凋亡大肠癌CT26细胞对小鼠T淋巴细胞增殖及活性的影响

目的 探讨凋亡大肠癌CT26细胞对小鼠T淋巴细胞增殖及活性的影响.方法 (1)取对数生长期大肠癌CT26细胞制备凋亡肿瘤细胞及坏死肿瘤细胞,取Balb/c小鼠脾脏分离T淋巴细胞,并制备细胞毒性T淋巴细胞(CTL).(2)将凋亡肿瘤细胞、坏死肿瘤细胞、正常肿瘤细胞分别与小鼠CTL共同培养(分别为凋亡肿瘤细胞组、坏死肿瘤细胞组、肿瘤细胞对照组),测定CTL的活性及增殖情况.(3)将小鼠T淋巴细胞分为对照组、刀豆蛋白A(ConA)组、凋亡肿瘤细胞+ ConA组、坏死肿瘤细胞+ConA组、活肿瘤细胞+ConA组,经相应处理后测定各组T淋巴细胞的活性及增殖情况.(4)选取30只Balb/c小鼠分为凋亡肿瘤细胞组、坏死肿瘤细胞组、肿瘤细胞对照组各10只,分别将凋亡、坏死、对数生长期CT26细胞通过皮下及静脉输注小鼠,检测各组小鼠血清调节性T细胞(Treg)的表达情况.(5)另取30只小鼠,按方法(4)分组及处理后,测定各组小鼠血清可溶性Fas(sFas)、白细胞介素(IL)-12、干扰素-γ(IFN-γ)、IL-4、IL-10、转化生长因子-β(TGF-β)的表达水平.结果 凋亡肿瘤细胞组中CTL的活性、A450值均低于其他两组(P＜0.05).凋亡肿瘤细胞+ ConA组的T淋巴细胞CD69、CD25、CD71的表达率以及G2、G3及G4期的T淋巴细胞数量均低于ConA组、活肿瘤细胞+ ConA及坏死肿瘤细胞+ConA(P ＜0.05).凋亡肿瘤细胞组Treg的表达水平、sFas相对表达量低于肿瘤细胞组、坏死肿瘤细胞组(P＜0.05).凋亡肿瘤细胞组IL-10、TGF-β表达水平高于其他两组(P＜0.05),而IL-12表达水平均低于其他两组(P＜0.05),IL4表达水平则高于肿瘤细胞对照组(P＜0.05).结论 凋亡小鼠大肠癌CT26细胞能够抑制小鼠T淋巴细胞增殖及活性,影响大肠癌小鼠正常免疫功能.     

复方高渗NaCl溶液治疗小鼠S_(180)的疗效及免疫状态观察

将S_(180)瘤组织接种于小鼠大腿外侧皮下.第6天时,用复方高渗NaCl溶液向治疗组小鼠肿瘤的边缘、基底和内部注射。对照组动物肿瘤不给任何注射。到接种后第13天(治疗组治疗后第7天)测定T淋巴细胞量、巨噬细胞吞噬功能及Fc受体活性,两组相比差异非常显著(P<0.01)。治疗组治愈率为86.7％,1个月存活率为93.4％.对照组无一自愈,94％在20天内死亡。     

胶质瘤细胞来源的exosome负载树突状细胞诱导肿瘤特异性杀伤的研究

背景与目的：对于各种恶性肿瘤细胞抗原致敏的树突状细胞疫苗研究目前已成为全球抗肿瘤治疗的热门话题。本研究旨在观察胶质瘤细胞来源外来体致敏的树突状细胞,诱导细胞毒性T淋巴细胞（CTL）杀伤胶质瘤细胞的活性。方法：采用差速离心法分离胶质瘤细胞分泌的外来体,制备胶质瘤冻融抗原;从胶质瘤患者外周血中获取单状核细胞和T淋巴细胞,培养并获取树突状细胞;将胶质瘤来源的外来体、胶质瘤冻融抗原分别冲击树突状细胞并与T淋巴细胞混合培养．通过MTT法检测两种方法制备的效应细胞对胶质瘤细胞的杀伤效应。结果：当效靶比分别为10：1、25：1、50：1时,冻融抗原-DC＋T细胞对恶性胶质瘤细胞杀伤率为（17．50±1．13）％、（21．18±1．36）%、（30．20±1．46）％。外来体-DC＋T细胞组对恶性胶质瘤细胞杀伤率分别是（32．25±1．36）％、I（43．04±1．22）％、（70．42±1．40）％,二者杀伤力的差异具有统计学意义（P〈0．05）。结论：胶质瘤细胞来源的外来体致敏树突状细胞,其特异性杀伤胶质瘤细胞的能力明显优于胶质瘤冷冻抗原。     

The effect of Asparagus officinalis polysaccharide on erythrocyte of S180 mice played in immunological reaction of lymphocytes

Objective To contrast the roles of Asparagus officinalis polysaccharide on erythrocyte of S180 mice played in immunological reaction of lymphocytes.To study the effect of Asparagus officinalis polysaccharide on the erythrocyte function of S180 mice.Methods Suspensions of lymphocytes(1×106/mL)and autologous plasma were respectively separated from anticoaguted whole blood of healthy mice with the lymphocyte separation medium.The erythrocytes(1×108/mL)were separated from whole blood of Asparagus officinalis polysaccharide mice.Using the autologous plasm as reactive medium,the role of erythrocytes in regulating the immunological reaction of lymphocytes was appraised.The expression of CD25 on lymphocytes was detected using flow cytometry.Results The immunogical regulating ability of erythrocyte in mice with control groups is much lower than that of normal groups,and the immunogical regulating ability of erythrocyte in mice with Asparagus officinalis polysaccharide groups is much higher than that of control groups.Conclusions According to the effects of erythrocyte CD35 on the immuno-response of lymphocyte and the different of the expression of CD25 on lymphocytes,we prove that Asparagus officinalis polysaccharide can improve the erythrocyte function of S180 mice.     

Folate Deficiency in Malnutrition, Malabsorption, and During Phenytoin Treatment Diagnosed by Determination of Serine Synthesis in Lymphocytes

Abstract A new method for the detection of folate deficiency by measuring the rate of incorporation of ¹⁴C-formate into serine in isolated lymphocytes has been applied to a group of 24 control subjects and 23 patients. 5 of the patients suffered from malnutrition, 14 had malabsorption and 4 were under treatment with phenytoin.
In 17 of these patients, functional folate deficiency was diagnosed by a decreased rate of serine synthesis in the lymphocytes. 11 cases of folate deficiency were found by measuring the folate concentration of erythrocytes by the L. casei method. The lymphocyte method failed in only one case out of 18.
In the 24 control subjects the average incorporation of ¹⁴C-formate into serine per 10⁹ lymphocytes per 4 h was 9.2 ± 0.4% (range 6.3–14.3 %) of added activity. For 16 patients with functional folate deficiency, the corresponding value was 4.1 ± 0.3% (range 1.0–5.9%). The folate activities of lymphocytes from 12 patients before and after 4 weeks of oral treatment with 15 mg folic acid daily were 3.4±0.3 and 7.5±1.0% respectively. The lymphocytes responded within 4 weeks to deprivation of or treatment with folic acid.
The ¹⁴C-formate incorporation into methionine, RNA sad protein showed the same pattern as the incorporation into serine.     

Role of 4-1BB in Allograft Rejection Mediated by CD8+ T Cells

Blockade of traditional costimulatory molecules fails to inhibit rejection in many models where CD8+ T cells are sufficient to mediate rejection. This observation demonstrates that in many settings CD8+ T cells are not dependent upon CD28 or CD154 signals to mediate rejection. 4-1BB (CD137) has been shown to be an important regulatory molecule for CD8+ T cells in a variety of nontransplant models. Here we show that blocking the 4-1BB pathway significantly inhibited rejection of intestinal allografts by CD8+ but not CD4+ T cells. This effect was associated with significantly decreased expression of the genes encoding TNFalpha and secondary lymphoid chemokine (SLC) within the spleens of recipient mice. Disruption of the 4-1BB pathway also impaired the priming of alloantigen-specific CD8+ T cells and the accumulation of recipient dendritic cells within the spleen. These data directly demonstrate an important role for 4-1BB in allograft rejection; particularly rejection mediated by CD8+ T cells. Our data suggest that in addition to providing a direct costimulatory signal to T cells, the 4-1BB pathway may regulate other important steps in the immune response such as the migration of T cells and dendritic cells.     

虫草素通过调节CD4~+T淋巴细胞PD-1受体促进肿瘤免疫及机制研究

目的探讨虫草素体外对肿瘤相关CD4~+T淋巴细胞肿瘤杀伤作用及其内在机制。方法通过移植瘤分离得到肿瘤相关CD4~+T淋巴细胞,运用MTS法研究虫草素对肿瘤相关CD4~+T淋巴细胞增殖作用的影响,运用LDH法研究虫草素处理后的肿瘤相关CD4~+T淋巴细胞对于其共孵育肿瘤细胞的杀伤作用,运用ELISA法对CD4~+T淋巴细胞与肿瘤细胞共孵育后的细胞上清中的细胞因子进行检测,同时运用Western blot法对虫草素处理后的CD4~+T淋巴细胞中的相关蛋白进行研究。结果虫草素能够促进肿瘤相关CD4~+T淋巴细胞的增殖和杀伤作用且呈现出一定的剂量依赖性,虫草素能够降低肿瘤相关CD4~+T淋巴细胞的PD-1蛋白表达,增强PI3K/AKT信号通路相关蛋白的表达且促进IL-2、IFN-γ的表达。结论虫草素能够促进肿瘤相关CD4~+T淋巴细胞的增殖和杀伤作用,其机制可能与抑制肿瘤免疫检查点PD-1有关。     

实弹射击考核对不同特质焦虑新兵T淋巴细胞亚群及ACTH的影响

目的：研究急性应激对不同特质焦虑新兵的T淋巴细胞亚群及ACTH的影响。方法：状态一特质焦虑问卷（STAI）将新兵分成高、低特质焦虑组各16人。用流式细胞仪及ELISA测定被试基础期（打靶前4d）、应激期（打靶时）、恢复期（打靶后5d）的CD4^＋、CCD8^＋及ACTH浓度。结果：①高特质组3个时期的CD4^＋、CD4^＋／CCD8^＋比值均低于低特质组。CD8＋高于低特质组；高特质组应激期CD4^＋显著下降（P〈0．05）、CD4^＋／CCD8^＋比值与恢复期有显著差异（P〈0．01）．而低特质组仅应激期CD4^＋定量与恢复期有显著差异（P〈0．01）。②实弹射击前、中、后的ACTH浓度，高特质组变化呈极显著差异（P〈0．001），而低特质组应激期与基础期呈显著差异（P〈0．05）。结论：①急性应激影响新兵的CD4^＋、CCD8^＋及其比值，其程度与特质焦虑有关。②应激时高特质组ACTH浓度下降显著大于低特质组，且随应激源消除不易恢复。     

Phenotypic and functional abnormalities of T lymphocytes in pathological hyperprolactinemia

The phenotype and function of T cells circulating in patients with pathological hyperprolactinemia were analyzed and compared to those in sex- and age-matched control subjects. Two-color immunofluorescence study revealed an increased number of CD4⁺ TQ1⁺ cells and the presence of phenotypically immature CD1⁺ T cells, also exhibiting transferrin surface receptor, in peripheral blood of the hyperprolactinemic patients. After chronic treatment with the dopamine agonist bromocriptine, T-cell abnormalities disappeared. In addition, some untreated patients showed enhanced T-cell suppressor activity in anin vitro pokeweed mitogen-driven B-cell transformation assay. These immunological findings confirm a link between neuroendocrine and immune systems in humans.     

魔芋葡甘聚糖免疫活性研究进展

魔芋葡甘聚糖是天然植物魔芋的主要成分,具有广泛生物学功能。该文从免疫器官、单核巨噬细胞与T、B淋巴细胞、细胞因子以及肠粘膜免疫等方面综述魔芋葡甘聚糖的免疫调节作用。