白细胞介素-12基因再修饰基因瘤苗免疫小鼠的脾细胞过继治疗肝癌

目的　探讨白细胞介素 (IL) 2和B7 1双免疫基因转染肝癌细胞瘤苗免疫小鼠后获得的脾淋巴细胞经mIL 12基因修饰后治疗小鼠肝癌的可行性和疗效。方法　用 2 0 0PFU 细胞滴度的重组腺病毒载体 (Adv)将人IL 2和B7 1基因共同转染小鼠肝癌Hepal 6细胞株 ,经 80mg L丝裂霉素 (MMC)处理制备成肝癌细胞瘤苗 ,免疫同系小鼠后分离其脾淋巴细胞 (SLC) ,转染mIL 12基因 (Adv滴度 2 0 0PFU 细胞 )后注射入直径 1cm的小鼠皮下移植肝癌内 ,观察抗瘤效果。结果　转mIL 12基因SLC治疗组小鼠瘤体增加值最小 ( 0 .0 8± 0 .0 5 )cm3,与各对照组比差异有显著性 [(转染BGFP基因SLC、未转染SLC和生理盐水注射组分别为 ( 3 .46± 0 .15 ) ,( 3 .5 6± 0 .2 3)和( 8.12± 0 .5 4)cm3,P <0 .0 5 ]。结论　IL 2和B7 1双基因修饰肝癌瘤苗诱导小鼠产生的免疫脾淋巴细胞可能成为一种新的过继免疫治疗效应细胞及携带IL 12基因的载体细胞 ;基因治疗、特异性主动免疫和过继性细胞免疫治疗结合将有更优越的抗瘤效果。     

IL－2LAK细胞局部治疗眼部恶性肿瘤的临床效果

应用γＩＬ－２体外诱导自身ＬＡＫ细胞（ｌｙｍｐｈｏｋｉｎｅａｃｔｉｖａｔｅｄｋｉｌｌｉｎｇｃｅｌｌ），对眼部早、中期恶性肿瘤病人进行瘤区注射，取得较好效果。２５例恶性肿瘤中，ＩＬ－２ＬＡＫ细胞治疗１０例；肿瘤完全消退９例，部分消退１例。手术＋ＩＬ－２ＬＡＫ细胞治疗１５例，肿瘤完全消退。随访中除１例肿瘤部分消退失访外，其他２４例经１～５年观察均未见复发。以上结果表明，自身ＬＡＫ细胞局部治疗是一种有效的肿瘤免疫治疗方法。     

Comparison of allergenicity and immunogenicity of an intact allergen vaccine and commercially available allergoid products for birch pollen immunotherapy

BACKGROUND: Specific immunotherapy with intact allergen vaccine is a well-documented treatment for allergic diseases. Different vaccine formulations are currently commercially available, the active ingredient either being intact allergens or chemically modified allergoids. The rationale behind allergoids is to decrease allergenicity while maintaining immunogenicity. However, data from the German health authorities based on reporting of adverse events over a 10-year period did not indicate increased safety of allergoids over intact allergens. OBJECTIVE: The objective of this study was to investigate the effect of chemical modification on allergenicity and immunogenicity comparing four commercial allergoid products for birch pollen immunotherapy with an intact allergen vaccine. METHODS: Solid-phase IgE inhibition and histamine release assays were selected as model systems for allergenicity, and a combination of human T cell proliferation and IgG titres following mouse immunizations were used to address the immunogenicity of the intact allergen vaccine and the four allergoids. In all assays, the products were normalized with respect to the manufacturer's recommended maintenance dose. RESULTS: IgE inhibition experiments showed a change in epitope composition comparing intact allergen vaccine with allergoid. One allergoid product induced enhanced histamine release compared to the intact allergens, while the other three allergoids showed reduced release. Standard T cell stimulation assays using lines from allergic patients showed a reduced response for all allergoids compared with the intact allergen vaccine regardless of the cell type used for antigen presentation. All allergoids showed reduced capacity to induce allergen-specific IgG responses in mice. CONCLUSION: While some allergoids were associated with reduced allergenicity, a clear reduction in immunogenicity was observed for all allergoid products compared with the intact allergen vaccine, and the commercial allergoids tested therefore do not fulfil the allergoid concept.     

树突状细胞负载前列腺癌细胞抗原后的抗肿瘤免疫作用

目的 探讨小鼠树突状细胞(DC)负载前列腺癌细胞株RM-1的裂解产物后，诱导特异性细胞毒T淋巴细胞(CTL)及其抗肿瘤的免疫作用。方法 将RM-1细胞的裂解产物(T-lysate)作为肿瘤抗原负载小鼠骨髓来源的DC，构建DC瘤苗(T-lysate／DC)，采用流式细胞术和四唑氮蓝(MTT)还原法检测其免疫活性；ELISA法检测其诱导细胞因子白介素(IL)-2和γ-干扰素(IFN-γ)的作用；体内实验检测DC瘤苗的抗肿瘤免疫治疗作用和免疫保护作用。结果 DC负载RM-1细胞的裂解产物后，其主要组织相容性复合物(MHC-Ⅰ、Ⅱ)及共刺激分子(B7-1、B7-2)的表达明显增高；T-lysate／DC能够诱导小鼠产生RM-1特异性CTL，使细胞上清液中IL-2和IFN-γ水平升高，对小鼠具有免疫保护作用，并能有效抵抗肿瘤细胞的攻击；经T-lysate／DC治疗的荷瘤小鼠瘤体生长减慢，存活期延长，瘤体出现坏死和炎细胞浸润。结论 DC负载前内腺细胞解产物后，能够有效诱导搞肿瘤免疫反应，为前列腺癌的临床治疗提供了新策略。     

主动免疫恢复T辅助细胞平衡治疗习惯性流产

目的:寻求安全、有效地治疗不明原因习惯性流产(UHA)的方法,并探讨主动免疫治疗前后的细胞因子的免疫功能变化及T辅助细胞的平衡状态。方法:①对35例UHA患者采用丈夫肘静脉血20 m l分离淋巴细胞行主动免疫治疗(妊娠前治疗两次,妊娠后治疗两次);②采用酶联免疫吸附方法对68例UHA患者及30例正常妇女外周血IL-2、IL-4、IL-10、IFN进行检测;③对29例主动免疫后妊娠成功者治疗前后的IL-2、IL-10、Th1/Th2进行检测。结果:①主动免疫组33例妊娠,妊娠成功并分娩29例,妊娠成功率87.88%(29/33),未经主动免疫治疗的对照组妊娠成功率26.67%(8/30),两组相比差异有显著性(P<0.010);②68例UHA患者较正常妇女外周血中IL-2明显升高(P<0.01),IL-10降低(P<0.05),Th1/Th2比值增高(P<0.05);③29例UHA患者主动免疫治疗后妊娠成功者IL-10显著上升(P<0.1),Th1/Th2比值下降(P<0.05)。结论:UHA患者Th1、Th2平衡异常,主动免疫治疗可诱导Th1向Th2转移,恢复Th1、Th2之间的平衡,可用于UHA的治疗。     

老年人免疫功能异常变化的临床观察及其评价(Ⅰ)

老年人生理适应与防卫能力进行性下降,对有害因子易伤性增加,导致多种老年病的发生。因此,免疫系统功能与衰老的关系问题,已是现代老年病学和免疫学进行多方深入研究的重要内容。我们近两年多来对老年人免疫功能变化做了临床观察检测与统计分析。现报告如下。 1.临床资料     

IL－10对大鼠佐剂性关节炎的治疗及免疫机理探讨

形成佐剂性关节炎（ＡＡ）的动物模型，研究ＩＬ－１０的抗炎作用及其机理。结果表明，经腹腔注射ＩＬ－１０后能改善ＡＡ大鼠病情，用ＩＬ－１０治疗取得显效后，ＡＡ大鼠血清及关节液内炎症细胞因子ＩＬ－１，６，８，ＴＮＦ水平均明显下降，腹腔巨噬细胞和脾淋巴细胞分泌细胞因子的能力均受到抑制。表明ＩＬ－１０是通过抑制炎症细胞因子的产生而发挥治疗作用的。此项研究为用ＩＬ－１０治疗人风湿性关节炎（ＲＡ）等难治性炎症性疾病提供了理论和实验依据     

Preservation and redirection of HPV16E7-specific T cell receptors for immunotherapy of cervical cancer

Human papilloma virus (HPV) type 16 infections of the genital tract are associated with the development of cervical cancer (CxCa) in women. HPV16-derived oncoproteins E6 and E7 are expressed constitutively in these lesions and might therefore be attractive candidates for T-cell-mediated adoptive immunotherapy. However, the low precursor frequency of HPV16E7-specific T cells in patients and healthy donors hampers routine isolation of these cells for adoptive transfer. To overcome this problem, we have isolated T cell receptor (TCR) genes from four different HPV16E7-specific healthy donor and patient-derived human cytotoxic T lymphocyte (CTL) clones. We examined whether genetic engineering of peripheral blood-derived CD8+ T cells in order to express HPV16E711-20-specific TCRs is feasible for adoptive transfer purposes. Reporter cells (Jurkat/MA) carrying a transgenic TCR were shown to bind relevant but not irrelevant tetramers. Moreover, these TCR-transgenic Jurkat/MA cells showed reactivity towards relevant target cells, indicating proper functional activity of the TCRs isolated from already available T cell clones. We next introduced an HPV16E711-20-specific TCR into blood-derived, CD8+ recipient T cells. Transgenic CTL clones stained positive for tetramers presenting the relevant HPV16E711-20 epitope and biological activity of the TCR in transduced CTL was confirmed by lytic activity and by interferon (IFN)-gamma secretion upon antigen-specific stimulation. Importantly, we show recognition of the endogenously processed and HLA-A2 presented HPV16E711-20 CTL epitope by A9-TCR-transgenic T cells. Collectively, our data indicate that HPV16E7 TCR gene transfer is feasible as an alternative strategy to generate human HPV16E7-specific T cells for the treatment of patients suffering from cervical cancer and other HPV16-induced malignancies.