgia Cornacchini , Giulia Di Maggio , Cristina Garanzini , Giovanni Grillo , Benedetta Mazzi , Silvano Rossini , Michela Tassara

The presence of donor-specific anti-HLA antibodies (DSAs) in patients who are candidates for allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a recognized risk factor for delayed engraftment and mortality. In the absence of an alternative donor, patient desensitization is indicated to reduce or eliminate DSAs; however, harmonization among transplant centers remains insufficient and reduction of DSAs is sometimes unsuccessful. Here, we present the feasibility and results of our recent monocenter experience on nine immunized HLA class I DSA+ patients undergoing pre-transplant desensitization using HLA-specific platelet transfusion, as a means of antibody adsorption through platelets expressing the same HLA specificities targeted by DSAs. All allo-HSCTs were from HLA-mismatched donors, both unrelated or related haploidentical. The approach appeared safe for donors and patients and potentially useful in mitigating the detrimental impact of class I DSAs, with successful engraftment observed in two of four patients with MFI > 10,000 and in all patients with MFI <10,000. Until harmonized and more risk-adapted desensitization strategies become available, sharing inter-center experiences between clinical and transfusion units will likely improve the management of hyperimmunized patients and enhance the availability of emerging desensitizing strategies.

异体造血干细胞移植（allogenic hematopoietic stem cell transplantation, alloo - hsct）的候选患者体内存在供体特异性抗hla抗体（dsa）是公认的延迟移植和死亡的危险因素。在没有替代供体的情况下，建议患者脱敏以减少或消除dsa；然而，移植中心之间的协调仍然不足，dsa的减少有时是不成功的。在这里，我们介绍了我们最近的单中心经验的可行性和结果，我们对9名免疫HLA I类DSA+患者进行移植前脱敏，使用HLA特异性血小板输注，作为一种通过表达相同HLA特异性的血小板吸附抗体的方法。所有同种异体造血干细胞均来自hla错配供体，无亲缘关系或有亲缘关系的单倍相同供体。对于供体和患者来说，该方法似乎是安全的，并且可能有助于减轻I类dsa的有害影响，在4名MFI患者中有2名 > 10,000和所有MFI患者中观察到成功的植入

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引用次数: 0

Macrophage polarization in organ transplantation rejection and targeted therapeutic strategies 巨噬细胞极化在器官移植排斥反应中的作用及其靶向治疗策略。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-17 DOI: 10.1016/j.trim.2025.102316

Changqing Qu , Yi Chen , Xiaoyu Xu , Xianduo Li , Dongdong Chen , Wenzhi Du , Minrui Zhang , Zhe Yang , Jianning Wang

Organ transplantation improves survival and quality of life in end-stage organ failure, but rejection remains a key barrier to long-term graft success. While current immunosuppression primarily targets adaptive immunity, its limitations and side effects necessitate alternative therapeutic strategies. Macrophages infiltrate grafts extensively, which are involved in antigen presentation function, ischemia-reperfusion injury, and both acute and chronic rejection processes. Notably, their polarization toward an anti-inflammatory phenotype may alleviate rejection and potentially contribute to graft immune tolerance. In this review, we will give an overview on macrophage phenotypes and their functional diversity in allograft rejection. Also, we will discuss emerging strategies to modulate macrophage polarization, including therapies exploiting their dual regulatory capacity, nanoparticle-based targeting systems, gene therapies, and microRNA-mediated regulation. A deeper understanding of macrophage biology in transplantation could enable more sophisticated anti-rejection approaches, whose integration with conventional immunosuppression may ultimately enhance long-term graft outcomes.

器官移植可以改善终末期器官衰竭患者的生存和生活质量，但排斥反应仍然是移植长期成功的关键障碍。虽然目前的免疫抑制主要针对适应性免疫，但其局限性和副作用需要其他治疗策略。巨噬细胞广泛浸润移植物，参与抗原呈递功能、缺血再灌注损伤以及急性和慢性排斥反应过程。值得注意的是，它们向抗炎表型的极化可能减轻排斥反应，并可能有助于移植物免疫耐受。在这篇综述中，我们将概述巨噬细胞表型及其在同种异体移植排斥反应中的功能多样性。此外，我们还将讨论调节巨噬细胞极化的新策略，包括利用其双重调节能力的治疗方法、基于纳米颗粒的靶向系统、基因治疗和微rna介导的调节。对移植中巨噬细胞生物学的更深入了解可以实现更复杂的抗排斥方法，其与传统免疫抑制的结合可能最终提高移植的长期效果。

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引用次数: 0

Natural killer cell KIR genotype and NKG2C phenotype in correlation to BKV infection following kidney transplantation 肾移植后自然杀伤细胞KIR基因型和NKG2C表型与BKV感染的相关性

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-06 DOI: 10.1016/j.trim.2025.102303

Shubo Tan , Xiaobin Lin , Xueqi Li, Jianhua Long, Yuan Luo, Yao Xiao, Jianjun Li

Purpose

To explore the correlation between the NK cell killer immunoglobulin-like receptor (KIR) genotype, NKG2C phenotype, and BK virus (BKV) infection following kidney transplantation.

Methods

Clinical data from 51 recipients who underwent allogeneic kidney transplantation were retrospectively analyzed. Recipients were grouped based on their KIR genotype (A/A or B/X genotype) and NKG2C⁺ NK cell ROC cut-off values. SPSS statistical analysis software was used to assess their association with BKV infection.

Results

Seventeen recipients with the KIR-A/A genotype and 34 with the KIR-B/X genotype were included in this study. Statistical analysis revealed a significant difference in the distribution of activating KIR gene numbers between the two groups. Univariate analysis revealed significant differences in survival curves between the groups according to KIR genotype and activating KIR gene number. Cox multivariate analysis identified the KIR genotype and delayed graft function as independent factors influencing BKV infection following kidney transplantation. ROC curve analysis revealed that the area under the curve for the NKG2C⁺ NK cell ratio in the infection-free survival (IFS) evaluation was 0.788, with an optimal cutoff value of 16.5 %. BKV infection rates in recipients with NKG2C⁺ NK cell ratios <16.5 % and ≥ 16.5 % were 66.7 % and 20 %, respectively. Recipients with NKG2C⁺ NK cell ratios <16.5 % had worse IFS than those with NKG2C⁺ NK cell ratios ≥16.5 %.

Conclusion

A correlation was found between NK cell KIR genotype, NKG2C⁺ NK cell phenotype, and BKV infection following kidney transplantation, providing a reference for selecting suitable donors for transplantation and reducing the risk of post-transplant BKV infection.

目的：探讨肾移植术后NK细胞杀伤免疫球蛋白样受体（KIR）基因型、NKG2C表型与BK病毒（BKV）感染的相关性。方法：回顾性分析51例异体肾移植患者的临床资料。根据受体的KIR基因型（A/A或B/X基因型）和NKG2C+ NK细胞ROC临界值对受体进行分组。采用SPSS统计分析软件评估其与BKV感染的相关性。结果：共纳入17例KIR-A/A基因型受体和34例KIR-B/X基因型受体。统计分析显示，两组之间激活KIR基因数量的分布有显著差异。单因素分析显示，根据KIR基因型和激活KIR基因数量，组间生存曲线存在显著差异。Cox多因素分析发现KIR基因型和移植延迟功能是影响肾移植后BKV感染的独立因素。ROC曲线分析显示，NKG2C+ NK细胞比例在无感染生存（IFS）评价中的曲线下面积为0.788，最佳截断值为16.5 %。NKG2C+ NK细胞比值+ NK细胞比值+ NK细胞比值≥16.5 %的受体BKV感染率。结论：NK细胞KIR基因型、NKG2C+ NK细胞表型与肾移植术后BKV感染存在相关性，为选择合适的移植供体、降低移植后BKV感染风险提供参考。

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引用次数: 0

Hematopoietic stem cell transplantation across ABO compatibility barrier in HLA genoidentical related donor: Is it an unfavorable factor for overall survival (OS) in Tunisian patients? 在HLA基因相同的相关供者中，跨越ABO相容性屏障的造血干细胞移植：这是突尼斯患者总生存（OS）的不利因素吗？

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-10 DOI: 10.1016/j.trim.2025.102311

Mohamed Hichem Sellami , Yesmine Boughzala , Eya Ghazouani , Nour Ben Abdeljelil , Ines Turki , Wafa Aissa , Manel Chaabane , Houda Kaabi , Tarek Ben Othman , Slama Hmida

Benchmarking of survival following Hematopoietic Stem Cell Transplantation (HSCT) is very useful in the field of transplant medicine. It has been reported that decreased survival rates after HSCT may be affected by the ABO mismatch in some cases. In this study, we aim to assess this relationship in a Tunisian cohort of adult patients who have received HSCT from HLA genoidentical related donors.

A total of 147 patients and their 147 respective related donors, were retrospectively analyzed in this study. HLA and ABO antigens typing was performed according to established routine methods. Overall Survival (OS) was measured from the date of the HSCT until death from any cause or the last follow-up examination. OS in the ABO-matched group and the ABO-mismatched groups was accessed using mean survival time, comparison of survival curves (using the Logrank test) and calculation of the hazard ratios (HR) with 95 % confidence interval (CI).

The comparison of survival curves showed no significant difference (chi2 = 0.898, p = 0.825) in the OS between the ABO-compatible graft recipients and those who are incompatible (Bidirectional, Major or Minor incompatibility). Survival analysis with HR showed no difference in mortality between the fourth targeted subgroups. This is the same result even when the source of the hematopoietic graft, i.e., peripheral blood stem cells (PBSCs) or bone marrow (BM) stem cells, was taken into consideration.

It is very likely that there is no effect of ABO mismatching on OS in Tunisian adult recipients of HSCs. This result is in good agreement with that of other recent reports that failed to establish a significant relationship between ABO incompatibility and OS after the HSCT.

造血干细胞移植（HSCT）后生存的基准测试在移植医学领域非常有用。据报道，在某些情况下，造血干细胞移植后生存率下降可能受到ABO不匹配的影响。在这项研究中，我们的目的是评估突尼斯一组接受HLA基因相同相关供者造血干细胞移植的成年患者的这种关系。本研究对147例患者及其147例相关供体进行回顾性分析。按常规方法进行HLA和ABO抗原分型。总生存期（OS）从造血干细胞移植之日起至任何原因死亡或最后一次随访检查。采用平均生存时间、生存曲线比较（采用Logrank检验）和计算风险比（HR）（95% %置信区间（CI））来获取abo匹配组和abo不匹配组的OS。生存曲线比较显示，abo血型相容与abo血型不相容（双向、主要或次要不相容）的移植受者OS无显著差异（chi2 = 0.898,p = 0.825）。HR的生存分析显示，第四个目标亚组之间的死亡率没有差异。即使考虑到造血移植物的来源，即外周血干细胞（PBSCs）或骨髓干细胞（BM），结果也是一样的。很可能ABO错配对突尼斯成年造血干细胞受者的OS没有影响。这一结果与最近其他未能建立造血干细胞移植后ABO不相容与OS之间的显著关系的报道很好地一致。

{"title":"Hematopoietic stem cell transplantation across ABO compatibility barrier in HLA genoidentical related donor: Is it an unfavorable factor for overall survival (OS) in Tunisian patients?","authors":"Mohamed Hichem Sellami , Yesmine Boughzala , Eya Ghazouani , Nour Ben Abdeljelil , Ines Turki , Wafa Aissa , Manel Chaabane , Houda Kaabi , Tarek Ben Othman , Slama Hmida","doi":"10.1016/j.trim.2025.102311","DOIUrl":"10.1016/j.trim.2025.102311","url":null,"abstract":"<div><div>Benchmarking of survival following Hematopoietic Stem Cell Transplantation (HSCT) is very useful in the field of transplant medicine. It has been reported that decreased survival rates after HSCT may be affected by the ABO mismatch in some cases. In this study, we aim to assess this relationship in a Tunisian cohort of adult patients who have received HSCT from HLA genoidentical related donors.</div><div>A total of 147 patients and their 147 respective related donors, were retrospectively analyzed in this study. HLA and ABO antigens typing was performed according to established routine methods. Overall Survival (OS) was measured from the date of the HSCT until death from any cause or the last follow-up examination. OS in the ABO-matched group and the ABO-mismatched groups was accessed using mean survival time, comparison of survival curves (using the Logrank test) and calculation of the hazard ratios (HR) with 95 % confidence interval (CI).</div><div>The comparison of survival curves showed no significant difference (chi2 = 0.898, <em>p</em> = 0.825) in the OS between the ABO-compatible graft recipients and those who are incompatible (Bidirectional, Major or Minor incompatibility). Survival analysis with HR showed no difference in mortality between the fourth targeted subgroups. This is the same result even when the source of the hematopoietic graft, i.e., peripheral blood stem cells (PBSCs) or bone marrow (BM) stem cells, was taken into consideration.</div><div>It is very likely that there is no effect of ABO mismatching on OS in Tunisian adult recipients of HSCs. This result is in good agreement with that of other recent reports that failed to establish a significant relationship between ABO incompatibility and OS after the HSCT.</div></div>","PeriodicalId":23304,"journal":{"name":"Transplant immunology","volume":"93 ","pages":"Article 102311"},"PeriodicalIF":1.4,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145275787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Recurrent sarcoidosis in a transplanted kidney: A case report and literature review 移植肾复发性结节病1例报告并文献复习。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-09-16 DOI: 10.1016/j.trim.2025.102300

Abd Assalam Qannus , H. Tahsin Özpolat , Dory Salazar , Erica Bracamonte , Bekir Tanriover , Venkatesh Ariyamathu

Sarcoidosis is a multisystem inflammatory disorder that primarily affects the lungs. However, extra-pulmonary involvement, including the kidneys and heart, is also observed. Kidney involvement may manifest as hypercalcemia or as acute or chronic kidney injury, which could progress to end-stage kidney disease. Patients with sarcoidosis have been reported to successfully undergo kidney transplants, although some have experienced recurrence of sarcoidosis after the transplant, affecting both the kidneys and other organs. Here, we present a case of a patient who underwent a kidney transplant followed by the recurrence of sarcoidosis, which manifested as hypercalcemia five months after the transplant.

结节病是一种多系统炎症性疾病，主要影响肺部。然而，肺外受累，包括肾脏和心脏，也被观察到。肾脏受累可表现为高钙血症或急性或慢性肾损伤，可发展为终末期肾脏疾病。有报道称，结节病患者成功地接受了肾移植，尽管有些患者在移植后结节病复发，影响肾脏和其他器官。在此，我们报告一例接受肾移植后结节病复发的患者，移植后5个月表现为高钙血症。

引用次数: 0

Diagnostic value of dd-cfDNA and CXCL-10 in kidney allograft recipients for identifying acute rejection dd-cfDNA和CXCL-10对肾移植受者急性排斥反应的诊断价值。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-05 DOI: 10.1016/j.trim.2025.102307

Qi Yu , Yan Zhang , Zijian Gao , Boqian Wang , Hongwei Yang , Long He

Background and hypothesis

Acute rejection (AR) remains a major challenge in kidney transplantation. Current methods used to diagnose acute rejection are either invasive or not sufficiently sensitive. Thus, in this study, we aimed to develop a novel and sensitive diagnostic tool for predicting acute rejection after kidney transplantation. We investigated if donor-derived cell-free DNA (dd-cfDNA) in the serum and C-X-C motif chemokine 10 (CXCL-10) levels were closely related to the occurrence of AR after renal transplantation.

Methods

We collected data and tested the serum levels of dd-cfDNA and CXCL-10 in AR patients and patients who underwent kidney transplantation during the same period. Logistic analysis was used to verify risk factors associated with AR. The analysis of the receiver operating characteristic curve(ROC) was used to reveal correlation between dd-cfDNA，CXCL-10 levels and AR patients.

Results

We included 13 AR patients and 18 patients without acute rejection. The AR group had a lower BMI (P = 0.006), and higher levels of eGFR (P = 0.006) and CysC (P = 0.009). Additionally, ln dd-cfDNA percentage and ln CXCL-10 content were higher in the AR group compared to the control group (P = 0.007, P = 0.004).The results of logistic analysis suggested CysC(OR:2.562[95 % CI:1.377–4.766]，P = 0.003),ln dd-cfDNA percent (OR: 10.521 [95 % CI: 2.091–52.938]，P = 0.004), and ln CXCL-10 content(OR:49.052 [95 % CI: 1.730–1391.073]，P = 0.023) can indicate the risk of AR and were independently correlated with the occurrence of acute rejection of the kidney graft. The AUC of serum dd-cfDNA (AUC = 0.723, P = 0.0348) percent and CXCL-10 content (AUC = 0.814, P = 0.0029) showed significant diagnostic performance. The combined dd-cfDNA and CXCL-10 diagnosis showed higher AUC and specificity.

Conclusions

Serum dd-cfDNA and CXCL-10 levels were significantly elevated in patients with acute rejection. Thus, dd-cfDNA and CXCL-10 can play suggestive roles in detecting graft injury after kidney transplantation.

背景和假设：急性排斥反应（AR）仍然是肾移植的主要挑战。目前用于诊断急性排斥反应的方法要么是侵入性的，要么不够敏感。因此，在这项研究中，我们旨在开发一种新的、灵敏的诊断工具来预测肾移植后的急性排斥反应。我们研究了血清中供体来源的无细胞DNA （dd-cfDNA）和C-X-C基序趋化因子10 （CXCL-10）水平是否与肾移植后AR的发生密切相关。方法：收集资料，检测AR患者和同期肾移植患者血清中dd-cfDNA和CXCL-10水平。采用Logistic分析验证与AR相关的危险因素。采用受试者工作特征曲线（receiver operating characteristic curve， ROC）分析dd-cfDNA、CXCL-10水平与AR患者的相关性。结果：我们纳入13例AR患者和18例无急性排斥反应的患者。AR组BMI较低（P = 0.006），eGFR （P = 0.006）和CysC （P = 0.009）水平较高。此外，AR组的ln dd-cfDNA百分比和ln CXCL-10含量高于对照组（P = 0.007,P = 0.004）。物流分析的结果提出CysC (OR: 2.562(95 %置信区间:1.377—-4.766),P = 0.003),ln dd-cfDNA百分比(OR: 10.521(95 %置信区间:2.091—-52.938),P = 0.004),和ln CXCL-10内容(OR: 49.052(95 %置信区间:1.730—-1391.073),P = 0.023)可以表明基于“增大化现实”技术的风险,独立与肾移植急性排斥反应的发生。血清dd-cfDNA的AUC (AUC = 0.723,P = 0.0348)%和CXCL-10含量（AUC = 0.814,P = 0.0029）具有显著的诊断价值。dd-cfDNA和CXCL-10联合诊断显示更高的AUC和特异性。结论：急性排斥反应患者血清dd-cfDNA和CXCL-10水平显著升高。因此，dd-cfDNA和CXCL-10在肾移植后移植物损伤检测中具有提示作用。

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引用次数: 0

Assessing the influence of isolation techniques on NK cell transcriptomic profiles 评估分离技术对NK细胞转录组谱的影响。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-09-14 DOI: 10.1016/j.trim.2025.102298

Rana ElNouty , Ahmed Moustafa , Maha Mostafa , Amged Ouf , Khaled Abou-Aisha , Mona Rady

Natural Killer (NK) cells are vital components of the innate immune system, playing a crucial role in defending the body against tumors and virally infected cells. While various methods exist for their isolation, the profound impact of these techniques on NK cell biology remains poorly characterized.This study presents a comprehensive analysis of the transcriptomic profiles of NK cells isolated using different positive selection methods; anti-CD56, anti-CD7 (with two distinct lineage depletion protocols), and a combination of anti-CD16 and anti-CD56 antibodies, compared to negative selection using immunomagnetic beads. Our integrated analysis of RNA-Seq datasets revealed that the isolation method is a dominant source of transcriptomic variation, accounting for 68.6 % of the total dataset variance, with technical factors being inextricably confounded with this biological signal. We identified extensive method-specific transcriptional signatures, with minimal overlap (<0.1 %) in differentially expressed genes (DEGs) across techniques. Functional enrichment analysis demonstrated that these signatures correspond to starkly different functional states: anti-CD16/anti-CD56 selection enriched for a highly activated, cytotoxically competent NK cell population with upregulated pathways in cytotoxicity and immune surveillance, while one anti-CD7-based method captured NK cells in a suppressed state, showing significant downregulation of lymphocyte activation and cytotoxicity pathways. Marker expression analysis further revealed extreme inter-study heterogeneity, with fold-changes in key cytotoxic genes exceeding 70,000-fold between methods. These findings highlight that the choice of isolation technique is not neutral but fundamentally determines the transcriptional and functional identity of the studied NK cell population. Our results highlight the critical importance of methodological standardization in NK cell research and provide essential guidance for selecting isolation strategies tailored to specific research or therapeutic applications.

自然杀伤细胞（NK）是先天免疫系统的重要组成部分，在保护身体免受肿瘤和病毒感染细胞的侵害方面起着至关重要的作用。虽然存在各种方法来分离NK细胞，但这些技术对NK细胞生物学的深远影响仍然缺乏表征。本研究对使用不同阳性选择方法分离的NK细胞的转录组学特征进行了全面分析；与使用免疫磁珠的阴性选择相比，抗cd56，抗cd7（有两种不同的谱系耗尽方案），以及抗cd16和抗cd56抗体的组合。我们对RNA-Seq数据集的综合分析表明，分离方法是转录组变异的主要来源，占总数据集变异的68.6% %，技术因素与这种生物信号不可避免地混淆在一起。我们发现了广泛的方法特异性转录特征，重叠最小(

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引用次数: 0

Fxyd5 downregulation protects the heart from ischemia/reperfusion injury by suppressing myocardial inflammation Fxyd5下调通过抑制心肌炎症保护心脏免受缺血/再灌注损伤。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-09-03 DOI: 10.1016/j.trim.2025.102282

Lei Zhang , Junjie Xu , Fujiang Cui , Jin Jin , Liwen Liu , Lei Wang , Yuxia Gao

Background

Myocardial ischemia/reperfusion (I/R) injury is a common cause of death. FXYD domain-containing ion transport regulator-5 (Fxyd5) is a type I membrane protein that plays a significant role in mediating cellular functions. However, the expression and function of Fxyd5 in myocardial I/R injury remain unclear.

Methods

Male C57BL/6 mice (8–10 weeks) were subjected to myocardial I/R by ligating the left anterior descending coronary artery. Preventive intervention was performed using adeno-associated virus 9 (AAV9)-mediated Fxyd5 knockdown. An in vitro hypoxia/reoxygenation (H/R) in H9c2 cardiomyoblasts was used for validation.

Results

Myocardial I/R injury significantly upregulated Fxyd5 expression (p < 0.01). Silencing Fxyd5 markedly improved cardiac function, as evidenced by a 58.0 % increase in EF (p < 0.01) and a 54.5 % reduction in infarct size (p < 0.01). Fxyd5 knockdown also attenuated myocardial apoptosis and inflammation, demonstrated by decreased cleaved caspase-3 expression and reduced levels of IL-6 (−44.2 %) and TNF-α (−42.7 %) (p < 0.01). Mechanistically, Fxyd5 silencing suppressed NF-κB activation, and these protective effects were confirmed in vitro in H/R-treated cardiomyocytes.

Conclusion

Fxyd5 silencing significantly attenuated I/R-induced myocardial injury, reduced infarct size, and improved cardiac function, while also decreasing apoptosis and inflammatory cytokine expression (p < 0.01). These findings indicate that Fxyd5 contributes to the pathogenesis of myocardial I/R injury and that its inhibition confers cardioprotective effects, partly through suppression of the NF-κB pathway.

背景：心肌缺血/再灌注（I/R）损伤是常见的死亡原因。FXYD结构域离子运输调节因子-5 （Fxyd5）是一种I型膜蛋白，在调节细胞功能中起重要作用。然而，Fxyd5在心肌I/R损伤中的表达和功能尚不清楚。方法：雄性C57BL/6小鼠（8 ~ 10 周）结扎左冠状动脉前降支进行心肌I/R。采用腺相关病毒9 （AAV9）介导的Fxyd5敲低进行预防性干预。采用体外缺氧/再氧化（H/R）法对H9c2型心肌细胞进行验证。结果：心肌I/R损伤可显著上调Fxyd5的表达（p ）结论：Fxyd5沉默可显著减轻I/R诱导的心肌损伤，减少梗死面积，改善心功能，同时减少细胞凋亡和炎症细胞因子的表达(p

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引用次数: 0

Pronase treatment enhances the sensitivity of B cell flow cytometry crossmatch: A single-center data analysis Pronase治疗提高B细胞流式细胞术交叉匹配的敏感性：单中心数据分析。

IF 1.4 4区医学 Q4 IMMUNOLOGY

Transplant immunology

Pub Date : 2025-12-01 Epub Date: 2025-10-30 DOI: 10.1016/j.trim.2025.102325

Yan Li , Dean Sylvaria , Madeleine Billingsley , Indira Guleria

The Flow Cytometry Crossmatch (FCXM) is a vital tool in HLA laboratories for assessing transplantation immunological risk. This study evaluates FCXM results using data from 2018 to 2023, with the aim to assess the efficacy of pronase treatment in FCXM assay by comparing control serum samples in the FCXM assays performed during this time period. Tests followed our standard protocol using the BD FACSLyric™ Flow Cytometry System and BD FACSCalibur™ Flow Cytometry System, employing normal human serum (NHS) as negative control and pooled patient serum samples with strong anti-HLA antibodies as positive control. Pronase and non-pronase tests were performed and compared to rule out any false T cell positive crossmatches that could be attributable to pronase treatment.

Results showed no significant difference in delta values for T cell crossmatches between pronase-treated (MCS 256 ± 112) and non-treated (MCS 247 ± 116) groups (p = 0.253). However, B cell crossmatches had significantly higher delta values with pronase treatment (MCS 353 ± 129) versus non-treatment (MCS 300 ± 117), indicating enhanced sensitivity with pronase (p < 0.000001). Pronase-treated B cell negative controls had lower signal (MCS 245 ± 61) compared to non-treated (MCS 300 ± 73), with positive control values consistent across both groups.

Anti-CD23 analysis demonstrated a reduction in all the tests performed (MCS 372 ± 64 vs 217 ± 50), confirming efficacy of pronase treatment. These findings highlight the importance of pronase in improving FCXM sensitivity for B cells, enhancing the accuracy and reliability of FCXM protocols for better transplantation outcomes.

流式细胞仪交叉配型（FCXM）是HLA实验室评估移植免疫风险的重要工具。本研究使用2018年至2023年的数据评估FCXM结果，目的是通过比较这段时间内FCXM检测中运行的各种血清样本，评估pronase治疗在FCXM检测中的疗效。测试采用我们的标准方案，使用BD FACSCalibur™流式细胞仪系统和BD FACSCalibur™流式细胞仪系统，以正常人血清（NHS）作为阴性对照，合并具有强抗hla抗体的患者血清样本作为阳性对照。进行Pronase和非Pronase测试并进行比较，以排除可能归因于Pronase治疗的任何假T细胞阳性交叉匹配。结果显示，pronase处理组（MCS 256 ± 112）与未处理组（MCS 247 ± 116）T细胞交叉匹配δ值无显著差异（p = 0.253）。然而，pronase处理（MCS 353 ± 129）与未处理（MCS 300 ± 117）相比，B细胞交叉配型的δ值显着更高，表明pronase增强了敏感性(p

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