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血细胞对异物及自身组织碎片的吞噬作用,是家蚕细胞性免疫的主要反应之一。通过比较家蚕体内和体外血细细胞对微孢子原虫的蚕噬作用,发现其体内与体外的吞噬作用有明显的差别,血细胞在蚕体内蚕噬作用迅速,在体外的吞噬作用缓慢,推测在血淋巴中存在信息因子,引起蚕体内血细胞的趋化反应及整个免疫系统的协同反应。 相似文献
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采用已建立的家蚕(Bombyx mori)造血器官(HPO)体外培养系统,研究了2种蜕皮激素ecdysone和20-hydroxyecdysone(20E)以及保幼激素类似物methoprene对体外培养的家蚕HPO造血功能的调节作用。添加0.005~0.5μg/mL20E可促进5龄第2天幼虫(V-2)HPO的血细胞增殖和释放,但会抑制熟蚕开始时期(W-0)HPO的血细胞增殖和释放;添加低浓度(0.036 5μg/mL)ecdysone可以促进V-2和W-0期HPO的血细胞增殖及释放,高浓度(0.365~3.650μg/mL)ecdysone对血细胞增殖无促进作用,但有促进血细胞释放作用;添加低浓度(2μg/mL)methoprene不会阻止HPO的血细胞增殖,但抑制血细胞的释放,高浓度(20~200μg/mL)methoprene可抑制HPO的血细胞增殖和释放。此外,0.036 5μg/mL ecdysone与2μg/mL methoprene共同处理时,ecdysone可以掩盖methoprene抑制血细胞释放的作用。研究结果表明,ecdysone和20E对家蚕HPO的造血功能起促进作用,而methoprene则起抑制作用,ecdysone可以拮抗methoprene抑制血细胞释放的作用。 相似文献
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双酚A对体外培养家蚕生精囊形成精子的影响 总被引：1，自引：0，他引：1
为了探讨环境有害物质对家蚕生殖发育的影响,利用家蚕生精囊(spermatocyst)体外培养系统,研究塑料工业广泛使用的化合物双酚A(BPA)对体外培养家蚕生精囊精子形成的影响。在5龄第4天家蚕生精囊体外培养体系中添加100μg/mLBPA,可使生精囊全部不能伸长,绝大多数在72 h内死亡;添加10μg/mL BPA时,生精囊培养72 h后表现出明显伸长障碍,有核精子束数量减少且长度短;添加1μg/mL BPA时,有核精子束数量减少且长度短,还出现一些退化膨大的生精囊。在上蔟第2天的家蚕生精囊体外培养体系中添加0.1μg/mL BPA,可对无核精子形成产生毒性效应,明显影响到有核精子束的形成及精子束的浓缩成熟。实验结果显示:较低浓度的BPA对家蚕精子形成具有明显的毒害作用;上蔟第2天的家蚕生精囊体外培养体系对低浓度BPA的毒性反应敏感,可应用于家蚕以及昆虫的生殖毒理体外研究实验。 相似文献
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为探讨家蚕血细胞的吞噬作用,对蚕体腔分别注射微粒子孢子M_(1 2)和核多角体,经30min、1h、2h后,用相差显微镜及扫描电镜进行观察。家蚕血细胞对微粒子M_(1 2)孢子及核多角体的捕食作用,主要为颗粒细胞,其吞噬作用较之其他血细胞显著,家蚕血细胞对微粒子及核多角体的吞噬作用主要为膜捕捉(active en-membranosis),注射病原微生物30min后,可观察到多数的颗粒细胞已在捕食病原微生物;经2h后,多数的颗粒细胞已吞噬了数个甚至10数个病原微生物。 相似文献
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《中国蚕业》2018,(4)
家蚕是鳞翅目重要的模式生物,但是目前家蚕培养细胞系少,给家蚕基础研究带来不便,也影响了家蚕作为模式生物的应用。组织体外培养在一定程度上可以代表体内环境,且操作相对简单。我们尝试并建立了一种采用TC-100培养基体外培养丝腺组织的方法,取健康家蚕5龄2 d的幼虫,用75%乙醇进行表面消毒,灭菌水冲洗,剪开腹部表皮固定,用镊子轻轻拉出丝腺组织,用75%乙醇快速漂洗、PBS缓冲液冲洗2～3次,放入含有800μL TC-100培养基的12孔细胞培养板的培养孔中,每孔4条丝腺组织,27℃培养;将0.64μg增强荧光蛋白基因表达载体pcDNA3.0[ie1-egfp-SV40]加入到25μL不完全TC-100培养基中孵育15 min后转染丝腺组织,48 h后用荧光显微镜观察,培养基清澈,丝腺组织外形完好、发出亮绿色荧光,表明egfp在丝腺中得到了表达。为进一步验证体外培养丝腺的转染效果,将家蚕miR-0031*的抑制物(inhibitor)和阴性对照(inhibitor negetive control)各10μL按照同样方法配成转染液,加入到放有丝腺组织的培养孔中,48 h收集丝腺组织,提取总RNA,荧光定量PCR检测靶基因BmFib-L的表达量,结果显示,inhibitor抑制miR-0031*的表达,促使BmFib-L的表达量上升,表明用体外培养丝腺进行转染等试验是可行的。该方法的建立为研究蚕丝蛋白基因表达调控的分子机制提供新的技术途径,也为家蚕其他组织的体外培养和研究提供借鉴。 相似文献
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丝氨酸蛋白酶抑制剂(serpin)对昆虫的免疫反应起重要调节作用。根据家蚕基因组数据库序列设计引物,从家蚕幼虫血细胞中克隆了Bmserpin6的编码基因,并构建重组表达载体,在大肠杆菌中成功表达了重组Bmserpin6蛋白。将纯化的重组Bmserpin6注射家蚕5龄幼虫后6 h提取家蚕血液制备血清,检测血清样品中的酚氧化酶原活性显著下降(P0.05)。体腔注射重组Bmserpin6后的家蚕5龄幼虫再通过微球菌诱导蚕体内抗菌肽gloverin2基因的表达,结果显示幼虫脂肪体和血细胞中的gloverin2基因表达显著下调(P0.05)。依据上述结果推测:体外表达的重组Bmserpin6可以调节蚕体内2个重要的免疫途径,即抑制蚕体酚氧化酶原的激活和外源细菌诱导的蚕体抗菌肽的产生。 相似文献
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环境激素阿特拉津对家蚕血细胞的影响 总被引：1，自引：0，他引：1
为了探讨环境激素阿特拉津(Atrazine,AT)对家蚕生理的影响,用添加AT的人工饲料饲养家蚕,调查了5龄幼虫血细胞数和血细胞形态的变化,同时调查了血液细胞DNA的损伤情况。结果表明:AT连续添食后,家蚕5龄第4日-6日家蚕血球细胞特征性释放高峰时,0.05mmol/kg低浓度AT区的血球细胞数显著增加(p<0.01),0.10mmol/kg浓度AT区血球细胞数则显著减少(p<0.05),而0.20mmol/kg浓度AT区则没有明显影响;实验浓度的AT添食,家蚕5龄幼虫血球细胞的形态没有发现畸形变化,也未见血球细胞DNA的遗传损伤。结论:家蚕食下能够完成幼虫生长发育的环境激素AT(0.20mmol/kg以下浓度)后,血球细胞形态和细胞核DNA没有受到明显不良影响,但AT会影响家蚕血球细胞数目的变化,可能是家蚕积极应对体外化学物质影响的生理反应。 相似文献
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家蚕血细胞系BmHc对脂多糖(LPS)和20-羟蜕皮酮(20E)的敏感性 总被引：1，自引：1，他引：0
为研究家蚕的细胞信号转导及外源基因在家蚕血细胞中的表达,用TC-199培养基建立起一个血细胞系BmHc。BmHc细胞系由原血球、小球细胞、颗粒细胞和浆细胞等血细胞组成,细胞倍增时间约30 h。脂多糖(LPS)刺激血细胞合成抗菌蛋白和多肽,启动细胞吸收Ca2+参与细胞的增殖。用Flu-3 AM处理细胞30 min并结合TCS-SP2激光共聚焦扫描显微镜技术对Ca2+的分布、定位进行研究的结果表明,Ca2+主要分布在原血球和颗粒细胞的膜表面,但有部分荧光分布深入到原血细胞核,显示细胞核参与了钙信号的转导途径。同等条件下,用20羟蜕皮酮(20E)处理仅可见轻微的荧光反应。LPS激发并打开了细胞膜上的Ca2+通道,引起细胞内Ca2+浓度的变化。在培养基内加入20E引起的Ca2+浓度变化相对较小,因此,20E不是细胞膜上Ca2+通道的有效激发剂。 相似文献
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家蚕有5种血细胞,可以通过吖啶橙(acridine orange,AO)和碘化丙啶(propidium iodide,PI)染色来区分。溴化噻唑蓝四氮唑[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide,MTT]是一种广泛应用于细胞增殖和活细胞鉴定的染色剂。通过对家蚕血细胞进行复染,即先用MTT染色,再用AO和PI染色,发现以下2种情况:很多颗粒细胞、浆细胞和所有的原血球细胞不能被MTT染色,但AO染色显示为活的细胞;被MTT染色的细胞表面有紫色纤维状物伸出,除了一部分具有伪足的颗粒细胞外,所有被MTT染色的血细胞也被PI染色,细胞核是红色。因此,利用这3种染料对家蚕血细胞复合染色来判断细胞的存活与死亡,需要仔细分析并分别对待。 相似文献
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Demonstration of Anaplasma marginale in hemolymph of Dermacentor andersoni by animal inoculation and by fluorescent-antibody technique 总被引：1，自引：0，他引：1
K M Kocan R D Oberst S A Ewing J A Hair S J Barron 《American journal of veterinary research》1983,44(5):798-801
Hemolymph was collected from adult Dermacentor andersoni Stiles that had been infected with Anaplasma marginale Theiler as nymphs. Before hemolymph was collected, the adult ticks were either incubated and unfed at 37 C for 2.5 days or fed for 6 days on sheep. Hemolymph collected from groups of 100 ticks was inoculated into susceptible splenectomized calves. Smears of hemolymph from the same groups of ticks were prepared for examination by fluorescent antibody technique. Hemolymph from incubated ticks caused anaplasmosis in 2 of 4 trials, and hemolymph from feeding ticks caused anaplasmosis in 4 of 4 trials. Moderately fluorescing bodies were demonstrated in some hemocytes from incubated ticks, whereas hemocytes from feeding ticks contained numerous clusters of brightly fluorescing bodies. Fluorescing bodies were not observed in hemocytes from control ticks. 相似文献
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N H Kumazawa T Tanigawa Y Tanaka H Osatake K Tanaka 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1991,53(2):297-300
Vibrio parahaemolyticus D-3 was observed to attach to hemocytes of a marine gastropod mollusc, Nerita albicilla, regardless of the presence of N. albicilla serum. The organism attached to hemocytes of an estuarine gastropod, Clithon retropictus, in the presence of C. retropictus serum while the attachment to the hemocytes was decreased significantly in the absence of the serum. These evidences suggest that N. albicilla hemocytes would facilitate the clearance of V. parahaemolyticus from the alimentary tract of the mollusc and that C. retropictus hemocytes would protect C. retropictus against the invasion of V. parahaemolyticus to hemocoel of the mollusc. 相似文献
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利用杆状病毒Bac-to-Bac系统在家蚕表达猪繁殖与呼吸综合征病毒的E蛋白和M蛋白 总被引：2，自引：0，他引：2
猪繁殖与呼吸综合征病毒(PRRSV)是一种严重危害养猪业的传染性病原。PRRSV病毒的E蛋白和M蛋白基因是开发PRRSV病毒新型疫苗的目标基因。利用杆状病毒Bac-to-Bac表达系统,将PRRSV病毒的E蛋白和M蛋白的基因亚克隆到杆状病毒转移载体pFastBacHTb中,获得重组转移质粒pFastBacHTb-E和pFastBacHTb-M,转化大肠杆菌Bm DH10 Bac感受态细胞,获得重组杆粒BmNPV Bacmid-E、BmNPV Bacmid-M,将这些重组杆粒转染家蚕培养细胞Bm5,获得重组病毒BmNPV-E和BmNPV-M。将2种重组病毒分别接种5龄起蚕,用SDS-PAGE和Western blotting方法在重组Bacmid DNA转染的Bm5细胞和感染重组病毒的家蚕幼虫血细胞中分别检测到分子质量约20 kD和18 kD的E蛋白和M蛋白,表明PRRSV病毒的E蛋白和M蛋白在家蚕培养细胞及幼虫体内获得了表达,为利用家蚕-杆状病毒表达系统研制PRRSV的新型疫苗与诊断试剂奠定了基础。 相似文献
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N H Kumazawa K Iwao N Morimoto 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1992,54(2):243-247
Hemocytes of two marine neritid gastropods, Nerita albicilla and Heminerita japonica, were attracted chemotactically to live Vibrio parahaemolyticus and Escherichia coli strains. Chemotactic attraction of N. albicilla hemocytes was enhanced in the presence of N. albicilla plasma, while that of H. japonica hemocytes was not enhanced in the presence of H. japonica plasma. Chemotactic activity of the hemocytes seems to participate in the rapid elimination of V. parahaemolyticus from these gastropods. 相似文献
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N H Kumazawa N Morimoto 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1992,54(5):851-855
Hemocytes from adult and juvenile specimens of a brackish-water clam, Corbicula japonica, were attracted chemotactically to live cells of Vibrio parahaemolyticus and Escherichia coli strains in a balanced salt solution, which was enhanced significantly in the presence of respective C. japonica plasma. Chemotactic attractions of adult's and juvenile's hemocytes were seen also in artificial seawater at a similar level to those in the balanced salt solution. Chemotactic attractions of juvenile's hemocytes to these strains were lower in level than those of adult's hemocytes. C. japonica plasma seems to facilitate for C. japonica hemocytes to recognize these organisms. 相似文献
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In the present study, 19 monoclonal antibodies (mAbs) against adult Ornithodoros moubata hemocytes were established, and the reactivity of the hemocytes to these mAbs was examined by an indirect fluorescent antibody test (IFAT), Western blot and immunoprecipitation analyses. It was shown that the reactivities of the hemocytes to the mAbs varied among morphologically similar hemocyte types, and most mAbs produced in the present study showed the multiple band reactivity. However, the presence of shared epitopes among peptide subunits of the same protein or entirely different proteins are not common, so their reactivity could not be explained in detail. These results suggest that there are morphologically similar but functionally differentiated hemocytes. Therefore, in addition to morphological classification, the molecular-based classification of the hemocytes is also required. In order to assess the lethal effect of blood meal containing each mAb, artificial feeding was performed. The OmHC 31 showed the strongest lethal effect on adult female O. moubata. In conclusion, anti-hemocyte mAbs produced in this study are useful not only for the immunological classification of hemocytes but also for the immunological control of the tick. 相似文献
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《Journal of aquatic animal health》2013,25(2):179-182
Abstract Phagocytosis of zymosan A particles by hemocytes of eastern oyster Crassostrea virginica stimulates the production of reactive oxygen species (ROS), which are easily quantified by luminol-augmented chemiluminescence (CL). Antimicrobial defense mechanisms of hemocytes may involve the activity of cytotoxic ROS. The CL response to phagocytosis of zymosan by hemocytes from C. virginica with advanced Perkinsus marinus infections is more elevated than that produced by zymosan in cells from uninfected oysters. This effect is perhaps akin to macrophage activation. Phagocytosis of P. marinus cells by hemocytes withdrawn from uninfected oysters produced no detectable CL response. Hemocytes withdrawn from oysters with P. marinus infections ranging from light to heavy were evaluated for CL responses after phagocytosis of zymosan or P. marinus. Increases in CL stimulation by zymosan were seen as the intensity of infection increased. Despite avid phagocytosis of P. marinus, CL activity of the hemocytes was not seen, regardless of the level of infection of the host. Lack of hemocytic ROS stimulation by ingestion of P. marinus cells may contribute to in vivo survival of this parasite. 相似文献
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中国对虾血淋巴细胞的原代培养及其与量子点标记的VP37p的作用 总被引：1，自引：0，他引：1
血淋巴细胞是对虾白斑综合征病毒(WSSV)靶细胞之一,WSSV结构蛋白与血淋巴细胞的作用在病毒感染中起重要作用。论文主要研究中国对虾血淋巴细胞的原代培养及在原代细胞培养条件下观察重组表达的对虾白斑综合征病毒VP37片段(VP37p)与对虾血淋巴细胞的作用。试验表明,中国对虾血淋巴细胞体外培养可存活12d,体外培养的对虾血淋巴细胞与量子点标记的VP37p具有相互作用。 相似文献