抗卵巢抗体对卵巢组织及其功能影响的实验研究

目的 研究抗卵巢抗体对卵巢组织及其功能的影响。方法 以正常小鼠卵巢匀浆抽提物作为抗原免疫兔,制备兔抗鼠卵巢抗血清,并提取抗卵巢抗体（ＡＯＡ）,将ＡＯＡ被动免疫小鼠,应用光镜,电镜观察不同剂量ＡＯＡ对卵巢组织的影响,测定不同剂量ＡＯＡ作用下的自然受孕率才超促排卵疗效。结果 ＡＯＡ作用下的小鼠卵巢多种组织成分增妻生不同程度的病理变化,其中尤为明显的是透明带和颗粒细胞,随ＡＯＡ剂量的增加而损伤加重。自然     

PM_(2.5)暴露对小鼠卵巢雌、孕激素受体表达的影响

目的:通过小鼠PM_(2.5)染毒模型,观察小鼠卵巢组织病理变化及雌激素受体(ER)、孕激素受体(PR)表达的变化,探讨PM_(2.5)对卵巢的毒性作用。方法:将小鼠随机分为对照组、低剂量组(0.2592μg/μl)、中剂量组(1.56695μg/μl)和高剂量组(3.4560μg/μl),采用改良型气管滴注法,分别给予PBS或PM_(2.5)混悬液30μl/次,每3天1次,间断滴注7次。观察其染毒期间的精神、体重变化。染毒结束后于动情间期称量小鼠体重及卵巢重量,计算卵巢脏器系数;观察小鼠卵巢病理变化。免疫组化法检测卵巢ER、PR的分布,Western blot法检测卵巢ER、PR蛋白的表达变化。结果:与对照组比较,高剂量染毒组小鼠第14~18天体重减轻,差异有统计学意义(P0.05);中剂量组的卵巢湿重增加,差异有统计学意义(P0.05);高剂量组的卵巢湿重和卵巢脏器系数增加明显,差异均有统计学意义(P0.01,P0.05)。随着PM_(2.5)染毒剂量的增加,卵巢皮质变薄,结构混乱,间质纤维化,颗粒细胞排列较对照组紊乱,卵泡数减少,卵泡闭锁相对增多,黄体细胞明显增大变空,炎细胞浸润增多等。免疫组化染色结果示,高剂量组卵巢组织中ER、PR表达显著低于PBS对照组(P0.05)。Western blot法检测结果示,与对照组比较,随着染毒剂量的增加,卵巢中ER、PR蛋白表达逐渐降低,高剂量组中ER、PR蛋白表达均明显减少,差异有统计学意义(P0.05)。结论:随着PM_(2.5)暴露剂量增加,小鼠卵巢组织损害明显,提示暴露于高浓度的PM_(2.5)环境可损害卵巢组织结构,进而可能破坏卵巢功能。     

卵巢早衰的病因

女性在 40岁以前发生的自然绝经称卵巢早衰 (prematureovarianfailure,POF) ,POF可能为多因素引起的综合征。特发性POF占高促性腺激素闭经的 81%,是POF的最常见类型 ,这些患者通常无明确病因可遁。其他类型的POF可能与下列因素有关 :①遗传学因素 :X染色体数量和结构异常 ,与生育相关的基因突变 ;②自身免疫性因素 :机体产生卵巢自身免疫性抗体 ,如抗卵巢抗体 ,抗颗粒细胞抗体 ,抗透明带抗体 ,抗核抗体 ;③高促性腺激素血症和卵泡刺激素 (FSH)受体缺陷 ;④酶学障碍 :如半乳糖 1磷酸尿嘧啶转移酶 (galactose l phosphateuridyl trans…     

T淋巴细胞亚群与MIF在自身免疫性卵巢早衰患者中的应用价值研究

目的探讨自身免疫性卵巢早衰患者中关于巨噬细胞移动抑制因子(MIF)与T淋巴细胞亚群在自身免疫性卵巢早衰患者中的应用价值。方法选取2015年7月至2017年2月在山东省巨野县人民医院妇科接受治疗的自身免疫性卵巢早衰患者45例作为观察组,并选择同期体检卵巢功能正常者43例为对照组。测定并比较两组血清自身免疫反应性指标水平、MIF水平以及T淋巴细胞亚群水平。结果两组血清自身免疫反应性指标检测阳性率相比,抗卵巢抗体、抗甲状腺球蛋白抗体、抗透明带抗体及MIF差异有统计学意义(P0.05);两组MIF水平相比,差异有统计学意义(P0.05);观察组与对照组CD4+T细胞差异无统计学意义(P0.05),CD8+T细胞明显高于对照组(P0.05),CD4+/CD8+明显下降,两组相比,差异有统计学意义(P0.05);MIF阳性组与MIF阴性组CD4+T细胞差异无统计学意义(P0.05),CD8+T细胞明显高于MIF阴性组(P0.05),MIF阳性组CD4+/CD8+比值明显低于MIF阴性组(P0.05)。结论 MIF在卵巢早衰患者血中浓度及检出率较高,提示MIF在卵巢早衰的发生发展中起重要的作用。CD8+T细胞升高及CD4+/CD8+比值下降与卵巢早衰患者的自身免疫功能紊乱密切相关。     

卵巢早衰患者自身免疫反应性指标的检测及其意义

目的:检测卵巢早衰患者自身免疫反应状态,寻求有意义指标,为临床诊治提供指导。方法:49例卵巢早衰并不孕症患者为实验组,20例输卵管性不孕症患者为对照组,分别检查血清抗核抗体(antinuclear antibodies,ANA)、抗卵巢组织抗体(antiovarianantibodies,AOAb)、抗心磷脂抗体(Anticardiolipin,ACA)、抗甲状腺微粒体抗体(antithyroidmicrosomal antibodies,A-TG)、抗双链DNA抗体(ds-DNA)、免疫球蛋白(IgG,IgA,IgM,IgE)和补体(C3,C4)含量,并进行统计学分析。结果:实验组ANA、AOAb、ds-DNA、A-TG、C3阳性率分别为40.8%、42.9%、36.7%、46.9%和26.5%,均显著性高于对照组(P<0.05);试验组ACA、IgM异常率较高,分别30.6%、24.5%,但与对照组无显著差别(P>0.05);实验组IgG、IgA、IgE和C4异常率较低,分别为6.1%、2%、12%、4.1%,与对照组无显著差异(P>0.05)。结论:卵巢早衰患者有多种自身免疫性抗体,如ANA、AOAb、ds-DNA、A-TG、C3等,检测上述抗体对诊断自身免疫性卵巢早衰有临床意义。     

抗卵巢抗体对体外受精-胚胎移植过程及结局的影响

目的 :研究抗卵巢抗体对体外受精 胚胎移植 (IVF ET)的影响。方法 :接受IVF ET治疗的患者 80例 ,在施行IVF治疗前抽取静脉血 3ml ,测定血清中抗卵巢抗体(AOAb)阳性率 ,并分别计算阳性组和阴性组患者的取卵数、受精率、卵裂率、优质胚胎率、冷冻胚胎率、临床妊娠率及胚胎种植率。结果 :AOAb的阳性率为 2 8.8% (2 3/ 80 )。AOAb阳性患者中平均取卵数、胚胎种植率和临床妊娠率均低于抗体阴性组。结论 :抗卵巢抗体的存在影响了IVF ET的诸过程 ,降低了临床妊娠率。     

妇科养荣胶囊改善化疗所致小鼠卵巢功能损伤的效果

目的:观察妇科养荣胶囊(Fu Ke Yang Rong capsule,FKYRC)对卵巢损伤小鼠卵巢储备功能和生育力的保护作用。方法:通过成年雌性小鼠一次性腹腔注射120 mg/kg环磷酰胺和10 mg/kg白消安建立卵巢损伤小鼠模型,于造模前7 d至造模后60 d每日对卵巢损伤小鼠用高、中、低剂量(6 g/kg、4 g/kg和2 g/kg)FKYRC(H组、M组、L组)连续灌胃给药;同时,于造模前7 d一次性皮下注射给予Gn RHa(38 mg/kg)作为阳性对照组(Gn RHa组),用生理盐水(0.2 m L/d)代替FKYRC连续灌胃作为卵巢损伤对照组(模型组),另设正常对照组:非卵巢损伤正常小鼠腹腔注射给予等体积DMSO(NC组)。分别于造模后30 d和60 d取材,计数卵巢组织中各级卵泡数量,检测血清E2、FSH、抑制素B(INHB)和抗苗勒氏管激素(AMH)水平,检测卵巢组织中翼状螺旋/叉头转录因子2(FOXL2)和AMH蛋白的表达水平。并观察造模60 d后各组妊娠率和窝仔数。结果:各给药组和模型组小鼠血清E2低于NC组(P0.05),但FSH水平组间无统计学差异(P0.05)。造模各组卵泡数明显低于NC组(P0.05),且H组和Gn RHa组造模后60 d时卵巢组织中窦前卵泡与窦卵泡数较高,但各干预组与模型组比较无统计学差异(P0.05)。模型组小鼠卵巢组织中FOXL2和AMH蛋白表达水平显著低于各FKYRC干预组(P0.05),H组的妊娠率(80.00%)显著高于模型组(36.36%)(P0.05)。结论:连续FKYRC(6 g/kg)给药60 d后可明显改善烷化剂所致卵巢损伤小鼠的卵巢储备功能和生育能力,其机制可能与上调颗粒细胞中FOXL2和AMH的表达水平相关。     

顺铂诱导化疗损伤性卵巢早衰大鼠模型的探讨

目的:探讨顺铂诱导大鼠化疗损伤性卵巢功能早衰大鼠模型的可行性。方法:成熟雌性SD大鼠腹腔注射低、高剂量顺铂4.5 mg/kg(A组)、6.0 mg/kg(B组)和生理盐水(C组),每周1次,共2次,建立大鼠化疗损伤性卵巢早衰模型。检测血清FSH水平及光学显微镜下计数卵巢最大切面原始卵泡、初级卵泡、闭锁卵泡,阴道涂片观察动情周期变化。结果:A、B组大鼠动情周期均明显长于C组(P<0.05),并呈现剂量相关性改变。B组动情周期天数明显长于注射前(P<0.01);血清FSH水平A组与C组相比无统计学意义(P>0.05),B组明显高于A组和C组(P<0.05)。各顺铂组血清FSH水平注射后均较注射前明显升高(P<0.05)。腹腔注射顺铂后,A组、B组大鼠卵巢最大切面原始卵泡数和初级卵泡数均明显降低(P>0.05),而闭锁卵泡数均明显增加(P<0.05),并呈现剂量相关性改变。结论:顺铂可诱导化疗损伤性卵巢早衰。此化疗损伤性卵巢早衰大鼠模型血FSH明显升高,卵巢组织学衰退性改变与人类化疗损伤性卵巢早衰病变过程相似。     

壬基酚孕期经口染毒对F1代昆明小鼠卵巢发育的影响

目的:观察壬基酚孕期暴露对F1代昆明小鼠卵巢发育的影响。方法:交配后7d起,对孕鼠分别每日灌胃壬基酚(NP)(分别为1.2mg/kg、12mg/kg、120mg/kg),对照组灌胃花生油,直至交配后17d,每组7只。子代小鼠出生14d时取卵巢组织病理学检查,计数各级卵泡。观察子代雌鼠观察阴道开口时间和规律动情周期出现时间。子代雌鼠成年3月龄后,于动情前期处死,放免法测定血清E2、FSH浓度,酶联免疫吸附法测定血清抑制素B浓度,取卵巢组织行病理学检查,计数各级卵泡。结果:NP低剂量组阴道开口出现时间提前(P<0.05);低、中剂量组规律动情周期出现时间提前并可见动情周期延长(P<0.05);高剂量组性成熟期始基卵泡数减少(P<0.05),血清抑制素B水平降低(P<0.05);各用药组动情前期血清E2水平降低(P<0.05),FSH水平无明显变化。用药组体质量较对照组明显升高(P<0.05)。结论:壬基酚胎仔宫内暴露,在较低剂量时可影响小鼠的动情起始及动情周期,在较高剂量时可降低卵巢储备,并可影响成年后小鼠雌激素水平。     

卵巢上皮性癌6B11抗独特型微抗体疫苗免疫方案的探讨

目的探讨卵巢上皮性癌6811抗独特型微抗体（6811微抗体）的最佳免疫剂量和免疫间隔时间,为用于临床治疗卵巢上皮性癌打下基础。方法（1）采用基因工程方法构建6811微抗体的重组表达载体,并将其转化到大肠杆菌BL21中进行表达。直接酶联免疫吸附试验（ELISA）法测定其免疫学活性。（2）将BALB／c小鼠随机分为A、B、C、D、E、F组,于第1、14、28天时均分别给予6811微抗体150、100、50μg及小鼠IgG 100μg、人IgG 100μg、磷酸盐缓冲液100μl,其中A、B、C组为实验组,D、E、F组为对照组。间接ELISA法测定各组小鼠血清中抗卵巢上皮性癌6811抗独特型抗体（6811抗体）的抗体（即Ab3）的水平,并观察其变化规律。（3）将小鼠脾淋巴细胞作为效应细胞,人卵巢上皮性癌细胞株SKOV3细胞作为靶细胞,设定效靶比为250：1、125：1、60：1、30：1,^51Cr释放实验检测各组小鼠脾淋巴细胞产生的抗体依赖的细胞毒效应（ADCC）;设定血清稀释度为1：2、1：25、1：50,^51Cr释放实验检测各组小鼠脾淋巴细胞产生的补体依赖的细胞毒效应（CDC）。分别于紫外光波长490nm时测定其吸光度（A）值。结果（1）大肠杆菌B121中成功表达6811微抗体,并证实其保留了6811抗体的免疫学活性。（2）用上述不同剂量6811微抗体免疫BALB／c小鼠,在第14天注射后1周,血清中可检测到较低水平Ab3;第28天注射后1周,Ab3水平达到最高值,A、B、C组平均A值分别为1．16、1．11、1．06;此Ab3水平持续6周,6周内A、B、C组平均A值分别为1．05、1．06、0．94,B组略高于A、C两组,但3组间比较,差异无统计学意义（P〉0．05）;第7周Ab3水平明显下降。此时,给予加强免疫,1周后Ab3水平很快升高,并持续至少2周。（3）在4个不同的效靶比条件下,实验组小鼠脾淋巴细胞诱导产生的ADCC明显高于对照组（P〈0．05）;效靶比为1：125时,A、B、C组^51Cr自然释放率分别为23％、17％、12％,3组间比较,差异有统计学意义（P〈0．05）。血清稀释度1：50时,A、B、C组^51Cr自然释放率分别为47％、39％、26％,A、B组BALB／c小鼠血清诱导产生的CDC明显高于C组（P〈0．05）,而A和B组相比,差异则无统计学意义（P〉0．05）。结论6811微抗体可作为卵巢上皮性癌疫苗,其较佳的动物免疫方案为：免疫剂量为100μg;基础免疫分别于第1、14、28天进行,共3次;间隔6周给予加强免疫。     

ZP3 peptides administered orally suppress murine experimental autoimmune ovarian disease

Experimental autoimmune ovarian disease (AOD) is a T cell-mediated chronic inflammatory disease that may lead to premature ovarian failure. Autoimmune disease can be suppressed by oral administration of autoantigens leading to tolerance. One of the major mechanisms of oral tolerance is induction of regulatory CD4+ T cells that can mediate active suppression by producing immunomodulatory cytokines. However, the role of oral tolerance as a treatment for experimental AOD has received little attention. Therefore, the purpose of this study was to examine the conditions necessary to produce oral tolerance in experimental AOD in B6AF1 female mice. In this study, mice received different doses of peptides of the mouse zona pellucida 3 (pZP3) via gastric intubation for 7 times. After 4 times of oral administration, AOD was induced by immunization with pZP3. The optimal tolerating regimen for oral administration of pZP3 in mice was 10 microg, which decreased morbidity of oophoritis compared to the control group. In this moderate-dose therapeutic group (MD), alterations in the estrous cycle were normalized and CD4+ T cells that were CD25+ increased while those that were CD25- decreased. The severity of autoimmune oophoritis and the titer of ZP autoantibodies were also significantly reduced. These findings suggest that oral administration of pZP3 may be successfully used as an oral tolerance strategy for suppression of AOD.     

Analysis of B cell epitopes of a glycoprotein porcine zona pellucida (pZP1)

The zona pellucida (ZP) of mammalian oocytes forms an extracellular matrix composed of three major glycoproteins and plays an important role in sperm-zona interactions. As ZP had a strong organ-specific but species-cross-reactive antigenicity and passive or active immunization with ZP antigens could impair fertilization, the possibility of developing a immunocontraceptive vaccine has been extensively studied. Studies on active immunization with porcine ZP (pZP) that contain B cell epitopes and T cell epitopes demonstrated that a temporary infertility could be induced along with the elevation of antibody titers, but it was always associated with ovarian failure. This could be due to the oophoritis by activation of pathogenic T cell immunity. It is the general consideration that any adverse effects by vaccination should be avoided for an immunocontraception. From this point of view, the analysis of B cell epitopes of pZP protein would be helpful for construction of a safe immunocontraceptive vaccine with zona antigens. We determined the amino acid sequence of the B epitope in the pZP1 protein by using a monoclonal antibody (MAb-5H4) that possesses a fertilization blocking ability. In addition, antiserum raised to the epitope sequence was revealed to block in vitro fertilization of homologous animal species.     

Circulating auto-antibodies against the zona pellucida and thyroid microsomal antigen in women with premature ovarian failure

Premature ovarian failure (POF) is a disorder of multicausal etiology leading to infertility in women. Development of ovarian auto-antibodies is a causative factor in most POF cases, but no consensus on the ovarian antigenic determinants has been reached till date. In the present study, sera from 15 POF cases, seven normally cycling women and eight menopausal women were studied by immunohistochemistry (IHC) for the presence of anti-ovarian antibodies. 10 of the 15 POF sera (66.6%) presented with anti-ovarian antibodies (Ao). Of these, two demonstrated antibodies to the zona pellucida (ZP) as well as strong immunoreactivity to granulosa cells (Azg), while the remaining eight exhibited anti-ZP antibodies with negligible staining in granulosa cells (Az). The antibodies showed cross-reactivity with ZP from various species such as human, sheep, marmoset, pig and mouse. Among various murine tissues, the antibodies cross-reacted only with thyroid and not with uterus, spleen, kidney, liver, adrenal, pancreas and pituitary. Five of the eight Az individuals presented with significant titres of anti-thyroid antibodies (Azt). In the control group, one menopausal control presented with reactivity to both ZP and GC, the autoimmunity possibly being a consequence of surgical trauma; while one normally cycling woman tested positive for anti-thyroid antibodies. The IHC results were confirmed by ELISA using heat-solubilized isolated ZP (SIZP) as the antigen. Out of seven Ao samples assessed by ELISA, five reacted with SIZP. Preincubation of these five samples with varying concentrations of SIZP demonstrated a dose-dependent decrease in reactivity in ELISA and abolished staining in IHC, confirming the specificity of auto-antibodies to ZP in the POF group. Our results thus suggest that ZP is an important ovarian antigen in autoimmune POF.     

人卵透明带蛋白huZP3a~(22～176)和huZP3b~(177～348)肽段在大肠杆菌中的表达及其纯化

目的:探索将huZP3a22~348蛋白编码基因拆分成两段,分别表达的可行性,并研究表达产物的免疫原性。方法:用PCR技术, 将去N-端信号肽和C-端跨膜区的huZP322~348蛋白编码基因拆成大小相近的两段, 以完整阅读框的形式分别重组插入热诱导型pBV221的多克隆区。结果:大肠杆菌分别特异地表达了可单独或复合应用的huZP3a22~176和huZP3b177~348,在经过抗人ZP3不同抗原区合成肽抗体的蛋白印迹鉴定后, 用改良的制备性PAGE方法分离纯化这两种表达产物。同时用兔抗猪ZP IgGs蛋白印迹试验表明, huZP3a和pZP3b的几个共同线性抗原表位都存在于其肽链的前半区域。结论:通过基因重组技术可获得足够量的huZP3a和huZP3b,这为开展huZP3a和huZP3b的免疫原性以及huZP3诱发人精子顶体胞吐的功能域等研究奠定了基础。     

Antifertility effect of active immunization with ZP4 glycoprotein family of porcine zona pellucida in hamsters.

Female golden hamsters were immunized with solubilized porcine zona pellucida (s-PZP) or ZP4 glycoprotein family isolated from s-PZP by preparative SDS-PAGE. Both antigen preparations induced production of antibodies which reacted not only with porcine zona pellucida but also with the hamster zona pellucida. The hamsters immunized with solubilized porcine zona pellucida mainly produced antibodies reactive to ZP3, while the hamsters immunized with ZP4 mainly produced antibodies reactive to ZP4. The former animals became permanently infertile but the infertility in the latter animals was temporary and they became pregnant later. Histological studies revealed that the ovarian follicles in hamsters immunized with s-PZP were completely destroyed leaving only atrophic follicle-like cell clusters, while in the ovaries of hamsters immunized with ZP4 a number of small follicles with oocytes remained intact. These observations are encouraging for the further characterization of the ZP4 antigens as candidates for the development of a contraceptive vaccine.     

Native zona pellucida reactivity and in-vitro effect on human sperm-egg binding with antisera against bonnet monkey ZP1 and ZP3 synthetic peptides

In mammals, zona pellucida glycoprotein-3 (ZP3) is the putative ligand for primary sperm binding and induces the acrosome reaction. Recent evidence suggests that zona pellucida glycoprotein-1 (ZP1) also play an important role, in some species, during fertilization. In order to identify synthetic peptide immunogens capable of inducing antibodies reactive with native zona and inhibiting sperm-oocyte interaction, peptide encompassing the amino acid (aa) residues 334-343 of bonnet monkey ZP3 (bmZP3) was synthesized co-linearly with a 'promiscuous' T-cell epitope of circumsporozoite protein (CSP, 378-398 aa) of Plasmodium falciparum. In addition, four peptides corresponding to bonnet monkey ZP1 (bmZP1((58-79 aa)), bmZP1((136-153 aa)), bmZP1((212-228 aa)) and bmZP1((251-273 aa))) were synthesized. The synthetic peptides corresponding to bmZP1 were conjugated with diphtheria toxoid. Immunization of female BALB/cJ mice with the above conjugates and CSP-bmZP3((334-343 aa)) peptide led to the generation of an adequate antibody response against the respective zona peptide. Antibodies against bmZP1((251-273 aa)) and CSP-bmZP3((334-343 aa)) recognized bonnet monkey and human zona pellucida in an indirect immunofluorescence assay. Further, these antibodies when tested independently or in combination also significantly inhibited the binding of human spermatozoa to zona pellucida in a hemizona assay. These studies will further help in the design of synthetic peptide immunogens comprising of multiple B cell epitope from different zona proteins for better immunocontraceptive efficacy.     

不同病因对卵母细胞透明带、胚胎发育和妊娠结局的影响

目的:探讨女性不同病因对卵母细胞透明带、胚胎发育和妊娠结局等的影响。方法:回顾性分析因单因素不孕体外受精治疗的197个周期,以输卵管因素为对照组(n=145),比较多囊卵巢综合征(polycystic ovarian syndrome,PCOS)(n=25)、子宫内膜异位症(endometriosis,EMs)(n=19)和不明原因不孕症(n=8)等各组的受精率、种植率、妊娠率以及透明带双折射性和透明带厚度。结果:平均获卵数PCOS组(17.9±1.9枚)显著高于其它各组(对照组:11.3±0.6枚,EMs组:9.4±1.2枚和不明原因组:13.1±2.9)(P<0.01),其它各组无统计学差异(P>0.05);PCOS组、EMs组和不明原因组受精率(83.66%、75.40%、77.12%)、优质胚胎率(57.26%、51.14%、59.30%)、种植率(25.53%、17.39%、29.40%)和妊娠率(52.38%、41.18%、42.86%)与对照组(80.13%、58.99%、28.76%、43.70%)间无统计学差异(P>0.05),但EMs组各项指标均低于对照组。透明带双折射值各组间差异极显著(对照组:7.13±0.28,PCOS组:4.97±0.55,EMs组:3.67±0.75,不明原因组:14.19±1.41)(P<0.01),其中EMs组透明带双折射值最低。受精当日透明带厚度EMs组显著高于其它各组(对照组:18.84±0.07,PCOS组:18.48±0.17,EMs组:19.75±0.24,不明原因组:18.98±0.26)。结论:EMs对受精率、妊娠率和透明带双折射值等的影响最明显,透明带的双折射值和厚度可在一定程度上反映不同病因对卵母细胞发育潜力的影响。     

The zona pellucida: using molecular genetics to study the mammalian egg coat

An extracellular matrix that mediates critical steps in fertilization and early development surrounds all vertebrate eggs. In mice and humans, this matrix is known as the zona pellucida and comprises three glycoproteins: ZP1, ZP2 and ZP3. Homologues of these proteins isolated from other vertebrates have conserved protein motifs that may be important for establishing a common fibrillar structure. However, specific but contradictory biological roles have been assigned to individual egg coat proteins based on assays in vitro in a wide range of species. Mouse lines lacking either ZP1 or ZP3 have been established with abnormal or absent zona matrices and varying degrees of infertility to examine zona structure and function in vivo. By crossing mouse lines lacking individual zona proteins with those expressing human homologues, the structural integrity of the zona matrix can be restored. Because mouse and human spermatozoa exhibit order-specific binding to the zona pellucida, mice with 'humanized' chimaeric zonae may provide an experimental system to elucidate the molecular basis of sperm-zona interaction.     

17D_3pZP单克隆抗体对卵透明带及卵巢等组织的免疫学反应

目的: 观察单克隆抗体17D3pZP对猪卵透明带,人卵透明带及人卵巢等组织的免疫学反应。方法: ABC酶免疫组织化学技术。结果: 17D3pZP单克隆抗体能特异性识别猪卵透明带及人卵透明带抗原,而与人卵巢及其他各重要脏器组织无反应。结论: 17D3pZP单克隆抗体能识别卵透明带抗原,而与机体其他组织抗原无交叉反应。有望通过它进一步提取并纯化相应靶抗原,为制备卵透明带免疫避孕疫苗奠定基础。     

Immunocontraceptive potential of recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-C expressed in Escherichia coli and its corresponding synthetic peptide

Zona pellucida (ZP) glycoproteins have been proposed as candidate antigens for development of immunocontraceptive vaccines. In this study, the efficacy to block fertility by immunization with recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-C (r-bmZPC) expressed in Escherichia coli and its synthetic peptide (P(4): KGDCGTPSHSRRQPHVVSQWSRSA, aa residues 324-347) conjugated to diphtheria toxoid (DT) has been evaluated in a homologous system. Female bonnet monkeys, immunized with P(4)-DT conjugate showed better immunocontraceptive potential as compared to an r-bmZPC-DT immunized group. In spite of high anti-P(4) antibody titres, animals continued to have ovulatory cycles and showed no disturbance in cyclicity (except summer amenorrhoea). No ovarian pathology was observed in the P(4) immunized group. These results suggest that immunization with the P(4) may lead to block in fertility without obvious ovarian dysfunction. However, further inputs are required to identify additional ZP based B-cell epitopes to enhance the contraceptive efficacy.