Immune Responses to Antigens of in vitro Reared Echinococcus granulosus Adult Worms in Balb/c Mice

Background: Cystic echinococcosis (CE), also known as echinococcosis/hydatidosis, is one of the most important parasitic diseases in the world. It enhances both humoral and cellular (Th1 and Th2) responses in infected host. Different antigens of the worm may favor the Th1 or Th2 immune responses in CE patients. Objective: To evaluate the humoral and cellular immune responses of Balb/c mice against the crude and excretory/secretory (E/S) antigens of in vitro reared Echinococcus granulosus adult worms. Methods: A total of 20 Balb/c mice divided into 5 groups of 4 mice each. Three groups of mice (n=4) were immunized with crude, E/S and an immunodominant antigen of in vitro reared Echinococcus granulosus adult worms on day 1 and 28. The fourth and the fifth groups were negative control groups and received PBS plus adjuvant, or nothing, respectively. Two weeks after the second injection, the mice were killed and their blood was collected for determining antibody responses, and their spleens were employed for proliferation assay. Total IgG was measured by indirect ELISA. Spleen cells of immunized mice were cultivated and exposed to different antigens of adult worms including E/S and crude antigens. Levels of IFN-γ, IL-12, IL-4 and IL-10 were measured in the recovered cell culture supernatants by capture ELISA. Results: Total IgG assay showed the highest level of antibody produced in mice immunized with crude antigens. Proliferation assay showed a statistically significant production of cytokines in the mice immunized with crude antigens (p<0.05). The highest levels of IFN-γ, IL-12 and IL-4 were produced in mice immunized with crude antigen of the in vitro reared Echinococcus granulosus adult worms followed by E/S antigens. Immunodomonant antigen induced the lowest levels of cytokines (IL-12, IFN-γ, IL-4 and IL-10) in immunized mice. Conclusion: Significant levels of Th1 related cytokines (IFN-γ and IL-12) were produced in Balb/c mice immunized with crude antigen of the in vitro reared Echinococcus granulosus adult worms.     

Effect of neutralizing antibodies to IL-10 and C5 on the renal damage caused by a pathogenic human anti-dsDNA antibody

OBJECTIVES: There is considerable evidence suggesting that anti-double-stranded deoxyribonucleic acid (anti-dsDNA) antibodies are involved in the pathogenesis of lupus nephritis. It was shown previously using severe combined immune deficient (SCID) mice that when the hybridomas secreting human immunoglobulin G (IgG) anti-dsDNA antibodies, RH14 and DIL-6, were implanted intraperitoneally the antibodies produced by RH14, but not DIL-6, deposited in the kidneys, caused pathological changes in the renal tissues and induced proteinuria. In this study we have further analysed the effect of activated terminal complement proteins and interleukin-10 (IL-10) in the pathogenesis of glomerulonephritis caused by the RH-14. METHODS: SCID mice implanted with RH-14 or DIL-6 cell lines were treated with neutralizing antibodies to IL-10 (mAb B-S10) or anti-complement factor 5 (anti-C5) (mAb BB5.1) intraperitoneally. Control groups received either an isotype control antibody (135.8) or phosphate-buffered saline (PBS). Serum human IgG levels and proteinuria were estimated and the extent of renal involvement was examined by histopathological and electron microscopic techniques. RESULTS: While there was a tendency to reduce proteinuria in the anti-IL-10 injected group the anti-C5 injected group showed a significant reduction in proteinuria (P<0.01) compared with the groups injected with either the control mAb or PBS. There was a considerable reduction in the serum human IgG levels in the anti-IL-10 but not in the anti-C5 treated animals. Both anti-IL-10 and anti-C5 treated groups showed significantly reduced renal impairment as revealed by histopathological examination and proteinuria assessment. CONCLUSION: The findings, while confirming the role of IL-10 and activated terminal complement component in the production of antibody at the cellular level and at the site of glomerular immune deposition in this model, respectively, also suggest the beneficial effect of a combined therapy using both anti-IL-10 and anti-C5 mAb to prevent or reduce the effect of the humoral immune response in lupus disease.     

广州市褐云玛瑙螺感染广州管圆线虫的调查分析

目的对广州市褐云玛瑙螺(Achatina fulica)感染广州管圆线虫(Angiostrongylus cantonensis)第Ⅲ期幼虫的情况进行调查和比较,填补广州市广州管圆线虫病的流行病学资料。方法分别于2000年7月和2004年7月对广州市天河区、海珠区和越秀区褐云玛瑙螺进行采样收集,消化法分离第Ⅲ期幼虫,计算感染率和感染度并与过去及国内其它地区进行比较,用灌胃法构建大鼠和小鼠动物模型,检测脑部和心肺晚期虫体。结果广州市褐云玛瑙螺广州管圆线虫感染率,2000年为44.2%(57/129),感染度401条/螺,而2004年为27.3%(33/121),感染度为72条/螺,2次感染率差异有显著性(χ2=7.5,P<0.01),各区褐云玛瑙螺的感染率差异较大,最高达65.9%。褐云玛瑙螺的感染率与其螺重呈正比。用分离的第Ⅲ期幼虫构建了小鼠和大鼠动物模型,分别从脑部和心肺检出较晚期虫体。结论广州市褐云玛瑙螺的广州管圆线虫感染率和感染度2004年较2000年有明显下降,2004年感染率低于曾发生过广州管圆线虫病暴发流行的地区;小鼠适于构建广州管圆线虫第Ⅳ、Ⅴ期模型,大鼠适于构建第Ⅴ期和成虫模型。     

IL-17对日本血吸虫感染抗体及Th应答的影响

目的观察IL-17在日本血吸虫感染小鼠中对抗体及Th细胞免疫应答的影响。方法在小鼠日本血吸虫感染前后,连续多次腹腔注射rm-IL-17细胞因子（实验组1）或PBS（对照组1）,连续多次腹腔注射anti-IL-17中和抗体（实验组2）或同型抗体（对照组2）。运用流式细胞术,采用表面分子、胞内因子同时染色的三色标记法,对日本血吸虫感染小鼠脾脏、淋巴结中的Th17、Th1和Th2细胞亚群占CD4＋T细胞的比例进行观察;同时采用ELISA法检测各组小鼠血清中血吸虫抗原特异性总IgG及亚类IgG1和IgG2a的水平。结果实验组1 CD4＋T细胞中的Th17、Th1和Th2细胞亚群的比例分别是对照组1的1.09倍（P（0.05）、1.17倍（P（0.05）和1.15倍（P（0.05）;实验组2 CD4＋T细胞中的Th17、Th1和Th2细胞亚群的比例分别是对照组2的1.01倍（P（0.05）、1.11倍（P（0.05）和0.98倍（P（0.05）。rm-IL-17注射后仅可溶性虫卵抗原（SEA）特异性IgG1水平显著下降（P（0.05）、可溶性成虫抗原（SWA）特异性IgG2a的水平升高（P（0.05）,其他抗体水平均无...     

Monoclonal antibody 12D5 inhibits eosinophil infiltration in the brain of Angiostrongylus cantonensis-infected BALB/c mice

Each of BALB/c mice was infected with 50 Angiostrongylus cantonensis larvae. One group of mice received an intraperitoneal injection of 50 μg 12D5 monoclonal antibody (mAb) against a 98 kDa antigen of adult worms at 10 days post-infection (dpi), with a booster injection of 25 μg at 12 dpi. Five mice from each group were sacrificed at 14 dpi for pathological examination and RNA extraction. The infiltration of eosinophils and severity of eosinophilic meningitis were reduced in 12D5 mAb-treated mice compared with the infected mice without 12D5 treatment. The levels of eotaxin mRNA expression in spleen significantly increased and the expression of the Th2-type cytokine IL-5 significantly decreased. However, the expression of IL-4 was not changed. 12D5 mAb can observably enhance the survival rate of infected mice and reduce symptoms of angiostrongyliasis. A. cantonensis infection is a major cause of eosinophilic meningoencephalitis. The results of this study could be helpful for the development of treatment of human angiostrongylosis.     

Local antibody production and immune complex formation in rats experimentally infected with Angiostrongylus cantonensis

The systemic humoral immune responses and tissue localization of worm-antigen, antibodies (IgG), and complement (C3) were examined in rats experimentally infected with Angiostrongylus cantonensis. While the worms remained in the subarachnoid space, it was infiltrated with plasma cells and lymphoid cells containing IgM and IgG. When the infiltration of these cells became more pronounced, the serum antibody titer began to increase. At the same time, deposits of IgM, IgG, and C3 were found in the glomeruli of the kidney. A number of eggs were observed in the lungs, enclosed in granulomatous tissues. Infiltrates of plasma cells including IgM and IgG, and deposits of IgM, IgG, and C3 were detected around the eggs and in the granulomatous tissues. A marked increase in serum antibody was observed. A. cantonensis larvae induce local antibody (IgM and IgG) production in the central nervous system prior to an increase of serum antibody titer. Measurement of cerebrospinal fluid antibody titer at an early stage of infection may confirm infection. The larvae showed no evidence of damage in spite of marked local antibody production in the central nervous system. The eggs in the lungs stimulated both local and systemic antibody production, and immune complexes were formed in the lung and the circulatory system. Immune complexes may participate in the formation of granuloma.     

曼氏裂头蚴病动物模型的建立及诊治技术研究Ⅱ原尾蚴灌胃法建立曼氏裂头蚴病小鼠模型

目的　探索通过感染原尾蚴的剑水蚤灌胃小鼠建立曼氏裂头蚴病小鼠动物模型的可行性。方法　实验室条件下用曼氏裂头蚴感染家猫,收集当日新产猫粪,用去氯水淘洗后过滤获得曼氏迭宫绦虫虫卵,制成虫卵悬浊液。将20只C57BL/6j小鼠随机分为实验组和对照组,实验组15只、对照组5只。以曼氏迭宫绦虫钩球蚴感染野生剑水蚤,使每只剑水蚤体内含有3 ~ 5条原尾蚴;再将感染原尾蚴的剑水蚤灌胃实验组小鼠,每只小鼠灌入剑水蚤15只,对照组灌胃等体积去氯水。6个月后处死小鼠,分离疑似裂头蚴虫体,并用间接酶联免疫吸附试验检测小鼠血清中抗曼氏裂头蚴特异性IgG抗体水平。提取疑似虫体基因组DNA,以曼氏裂头蚴细胞色素C氧化酶1（CO1）基因片段为目的序列进行PCR扩增以鉴别疑似虫体。结果　实验组15只小鼠中,6只小鼠血清抗曼氏裂头蚴特异性IgG抗体检测呈阳性;于其中4只小鼠皮下发现并分离出乳白色虫体。每只小鼠均仅检出1条虫体,虫体形态与曼氏裂头蚴相似。基于CO1基因的疑似虫体PCR扩增产物在毛细管电泳上于151 bp处显示特异性条带,测序结果与已知曼氏裂头蚴序列100%符合。结论　本研究成功建立了一种曼氏裂头蚴病小鼠动物模型。     

抗广州管圆线虫成虫单克隆抗体的研制及初步应用

目的制备抗广州管圆线虫成虫单克隆抗体并研究其初步应用。方法用广州管圆线虫成虫可溶性抗原免疫BALB/c小鼠,经融合,筛选分泌高滴度、高特异性单克隆抗体的杂交瘤细胞株。使用所得的单克隆抗体以Western blotting检测广州管圆线虫病患者血清,并建立双抗体夹心ELISA法检测感染大鼠血清。结果获得3株分泌高滴度抗广州管圆线虫成虫可溶性抗原的单克隆抗体杂交瘤细胞株(2A2、3F1、4H2),其分泌的抗体与日本血吸虫、卫氏并殖吸虫、猪囊尾蚴、旋毛虫抗原均不发生交叉反应。3株单克隆抗体均可识别广州管圆线虫成虫可溶性抗原蛋白Mr 15 000,以及患者血清中两种循环抗原Mr24 000和Mr15 000。双抗体夹心ELISA检测阳性率为76.5%。结论制备的抗广州管圆线虫成虫可溶性抗原的杂交瘤细胞株能分泌高滴度、高特异性的单克隆抗体,且在循环抗原的检测方面有应用前景。     

Mechanisms of effects of complement inhibition in murine collagen-induced arthritis

OBJECTIVE: To determine the mechanisms of amelioration of collagen-induced arthritis (CIA) in DBA/1J mice by inhibition of complement activation. METHODS: Mice received 2 intradermal injections of bovine type II collagen (CII), on days 0 and 21. From day 21 (immediately after the second injection of CII) through day 35, mice received intraperitoneal injections of either phosphate buffered saline (PBS), a monoclonal mouse antibody to murine C5 (anti-C5 antibody), or the C3 convertase inhibitor Crry-Ig. RESULTS: On days 30 and 32, the clinical disease activity score was lower in mice treated with anti-C5 antibody than in those treated with Crry-Ig. Histopathologic evidence of joint damage was 75% lower in the mice treated with anti-C5 antibody than in those treated with either PBS or Crry-Ig. Spleen cells from mice receiving either form of complement inhibition exhibited decreased CII-stimulated proliferation, whereas increased proliferative responses were exhibited by lymph node cells from mice treated with Crry-Ig. Treatment with anti-C5 antibody decreased production of IgG1 anticollagen antibody, while production of IgG2a antibody was inhibited by both complement inhibitory treatments. CII-stimulated spleen cells from anti-C5-treated mice produced lower levels of tumor necrosis factor alpha (TNFalpha) and interleukin-10 (IL-10) compared with those from mice treated with Crry-Ig. Lower steady-state messenger RNA (mRNA) levels for TNFalpha, interferon-gamma (IFNgamma), IL-18, and IL-6 were observed in the joints of anti-C5-treated mice, and for IFNgamma and IL-6 in mice receiving Crry-Ig, all in comparison with PBS-treated mice. However, mRNA levels for IL-1beta and TNFalpha were lower in the joints after treatment with anti-C5 compared with Crry-Ig. CONCLUSION: These results indicate that inhibition of complement in CIA leads to decreased production of IgG2a antibody and suppressed CII-induced spleen cell proliferation. The greater inhibitory effects on CIA of anti-C5 antibody in comparison with Crry-Ig may be attributable primarily to decreased levels of IL-1beta and TNFalpha mRNA in the joints.     

Pretreatment with cyclosporine and anti-interleukin 2 receptor antibody abrogates the anti-idiotype response in rat recipients of cardiac allografts.

A 10-day course with ART-18, a mouse monoclonal antibody (mAb) directed against the rat interleukin 2 receptor (IL-2R), prolongs the survival of (LEW x BN)F1 cardiac allografts in LEW recipients to approximately 3 weeks (acute rejection = 8 days, P less than 0.001). We examined host responses against ART-18 idiotype (Id) and mouse immunoglobulin in recipients immunomodulated with ART-18 mAb. Treatment with ART-18 elicited high titers of anti-Id antibodies 14 days after transplantation. However, naive rats given ART-18 before transplantation showed strong anti-Id responses as early as day 4 after engraftment, coinciding with abrogation of the treatment effect (graft survival, approximately 10 days). Preimmunization with irrelevant mouse IgG, which elicited high titers of anti-IgG, did not influence the efficacy of ART-18 upon graft survival (17 days). The use of cyclosporin A (CsA) in conjunction with ART-18 prior to transplantation suppressed the anti-Id response and led to dramatic graft prolongation (greater than 58 days), with two of five grafts surviving indefinitely. This striking effect of CsA plus ART-18 pretreatment did not depend upon CsA per se, as grafts were rejected within 12 days in animals pretreated with CsA alone; in both groups CsA trough levels were comparable. Moreover, administration of CsA before transplantation in concert with control IgG (instead of ART-18) prompted rejection within 2-4 weeks. Thus, discrete interaction(s) between anti-IL-2R mAb and CsA prior to engraftment induces partial host unresponsiveness/tolerance to anti-IL-2R mAb treatment following transplantation and suppresses the neutralizing anti-Id responses, which results in long-term/permanent graft acceptance. This study provides a strategy to overcome the anti-Id response mounted by graft recipients that otherwise limits the efficacy of anti-IL-2R mAb treatment.     

Immunological relationships during primary infection with Heligmosomoides polygyrus (Nematospiroides dubius): parasite specific IgG1 antibody responses and primary response phenotype

IgG1 antibody responses to Heligmosomoides polygyrus were measured in eight mouse strains supporting acute (< 8 weeks, SJL, SWR), intermediate (10–20 weeks, NIH, BALB/c) or chronic (> 25 weeks, C57BL/0, CBA, C3H, AKR) primary infections. Mice supporting acute or intermediate infections produced more intense antibody responses and total serum IgG1 concentrations were higher than in mice tolerating chronic infections. Positive correlations across mouse strains between the intensity of the antibody response and the percentage loss of worms in weeks 6 and 10 were established. No correlation was found between the response within mouse strains and loss of worms by individual mice. Heavy infections gave marginally higher antibody titres than low intensity infections, but few significant differences were detected and it was concluded that infection intensity did not markedly influence the magnitude of the antibody response. Male and female mice responded similarly despite the earlier loss of worms from females. No association was found between the primary response phenotype and recognition of particular antigens in Western blot analysis, nor did intensity of infection or host gender affect recognition. The possibility that immunomodulatory properties of adult worms may have had a differential influence on ability of strains of contrasting response phenotype to mount IgG1 responses was discussed.     

Ultrastructural and morphometric analyses of eosinophils from the cerebrospinal fluid of the mouse and guinea-pig infected with Angiostrongylus cantonensis

Guinea-pigs infected with Angiostrongylus cantonensis developed pleocytosis and eosinophilia in the cerebrospinal fluid (CSF) at day 12 post-infection (p.i.), showing a peak response at day 20 p.i., followed by a gradual reduction. Ultrastructural observations on CSF eosinophils from infected mice and guinea-pigs revealed various signs of eosinophil degranulation after day 14 p.i., suggesting the exocytosis of lysosomal material. Morphometric analysis indicated that CSF eosinophils after day 22 p.i. contained fewer granules as well as smaller granules than those at days 14-20 p.i. These data suggest that CSF eosinophils release granule constituents into the outside of the cells and these secretion products could interact with the intracranial worms and are probably related to worm death. As degenerative atrophy or partial loss of Purkinje cells and the spongy vacuolation of the white matter were noted in the cerebellum of infected mice, it was suggested that CSF eosinophils could be a possible cause of neurological disorders in angiostrongyliasis cantonensis.     

Suppression of in vivo polyclonal IgE responses by monoclonal antibody to the lymphokine B-cell stimulatory factor 1.

The lymphokine B-cell stimulatory factor 1 (BSF-1) has been shown to greatly enhance the differentiation of lipopolysaccharide-activated B cells into IgG1- and IgE-secreting cells in vitro. To determine whether in vivo IgG1 and IgE antibody responses are BSF-1 dependent, the ability of a monoclonal rat IgG1 anti-BSF-1 antibody, 11B11, to affect polyclonal IgG1 and IgE production in mice infected with the nematode parasite Nippostrongylus brasiliensis or injected with a purified goat antibody to mouse IgD was studied. 11B11-containing ascites fluid or purified 11B11 strongly inhibited IgE production in both systems but did not affect IgG1 production, while control ascites or normal rat IgG1 had no IgE-inhibitory activity. These results indicate an important physiologic role for BSF-1 in the generation of IgE antibody responses and suggest means for limiting the production of antibodies responsible for allergic reactions without inhibiting protective antibody responses.     

BALB/c小鼠感染广州管圆线虫后脑内的动态变化

目的观察广州管圆线虫感染BALB/c小鼠后其虫数在小鼠脑内的动态变化,探讨其感染后对人体可能的致病机制。方法以广州管圆线虫Ⅲ期幼虫感染实验BALB/c小鼠,按感染的不同时间分批处死,镜下及肉眼观察脑内虫体数量及分布情况。结果在BALB/c小鼠大、小脑内,广州管圆线虫的分布符合寄生虫感染宿主后先增后减的一般基本变化规律,虫体寄生的部位因感染虫体而受损。引起症状的时间变化与虫体在脑内变化规律基本相符。结论小鼠感染广州管圆线虫以后,出现共济失调,抽搐等症状的时间与虫体在小鼠脑内的变化规律基本相符。为临床研究该病的相关临床症状及治疗提供了一定的实验依据。     

First report of Angiostrongylus cantonensis in Puerto Rico

From January to April 1984, 63 Rattus rattus and 40 R. norvegicus were trapped in northeastern Puerto Rico and examined for Angiostrongylus cantonensis adults. Nineteen (47.5%) of the R. norvegicus and 10 (15.9%) of the R. rattus were infected, giving an overall infection rate of 28.2%. Four species of terrestrial snails and one species of brown slug were examined for A. cantonensis larvae. Two snail species, Subulina octona and Aquebana belutina, were found infected with third stage larvae of A. cantonensis. These larvae were harvested and inoculated per os into adult white mice. Immature adult worms were found in the brain tissue of all mice inoculated. This is the first report in the rat and snail populations of Puerto Rico.     

两种药物治疗广州管圆线虫病的效果观察

目的　观察阿苯哒唑和甲苯咪唑治疗广州管圆线虫病的效果。　方法　以大白鼠为动物模型 ,用不同剂量和不同疗程的阿苯哒唑和甲苯咪唑进行治疗 ,然后解剖检虫、计数、观察 ,同时对广州管圆线虫病病人进行治疗。　结果　阿苯哒唑和甲苯咪唑总剂量达 12 0 mg/ kg时可杀死鼠体内的虫体 ;总剂量为 6 0 mg/ kg,疗程达 6 d时 ,也可杀灭虫体。但总剂量也为 6 0 mg/ kg而疗程只有 3d或总剂量 <6 0 mg/ kg时 ,部分虫体仍可存活 ,但虫体的发育受到严重影响 ,无法达到性成熟 ,不能产卵。对部分确诊的广州管圆线虫病病人进行治疗也收到很好的效果。　结论　阿苯哒唑和甲苯咪唑两种药物对广州管圆线虫病有显著的疗效。     

双抗体夹心ELISA法检测广州管圆线虫循环抗原的研究

目的建立双抗体夹心ELISA法检测广州管圆线虫循环抗原(Cag)。方法采用两株抗广州管圆线虫成虫可溶性抗原单克隆抗体,应用双抗体夹心ELISA法检测广州管圆线虫CAg,并摸索该法的最佳实验条件。对感染大鼠、小鼠、疑似和确诊广州管圆线虫病病例血清进行检测,评价方法的敏感性;对日本血吸虫、肺吸虫、旋毛虫、囊虫病人血清进行交叉反应试验,评价方法的特异性。结果10μg/ml单抗包被,酶标记单抗1∶1 600稀释为最佳工作浓度。利用该法检测不同血清广州管圆线虫循环抗原的敏感性为84.2%~87.2%,特异性为100%。结论建立的双抗体夹心ELISA法检测广州管圆线虫循环抗原具有敏感性高、特异性强和经济、简便易行等优点。     

感染广州管圆线虫大白鼠血清抗体动态的初步研究

应用广州管圆线虫雌虫抗原,采用PVC-FLISA检测大鼠感染广州省圆线虫后血清抗体的动态。抗体于感染后第三周明显增多,第四周和第五周继续增高,第八、九周达高峰,第21至第25周下降,但仍为阳性。每鼠分别感染50、100及200条此虫第三期幼虫的3个感染组OD均值间无显著差异。感染大鼠血清抗体的出现时间与此虫产卵及鼠粪中幼虫出现时间未见一致。报告对部分实验鼠解剖查虫的结果。     

日本血吸虫硫氧还蛋白谷胱甘肽还原酶免疫功能的研究

目的 目的 研究日本血吸虫硫氧还蛋白谷胱甘肽还原酶 （Thioredoxin Glutathione Reductase of Schistosoma japoni? cum,SjTGR） 的免疫原性及抗日本血吸虫感染的免疫保护性作用。 方法 方法 75只小鼠随机分为5组： 空白组、 PBS组、 CpG2免疫组、 TGR免疫组和TGR + CpG2联合免疫组, 免疫3次。首次免疫前及第3次免疫后2周经尾静脉采血, 采用酶联免疫法分别检测血清中抗SjTGR蛋白IgG、 IgG1和IgG2a的抗体水平。第3次免疫后2周, 每只小鼠经腹部感染日本血吸虫尾蚴40±2条, 42 d后剖杀, 收集成虫和虫卵, 计算减虫率和减卵率; 制备各组小鼠单个脾淋巴细胞, 采用流式细胞术检测脾细胞表面标志物CD44high 、 CD4+ CD44high 或CD8+ CD44high 水平; 用SjTGR刺激免疫小鼠脾细胞72 h, 采用酶联免疫法检测脾细胞的IL?2、 IL?4、 IL?10和IFN?γ的水平。 结果 结果 第3次免疫后2周TGR与CpG2 + TGR免疫组小鼠血清抗SjTGR 抗体的IgG滴度均达1： 200 000以上, IgG2a与IgG1的比值 （IgG2a/ IgG1） 随时间的延长缓慢增高。TGR免疫组和TGR + CpG2联合免疫组细胞上清中的IFN?γ和IL?2水平与空白组、 PBS组、 CpG2免疫组相比明显增高 （P < 0.05）。TGR免疫组与TGR + CpG2免疫组小鼠脾细胞的CD44high 、 CD8+ CD44high 及CD4+ CD44high 与空白组和PBS组相比细胞比值增加 （P < 0.05）。TGR免疫组和CpG2 + TGR免疫组的减虫率分别为9.4%, 10.5%, 减卵率分别为9.2%和32.8%。 结论 结论 SjTGR具有较强的免疫原性, 可诱导小鼠免疫反应向Th1型极化, 但不足以产生抗日本血吸虫感染的保护效应。CpG2 ODN可能是一种有效的Th1型免疫佐剂。     

Mechanisms of effects of complement inhibition in murine collagen‐induced arthritis

Objective

To determine the mechanisms of amelioration of collagen‐induced arthritis (CIA) in DBA/1J mice by inhibition of complement activation.

Methods

Mice received 2 intradermal injections of bovine type II collagen (CII), on days 0 and 21. From day 21 (immediately after the second injection of CII) through day 35, mice received intraperitoneal injections of either phosphate buffered saline (PBS), a monoclonal mouse antibody to murine C5 (anti‐C5 antibody), or the C3 convertase inhibitor Crry‐Ig.

Results

On days 30 and 32, the clinical disease activity score was lower in mice treated with anti‐C5 antibody than in those treated with Crry‐Ig. Histopathologic evidence of joint damage was 75% lower in the mice treated with anti‐C5 antibody than in those treated with either PBS or Crry‐Ig. Spleen cells from mice receiving either form of complement inhibition exhibited decreased CII‐stimulated proliferation, whereas increased proliferative responses were exhibited by lymph node cells from mice treated with Crry‐Ig. Treatment with anti‐C5 antibody decreased production of IgG1 anticollagen antibody, while production of IgG2a antibody was inhibited by both complement inhibitory treatments. CII‐stimulated spleen cells from anti‐C5–treated mice produced lower levels of tumor necrosis factor α (TNFα) and interleukin‐10 (IL‐10) compared with those from mice treated with Crry‐Ig. Lower steady‐state messenger RNA (mRNA) levels for TNFα, interferon‐γ (IFNγ), IL‐18, and IL‐6 were observed in the joints of anti‐C5–treated mice, and for IFNγ and IL‐6 in mice receiving Crry‐Ig, all in comparison with PBS‐treated mice. However, mRNA levels for IL‐1β and TNFα were lower in the joints after treatment with anti‐C5 compared with Crry‐Ig.

Conclusion

These results indicate that inhibition of complement in CIA leads to decreased production of IgG2a antibody and suppressed CII‐induced spleen cell proliferation. The greater inhibitory effects on CIA of anti‐C5 antibody in comparison with Crry‐Ig may be attributable primarily to decreased levels of IL‐1β and TNFα mRNA in the joints.