新型白血病疫苗对小鼠巨噬细胞作用的研究(英文)

目的探讨自制的一种白血病疫苗对 C57/BL6小鼠 M(?)杀伤功能的影响。方法建立白血病荷瘤小鼠模型,用自制的3种不同的白血病疫苗进行预防或主动免疫治疗,用 MTT 比色法检测预防或治疗2周、4周后小鼠 M(?)杀伤活性,并与对照组比较。结果 (1)随着白血病细胞在小鼠体内的生长,小鼠的 M(?)免疫功能受到严重抑制。(2)灭活肿瘤细胞+IFA(不完全福氏佐剂)+细胞因子(rGM-CSF+rIL-2+rIL-6)的白血病疫苗在提高小鼠的 M(?)免疫功能方面,优于灭活白血病细胞+IFA 疫苗,而仅含灭活白血病细胞的疫苗作用不明显。结论灭活白血病细胞+IFA+细胞因子(rGM-CSF+rIL-2+rIL-6)的肿瘤疫苗可以激活以 M(?)为代表的非特异性细胞免疫功能,如非特异性细胞毒、免疫监视、抗原递呈等功能,为进一步活化 T 淋巴细胞打下基础,此种疫苗在血液恶性肿瘤的特异性主动免疫治疗中很有潜力。     

白血病瘤苗对荷瘤小鼠细胞免疫功能影响的研究

目的：探讨白血病细胞疫苗主动免疫治疗在血液恶性肿瘤中的作用。方法：建立红白血病荷瘤小鼠,用自制的3种不同的白血病细胞瘤苗进行主动免疫治疗,用MTT比色法测其治疗2周和4周的Mφ和CTL的杀伤活性,并与对照组相比较。结果：（1）随着肿瘤细胞在小鼠体内的增长,小鼠的细胞免疫功能受到严重抑制,特异性细胞免疫功能的抑制滞后于非特异性细胞免疫功能。（2）灭活肿瘤细胞＋IFA＋细胞因子（rGM-CSF rIL-2 rIL-6)的肿瘤疫苗在提高荷瘤小鼠的细胞免疫功能方面,尤其是特异性细胞免疫功能方面优于灭活肿瘤细胞＋IFA肿瘤疫苗,而仅含灭活肿瘤细胞的疫苗作用不明显,结论：灭活肿瘤细胞＋IFA＋细胞因子（rGM-CSF rIL-2 rIL-6)的肿瘤疫苗在血液恶性肿瘤的特异性主动免疫治疗中很有潜力。     

细胞瘤苗对红白血病荷瘤小鼠抗瘤作用的研究

目的：观察三种瘤苗抗肿瘤作用。方法：用不同佐剂的三种瘤苗，免疫预防及治疗红白血病荷瘤小鼠。观察肿瘤生长情况，小鼠寿命，局部肿瘤、全身脏器反应及病理变化。结果：由灭活细胞、不完全福氏佐剂及细胞因子组成的瘤苗，预防后小鼠成瘤率低，预防及治疗后肿瘤生长速度减慢，存活期延长，与灭活细胞，灭活细胞加不完全福氏佐剂瘤苗相比，有显著性差异；病理可见瘤组织坏死及以单个核细胞为主的炎细胞浸润，各脏器无异常，结论：包含细胞因子的瘤苗是安全，方便，经济、有效的瘤苗。     

急性白血病的新型主动免疫治疗研究

目的：应用化疗不能治愈大多数急性白血病,近年人们对基因工程疫苗寄以厚望,动物实验已展现其具有明显疗效,但迄今尚未见单独应用白血病疫苗主动免疫及治疗急性白血病患取得明显疗效的报道。本应用基因工程生产的细胞因子及白血病细胞制备的疫苗,主要免疫治疗21例急性白血病,以探讨提高主动免疫治疗急必白血病疗效的方法。方法：1．新型白血病疫苗制备;取化疗前或化疗末缓解的白血患自身骨髓15ml肝素抗凝,应用淋巴细胞分离液分离出富含白血病细胞的骨髓单个核细胞,经丝裂霉素处理后,加入含基因工程生产的粒单细胞集落刺激因子,白细胞介素2及白细胞介素6等的福氏佐剂中。2．主动免疫治疗程序：对所有接受免疫治疗患在停用化疗两周后进行骨髓检查并开始接种疫苗,每次用含10^8个白血病细胞的疫苗1．6ml,分两点前臂皮下注射,每隔7天重复一次,共4次,然后长期随访。如无效时再改用其它治疗;如有效,每隔6个月加强注射一次。结果：21例急性白血病患均因化疗无效或不能耐受耐改用疫苗治疗。免疫治疗达到完全缓解（CR）5例,为24％,部分缓解（PR）4例,为19％,微效（MR）3,为14％,总有效率达到575。有效多为女性,低增生性急性单核细胞白血病（M5）。疗效平均持续时间约7个月后易复发;而例有效率患在每6个月时应用此疫苗加强免疫治疗,已分别存活18个月及24个月。用此疫苗治疗时未见明显毒副作用,对5例疫苗治疗有效及3例无效患应用免疫组化方法检测其白血病细胞膜上MHC1．2类分子及共刺激分子（B7－1）,发现5例有效病例的MHC1．2类分子及B7－1均为阳性,而3例无效病例虽MMHC1．2类分子阳性,但B7分子均为阴性。结论：结果表明应用此新型白血病疫苗主动免疫治疗化疗无效及不能耐受化疗的某些急性白血病,具有肯定疗效,且经济、简便、无明显毒副作用,值得进一步研究,特别适用于单核细胞性的、B7阳性的、低增生性的白血病患。     

新型白血病疫苗对小鼠巨噬细胞作用的研究

The study is to evaluate the cytotoxicity of macrophage of C57BL/6 mice treated with the prepared leukemia vaccine. Establish the leukemia burden mice and prepare three types of leukemia vaccine, before they are administered on the mice respectively. Measure the cytotxicity of M derived from the mice immunotherapy or prevention after 2–4 weeks, adoption with MTT colorimetric method and compare with control group accordingly. The finding of the study include: (1) Seriously depressed cytotoxicity of M, with the growth of leukemia cells in the mice, (2) More effective and efficient cellular immunity of tumor burden mice with the administration of tumor vaccine prepare from inactivated tumor cells, IFA and cytokines, such as rGM-CSF, rIL-2 and rIL-6, than tumor vaccine from either inactivated tumor cells and IFA or inactive tumor cells per se. The tumor vaccine prepared with inactive tumor cells, IFA and cytokines can activate the non-specific cellular immunity such as M, as well as, antigen present, immune surveillance and etc. and this activation forms a base for further activate T lymphocyte. Naturally, this will certainly have a promising future in the therapy against hematopietic tumor.     

特异性抗髓性白血病多肽疫苗的研制和应用

化疗和造血干细胞移植是治疗白血病的主要方法，但是由于受供者来源和年龄的限制，化疗后微小残留病灶的存在使白血病容易复发。对白血病化疗缓解后给予疫苗治疗是消除残留病灶的理想方法。白血病相关抗原（LAAs）是目前研究最多的免疫治疗的靶抗原，对于髓性白血病，大多数LAAs既是白血病细胞过度表达的自身抗原，也是染色体易位的融合蛋白。由于许多LAAs不是膜蛋白，不能作为抗体治疗的对象。     

治疗急性髓系白血病的树突状细胞疫苗的研究进展

摘 要 本文综述了免疫治疗急性髓系白血病（acute myeloid leukemia, AML）的树突状细胞（dendritic cells, DC）疫苗的近期研究进展，主要涉及各种类型的白血病肿瘤抗原的确认与制备，包括AML特异融合蛋白及相关多肽、癌症睾丸抗原、酸洗脱多肽、白血病细胞冻融抗原/凋亡细胞、AMLDC融合细胞、白血病细胞来源的DC等；DC的制备与特性以及优化方法；临床试验研究的初步结果等，还对今后的研究方向进行了分析和展望。     

利播仃(Lipostn)肿瘤疫苗诱发BALB/c小鼠的抗肿瘤免疫作用Ⅱ——细胞免疫

前文阐述了利播仃肿瘤诱发BALB／c小鼠的体液免疫．激活B淋巴细胞，使其分泌高滴度、高特异性及有细胞毒的抗体。对于主动特异性免疫治疗肿瘤的疫苗来说，仅有此作用是不够的，必须能激活机体的免疫系统，发挥免疫功能，产生细胞免疫，对肿瘤细胞发起多方位的主动攻击。这就是肿瘤疫苗的主动特异性免疫治疗肿瘤的独特之处。     

新城疫病毒瘤苗对荷瘤小鼠的主动免疫治疗作用

将小鼠原发性肝癌细胞（Ｈ２２）经γ射线照射灭活后，与新城疫病毒（ＮＤＶ）共同孵育，制备病毒瘤苗，透射电镜下见ＮＤＶ吸附于Ｈ２２细胞表面。以病毒瘤苗皮内注射治疗不同负荷的Ｈ２２肿瘤荷瘤小鼠，连续２次，间隔１周，每次每只小鼠免疫剂量为１．５×１０７个肿瘤细胞。结果表明病毒瘤苗能抑制肿瘤生长，对荷瘤小鼠有主动特异性免疫治疗作用。各免疫组小鼠存活时间较对照组明显延长，肿瘤重量较对照组轻，肿瘤的平均直径也较对照组小，免疫组荷瘤存活小鼠肿瘤病理切片显示肿瘤细胞呈中、重度坏死，坏死的肿瘤细胞之间及其周围可见大量淋巴细胞及单核样细胞浸润，并有纤维组织增生。     

CpG ODN对急性粒-单核细胞白血病荷瘤小鼠免疫治疗作用研究

目的:探讨两种CpG ODN序列对急性粒-单核细胞白血病荷瘤小鼠模型的免疫治疗作用。方法:通过注射WEHI-3细胞构建急性粒-单核细胞白血病荷瘤小鼠模型，分别皮下注射PBS无菌缓冲液、CpG 1826、CpG Seq14，观察治疗后各组小鼠的一般状况、生存时间、瘤结节生长、病理切片等，综合评价两种CpG ODN的治疗效果。结果:皮下接种1×106个WEHI-3细胞，成功建立急性粒-单核细胞白血病荷瘤小鼠模型。肿瘤组平均生存时间20.87天，CpG 1826组 32.60天，CpG Seq14组 28.35天，两种CpG治疗组小鼠平均存活时间明显较肿瘤组长，P＜0.05；实验初期，肿瘤组及两CpG治疗组小鼠较正常组小鼠体重增长明显，终末期体重明显下降；肿瘤组小鼠瘤结节较同期两CpG治疗组小鼠大，且增长速度快；肿瘤组小鼠肝脏、脾脏、肿瘤组织中均可见广泛瘤细胞浸润，CpG 治疗组肝脏、脾脏中瘤细胞浸润明显减少，脾脏白髓区明显较对照组扩大，小鼠免疫功能增强，减少了肿瘤细胞的器官浸润；两实验组小鼠注射CpG部位均未见红肿、硬结、坏死等不良反应。结论:CpG 1826、CpG Seq14直接用于急性粒-单核细胞白血病荷瘤小鼠模型的免疫治疗，均可增强小鼠抗肿瘤作用，延长生存时间，且无明显毒副作用。     

不同时期应用细胞瘤苗对红白血病荷瘤小鼠抗瘤作用的研究

目的：探讨不同时间应用细菌瘤苗对红细胞病荷瘤小鼠的抗瘤作用。方法：在不同时间内应用瘤苗免疫治疗红白血病荷瘤小鼠，观察小鼠生存期，皮下肿瘤大小及肿瘤，注射部位病理变化。结果：3天，7天瘤苗治疗的小鼠生存期延长，皮下肿瘤生长缓慢，病理可见瘤组织坏死及大量以单个核细胞为主的炎细胞浸润，与PBS组及10天瘤苗组相比，有显著性差异。结论：3天，7天瘤苗比10天瘤苗组抗肿瘤作用强。     

Effect of Human Macrophage Colony-stimulating Factor on Granulopoiesis and Survival in Bone-marrow-transplanted Mice

Human macrophage colony-stimulating factor (hM-CSF) has been isolated from normal human urine and purified to a homogenous protein. The effect of hM-CSF on granulopoiesis was investigated in BALB/c mice transplanted with a suboptimal number of bone marrow cells. Lethally irradiated (7.8 Gy) mice were transplanted with 1 × 10⁶ syngeneic mouse bone marrow cells and treated with a daily intraperitoneal dose of 64 μg/kg of hM-CSF for 5 days following the transplant. The hM-CSF injection resulted in stimulation of the recovery of blood neutrophils as well as an increase in the number of granulocyte-macrophage progenitor cells (CFU-GM) in the femur and spleen. The survival of lethally irradiated mice was dependent on the cell number transplanted; most mice transplanted with 2 × 10⁴ cells died within 2 weeks. The recovery of hematopoiesis in mice transplanted with 2 × 10⁴ cells was modestly but significantly stimulated by hM-CSF administration initiated from 5 days before or 1 day after transplantation for a 5-day period. Furthermore, the hM-CSF administrations markedly reduced the mortality in these mice during the early period after the transplantation. Since anaerobic bacteria were frequently detected in arterial blood immediately before the deaths but were not found in the surviving mice, it is speculated that early deaths occurring within 2 weeks after the transplant may be caused by opportunistic infections, and hM-CSF injection may prevent these mortal infections through its stimulating effect on monocyte-macrophage functions that are responsible for the production of hematopoietic regulators.     

S311抗癌菌苗对腹水癌小鼠单核巨噬细胞抗腹水癌作用的影响

目的：探讨Ｓ３１１抗癌菌苗能否增强腹水癌小鼠腹腔巨噬细胞和其他几种单核巨噬细胞的得水癌作用，以及Ｓ３１１抗癌菌苗与ＩＦＮ－γ、ＩＬ－２在抗肿瘤增殖上是否具有协同作用，为Ｓ３１１抗癌菌苗免疫治疗胸腹水癌提供依据。方法：对腹水癌小康习腹腔内注射Ｓ３１１抗癌菌苗、Ｓ３１１抗癌菌苗和细胞因子，连续３周，分别珩第１、２、３周从小鼠体内分离腹腔巨噬细胞、脾巨噬细胞、肺巨噬细胞、血液单核细胞，并分别与鼠腹水癌细     

Priming Effects of Macrophage Colony-Stimulating Factor on Monocytic Leukemia Cells in Combination with Chemotherapy: Induction of Programmed Cell Death In Vivo

Two elderly patients with chronic myelomonocytic leukemia were treated with cytosine ara-binoside (Ara-C) and aclarubicin (ACR) under simultaneous administrations of macrophage colony-stimulating factor (M-CSF) (CAM), and both obtained good responses. Examination of apoptosis using flow cytometry revealed induction of apoptotic death of leukemia cells by CAM in Patient 2, while neither induction of apoptotic death of leukemia cells nor clinical response were seen with CAG (Ara-C, ACR, and granulocyte colony-stimulating factor) given prior to CAM in Patient 1. These findings suggested that chemotherapy combined with simultaneous administration of M-CSF could effectively reduce monocytic leukemia cells by inducing programmed cell death.     

双歧杆菌DNA对小鼠腹腔巨噬细胞分泌和杀瘤功能的影响

[目的]探讨双歧杆菌DNA对小鼠腹腔巨噬细胞的激活作用。[方法]纯化提取双歧杆菌基因组DNA ,并鉴定DNA制品CpG基序甲基化程度。收集双歧杆菌DNA(10μg/ml)体外诱导培养24h的小鼠腹腔巨噬细胞培养上清 ,采用ELISA法检测培养上清中IL 1β、IL 6的含量 ,用Griess试剂检测其NO的含量 ;采用MTT间接法观察了双歧杆菌DNA诱导的巨噬细胞体外杀伤活性的变化。[结果]实验组(双歧杆菌DNA组)巨噬细胞产生IL 1β、IL 6及NO的水平分别为(270.12±31.51)pg/ml,(578.81±47.30)pg/ml和(11.12±1.01)μmol/L,其巨噬细胞杀伤活性为(31.02±1.91)% ;对照组(PBS组)分别为(138.12±9.24)pg/ml、(78.80±10.21)pg/ml、(2.14±0.82)μmol/L和(18.69±1.70) % ,两组间各项指标比较均有显著性差异(均P<0.01)。[结论]双歧杆菌DNA能激活巨噬细胞 ,可增强巨噬细胞IL 1β、IL 6及NO产生的水平及杀伤活性     

Serial Transplantation of Adult T Cell Leukemia Cells into Severe Combined Immunodeficient Mice

The precise mechanism of the neoplastic cell growth of adult T cell leukemia (ATL) still remains unclear. In the present study, we have succeeded in serial transplantation of ATL cells from a patient into severe combined immunodeficient (SCID) mice. In this model, we found that only a leukemic cell clone from an ATL patient could be successively transplanted into SCID mice, although it was difficult to maintain leukemic cell clones in vitro , suggesting that the microenvironment provided by SCID mice is suitable for leukemic cell growth. We could not detect human T cell leukemia virus type I (HTLV-I) mRNA or interleukin 2 (IL-2) mRNA in either the tumor cells growing in mice or the original leukemic cells. Thus, it appears that neither HTLV-I viral expression nor the IL-2 autocrine mechanism is directly involved in the neoplastic cell growth of fresh ATL cells as well as HTLV-I-infected cell lines, at least in SCID mice. In addition, we could passage frozen cells and obtain a large number of expanded leukemic cells in this model. Such a serial transplantation model, which can avoid the changes in the nature of leukemic cells that are frequently observed in in vitro culture, and which can propagate leukemic cell clones, would be very suitable not only to study the mechanism of neoplastic cell growth, but also to test potential therapeutic agents for ATL.