AbstractThe aim of this study was to observe the effects of different conditioning regimens on fine immune indices after microtransplantation (MST) in patients with acute myeloid leukaemia (AML). This article discusses the possible immune mechanism of microtransplantation and describes a more optimized conditioning regimen.

本研究的目的是观察不同预处理方案对急性髓系白血病（AML）患者微移植（MST）后精细免疫指标的影响。本文讨论了微移植可能的免疫机制，并描述了一种更优化的预处理方案。

A total of 55 AML patients who received MST treatment at the Second Hospital of Lanzhou University from August 2015 to October 2023 were included in this study, and 13 AML patients who did not receive MST but directly received the maintenance therapy were included as the control group (C). The MST patients were divided into a conditioning regimen with venetoclax group (A) and a conditioning regimen without venetoclax group (B).

本研究纳入了2015年8月至2023年10月在兰州大学第二医院接受MST治疗的55例AML患者，其中13例未接受MST但直接接受维持治疗的AML患者作为对照组（C）。MST患者分为使用venetoclax组（a）的预处理方案和不使用venetoclax组（B）的预处理方案。

The fine immune indices were detected by flow cytometry and cytometric bead array analysis. Changes in the immune indices before and after treatment were observed, and the progression-free survival (PFS) and overall survival (OS) of patients in the MST group were analysed. Compared with those in Group B, patients in Group A had better PFS and OS.

通过流式细胞术和细胞计数珠阵列分析检测精细免疫指标。观察治疗前后免疫指标的变化，分析MST组患者的无进展生存期（PFS）和总生存期（OS）。与B组相比，A组患者的PFS和OS更好。

The proportion of Treg cells and the expression level of IL-2 were increased, while TNF-α and IFN-α were decreased after MST (P < 0.05). In Group B, total T cells, CD4+T cells and CD4+/CD8+T cells decreased; NK cells and total B cells increased; and IL-17A first increased and then decreased during the MST (P < 0.05).

MST后Treg细胞比例和IL-2表达水平升高，TNF-α和IFN-α降低（P<0.05）。在B组中，总T细胞，CD4+T细胞和CD4+/CD8+T细胞减少；NK细胞和总B细胞增加；IL-17A在MST期间先升高后降低（P<0.05）。

There were significant differences in total B cells, IL-4 and IFN-γ between Group A and Group B during MST. Moreover, there were significant differences in total T cells, CD4+T cells, Treg cells, IL-17A, IFN-γ and IL-2 between the patients in the MST group and those in the control group (P < 0.05). The MST conditioning regimen containing venetoclax significantly changed t.

MST期间，A组和B组的总B细胞，IL-4和IFN-γ有显着差异。此外，MST组患者与对照组患者的总T细胞，CD4+T细胞，Treg细胞，IL-17A，IFN-γ和IL-2有显着差异（P<0.05）。含有venetoclax的MST预处理方案显着改变了t。

IntroductionCurrently, the main treatment for acute myeloid leukaemia (AML) is chemotherapy and haematopoietic stem cell transplantation1,2,3. Although new chemotherapeutic drugs and targeted drugs have made good progress, they still cannot significantly prolong the survival of patients4,5. Although allogeneic haematopoietic stem cell transplantation (allo-HSCT) can prolong the survival of AML patients, this effect is often accompanied by severe graft-versus-host disease (GVHD) and infection6.

引言目前，急性髓细胞白血病（AML）的主要治疗方法是化疗和造血干细胞移植1,2,3。虽然新的化疗药物和靶向药物取得了良好的进展，但它们仍然不能显着延长患者的生存期4,5。虽然异基因造血干细胞移植（allo-HSCT）可以延长AML患者的生存期，但这种效应通常伴有严重的移植物抗宿主病（GVHD）和感染6。

After transplantation, patients receive GVHD prophylaxis6,7. Traditional bone marrow transplantation is not only highly toxic but also limited by many factors, such as economic status and age8,9. In recent years, microtransplantation (MST) technology has eliminated the toxicity of traditional transplantation, reduced the side effects of transplantation, and retained the graft-versus-leukaemia effect, making it highly efficient and minimally toxic.

移植后，患者接受GVHD预防6,7。传统的骨髓移植不仅毒性高，而且受到许多因素的限制，例如经济状况和年龄8,9。近年来，显微移植（MST）技术消除了传统移植的毒性，减少了移植的副作用，并保留了移植物抗白血病作用，使其高效且毒性最小。

Haematopoietic stem cell transplantation is a treatment method that preserves the normal immune function of the recipient.The first MST clinical trial study published in Blood in 2011 confirmed the efficacy and safety of MST in the treatment of AML10. The results of a multicentre study on MST in elderly AML patients in JAMA Oncology showed that MST reduced the recurrence rate and prolonged the survival time of elderly AML patients11.

造血干细胞移植是一种保留受体正常免疫功能的治疗方法。2011年发表在《血液》上的第一项MST临床试验研究证实了MST治疗AML10的有效性和安全性。。

Guo et al.12 reported that MST also has a good therapeutic effect on young and middle-aged AML patients, with these treatment results characterized by rapid haematopoietic recovery, low nonrelapse mortality and no occurrence of GVHD during therapy. The main purpose of MST conditioning is to kill malignant cells and tumour cells while preserving the patient's immune system as much as possible.

Guo等[12]报道，MST对中青年AML患者也有良好的治疗效果，这些治疗结果的特点是造血功能恢复快，无复发死亡率低，治疗期间无GVHD发生。MST调理的主要目的是杀死恶性细胞和肿瘤细胞，同时尽可能保护患者的免疫系统。

The traditional.

传统的。

Table 1 Patient characteristics.Full size tableThere were 16 patients in Group A, whose age distribution ranged from 31 to 69 years, with a median age of 54.5 years; these patients included 6 males (37.5%) and 10 females (62.5%). The AML classification was as follows: M2: 5 patients, M4: 1 patient, M4EO: 4 patients, and M5: 6 patients.

表1患者特征。全尺寸表A组有16例患者，年龄分布在31至69岁之间，中位年龄为54.5岁；这些患者包括6名男性（37.5%）和10名女性（62.5%）。AML分类如下：M2:5例，M4:1例，M4EO:4例，M5:6例。

The risk stratification revealed 5 patients at low risk, 5 patients at medium risk and 6 patients at high risk.There were 39 patients in Group B; the age distribution was 8–56 years, and the median age was 38 years. There were 17 males (42.9%) and 22 females (56.41%). The AML classification was as follows: M0, 1 patient; M2, 22 patients; M4, 3 patients; M4EO, 1 patient; and M5, 12 patients.

风险分层显示5例低风险患者，5例中风险患者和6例高风险患者。B组39例；年龄分布为8-56岁，中位年龄为38岁。男性17例（42.9%），女性22例（56.41%）。；M2，22名患者；M4，3名患者；M4EO，1名患者；和M5，12名患者。

The risk stratification revealed low risk in 20 patients, medium risk in 12 patients, and high risk in 7 patients.Twenty-four patients achieved CR after one course of induction chemotherapy, 15 patients achieved CR after two courses of induction chemotherapy, 10 patients achieved CR after three or more courses of induction chemotherapy, 5 patients did not achieve CR, and 1 patient underwent MST directly without induction chemotherapy.

风险分层显示20例患者风险较低，12例患者风险中等，7例患者风险较高。24例患者在一个疗程的诱导化疗后达到CR，15例患者在两个疗程的诱导化疗后达到CR，10例患者在三个或更多疗程的诱导化疗后达到CR，5例患者未达到CR，1例患者直接接受MST而不进行诱导化疗。

A total of 49 patients achieved CR, and 6 patients did not achieve CR before MST.Thirteen AML patients who did not receive MST and who received maintenance therapy after achieving CR were selected as the control group. The clinical characteristics of the patients in the control group are shown in Table 1; the age distribution ranged from 11 to 68 years, and the median age was 54 years.

共有49例患者达到CR，6例患者在MST前未达到CR。选择13名未接受MST且在达到CR后接受维持治疗的AML患者作为对照组。对照组患者的临床特征如表1所示；年龄分布范围为11至68岁，中位年龄为54岁。

There was 1 female and 12 males. According to the FAB classification, 8 patients had M2, 2 patients had M4, and 3 patients had M5.Safety analysis of MSTA total of 178 MSTs were performed in 55 patients. The overall safety was high, with no .

有1名女性和12名男性。根据FAB分类，M2型8例，M4型2例，M5型3例。55例患者共进行了178例MST的安全性分析。总体安全性很高，没有。

Table 2 Experimental Instruments.Full size tableTable 3 Experimental reagents.Full size tableThe expression levels of twelve cytokines were detected by cytometric bead array (CBA)A serum tube was used to collect 2 mL venous blood samples from the included microtransplant patients and the control patients in the morning and without eating.

表2实验仪器。全尺寸表表3实验试剂。全尺寸表通过细胞计数珠阵列（CBA）检测12种细胞因子的表达水平。血清管用于在早晨和不进食的情况下从包括的微移植患者和对照患者收集2 mL静脉血样品。

After centrifugation, the upper layer of serum was extracted, and the standard substance was diluted to prepare the capture pellet suspension, which was incubated in the dark after adding fluorescent reagent. Then, PBS was added, and the mixture was centrifuged. The supernatant was discarded, and then PBS was added to resuspend the cells.

离心后，提取上层血清，稀释标准物质制备捕获颗粒悬浮液，加入荧光试剂后在黑暗中孵育。然后，加入PBS，并将混合物离心。弃去上清液，然后加入PBS以重悬细胞。

Experimental equipment and experimental reagents are shown in Tables 2 and 3.Follow-up visitThe patients were followed up mainly by consulting inpatient and outpatient medical records and by communicating via telephone. The follow-up period ended on May 1, 2024. Progression-free survival (PFS) was defined as the time from the initiation of treatment to disease progression or death from any cause.

实验设备和实验试剂如表2和表3所示。随访患者主要通过查阅住院和门诊病历以及通过电话沟通进行随访。随访期于2024年5月1日结束。无进展生存期（PFS）定义为从开始治疗到疾病进展或因任何原因死亡的时间。

Overall survival (OS) was defined as the overall time from randomization (i.e., disease diagnosis) to death from any cause or the last follow-up.Statistical analysisPython 3.11 was used for statistical analysis in this study. Lymphocyte subsets and twelve cytokines were used as measurement data, and the measurement data were normally distributed.

。统计分析本研究使用Python 3.11进行统计分析。淋巴细胞亚群和十二种细胞因子用作测量数据，测量数据呈正态分布。

One-way ANOVA was used for comparisons between groups. The Kaplan‒Meier method was used for survival analysis, mainly to observe OS and PFS, and the log-rank test was used for survival analysis. P < 0.05 was considered to indicate statistical significance.Ethical approvalThe present study was approved by the Ethics Committee of the Lanzhou University Second Hospital .

单因素方差分析用于组间比较。Kaplan-Meier方法用于生存分析，主要观察OS和PFS，对数秩检验用于生存分析。P<0.05被认为表示统计学显着性。伦理认可本研究经兰州大学第二医院伦理委员会批准。

Data availability

数据可用性

The data that support the findings of this study are available from the corresponding author upon reasonable request.

根据合理的要求，通讯作者可以提供支持本研究结果的数据。

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Download referencesAcknowledgementsWe are grateful to Xu Changhong for his effort in this study.FundingThis work was supported by the National Natural Science Foundation of China (82360029), the Commissioned Project of National Clinical Medicine Research Center for Hematological System Diseases (2021WWA01), the Clinical Medical Research Center of Hematological Diseases in Gansu Province (21JR7RA435), the Natural Science Foundation of Science and Technology Program of Gansu Province (21JR7RA394) and the Natural Science Foundation of Science and Technology Program of Gansu Province (21JR11RA104).Author informationAuthor notesThese authors contributed equally: Zhang Liansheng and Li Lijuan.Authors and AffiliationsDepartment of Hematology, Lanzhou University Second Hospital, Lanzhou University, Lanzhou, 730000, ChinaZheng Hanxue, Meng Zilu, Zhang Liansheng & Li LijuanKey Laboratory of the Hematology of Gansu Province, Lanzhou University Second Hospital Lanzhou University, Lanzhou, 730000, ChinaZheng Hanxue, Meng Zilu, Zhang Liansheng & Li LijuanAuthorsZheng HanxueView author publicationsYou can also search for this author in.

下载参考文献致谢我们感谢徐长虹在这项研究中的努力。资助这项工作得到了国家自然科学基金（82360029），国家血液系统疾病临床医学研究中心（2021WWA01）委托项目，甘肃省血液疾病临床医学研究中心（21JR7RA435），甘肃省自然科学基金科技计划（21JR7RA394）和甘肃省自然科学基金科技计划（21JR11RA104）的支持。作者信息作者注意到这些作者做出了同样的贡献：张连生和李丽娟。作者和附属机构兰州大学第二医院血液科，兰州大学，兰州，730000，中国甘肃省血液学实验室，兰州大学第二医院，兰州大学，兰州，730000，中国郑汉学，孟子路，张连生和李丽娟正汉学观点作者出版物你也可以在中搜索这位作者。

PubMed Google ScholarMeng ZiluView author publicationsYou can also search for this author in

PubMed Google ScholarMeng ZiluView作者出版物您也可以在

PubMed Google ScholarZhang LianshengView author publicationsYou can also search for this author in

PubMed谷歌学者张连生查看作者出版物您也可以在

PubMed Google ScholarLi LijuanView author publicationsYou can also search for this author in

PubMed Google ScholarLi LijuanView作者出版物您也可以在

PubMed Google ScholarContributionsL.L.: Conceptualization. Z.H.: Writing—original draft. L.L., Z.L., M.Z.: Writing—review & editing. L.L. and Z.L.: Supervision. All authors have read and agreed to the published version of the manuscript.Corresponding authorsCorrespondence to

PubMed谷歌学术贡献l。五十、 ：概念化。Z、 H.：撰写初稿。五十、 L.，Z.L.，M.Z.：写作评论和编辑。五十、 L.和Z.L.：监督。所有作者都阅读并同意手稿的出版版本。通讯作者通讯

Zhang Liansheng or Li Lijuan.Ethics declarations

张连生或李丽娟。道德宣言

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相互竞争的利益

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Reprints and permissionsAbout this articleCite this articleHanxue, Z., Zilu, M., Liansheng, Z. et al. Effects of different conditioning regimens on HLA-mismatched microtransplantation and changes in fine immune indices in acute myeloid leukaemia.

转载和许可本文引用本文Hanxue，Z.，Zilu，M.，Liansheng，Z。等人。不同预处理方案对HLA错配微移植的影响以及急性髓性白血病精细免疫指标的变化。

Sci Rep 14, 19301 (2024). https://doi.org/10.1038/s41598-024-70332-7Download citationReceived: 15 March 2024Accepted: 14 August 2024Published: 20 August 2024DOI: https://doi.org/10.1038/s41598-024-70332-7Share this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

科学报告1419301（2024）。https://doi.org/10.1038/s41598-024-70332-7Download引文接收日期：2024年3月15日接受日期：2024年8月14日发布日期：2024年8月20日OI：https://doi.org/10.1038/s41598-024-70332-7Share本文与您共享以下链接的任何人都可以阅读此内容：获取可共享链接对不起，本文目前没有可共享的链接。。

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KeywordsAcute myeloid leukaemiaMicrotransplantationFine immune indicesImmunologic mechanismImmunotherapy

关键词急性髓系白血病显微移植精细免疫指标免疫机制免疫治疗

Subjects

主题

Cancer immunotherapyCytokinesImmunotherapyLymphocytesTransplant immunology

癌症免疫治疗细胞因子免疫治疗淋巴细胞移植免疫学

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