旋毛虫感染小鼠IgG、IL-2和T淋巴细胞亚群动态变化的观察

目的　了解小鼠感染旋毛虫后 Ig G抗体水平、IL - 2分泌量和 T淋巴细胞亚群的动态变化。方法　分别于旋毛虫感染后第 7、14、2 1、2 8和 35天 ,采用 EL ISA方法检测血清特异性 Ig G抗体水平和 IL- 2含量 ,采用流式细胞仪检测 CD4+、CD8+T细胞百分率。结果　 Ig G抗体水平在感染后逐渐上升 ,感杂后 35天达最高值 ;T淋巴细胞的变化表现为 CD4+T细胞减少、CD8+T细胞增多 ,CD4+/ CD8+细胞比值下降 ,以感染后第 14天最为显著 ,直到感染后第 35天仍未见恢复。IL - 2分泌量以感染后第 7天达高峰 ,然后迅速下降 ,到感染后第 35天低于正常组。讨论　旋毛虫感染的急性期 ,小鼠出现免疫抑制现象。抗旋毛虫感染的保护性免疫是由细胞免疫与体液免疫协同完成的。     

旋毛虫感染小鼠IgG,IL—2和T淋巴细胞亚群动态变化的 …

目的 了解小鼠感染旋毛虫后ＩｇＧ抗体水平、ＩＬ－２分泌量和Ｔ淋巴细胞亚群的动态变化。方法 分别于旋毛虫感染后第７、１４、２１、２８和３５天,采用ＥＬＩＳＡ方法检测血甭特异性ＩｇＧ抗体水平和ＩＬ－２含量,采用流式细胞仪检测ＣＤ４＾＋、ＣＤ８＾＋Ｔ细胞百分率。结果 ＩｇＧ抗体水平在感染后逐渐上升,感染后３５天达最高值;Ｔ淋巴细胞的变化表现为ＣＤ４＾＋细胞减少、ＣＤ８＾＋Ｔ细胞增多,ＣＤ４＾＋／ＣＤ８＾     

旋毛虫感染过程中小鼠T淋巴细胞亚群的变化

应用酸性α醋酸萘酯酶(ANAE)胞浆标记法观察实验感染旋毛虫小鼠外周血T淋巴细胞及其亚群的变化。结果表明感染后d_3淋巴细胞和T淋巴细胞绝对数平行地增多,于d_(14)达高峰,d_(60)仍高于正常水平。而斑块颗粒型细胞(辅助性T细胞,T_h)和分散颗粒型细胞(抑制性T细胞,T_a)呈反向变化,前者减少,后者增多,T_h/T_a比值降低,表明旋毛虫感染过程中宿主免疫功能受抑制。     

旋毛虫感染小鼠血清干扰素-γ、白介素-4含量的变化

目的 通过检测血清干扰素(IFN)-γ、白介素(IL)4的含量,探讨小鼠感染旋毛虫后免疫功能的变化.方法 将70只小鼠随机分为两组,模型组每只小鼠喂食感染旋毛幼虫200条,对照组小鼠不做处理,分别于小鼠感染旋毛虫后第7、14、21、28、35天,随机取模型组和对照组小鼠,常规眼眶静脉取血分离血清,用ELISA法测定IFN-γ、IL-4的值.结果 模型组IFN-γ分泌水平在感染后7d达峰值,感染后14和21 d水平虽有下降但仍高于对照组(P＜0.05),至感染后28、35 d与对照组相比无差异.模型组IL-4分泌水平于感染后14 d达峰值,随后缓慢下降但始终高于对照组(P＜0.01).结论 旋毛虫感染小鼠细胞免疫和体液免疫均成功地建立和转化,体液免疫在清除旋毛虫过程中至关重要.     

感染旋毛虫大鼠免疫功能的动态观察

目的　观察感染旋毛虫大鼠免疫功能的动态变化。　方法　采用免疫组化的方法分别检测感染旋毛虫后 7、14、21、28、35、60 d大鼠外周血CD4+、CD8+T淋巴细胞的百分率,并同时用酶联免疫吸附试验测定IgE水平。　结果　实验组较对照组CD4+T细胞减少, CD8+T细胞增多,CD4+/CD8+比值下降(P<0.01),以感染后第14 d最明显,直到感染后60 d仍未见恢复;大鼠外周血中IgE水平在感染旋毛虫后7 d增加,21 d达到高峰。　结论　感染旋毛虫的大鼠出现免疫抑制现象,感染急性期的免疫抑制程度最高,感染大鼠的免疫抑制主要发生在成虫产新生蚴及新生蚴移行期。IgE是大鼠旋毛虫感染急性期重要的抗体。     

环孢素A对感染旋毛虫小鼠的免疫调节作用

目的观察环孢素A(CsA)对感染旋毛虫小鼠免疫功能的影响。方法小鼠腹腔注射环孢素A感染旋毛虫后,分别采用ELISA和流式细胞仪检测小鼠不同时期外周血中IL-2含量、CD4+及CD8+T淋巴细胞的百分率。结果(1)单纯感染旋毛虫小鼠感染后1~5周IL-2的含量较正常组明显增加;注射CsA组小鼠感染后1周IL-2的含量较单纯感染组明显降低,感染后2~4周IL-2的含量与单纯感染组相比无明显差异,但仍高于正常组。(2)单纯感染旋毛虫小鼠感染后1~4周,CD4+T淋巴细胞较正常组明显减少,CD8+T淋巴细胞明显增多,CD4+/CD8+比值明显下降;注射CsA组小鼠CD4+T淋巴细胞细胞较单纯感染组明显减少,而CD8+T淋巴细胞无明显变化。结论CsA对感染旋毛虫小鼠具有一定的免疫调节作用。     

感染旋毛虫小鼠外周血T淋巴细胞及其亚群和血清IgG抗体的变化

本实验用酸性α醋酸萘脂酶染色法观察实验感染旋毛虫小鼠外周血T淋巴细胞及其亚群的变化,以斑点-ELISA试验观察血清IgG抗体的变化。小鼠感染后第3d起外周血T淋巴细胞开始增高,第14d达最高水平,此后开始降低,第77d仍未降至正常水平,而其中斑点型细胞(辅助性T细胞,T_h)和弥散型细胞(抑制性T细胞,T_s)呈反向变化,前者减少,后者增多,T_h/T_s比值降低,表明旋毛虫感染过程中宿主免疫功能受抑制。同时观察到感染后第7d外周血清IgG抗体滴度开始增高,第28-42d达最高水平,第140d仍维持较高水平。     

旋毛虫成囊前期幼虫的收集

目的 探索旋毛虫成囊前期幼虫的收集时间与方法.方法 20只成年大鼠,每只大鼠经口感染3000条旋毛虫脱囊幼虫,分别于第14、15、16、17、18、19、20天将大鼠处死,用人工胃蛋白酶消化肌肉收集旋毛虫.结果 感染后第14、15、16大膈肌均未见旋毛虫,第17、18、19、20天膈肌有旋毛虫的侵入并逐渐增多;第14、15、16天均未收集到旋毛虫,第17、18、19、20天平均每只大鼠分别收集5000、8000、10000、30000条旋毛虫.结论 旋毛虫在感染后第20天是旋毛虫成囊前期幼虫收集的最佳时期.人工胃蛋白酶消化肌肉可以收集成囊前期幼虫.     

都匀亚洲带绦虫和从江牛带绦虫实验感染小鼠免疫指标动态观察

目的 观察小鼠感染都匀亚洲带绦虫和从江牛带绦虫后血清免疫学指标的动态变化。方法 取新鲜都匀亚洲带绦虫和从江牛带绦虫孕节，分别用生理盐水洗净，置37℃水浴箱孵化成六钩蚴、经皮下或腹腔接种小鼠。分别于感染后40、60、75d取血作免疫学检查。结果 两实验组小鼠感染六钩蚴后血清免疫球蛋白均升高，其中IgM在感染后40d达高峰，之后开始下降；IgG、IgA在感染后60d达高峰，之后下降；补体C3、C4水平下降，至感染后75d，都匀亚洲带绦虫感染组和从江牛带绦虫感组分别下降82．8％和77．5％；检查两组小鼠血白细胞总数及分类，除嗜酸性粒细胞比例升高外其余无明显改变。结论免疫抑制小鼠感染都匀亚洲带绦虫和从江牛带绦虫后血清免疫学指标均发生显著改变，符合免疫应答规律。但从江牛带绦虫感染小鼠后，免疫学指标变化更明显。     

小鼠实验感染不同种旋毛虫后血清IgG抗体水平的变化

目的观察小鼠感染不同种旋毛虫后血清IgG抗体水平的变化。方法将50只小鼠随机分成5组（每组10只），分别感染旋毛虫（T1）、乡土旋毛虫（T2）、布氏旋毛虫（T3）、伪旋毛虫（T4）及纳氏旋毛虫（T7），每只感染300条幼虫，感染后1～6周每周尾部静脉采血，6周后每2周尾部静脉采血，至感染后20周，用旋毛虫肌幼虫ES抗原ELISA检测血清中抗旋毛虫IgG抗体水平。结果小鼠感染旋毛虫、乡土旋毛虫、布氏旋毛虫及纳氏旋毛虫后3～5周，血清IgG抗体水平快速升高，至感染后第8周达高峰，此后旋毛虫和乡土旋毛虫感染小鼠的血清IgG抗体水平缓慢下降，布氏旋毛虫及纳氏旋毛虫感染小鼠的血清IgG抗体水平迅速下降；小鼠感染伪旋毛虫后3～5周血清IgG抗体水平快速升高，至第16周达高峰，之后缓慢下降。5种旋毛虫感染小鼠的血清IgG抗体水平差异具有显著性（P〈0．05）。结论5种旋毛虫感染小鼠的血清IgG抗体水平和动态变化不同；旋毛虫肌幼虫ES抗原可用于其他4种旋毛虫（乡土旋毛虫、布氏旋毛虫、伪旋毛虫、纳氏旋毛虫）感染的血清学诊断及流行病学调查。     

旋毛虫DNA疫苗诱导小鼠免疫应答的研究

目的　观察旋毛虫DNA疫苗在小鼠体内诱导的免疫应答。方法　将旋毛虫肌幼虫编码 31kDa抗原结构基因真核表达质粒pcDNA3-TspE1分别用肌肉注射和基因枪免疫BALB/c小鼠 ,免疫血清用旋毛虫肌幼虫冰冻切片及石蜡切片抗原进行IFAT检测 ,并用旋毛虫肌幼虫可溶性抗原进行Westernblot分析。结果　IFAT表明 pcDNA3-TspE1接种BALB/c小鼠后产生的免疫血清与旋毛虫肌幼虫可产生阳性反应 ,在肌幼虫冰冻及石蜡切片上均显示黄绿色荧光。Westernblot检测结果显示 ,pcDNA3-TspE1接种小鼠后产生的免疫血清只能识别旋毛虫肌幼虫可溶性抗原中的 31kDa抗原组分 ,而 pcDNA3质粒接种小鼠后的血清不能与旋毛虫肌幼虫可溶性抗原发生免疫反应。结论　旋毛虫DNA疫苗 pcDNA3-TspE1能够诱导小鼠产生特异性体液免疫应答。     

人工消化法检验旋毛虫效果及其对肌幼虫的影响

目的观察人工消化法检验旋毛虫的效果及对肌幼虫活性、感染性的影响。方法将小鼠感染100条肌幼虫后30 d剖杀,分别应用3组消化法检验,即：消化2 h组（磁力搅拌法）、消化12 h组、消化20 h组,每组取肉样10份,每份含300个囊包,消化后分别计数每份肉样的幼虫数;同时对幼虫活性进行观察。40只昆明小鼠随机分为对照组和3个消化组（消化2、12、20 h组）,共4组,每组10只。对照组：每鼠感染100个囊包;消化组：将3组方法收集的幼虫分别感染小鼠,每鼠经口感染100条幼虫。感染后30 d剖杀,取膈肌压片镜检,并用磁力搅拌法收集幼虫、计数。结果消化2、12、20 h组的幼虫检出率分别为78.47%、76.73%、68.63%,死亡率分别为0.59%、4.60%、7.43%。3组收集的幼虫分别感染小鼠,30 d后剖杀,与对照组相比,消化2、12、20 h组的减虫率分别为60.56%、61.94%、73.07%。结论磁力搅拌法（消化2 h）在幼虫检出率、幼虫活性、幼虫感染性等方面均优于消化12、20 h实验组,为进行旋毛虫检验和动物实验,优先选择的方法。     

Immunological relationships during primary infection with Heligmosomoides polygyrus (Nematospiroides dubius): downregulation of specific cytokine secretion (IL-9 and IL-10) correlates with poor mastocytosis and chronic survival of adult worms

Mice were infected either with Trichinella spiralis (day 0). Heligmosomoides polygyrus (day – 14) or concurrently with both species and were killed in groups, together with naïve control mice, on 2 occasions (day 8 and 15 post infection with T. spiralis, corresponding to days 22 and 29 p.i. with H. polygyrus). The expulsion of T. spiralis was slowed significantly in concurrently infected mice and this was associated with a reduced mastocytosis and lower serum mucosal mast cell protease levels. Mesenteric lymph node (MLN) lymphocytes from all three experimental groups secreted IL-3 and IL-4 in copious amounts when stimulated in vitro by Concanavalin A (Con-A), but the secretion of high levels of IL-9 and IL-10 was essentially confined to mice infected with T. spiralis alone. It is suggested that adult H. polygyrus selectively modulate cytokine secretion by Th2 cells within the MLN during infection and that this is brought about as a direct consequence of the mechanism employed by H. polygyrus to depress mucosal inflammatory responses in order to facilitate its own survival.     

Immunological profile after splenectomy in children with beta-thalassaemia major

The in vitro immune functions of peripheral blood lymphocytes have been studied in 12 children with beta-thalassaemia major and hypersplenism. The study was performed prior to splenectomy and on the 2nd, 6th, 15th and 30th day after splenectomy. It was found that before splenectomy, patients had low numbers of blood leucocytes, normal rosette and T3 lymphocyte counts, low T4 and normal T8 lymphocyte counts with a T4/T8 ratio below 2, impaired T lymphocyte mitogenic responses induced by PHA, increased numbers of polyvalent and monovalent B lymphocytes and normal immunoglobulin levels of IgG and IgA. After splenectomy, especially on the 2nd day, leucocytosis, a significant decrease of T cells and their subsets and a reduction of the IgM levels were found. These parameters, except the IgM levels, increased until the 30th day after splenectomy and reached presplenectomy values. On the 2nd day after splenectomy, large mononuclear cells behaving like immunocytes appeared in the peripheral blood. They had the phenotype of T3, T4, T8, B lymphocytes and OKM1 monocytes. All the large mononuclear cells increased significantly on the 6th day after splenectomy and remained elevated during the whole study (30 days). The T4/T8 ratio was increased, but no increase was found in the functional responses of T lymphocytes.     

旋毛虫编码新生幼虫p46 kDa抗原基因重组融合蛋白对小鼠的免疫保护性研究

目的 探讨旋毛虫编码新生幼虫p46 kDa抗原基因重组融合蛋白WN10对小鼠的免疫保护性。方法 将纯化的重组融合蛋白WN10以20μg/只的剂量分3次免疫小鼠后，攻击感染旋毛虫肌幼虫200条／只，检查旋毛虫7日龄成虫数、雌虫体外产生新生幼虫数、感染35d的肌幼虫数并计算减虫率，同时用ELISA法检测血清中抗WN10抗体IgG滴度。结果WN10免疫小鼠后获得旋毛虫7日龄成虫、肌幼虫的减虫率分别为64．28％和61．21％，均与感染对照组和佐剂对照组差异显著；获得雌虫产新生幼虫的减虫率为16．46％，与感染对照组和佐剂对照组差异不显著；WN10免疫组小鼠在第3次免疫后1周可检测到高滴度的抗WN10抗体IgG，在攻击感染旋毛虫9周后仍维持在较高水平。结论 编码旋毛虫新生幼虫p46 kDa抗原基因重组融合蛋白可诱导小鼠产生较强的抗旋毛虫保护性免疫。     

Effects of trichinellosis on levels of eosinophils, eosinophil major basic protein, creatine kinase and basophils in the guinea pig

Summary The effects of multiple infections with Trichinella spiralis on the levels of eosinophil major basic protein (MBP), creatine kinase (CK), and leucocytes were studied in the guinea pig. Plasma MBP levels increased after each of four successive infections with T. spiralis. The times of peak elevations in MBP levels and eosinophilia correlated significantly, and both peaks occurred earlier with increased infection dose. CK levels were elevated most dramatically in the primary infection. A direct correlation between peak CK values and larval dose was observed, and the peak occurred earlier with larger larval doses. A dose-dependent basophilia was observed in the primary infection and, like the eosinophil response, basophilia occurred earlier and was markedly enhanced after the second infection. The elevations in MBP levels and eosinophil counts persisted into the third and fourth infections; the basophil response persisted only through the third infection.     

Early response of guinea-pig lungs to Trichinella spiralis infection

In order to assess immunological response, induced in guinea-pig lungs by Trichinella spiralis, cellular infiltration into pulmonary alveolar space and production of O(2)(-) and NO in alveolar macrophages obtained from bronchoalveolar lavage fluid (BALF), as well as accumulation of nitric oxide (NO) metabolites in BALF and serum, were evaluated during the early period of primary T. spiralis infection (from 4th to 8th and on 14th day after oral administration of larvae) and on 6th day after secondary infection. Primary infection caused increased infiltration of lymphocytes, macrophages, neutrophils and eosinophils, while secondary infection resulted in raised lymphocyte and eosinophil numbers. In spite of marked cellular infiltration of alveolar space, only very limited activation of effector cells, pointing to a suppressed innate response, was apparent, as (i) BALF supernatant phospholipid/protein concentration ratio, and lung levels of phospholipid peroxidation markers, conjugated dienes and malondialdehyde, did not change during 7 days following infection; (ii) primary, but not secondary, infection caused only a transient increase of superoxide anion production by alveolar macrophages; (iii) despite expression of inducible nitric oxide synthase in macrophages of control, infected and BCG-treated animals, and of interferon (IFN)-gamma-like activity in sera of infected animals, macrophage nitric oxide production was not affected by infection, even after additional stimulation in vitro (lipopolisaccharide + hrIFN-gamma) or in vivo (BCG or secondary T. spiralis infection); and (iv) increased nitrate concentrations were found in BALF supernatant and serum, but not in lung homogenates, of infected animals.     

Influence of nalcrom (sodium cromoglycate) on the course of the intestinal phase of trichinellosis in mice]

The influence of Nalcrom (sodium cromoglycate) on the course of the intestinal phase of trichinellosis in mice was investigated. The animals infected with 200 Trichinella spiralis larvae were treated with Nalcrom between 7-20 or 3-20 days after infection (d.a.i.). The drug was administered in two doses: 0.6 or 1.7 mg/mouse/day. In the all groups of animals received Nalcrom higher number of mast cells and eosinophils than in the control groups was observed. These results are the opposite of those obtained with Nalcrom in ulcerative colitis in man.