吞噬自身抗原对骨髓来源树突状细胞体外分化发育及功能的影响

目的:观察小鼠骨髓来源树突状细胞吞噬自身抗原后体外分化发育及功能的变化,研究其对免疫状态的影响.方法:小鼠骨髓细胞经GM-CSF+IL-4诱导分化为DCs,采用荧光显微镜和流式细胞仪检测DCs的纯度;运用免疫组化法及透射电镜鉴定DCs能否吞噬自身抗原;流式细胞仪检测吞噬自身抗原后DCs的CD86、MHCⅡ的变化; MTT法检测吞噬自身抗原后DCs刺激同基因背景小鼠脾淋巴细胞增殖能力的改变;ELISA法检测脾淋巴细胞经DCs刺激后,分泌IL-4、IFN-γ量的改变.结果:所培养DCs的纯度为90.6%,DCs能够吞噬自身抗原,吞噬自身抗原后的DCs体外分化发育异常,抑制同基因背景小鼠脾淋巴细胞的增殖,刺激同基因背景小鼠脾淋巴细胞分泌IL-4增加,分泌IFN-γ减少.结论:DCs吞噬自身抗原后能够抑制自身成熟,促进淋巴细胞免疫耐受,促进体液免疫.     

猪纤维蛋白原免疫原性研究

目的 研究猪纤维蛋白原的免疫原性和免疫效果,为该药物安全评价和进一步优化提供实验依据.方法 建立生物素-亲和素系统改良的ABC-ELISA检测抗体;新西兰白兔创伤试验考察血清抗体效价及其动态变化;免疫器官质量及脏器指数分析、噻唑蓝(MTT)比色法检测脾淋巴细胞增殖、氯化硝基四氮唑蓝(NTB)还原法检测中性粒细胞吞噬能力,揭示猪纤维蛋白原对小鼠免疫系统的影响.结果 1周后新西兰白兔血清即可检测出抗体,且第2周抗体含量达到峰值后下降至较稳定水平,抗体效价也呈现相同的变化趋势;小鼠免疫器官质量及脏器指数均未发生明显改变;与对照组相比,各实验组小鼠淋巴细胞增殖指数无明显差异;实验组小鼠中性粒细胞吞噬能力显著性增强.结论 猪纤维蛋白原刺激新西兰白兔机体后产生体液免疫应答,但在小鼠体内未表现出特异性免疫反应.     

北豆根提取物对小鼠和人淋巴细胞及巨噬细胞作用的体外实验研究

目的：探讨中药北豆根提取物的免疫学调节作用及其作用机制；比较北豆根水提物、醇提物和多糖成分的生物学活性。方法：采用水提取、醇提取及水回流等方法对北豆根进行成分分离。采用MTT法、中性红法检测了北豆根提取物对淋巴细胞增殖的作用以及对小鼠巨噬细胞代谢和吞噬功能的影响。结果：北豆根多糖成分(RMP)具有丝裂原样作用，可刺激小鼠脾细胞及人淋巴细胞增殖，对小鼠巨噬细胞的吞噬功能有一定的促进作用。而北豆根水提物(RMW)和醇提物(RME)对小鼠脾细胞、人淋巴细胞的增殖具有抑制作用，对小鼠巨噬细胞的代谢及吞噬功能也有抑制作用。结论：不同的北豆根成分对免疫细胞有不同的作用，水提物和醇提物对免疫细胞的增殖、代谢和功能具有一定的抑制作用；北豆根多糖成分对淋巴细胞具有丝裂原样作用，能增强小鼠巨噬细胞的吞噬功能，该活性可用于临床辅助治疗肿瘤。     

戴氏虫草和粉被虫草多糖对巨噬细胞等活性的影响

目的 研究虫草属新种-戴氏虫草菌丝体胞外水溶性多糖（CDP1）和粉被虫草菌丝体多糖（CPP1）在体外对小鼠腹腔巨噬细胞（Mφ）吞噬功能和脾细胞免疫功能的影响。方法 MTT比色法测定淋巴细胞增殖,孔雀绿比色法测定小鼠腹腔巨噬细胞吞噬功能,中性红比色法测定细胞毒淋巴细胞（CTL）活性,结果 CDP1在正常情况下不能促进小鼠腹腔巨噬细胞的吞噬功能,而CPP1却有促进作用;在免疫抑制的情况下,一定浓度的戴氏虫草菌丝体胞外水溶性多糖和粉被虫草多糖能恢复提高吞噬细胞的吞噬功能,而CPP1却有促进作用;在免疫抑制的情况下,一定浓度的戴氏虫草菌丝体细外水溶性多糖和粉被虫草多糖能恢复提高吞噬功能,它们不仅能促进正常脾活化T淋巴细胞的增殖,而且能恢复环磷酰胺和氢化可的松抑制免疫小鼠脾活化T淋巴细胞的增殖,在合适的浓度下能增高小鼠脾CTL活性,同时在一定浓度下能恢复免疫抑制小鼠脾CTL活性。结论 两者具有增强免疫功能的作用。     

六君子汤上调小鼠免疫功能的机制

目的:探讨六君子汤(LD)对小鼠的免疫调节作用。方法:用一定剂量的LD给正常小鼠和免疫抑制小鼠灌胃7d,用ELISA法测定血清溶血素的水平;采用中性红吞噬试验检测腹腔巨噬细胞的吞噬功能;应用重氮化反应测定NO的水平;采用MTT比色法测定淋巴细胞增殖反应。结果:中剂量的LD可提高正常小鼠的免疫功能,明显提高免疫抑制小鼠的免疫功能。结论:LD对小鼠的免疫功能具有上调作用。     

腹腔注射胶原蛋白肽增强糖皮质激素免疫抑制模型小鼠的免疫功能

目的观察腹腔注射胶原蛋白肽对糖皮质激素诱导的免疫抑制小鼠免疫功能的调节作用。方法采用糖皮质激素诱导的免疫抑制小鼠模型,连续7 d腹腔注射给予100、300、600 mg胶原蛋白肽/kg体质量。分别测量小鼠体质量、胸腺和脾脏质量,计算胸腺和脾脏指数;采用全自动血球计数仪检测外周血白细胞总数、中性粒细胞和淋巴细胞数、中性粒细胞和淋巴细胞百分比,采用流式细胞术检测CD4+/CD8+比值;采用MTT法检测刀豆蛋白A(ConA)诱导的脾脏T淋巴细胞增殖能力。结果与对照组相比,模型组小鼠中性粒细胞数及其百分比显著升高(P0.05),体质量、胸腺指数、脾脏指数、淋巴细胞数及其百分比、CD4+/CD8+比值、脾脏T淋巴细胞增殖能力显著降低(P0.05);腹腔注射胶原蛋白肽后,这些指标均向对照组水平恢复,并呈剂量依赖性关系;其中剂量最高600 mg/kg组的小鼠体质量、脾脏指数、淋巴细胞百分比、CD4+/CD8+比值显著高于模型组,中性粒细胞百分比显著低于模型组(P0.05),与对照组相比无显著差异。结论腹腔注射胶原蛋白肽能够增强小鼠的免疫功能。     

家蝇蛹血淋巴及其提取物的免疫调节作用

目的:观察家蝇蛹血淋巴及其提取物对小鼠脾淋巴细胞增殖的影响.方法:以不同浓度血淋巴及其提取物与脾淋巴细胞、T细胞和B细胞共孵育72小时,采用MTT法测定淋巴细胞的吸光度值,观察它们对淋巴细胞增殖的促进作用,采用免疫印迹法研究提取物对B淋巴细胞增殖的信号转导通路.结果:与对照组相比,血淋巴冻干粉、分子量在50 kD以上的冻干粉及其提取物对混合淋巴细胞和B细胞增殖有显著促进作用,然而对于T细胞的增殖却没有明显的促进作用.MTT实验和免疫印迹结果表明血淋巴提取物激活了B淋巴细胞的ERK通路,促进了B淋巴细胞的增殖.结论:家蝇蛹血淋巴及其提取物具有免疫调节作用,为天然免疫增强剂的开发提供了一定的参考依据.     

乳酸菌Z222产胞外多糖(EPSⅠ)对免疫细胞功能的影响

目的　检测乳酸菌Z2 2 2 产胞外多糖EPSⅠ对体外免疫细胞功能的调节作用。方法　不同浓度的EPSⅠ作用于小鼠的腹腔巨噬细胞和脾细胞 ,分别用中性红染色法检测巨噬细胞的吞噬能力 ,用MTT法检测淋巴细胞在体外的转化能力、NK细胞杀伤肿瘤细胞Yac 1的能力和产细胞因子IL 2的活性。结果　 (1)EPSⅠ单独作用小鼠脾细胞就能促进体外淋巴细胞增殖 ,且表现出剂量依赖关系。EPSⅠ亦可促进ConA诱导的体外小鼠淋巴细胞转化。 (2 )EPSⅠ体外作用于小鼠腹腔巨噬细胞均可提高MΦ的吞噬能力 ,与对照组比较差异都有显著性。 (3)与对照组相比EPSⅠ能增加体外NK细胞的活性 ,杀伤率最大值达 6 2 .4 1%± 2 .5 4 %。 (4 )EPSⅠ使小鼠脾细胞产IL 2的能力与对照组比较 ,差异有显著性。结论　乳酸菌多糖EPSⅠ对小鼠的免疫功能有一定的调节作用。     

古尼虫草多糖及其解聚物的免疫活性

目的　比较研究古尼虫草纯多糖组分FP及其解聚物的免疫活性。方法　采用超声波解聚多糖 ,并用葡聚糖凝胶分离获得解聚级分 ,通过MTT法和中性红比色法测定其对小鼠脾淋巴细胞增殖、腹腔巨噬细胞吞噬功能和细胞毒T淋巴细胞杀伤功能的影响。结果　FP的解聚级分FP 1、FP 2和FP 3有更强的免疫活性 ,在中、低剂量 (1和 10 μg mL)与对照相比有显著差异 ,而其中以FP 2的活性最明显。结论　古尼虫草多糖解聚后能明显提高免疫活性 ,为多糖生物制品研究开发提供了一条新途径     

炭疽中性蛋白酶部分片段的重组表达、纯化及免疫效果研究

构建表达炭疽杆菌中性蛋白酶基因部分片段N蛋白(位于248~532氨基酸),并对其免疫效果进行研究;设计引物采用PCR法自炭疽杆菌扩增中性蛋白酶部分片段N,T-A克隆后构建原核表达质粒pET28a(+)-N,通过Western blot检测重组蛋白的免疫原性。以Al(OH)3为佐剂皮下注射免疫BALB/c小鼠,用ELISA方法测血清总IgG水平,MTT比色法测定脾T淋巴细胞增殖,采用流式分选法对脾T淋巴细胞亚群分类;Western blot显示重组蛋白具有免疫原性,rN免疫BALB/c小鼠后,ELISA检测到免疫鼠体内有特异性抗体产生;T淋巴细胞亚群试验表明,重组N蛋白以诱发Th2型免疫反应为主;MTT试验表明,rN蛋白对小鼠脾细胞在体外有促进生长和增殖能力,说明该重组蛋白具有生物学活性。纯化的rN可诱导高水平的抗体反应,为进一步的功能研究奠定了基础。     

Sedative effects of the jasmine tea odor and (R)-(−)-linalool, one of its major odor components, on autonomic nerve activity and mood states

We investigated the effects of the odor of jasmine tea on autonomic nerve activity and mood states in a total of 24 healthy volunteers. We used the odor of jasmine tea at the lowest concentration that could be detected by each subject but that did not elicit any psychological effects. R–R intervals and the POMS test were measured before and after inhalation of the odors for 5 min. Both jasmine tea and lavender odors at perceived similar intensity caused significant decreases in heart rate and significant increases in spectral integrated values at high-frequency component in comparison with the control (P < 0.05). In the POMS tests, these odors produced calm and vigorous mood states. We also examined the effects of (R)-(–)-linalool, one of its major odor components, at the same concentration as in the tea, and (S)-(+)-linalool. Only (R)-(–)-linalool elicited a significant decrease in heart rate (P < 0.05) and an increase in high-frequency component in comparison with the controls, and produced calm and vigorous mood states. Thus, the low intensity of jasmine tea odor has sedative effects on both autonomic nerve activity and mood states, and (R)-(–)-linalool, one of its components, can mimic these effects.     

Nuclear morphology and morphometry of B-lymphocyte transformation. Implications for follicular center cell lymphomas

One of the major tenets of current non-Hodgkin's lymphoma classifications is the relationship of morphologic subtypes to stages in the sequence of normal B-lymphocyte transformation occurring in the germinal follicle. To test this hypothesis, quantitative morphometric image analysis was carried out on in vivo and in vitro samples of mouse splenic lymphocytes in which transformation was induced by bacterial lipopolysaccharide (LPS), a specific B-cell mitogen. The results were compared with a similar analysis of germinal center lymphocyte populations of normal human spleen. In the in vivo mouse model, initial stages of B-cell transformation were detectable as early as 4 hours after LPS injection, and the process was essentially fully developed by 48-72 hours. Quantitative evaluation revealed that the majority of nuclear profiles were nearly spherical or only slightly eccentric and that no major alteration in nuclear contour occurred during any phase of the progressive increase in nuclear size following mitogen-induced lymphocyte transformation. In fact, in this system, B lymphocytes with a nuclear profile cleft of greater than or equal to 0.4 mu accounted for only 3% of the combined unstimulated and LPS-activated population assessed (N = 9936). This compared with normal human spleen, in which 16% of germinal center lymphocyte populations had similarly cleft nuclear profiles. Sequential alterations in the organization of condensed chromatin occurred concomitant with gradual nuclear enlargement during mitogen-induced mouse spleen lymphocyte transformation. A comparative morphologic and morphometric assessment of nuclear profiles of lymphocyte populations in germinal centers of normal human spleen provides indirect evidence for a similar pattern of nuclear alterations in human B lymphocytes. Autoradiographic data obtained from LPS-activated mouse splenic lymphocytes indicate that nuclear morphologic aspects of the transformation process can occur entirely within the G1 phase of the cell cycle. The results of this study suggest that subtypes of non-Hodgkin's lymphoma largely composed of neoplastic lymphocytes with extensively convoluted or cleft nuclei do not reflect a morphologic stage in the transformation of normal lymphocytes. In addition, the heterogeneous nuclear forms of follicular center cell lymphocytes would appear to result from parallel transformation processes involving cleaved and noncleaved nucleated lymphocytes and not the sequential pathway proposed by Lukes and Collins.     

MTT法与流式细胞术检测肺炎合剂促进T淋巴细胞转化的实验研究

目的：探讨肺炎合剂对小鼠T淋巴细胞转化的影响，并试图为药研究淋巴细胞转化提供检测方法依据。方法：通过病毒性肺炎患儿淋巴细胞转化率降低的机体，用氢化可的松建立免疫低下小鼠模型，以不同剂量药液灌胃给药后，采用MTTI法和流式细胞仪检测法对比观察肺炎合剂对ConA诱导的小鼠脾T淋巴细胞转化的作用。结果：肺炎合剂使正常和免疫低下小鼠脾T淋巴细胞增殖能力显著增强。结论：肺炎合剂对小鼠脾T淋巴细胞增反应的促进作用，可能是该药作用机理之一。并认为流式细胞术是测定淋巴细胞转化的一种准确而可靠的方法。     

Nematospiroides dubius: Modulation of the host immunological responsive

Altered immunological reactivity occurred in mice infected with the larval stages ofNematospiroides dubius. This was shown by an increased responsiveness of mouse spleen lymphocytes and suppressed responsiveness of mesenteric lymph node (MLN) lymphocytes to phytohaemagglutinin (PHA) and concanavalin A (Con A) during the first 10–15 days after infection while at the same time spleen, MLN, and cervical lymph node (CLN) lymphocyte reactivity to lipopolysaccharide (LPS) was suppressed. Splenomegaly, with a marked increase of white pulp in the spleen, began 3 days after infection and there was a concomitant decrease in the numbers of cells in the MLN with depletion of the paracortical and medullary areas in particular.N. dubius larvae cultured in vitro produced a dialysable immunosuppressive factor which inhibited the proliferation of spleen lymphocytes from normal and immune mice and sheep in the presence of mitogens. Serum collected from infected mice also suppressed mitogen-induced transformation of normal mouse lymphocytes.     

Effect of normal mouse serum on mouse lymphocyte transformation in vitro

Mouse spleen cells were cultured in the presence or absence of one of the following mitogens: phytohemagglutinin (PHA), concanavalin A, allogeneic spleen cells or bacterial lipopolysaccharide. The addition of normal mouse serum (NMS) to the cultures usually depressed mitogen-induced lymphocyte transformation as measured by DNA, RNA or protein synthesis, whether calculated as gross or net synthesis. The degree of depression was increased as the concentration of NMS serum increased. DNA and RNA synthesis in unstimulated cultures were also usually depressed by NMS. NMS added 24 hours or more after the start of cultures with PHA or allogeneic lymphocytes was less depressive than serum present from the start and in some cases, actually increased DNA synthesis. The presence of NMS in cultures containing supra-optimal doses of concanavalin A could also increase DNA synthesis. Cells incubated with NMS for 24 hours survived as well as cells incubated without NMS, but subsequently responded less well to PHA. Cells passed through a column of glass wool had a lower baseline DNA synthesis, which was not inhibited by NMS; they responded less well to mitogens and this response was further depressed by NMS. It is suggested that NMS contains a factor which damps the proliferation of mouse T and B cells, without exerting an overt cytotoxic effect, and that it acts directly on lymphocytes.     

青蒿琥酯对迟发型超敏反应小鼠脾脏T淋巴细胞的免疫调节作用***☆◆

背景：青蒿琥酯是青蒿素低毒、高效的衍生物,具有免疫调节功能,但其具体机制仍需研究阐明。 目的：初步探讨青蒿琥酯对迟发型超敏反应小鼠脾脏T淋巴细胞的免疫调节作用。 方法：建立迟发型超敏反应小鼠模型,40只小鼠随机数字表法均分正常对照组、青蒿琥酯干预组、p-38MAPK抑制剂组、基质对照组进行实验观察。T淋巴细胞转化实验检测小鼠T淋巴细胞增殖水平;Western blot方法检测p38丝裂酶原激活蛋白激酶(p38MAPK)蛋白的活性表达。 结果与结论：局部给药后青蒿琥酯明显减轻迟发型超敏反应小鼠耳肿胀、降低脾指数、抑制刀豆蛋白A诱导的T淋巴细胞增殖;同时减弱p38 MAPK的磷酸化活性表达。提示青蒿琥酯可以有效抑制小鼠迟发型超敏反应,其作用途径可能与抑制p38 MAPK信号通路有关。     

Mycoplasma neurolyticum: a potent mitogen for rat B lymphocytes.

A mitogen prepared from Mycoplasma neurolyticum has been demonstrated to induce extensive transformation of in vitro cultured rat B lymphocytes. The data summarized in this report show that rat thymus cells as well as hydrocortisone-resistant thymocytes were not activated by this mitogenic agent. On the other hand, spleen cells obtained from thymectomized, lethally irradiated and bone marrow-reconstituted rats were extensively activated by M. neurolyticum. Furthermore, M. neurolyticum was shown to induce the development of antibody-producing cells, as attested by the appearance of direct plaque-forming cells against sheep red blood cells and trinitrophenylated sheep red blood cells in spleen cell cultures exposed to this mitogen. It was also demonstrated that stimulation of rat lymphocytes by this mitogen was inhibited by anti-rat immunoglobulin antibodies. In view of these data, it was suggested that M. neurolyticum, which activates mouse B lymphocytes, is a potent mitogen for rat B lymphocytes as well. This mitogen is a significantly more powerful mitogen for rat B lymphocytes then any other known mitogens. The availability of such mitogenic material in the rat system will enable studies on control mechanisms of action and differentiation of rat B lymphocytes.     

Mitogenic activity of staphylococcal peptidoglycan.

Staphylococcus aureus peptidoglycan displayed a marked dose-dependent mitogenic activity for mouse splenocytes and human peripheral blood lymphocytes in vitro, as measured by increased [3H]thymidine incorporation. Similarly it was mitogenic for athymic nude mouse spleen cells, whereas no blastogenic effect was observed in T cell-enriched and B cell-depleted mouse lymphocyte cultures. These data demonstrate that peptidoglycan-responding cells in mouse spleen cell cultures are B lymphocytes.     

积雪草免疫抑制活性成分的分离纯化及效应研究

 目的：对积雪草的免疫抑制活性成分进行分离、纯化及鉴定,并观察其对小鼠脾淋巴细胞增殖和凋亡的影响,探讨其免疫调节机制。方法：利用半制备型液相色谱对积雪草提取物进一步分离和纯化。运用流式细胞术观察积雪草活性成分对脾淋巴细胞凋亡和线粒体膜电位的影响。利用紫外光谱、电喷雾电离负离子模式质谱、［1H ］核磁共振（NMR）、［13C］ NMR等数据鉴定所得活性成分结构。结果：（1）所得积雪草活性成分化合物Ⅰ、Ⅱ的分子量分别为302和286。（2）积雪草中化合物Ⅰ、Ⅱ能显著抑制小鼠脾淋巴细胞增殖。（3） 积雪草中化合物Ⅰ、Ⅱ能促进小鼠脾淋巴细胞凋亡,降低线粒体膜电位。（4）［1H］ NMR和［13C］ NMR鉴定化合物Ⅰ、Ⅱ分别为槲皮素和山奈酚。结论：从积雪草中提取获得的黄酮化合物Ⅰ、Ⅱ分别为槲皮素及山奈酚,具有抑制小鼠脾淋巴细胞增殖的效应,其作用途径可能是促进脾淋巴细胞凋亡并降低线粒体膜电位。