感染旋毛虫小鼠外周血T淋巴细胞及其亚群和血清...

Acid alpha-naphthyl esterase (ANAE), a cytoplasmic marker, was used to identify the T lymphocytes and their subpopulations in peripheral blood of mice infected with Trichinella spiralis, and Dot-ELISA was used to identify the serum IgG antibody of the infected mice. The results showed that T lymphocytes increased on the d3 after infection, reaching the peak on d14, and then remained in number greater than normal up to d77. The spotted granular ANAE positive cells (Help T cells, Th) decreased and the scattered granular ANAE positive cells (Suppressor T cells, Ts) increased, leading to a drop of the Th/Ts ratio. The reduction in host immune function during T. spiralis infection might be related to the drop of Th/Ts ratio. At the same time, the serum IgG antibody increased on d7 after infection, reaching the peak on d28, and remained higher up to d140. The results indicated that the host's cellular immunological level was lower while the host's IgG antibody level was higher during T. spiralis infection.     

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。     

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.     

旋毛虫感染小鼠膈肌虫荷与血清及肉汁抗体水平的研究

目的观察旋毛虫感染小鼠膈肌虫荷(larvae per gram diaphragm,lpgd)与血清及肉汁抗体水平的关系。方法将32只雄性昆明小鼠随机分成4组(每组8只),每只分别感染50(A组)、100(B组)、300(C组)、500条(D组)旋毛虫幼虫,感染后42d剖杀,收集血清及肉汁,观察膈肌虫荷并用旋毛虫肌幼虫ES抗原ELISA检测血清及肉汁抗体。另将50只小鼠随机分成5组(每组10只),每组分别感染旋毛虫(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500条幼虫,感染后42d剖杀,观察感染不同种旋毛虫小鼠的膈肌虫荷与血清及肉汁抗体水平。结果感染旋毛虫的A、B、C、D4组小鼠的膈肌虫荷与血清及肉汁抗体水平均无相关性(P>0.05),但与感染剂量呈正相关(P<0.05),每组小鼠的血清与肉汁抗体水平也均具有相关性(P<0.05)。5种旋毛虫感染小鼠的膈肌虫荷与血清及肉汁抗体水平相比均无相关性(P>0.05),但血清与肉汁抗体水平相比均具有相关性(P<0.05)。旋毛虫感染小鼠的血清及肉汁抗体水平明显高于其他4种旋毛虫(乡土旋毛虫、布氏旋毛虫、伪旋毛虫及纳氏旋毛虫)感染小鼠的血清和肉汁抗体水平(P<0.05)。结论旋毛虫肌幼虫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)感染小鼠血清及肉汁中抗旋毛虫抗体的检测。     

Influence of cadmium on the course of intestinal and muscular phases in mice infected with Trichinella spiralis]

At the first experiment 20 Swiss male mice were infected with 100 larvae and 20 mice with 500 larvae T. spiralis per mouse. Two days after infection (d.a.i.) mice orally received 1.5 mg Cd (water solution CdCl2) each. 40 mice were infected only T. spiralis as control. At the second experiment muscle larvae used were isolated from mice (from the first experiment) which received Cd. 20 mice were infected with 100 larvae and 20-with 500 larvae per mouse. Two d.a.i. mice received 1.5 mg Cd. Mice from both experiments were killed at 5, 10, 20 and 42 d.a.i. Total number of adult T. spiralis worms present in the small intestine and muscle larvae were recovered by conventional technique. Results of the first experiment: the number of adult worms and muscle larvae recovered from mice received Cd. were statistically significant lower as in control. Results of the second experiment: the mean number of adult worms in experimental group and in control were the same but the mean number of larvae per gram of mice muscle were significant higher as in control.     

Enhanced resistance to Plasmodium berghei in mice previously infected with Trichinella spiralis

Infection with Trichinella spiralis larvae greatly enhanced the resistance of adult mice against fatal infection with Plasmodium berghei given 10 and 30 days after T. spiralis infection. Mice infected with T. spiralis had a markedly activated mononuclear phagocytic system and significantly low reticulocyte levels at the time the mice were challenged with P. berghei. Therefore, the partially subdued parasitaemia and prolonged survival of Trichinella—Plasmodium -infected mice may be attributed, in part, to macrophage activity and reticulocytopenia exerting a specific anti- P. berghei effect. This study suggests the role of T. spiralis induced reticulocytopenia and activated macrophages as potential mechanisms in resistance to P. berghei infection.     

Lymphocyte proliferation and cytokine production in mice infected with different geographical isolates of Trichinella spiralis

Antigen-specific proliferative responses and cytokine production were measured in lymphocytes taken from inbred NIH mice infected with different isolates of Trichinella spiralis. Stimulation with muscle larval homogenate antigen produced high proliferative responses, cells responding more strongly to antigen from one isolate (London-L) than to antigens of the other two isolates (Spanish-S and Polish-P). Antigen from the P-isolate elicited relatively poor responses. There was considerable, although variable, cross reactivity between isolate antigens and this was reflected in proliferation to heterologous stimulation. When pulsed with the mitogen Concanavalin A (Con-A) mesenteric lymph node cells (MLNC) produced IFN-gamma on day four post infection but not on day eight. In contrast, production of IL-4 and IL-5 was greatest on day eight. Differences were seen in levels of cytokine production between cells taken from mice infected with different isolates. These data show a sequential activation of Th1 and Th2 cells during infection with T. spiralis isolates, and suggest that the level of activation of each Th subset is influenced by the antigenic characteristics of each isolate.     

感染旋毛虫小鼠免疫能力的性别差异研究

目的 研究感染旋毛虫早期雌雄小鼠免疫能力的性别差异。方法 不同剂量的旋毛虫感染雌雄小鼠7 d后,测定脏器指数和白细胞组成比例,脾脏和胸腺组织进行HE染色分析,ELISA法测定血清中IL-2、IL-4、IL-10的含量,流式细胞仪检测脾脏免疫调节性T细胞比例。结果 与感染旋毛虫的雌性小鼠相比,感染旋毛虫雄性小鼠的胸腺指数明显增大(t=4.595, P<0.05);淋巴细胞百分比明显下降而单核细胞和粒细胞百分比明显上升(t=2.604, P<0.05);胸腺组织变化不明显而脾脏组织变化明显;免疫调节性T细胞比例上升,但无统计学意义。血清中细胞因子的变化也存在雌雄差异,与正常小鼠相比,感染旋毛虫的雌性小鼠IL-2含量上升而雄性小鼠下降(F=5.664,P<0.05);IL-4、IL-10含量均上升(F=10.461,P<0.05;F=1.170,P>0.05),且雄性高于雌性。结论 感染旋毛虫7 d后,雄性小鼠的免疫应答强于雌性小鼠。     

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。     

外源性一氧化氮对旋毛虫感染小鼠抗氧化能力的影响

目的探讨外源性NO供体亚硝基铁氰化钠(SNP)对旋毛虫病小鼠肝脏、外周血中抗氧化酶活性及脂质氧化的影响。方法采用消化法分离旋毛虫肌幼虫,感染BALB/c小鼠,400条/只。42 d后,收集感染鼠及正常小鼠外周血、肝脏。设A组(感染鼠肝脏匀浆物+SNP)、B组(正常鼠肝脏匀浆物+SNP)、C组(感染鼠外周血+SNP)、D组(正常鼠外周血+SNP)。每组内设5个SNP反应终浓度,分别为0(空白对照)、2、5、10、30μmol/L,37℃反应30 min,测定各组超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)及脂质氧化(MDA)含量。结果与正常鼠相比,旋毛虫感染鼠肝脏、外周血中SOD、CAT、GSH-Px活性上升,MDA含量增加,差异均有统计学意义(P均0.05);在A、B组中,10、30μmol/L SNP作用下SOD、CAT、GSH-Px活性较空白对照降低(P均0.05);与2~30μmol/L SNP反应后,MDA含量较空白对照明显升高(P均0.05)。在C、D组中,30μmol/LSNP作用下SOD、CAT、GSH-Px活性显著下降,MDA浓度升高,与空白对照比较差异均有统计学意义(P均0.01)。在2~30μmol/L浓度区间,SNP浓度与旋毛虫感染鼠肝脏、外周血中SOD、CAT、GSH-Px活性呈负相关(r肝SOD=-0.901,r肝CAT=-0.802,r肝GSH-Px=-0.847,r血SOD=-0.899,r血CAT=-0.685,r血GSH-Px=-0.635,P均0.05),与MDA含量呈正相关(r肝MDA=0.697,r血MDA=0.764,P均0.05)。结论旋毛虫感染可致小鼠肝脏、外周血中活性氧自由基产生增加,SOD、CAT、GSH-Px活性升高;外源性NO可降低旋毛虫感染小鼠SOD、CAT、GSH-Px活性,抑制小鼠的抗氧化能力。     

Suppression of mucosal mastocytosis by Nematospiroides dubius results from an adult worm-mediated effect upon host lymphocytes

Infections with the nematode Nematospiroides dubius fail to elicit mucosal mast cell (MMC) responses in the intestines of host mice, and suppress MMC responses generated by heterologous infection. Larval N. dubius have the capacity to prime for mastocytosis, and to elicit this response in primed mice during a challenge, but only if adult worms are prevented from developing, either by anthelmintic treatment or by irradiation of the larvae themselves. The suppressive effect of the adult stage was confirmed in experiments where such worms were implanted directly into the intestines of mice primed by exposure to irradiated N. dubius larvae or concurrently infected with Trichinella spiralis. Data on the mechanisms underlying this suppressive effect were obtained from experiments involving the adoptive transfer of mastocytosis by mesenteric lymph node cells (MLNC) from T. spiralis infected mice. When MLNC were taken from mice infected concurrently with both T. spiralis and N. dubius no enhanced mastocytosis was seen in recipients after challenge with T. spiralis. Exposure of MLNC from T. spiralis infected donors to the presence of adult N. dubius after transfer did not reduce the adoptively transferred response. The response was also unaffected when MLNC from adult N. dubius infected mice were simultaneously transferred with MLNC from T. spiralis donors. It is concluded that the suppressive effect of adult N. dubius upon the expression of mucosal mastocytosis acts upon the generation of lymphocytes capable of promoting the development of MMC from precursor cells.     

Influence of Trichinella spiralis infection on development of sarcoma-180 ascites tumors

Long-term infection of mice with Trichinella spiralis has been shown to stimuate increased host protection aginst transplantable solid tumors. The present investionation was initiated to determine whether parasitized mice were similarly protected from induction and progression of an ascites tumor. ICR/CD-1 mice were orally infected with 50, 100, 200, 300, or 400 T. spiralis larvae, and subsequently challenged intraperitoneally with 5 X 10(4) Sarcoma-180 (S-180) ascites cells. Animals were observed and weighed daily for development and progression of malignancy. Protection from fatal ascites neoplasia was found to be statistically significant under selected conditions of larval dose and challenge interval. Mice infected with 100 or 200 larvae were more resistant to S-180 progression than both uninfected controls or other infected groups. Protection was observed in groups challenged 2 weeks after nematode intubation but not at 6, 8 or 34 weeks. This finding is in contrast with long-term protection seen previously with B-16 melanoma tumors, and suggests that antineoplastic effects of T. spiralis infection differ for ascitic and solid murine tumors.     

A role for elevated plasma corticosterone in modulation of host response during infection with Trichinella pseudospiralis

Suppression of host inflammatory response in mice infected with Trichinella pseudospiralis was associated with host plasma corticosterone levels significantly higher than those seen in uninfected mice or in mice infected with T. spiralis. Increases in the population of mitochondria and depletion of lipid droplets in cells of the zona fasciculata were seen in the adrenals of mice infected with T. pseudospiralis. Elevations in enteritis, myositis and myocarditis accompanied 100% mortality in adrenalectomized mice infected with T. pseudospiralis, while lower levels of inflammation and no mortality were observed in sham operated or intact animals infected with this parasite. The severe myositis normally accompanying infection with T. spiralis was suppressed by concurrent infection with 1000 or 2000 T. pseudospiralis to levels equivalent to those seen in animals receiving 0.15 and 0.41 mg cortisone acetate/25 g mouse/day, respectively.     

Quantitative study of the effect of previous Trichinella spiralis infection on sarcoma 180 ascitic tumor formation in mice.

Tumor development in HaM/ICR mice inoculated intraperitoneally with Sarcoma 180 (S-180) can be easily quantitated by linear regression analysis of non-destructive measurement of total body mass. This procedure readily separated the postinoculation period into incubation and clinical phases. The former was dependent on S-180 dose, while the latter was not. Trichinella spiralis infection 28 days prior to S-180 challenge produced small but statistically significant increases in the length of both the incubation period and survival time, but did not affect the clinical phase. Administration of tumor cells 56 days followoing the helminthic infection had no detectable effect on any parameter of tumor development studied. The results are consistent with the hypothesis that T. spiralis infection temporarily altered the host reaction to S-180, possibly at the level of non-specific macrophage activity.     

旋毛虫肌幼虫在感染小鼠体内的分布

目的观察旋毛虫肌幼虫在感染小鼠体内不同部位横纹肌内的分布和密度。方法分别取感染旋毛虫小鼠的舌肌、咬肌、胸肌、腹肌、前肢肌、后肢肌、膈肌和背肌各50mg,肌肉压片镜检。结果膈肌幼虫密度最高,其次为舌肌、胸肌;前肢肌、后肢肌、咬肌、背肌幼虫密度无明显差异,均低于胸肌,腹肌幼虫密度最低。结论感染旋毛虫的小鼠从膈肌取材,检出肌幼虫的阳性率较其他部位高。     

旋毛虫成虫可溶性抗原免疫小鼠攻击感染后的免疫应答

目的　探讨旋毛虫成虫可溶性抗原 (AWSAg)接种小鼠后诱发的免疫应答。　 方法　制备旋毛虫AWSAg免疫小鼠 ,分别于攻击感染后 7、14、2 1、2 8和 3 5d ,动态观察免疫小鼠特异性IgG抗体水平、IL 2水平和T淋巴细胞亚群的变化。　结果　与非免疫鼠相比 ,免疫鼠血清特异性IgG抗体OD值在攻击感染后 7d明显升高 ,在观察期内一直高于非免疫鼠。感染后 7dIL 2水平明显增高 ,达观察期内最高值 ,2 1d后才逐渐减少 ,在感染后 2 8～ 3 5d仍高于正常组。免疫鼠CD4+ T细胞在攻击感染 7d明显增加并保持不变 ,非免疫鼠CD4+ T细胞及CD4/CD8比值明显较正常鼠减低。　结论　旋毛虫AWSAg免疫小鼠攻击感染后 ,出现细胞、体液免疫应答。     

Trials of penetration modification by inflammatory infiltrating cells into Trichinella spiralis larva capsules

The inflammatory infiltration cells penetrating through the larval capsule walls were observed. The observation were carried out in the masseter muscle of the mice infected with Trichinella spiralis larvae only (control) or infected and treated with PHA-P, TFX or dexamethason. The more numerous cells, than in the control, in the mice treated with PHA-P and TFX were seen. Opposite, the fever numerous cells inside the capsula walls in the mice that received dexamethason were found. The authors supposed that the phenomenon of inflammatory cells penetration through larval capsule play an important role in the defence mechanisms of the host.     

Activity of natural killer (NK) cells in the course of experimental trichinellosis in mice.

B6C3F1 mice were infected with 200 or 500 larvae of Trichinella spiralis per mouse and pulmonary NK cell-mediated clearance of semisyngeneic tumour cells was determined in vivo on days 10, 20, 30, and 60 after the infection. Cytotoxic activity of NK cells in the lungs was substantially elevated on days 20 and 30 after challenge with both "doses" of the parasite. At the same time large granular lymphocytes (LGLs) as well as cells expressing surface asialo-GM1 molecules were isolated in elevated numbers from spleens of the infected as opposed to the normal mice. Expression of other markers of differentiation, such as THy 1, CD4, and CD8 was also enhanced on splenocytes isolated from the infected mice on day 30 but not 20 after administration of the larvae. The present results indicate that NK cell-mediated activity in vivo is stimulated above the baseline level during migration and early muscle phases of the infection with T. spiralis in mice. The possible impact of this effect upon the course of trichinellosis as well as upon the growth of tumours in the infected host is discussed.     

田鼠巴贝虫感染BALB/c小鼠血细胞动态变化

目的 观察小鼠感染田鼠巴贝虫（Babesia microti）后体重、脾脏及血液细胞的变化情况。方法 取田鼠巴贝虫感染种鼠外周血（虫密度为20%）,腹腔接种6周龄健康BALB/c小鼠,100 μL/只,并设立健康小鼠对照组。感染组小鼠于感染后0～28 d隔日尾尖采血,涂薄血片,吉氏染色后油镜下观察田鼠巴贝虫增殖情况,并分别于感染后0、7、14、21、28 d测定小鼠体重、脾重,采用全自动血细胞分析仪分析小鼠血细胞。结果 小鼠在感染后第1天即可在外周血中查见田鼠巴贝虫虫体,第7天染虫率最高（55%）,随后感染率逐渐下降,至第28天外周血虫体镜检阴性,进入隐性感染阶段。小鼠于感染后第7天体重降至最低,随后逐渐恢复;脾脏重量在感染后第14天最重,后逐渐降低,但至第28天脾重仍高于正常组。全血细胞分析发现,感染组小鼠白细胞数量及淋巴细胞、嗜酸性粒细胞比例无明显变化,波动范围在正常小鼠参考值范围内。感染组小鼠红细胞计数、血红蛋白含量及血小板计数在感染后虫密度高峰期降至最低,后随着虫密度降低而逐渐恢复至正常水平。结论 田鼠巴贝虫感染可引起小鼠体重下降、脾脏增重、红细胞及血小板数量降低,全血细胞分析仪对巴贝虫病诊断具有参考价值。