定居因子和不耐热肠毒素经皮免疫诱导抗ETEC的保护性免疫

产肠毒素性大肠杆菌 ( ETEC)引起的腹泻常见 ,对 ETEC定居因子 ( CF)和不耐热肠毒素 ( L T)的免疫应答与保护性免疫相关。此项研究探讨了 CF与佐剂 L T或霍乱毒素( CT)一起经皮免疫 ( TCI)的效果。　　实验以重组 CS6 ( r CS6 ,CF的组分之一 )为抗原 ,CT为佐剂。剃去 BABL/c和C57BL /6小鼠背毛 ,将含有抗原或抗原加佐剂的溶液涂于去毛的皮肤 1小时。结果显示 ,r CS6加 CT TCI能诱生高滴度的抗 CS6和CT抗体。而单纯 r CS6 TCI不能诱生持久的抗 CS6抗体应答。对不同免疫方案的小鼠用CT口服攻击 ,对照小鼠口服 1 0 %碳…     

抗蠕虫药物涂肤霜剂对旋毛虫病的实验疗效

本文报道抗蠕虫药物涂肤霜剂对小鼠旋毛虫病的实验疗效。这些霜剂分别含甲苯达唑(MD)、阿苯达唑(AD)、左旋咪唑(LVM)及HD+LVM。对肠道内成虫、移行期幼虫及肌肉内成囊期幼虫的减虫率(％)分别为:MD78.35,74.05,及32.50;AD68.11,98.72,及100;LVM38.98,6.31,及39.19;MD+LVM74.40,85.34,及99.56。     

有关乙型肝炎疫苗的一些问题

本文指出对乙型肝炎疫苗的一些问题应作较为细致的研究 :注射途径 (皮下或肌肉 )、抗原 (Ag)量、佐剂 (需要与否 )、HBs Ag的抗原性和分子结构。重要的是获得高的血清阳转率 ,而不是高滴度抗体 (保护性免疫力并非取决于抗体滴度 )。两种疫苗可以使用 ,并作进一步研究。何不在健康成人和儿童中使用小剂量疫苗 (1.5～ 2 .5 μg HBs Ag) ?它可能无需佐剂并作皮下注射。2 0 μg疫苗可能仅对高危人群中的弱或无应答者作肌肉注射     

茯苓总多糖对H1N1流感疫苗和乙肝疫苗抗原的佐剂作用

目的研究茯苓总多糖(PCP)对人用H1N1流感疫苗抗原和乙肝疫苗抗原的佐剂作用。方法在50℃条件下,采用水提、醇沉、透析和冷冻干燥方法制备PCP。苯酚-硫酸法测定总多糖含量,凝胶渗透色谱法测定多糖相对分子质量分布,毛细管电泳法测定单糖组成。H1N1流感病毒裂解液为抗原(每鼠3μg),与PCP(每鼠0.2或1.0 mg)联用,肌内注射免疫小鼠1次,免疫后14 d采用ELISA法测定小鼠血清特异性抗体Ig G滴度。乙肝表面抗原(HBs Ag)蛋白为抗原(每鼠2μg),与PCP(每鼠1.0 mg)联用,肌内注射免疫小鼠2次,于第2次免疫后14,21,28和35 d,采用ELISA法测定小鼠血清特异性抗体Ig G滴度。结果PCP中多糖含量为40.8%,由岩藻糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为1.00∶1.36∶0.48∶2.67。PCP与H1N1流感抗原联用免疫小鼠1次,能明显提高小鼠抗原特异性抗体Ig G滴度(P<0.05),效果与铝佐剂相当。与单用HBs Ag抗原组相比,PCP与HBs Ag联用初次免疫即能显著提高小鼠血清抗原特异性抗体滴度(P<0.01),且与铝佐剂组无显著性差异;二次免疫后14~21 d,PCP与HBs Ag联用组小鼠抗体水平进一步升高,且高于铝佐剂组(P<0.05),28~35 d PCP与HBs Ag联用组小鼠抗体滴度与铝佐剂组相当。结论PCP对人用H1N1流感疫苗和HBs Ag疫苗具有良好的佐剂活性,该多糖能显著提高小鼠体液免疫功能。     

嵩乙醚的抗血吸虫作用

实验感染日本血吸虫的小鼠,用蒿乙醚或蒿甲醚100～200mg·kg^-1·d^-1×2d灌胃治疗,显示两药对小鼠体内不同发育期血吸虫的减虫率相仿。特别是d7童虫和d35成虫组的减虫率较高,分别达77.5～87.2%和51.7～61.3%。经蒿乙醚作用后,d7童虫和d35成虫的糖原明显减少或消失,虫的皮层和实质组织中的碱性磷酸酶活力亦明显受抑制,表明蒿乙醚具有抗日本血吸虫童虫和成虫的作用。     

小鼠口服微囊化可溶性抗原后的B细胞激活型式

口服可溶性蛋白抗原易造成免疫耐受,为此作者采用可生物降解的、抗酸性丙烯酸聚合物包裹可溶性抗原卵白蛋白(OVA),并经口免疫BDF_1小鼠.结果显示,小鼠口饲5mg微囊化OVA后产生强免疫应答,IgA、IgG和IgG1同种型抗体滴度显著升高;而免疫5mgOVA溶液的小鼠的抗体水平则很低.对粘膜免疫系统激活的研究显示,在口饲微囊化OVA小鼠的集合淋巴结、肠系膜淋巴结、脾脏和固有层中,可见OVA特异性IgA、IgG和IgG1同种型抗体分泌细胞的激活.免疫第7天,在集合淋巴结和肠系膜淋巴结中出现抗原特异性抗体分泌细胞的激活     

肿瘤患者人巨细胞病毒活动性感染检测方法的比较

目的探讨肿瘤患者化疗后人巨细胞病毒感染检测方法的应用价值。方法使用免疫组化法检测人巨细胞病毒(HCMV)pp65抗原、酶联免疫吸附试验检测IgG/M抗体,以及实时荧光定量(FQ-PCR)检测HCMV DNA。结果47份全血标本中HCMVpp65抗原阳性率为48.9%,平均抗原阳性细胞数7.9±8.1(1-65)/5×104WBC,HCMV DNA阳性率19.1%(10/47),HCMV DNA含量均值为6.320×105拷贝,白细胞HCMV-DNA阳性率51%(25/47),HCMV DNA含量均值为3.830×107拷贝,HCMVpp65抗原阳性率为48.9%(23/47),IgG抗体均阳性,IgM抗体阳性率为23.4%(12/47),以pp65抗原阳性为对照,IgM抗体检测的敏感率仅为49.3%。结论在连续动态检测HCMV多种指标时,结合DNA及抗原动态检测具有更高临床应用价值。     

青霉素过敏病人血清特异性IgG抗体

目的探讨IgG抗体与青霉素过敏反应的关系,从而进一步完善青霉素类抗生素过敏反应的诊断。方法化学合成8种青霉素抗原决定簇与HSA结合的全抗原,采用酶联免疫吸附试验(ELISA)检测241例青霉素过敏病人血清中IgG抗体。结果241例过敏病人中,除了BPA-IgG抗体外,其他7种特异性IgG抗体血清水平均高于正常对照组,特异性IgG抗体阳性率为46.5%,其中皮试阴性组IgG抗体的阳性率(64.0%)高于皮试阳性组(27.6%)。主要抗原决定簇IgG抗体的阳性率(41.5%)高于次要抗原决定簇IgG抗体的阳性率(9.1%),并且在皮试阴性组及不同症状组也同样如此。特异性IgG抗体检出阳性率随检测抗体的增多而增加,且检测3种和检测8种特异性IgG抗体所得阳性率差异无显著性。结论特异性IgG抗体参与了青霉素过敏反应的发生和发展,青霉素过敏反应与主要抗原决定簇IgG抗体关系更为密切。     

河豚毒素实验疫苗对小鼠口服河豚毒素中毒的免疫保护效应

目的　开发河豚毒素 (TTX)的抗毒疫苗。方法　TTX与载体蛋白中国鲎血蓝蛋白 (TTH)、破伤风类毒素 (TT)和牛血清白蛋白 (BSA)化学偶联 ,分别制成免疫抗原TTX TTH和TTX TT ,检测抗原TTX BSA。经腹腔或皮下注射免疫原 (用Freund佐剂 ) ,免疫BALB/c小鼠 ;每组 12只 ;5个月内接受免疫原累计量 375 μg/鼠。定期采集动物血清 ,ELISA法监测血清中抗体质量 (滴度及亲和力 )。经igTTX的生理盐水攻击免疫鼠 ,以检验抗毒效价 ;首次剂量6 30 μg·kg- 1,间隔 2～ 3周后提高剂量再次攻击活存动物 ,观察攻毒实验后症状 ,记录 2 4h存活率及死亡鼠的存活时间。结果　所试两种人工抗原的抗毒效应无明显差异。免疫鼠经ig 6 30 ,80 0 ,12 0 0 ,15 0 0 ,2 0 0 0 μg·kg- 1TTX攻毒时 ,存活率分别为10 0 % ,95 % ,90 % ,70 %和 4 5 % ;测得半数死亡剂量约为 2 0 0 0 μg·kg- 1。免疫鼠经 2～ 5次igTTX重复攻毒 ,86 %动物累积耐受剂量高于 3.5mg·kg- 1;4 3%动物累积耐受剂量高于 5 .5mg·kg- 1。中毒对照动物ig 6 0 0 μg·kg- 1TTX全部死亡。结论　研制的TTX的化学实验疫苗可高效预防TTX口服攻毒 ,免疫预防是对抗TTX中毒很有希望的途径     

大蒜汁对旋毛虫幼虫杀灭效果研究

目的通过比较经不同浓度大蒜汁浸泡含旋毛虫肌幼虫的肉块对小鼠感染力的影响,来评估大蒜汁对旋毛虫幼虫的杀灭效果。方法 30只昆明小鼠分为5组,喂食经不同浓度的大蒜汁(浓度分别为100.00%、50.00%、25.00%、12.50%)和生理盐水浸泡半小时含有旋毛虫肌幼虫的肉,饲喂30d后剖杀小鼠,观察和计数肌幼虫数。结果小鼠饲喂经100.00%、50.00%、25.00%、12.50%浓度大蒜汁和生理盐水浸泡半小时含有旋毛虫肌幼虫的肉后,在单位肌肉中检出旋毛虫肌幼虫数为0条、10条、60条、140条和235条。结论含旋毛虫肌幼虫的肉经一定浓度的大蒜汁浸泡后其旋毛虫肌幼虫的感染力会降低。     

香肠腌制法对旋毛虫感染性影响的实验研究

目的观察香肠腌制法对旋毛虫肌幼虫感染性的影响。方法30只昆明小鼠被随机分为对照组和实验组,实验组又分5组,共6组,每组5只。对照组每鼠经口感染300条收集的肌幼虫。实验组分5组,4℃,阳性鼠肉香肠配料分别腌制24、48、72、96、120h,然后将5组小鼠每鼠经口感染300条处理好的肌幼虫。感染后28d处死小鼠,取膈肌压片镜检,并将全部肌肉人工消化后计数幼虫数。结果24h组、48h组、72h组、96h组压片法和人工消化法镜检,感染小鼠的幼虫检出率均为100％;4组的幼虫均数均显著低于对照组（P〈0．01）;48h组、72h组、96h组的幼虫均数均显著低于24h组（P〈0．01）。120h组两种方法镜检,感染小鼠均未检出幼虫。结论使用香肠腌制法腌制肉类,随着腌制时间的延长,肉内旋毛虫幼虫的活性和感染性逐渐下降。     

A spectrum of antibody (IgG. IgG1, IgM) response in mice infected with trichinella spiralis treated with L-mimosine

The purpose of this study was to demonstrate the anti-inflammatory effects of L-mimosine on chronic inflammation, by investigating its effect on the immunological response of BALB/c mice infected with the nematode parasite Trichinella spiralis. Specific anti-parasite immunoglobulins (IgG, IgG1 and IgM) were detected by the ELISA method in the serum of both the treated and the untreated animals at different periods of time for 60 days post infection. Two groups consisting of 18 mice each were used. The mice were 6 weeks of age. Both groups were infected with 220 larvae (L1-T. spiralis) per os: one group was administered an intraperitoneal injection of L-mimosine (200 &mgr;g/100 ml/dose) for 27 days (the first injection started 7 days before infection) and the second group was administered an intraperitoneal injection of saline solution (100 &mgr;l/dose). Parasite specific IgG, IgG1 and IgM levels were determined in the sera of infected, untreated mice. The levels of IgG and IgG1 were increased following infection and remained elevated throughout the experimental period, while IgM was significantly decreased on the 50th day post-infection. These levels were found to be lower in the L-mimosine treated infected mice, compared to the untreated mice. The inhibition started from day 10 and continued until day 60. In healthy animals, the production of immunoglobulins was not measurable. Non-infected animals treated with L-mimosine also showed no detectable anti-parasite specific immunoglobulins.     

Microcapsules formulated in the enteric coating copolymer Eudragit L100 as delivery systems for oral vaccination against infections by gastrointestinal nematode parasites

Microcapsules using the copolymer of methacrylic acid (Eudragit L100) were formulated for oral delivery of vaccines against the enteral/parenteral nematode parasite Trichinella spiralis. Antigenic preparations from first stage larvae (L1) of T. spiralis were microencapsulated in Eudragit L100. The microcapsules prepared by the spray drying method were resistant to acid pH, although the antigen was rapidly released under neutral and basic environmental conditions. The native protein conformation and biological activity was preserved in the microcapsules, as assessed by SDS-PAGE and ELISA. When administered to NIH mice, the antigen loaded microcapsules protected against infection by T. spiralis at both the intestinal and muscular levels, the worm burden diminishing by 45.58 and 53.33%, respectively. Furthermore, following administration of the microparticles an increase of the serum IgG1 response, a marker for the Th2 type response, was evident. These results indicate that microcapsules formulated with anionic biocompatible polymers such as Eudragit may be useful for oral vaccination against nematode infections.     

Effects of aging on resistance to Trichinella spiralis infection in rodents exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Immune function, including resistance to infection, decreases as humans and rodents age. We have shown that preinfection exposure of young (9-11 weeks) mice or rats to TCDD decreased resistance to Trichinella spiralis (Ts) infection, expressed as delayed onset or completion of parasite elimination and as increased muscle burdens of larvae. It has also been shown that aged mice express lower constitutive levels of resistance to Ts infection, compared to young adult animals. This study tested the hypothesis that the age-related decrease in constitutive levels of resistance to Ts infection exacerbates the decreased resistance to infection that follows TCDD exposure. This hypothesis addresses the concern that TCDD may pose a greater threat to the elderly than to the population at large. Animals were given a single oral dose of 1, 10, or 30 microg TCDD/kg, 7 days before infection. Eleven days later, young (approximately 10 weeks) control rodents had eliminated a greater proportion of the original parasite burden from the intestine than aged control animals. Nevertheless, parasite elimination was decreased by TCDD exposure only in young rodents. The effect of TCDD exposure on numbers of encysted larvae was evaluated only in rats. Increased larvae burdens occurred in young rats at 30 microg TCDD/kg and at 10 or 30 microg TCDD/kg in aged rats. Parasite-specific splenocyte and lymph node cell proliferation was suppressed following dioxin exposure in young mice; cells from aged mice were markedly less responsive to stimulation, yet less sensitive to TCDD exposure. The response to parasite antigens was not affected in aged rats exposed to TCDD, although elevated mitogen-driven B-cell proliferation was observed. These results indicate that age-related constitutive immunosuppression did not exacerbate TCDD-induced suppression of T-cell mediated adult parasite expulsion; rather, advanced age provided some degree of protection. On the other hand, a lower dose of TCDD was required in aged rats to suppress the combined humoral and cellular responses that limit the burden of encysted larvae, compared to young rats. These model-dependent results preclude acceptance or rejection of the tested hypothesis.     

Effect of the broad spectrum anthelmintic drug flubendazole upon Schistosoma mansoni experimentally infected mice

The effect of the broad spectrum anthelmintic drug flubendazole (methyl 5-(p-fluorobenzoyl)-2-benzimidazolecarbamate, CAS 31430-15-6), a mebendazole derivative with a molecular weight of 313.29, on Schistosoma mansoni infection in mice was evaluated. Moreover, the relationship between the posttreatment worm burden, hepatic granuloma volume, and serum immunoglobulin profile (immunoglobulin G and immunoglobulin M, IgG and IgM), was also investigated. Two main groups of Swiss albino mice infected with Schistosoma mansoni cercariae were used in the experiment. Group I consisted of infected untreated control mice. The mice of group II were submitted to treatment with flubendazole 100 mg/kg body weight as single oral dose at different time intervals: Group IIa received treatment 24 h before infection. Group IIb received treatment 4 h after infection. Group IIc received treatment 25 days after infection. Mice treated 25 days after infection, compared to those treated in other time intervals, revealed a significant reduction in the recovery of adult schistosomes after portal perfusion (79.5%), a lower immunoglobulin level (IgG and IgM), and the smallest granuloma mean diameter (220.0 +/- 10.3 microns). These data were less salient in mice treated 4 h after, and 24 h before infection.     

Impact of interleukin-1 and interleukin-6 in murine primary schistosomiasis

BACKGROUND: Immunization with schistosome antigens invariably elicits a plethora of cytokines and, hence, it is reasonable to assume that these cytokines influence host responses to challenge lung-stage larvae and, consequently, the adult worm burden, and may be responsible for the erratic data generally observed in protection studies against schistosome infection. METHODS: Schistosoma mansoni-infected mice were administered with recombinant interleukin (IL)-1beta or IL-6 to evaluate the impact of cytokines in host responses to lung-stage schistosomula, and subsequent effects on adult worm parameters. Plasma lipid levels were assayed by colorimetric enzymatic tests and antibody responses by ELISA. Cytokine profile in peripheral blood mononuclear cells was evaluated by RT-PCR. RESULTS: S. mansoni infection elicited, at the time of parasite residency in the lung, significant increase in free fatty acids (FA) and decrease in cholesterol plasma levels in C57BL/6 and CD1 mice, and stimulation of mRNA expression for cytokines of T helper type (Th) 2 in BALB/c, Th1 in C57BL/6, and Th1/Th2 in CD1 mice. However, no specific antibody production was evident in any mouse strain. In BALB/c mice, exogenous IL-1beta-related plasma free FA level significant increase, stimulation of expression of IL-1 and IL-12 mRNA, and considerable increase in percent of specific antibody-producing mice were associated with significant reduction in adult worm burden and egg load. In contrast, exogenous IL-1beta elicited decrease in free FA plasma levels, and down-regulation of cytokines' mRNA expression in C57BL/6 and CD1 mice, changes associated with aggravation of the worm burden. Likewise, exogenous IL-6 failed to stimulate increase in plasma free FA levels or percent of antibody-producing mice except in BALB/c mice, effects that were protective for the host in BALB/c and for the parasite in C57BL/6 and CD1 mice. CONCLUSION: These findings were discussed in relation to the erratic data of protection experiments with schistosome subunit antigens in different mouse strains.     

Sufficiency of a single administration of filarial antigens adsorbed on polymeric lamellar substrate particles of poly (L-lactide) for immunization

A majority of antigens require repeated administration to ensure development of adequate humoral and cell mediated immune response. To minimize the number of administrations required, we investigated the utility of biodegradable polymeric lamellar substrate particles of poly (l-lactide) (PLSP) as adjuvant for filarial antigen preparations. PLSP was prepared and characterized and Brugia malayi adult worm extract (BmA) and its SDS-PAGE resolved 54-68 kDa fraction F6 were adsorbed on to PLSP. Swiss mice received a single injection of PLSP-F6, PLSP-BmA, FCA-F6, FCA-BmA and two doses of the plain antigens. Specific IgG, IgG1, IgG2a, IgG2b and IgE levels in serum, IFN-γ, TNF-α and nitric oxide (NO) release from cells of the immunized animals in response to antigen challenge were studied. The average size of PLSP particles was <10 μm and its % antigen adsorption efficacy was 60.4, 55.2 and 61.6 for BSA, BmA and F6, respectively. Single injection of PLSP-F6 or PLSP-BmA produced better immune responses compared to one injection of FCA-F6/BmA or two injections of plain F6 or BmA. Moreover, PLSP-F6 produced much better response than PLSP-BmA. These data demonstrate for the first time that PLSP is a superior immunoadjuvant for enhancing the immune response to filarial BmA and F6 molecules and obviates the need for multiple immunization injections.     

不同日龄的旋毛虫成虫感染小白鼠的实验研究

实验结果显示,每只小鼠由200条3日龄旋毛虫成虫感染的20只小鼠的肠内成虫数值范围6—57条,肌肉内幼虫数值范围17—33条。7日龄成虫感染的20只小鼠的肠内成虫数值范围2—34条,肌肉内幼虫数值范围2—55条。15日龄成虫感染的20只小鼠的肠内成虫数值范围0—8条,肌肉内幼虫数值范围0—3条。30日龄成虫感染的20只小鼠的肠内和肌肉内无成虫和幼虫所见。     

硝硫氰胺抗日本血吸虫病的研究

硝硫氰胺是抗血吸虫病的一种新药物。本文进一步研究该药物对实验性日本血吸虫病的预防和治疗的作用。实验动物在感染血吸虫尾蚴前后,即使口服500mg/kg/天×2或4天的硝硫氰胺,血吸虫组比对照组仅减少32.3～59%。药物对童虫几乎无作用,而对成虫则有明显的疗效。病鼠一次口服硝硫氰胺40mg/kg,停药后1～8天的肝移率达93.6～100%。各实验治疗组的减虫率为74～100%,但此疗效的高低与剂量和疗程有关。     

F30385实验治疗血吸虫病的初步研究

实验发现F30385是一个兼具明显杀日本血吸虫童虫与成虫的硝基呋喃丙烯酰胺类的口服药物。小白鼠一次口服F30385的半数致死置为979±98毫克/公斤(P=0.95)。小白鼠感染尾蚴后4-11天,一次口服F30385 11.4毫克/鼠,减虫率高达90-99%,显著比对32天成虫的杀虫作用强。按等毒性剂量用F30385及F30066治疗小白鼠与兔血吸虫病的结果,F30385的疗效比F30066高。7只感染血吸虫病的犬用总剂量为700毫克/公斤的7-14天疗法治疗后,减虫率为95%。动物口服F30385后的毒性反应主要为胃肠道刺激与肾和肝的受损。小白鼠病理观察结果认为,停药后病变均渐恢复。