过表达脑红蛋白通过激活PI3K/Akt信号通路防治AD的体外研究

 目的:探索过表达脑红蛋白（neuroglobin,NGB）对转染了pAPPswe的SH-SY5Y细胞的神经保护作用及机制。方法: 成功构建过表达NGB的质粒pEGFP-NGB并转染入已预先转染了pAPPswe的SH-SY5Y细胞,MTT法检测过表达NGB对该细胞存活率的影响;JC-1法检测其对细胞线粒体膜电位的影响;流式细胞术检测过表达NGB对细胞凋亡的影响;Western blotting法检测其对细胞中p-Akt、 Akt和caspase-3/9表达的影响;ELISA法检测其对细胞内Aβ42生成的影响。结果: MTT结果显示,与对照组和空质粒组比较,转染NGB后,pAPPswe-SH-SY5Y细胞的存活率明显提高,差异有统计学意义（P<0.05）。JC-1染色结果显示过表达NGB能够明显抑制转染pAPPswe对SH-SY5Y细胞线粒体膜电位的降低作用（P<0.05）。流式细胞术结果显示过表达NGB能够抑制早、晚期细胞的凋亡。而Western blotting显示过表达NGB不仅能抑制细胞内caspase-3和caspase-9蛋白水平的表达,而且还能够促进细胞内p-Akt蛋白的表达,而这种促进作用能够被PI3K/Akt的抑制剂LY294002所抑制。ELISA结果显示过表达NGB能够明显抑制细胞内Aβ42的生成。结论: 过表达NGB能够显著抑制pAPPswe诱导的细胞损伤,而且还抑制与细胞凋亡密切相关的caspase-3和caspase-9等蛋白表达。NGB的神经保护作用可能是通过激活PI3K/Akt信号通路来实现的。     

PI3K/Akt信号通路及其抑制剂的研究进展

磷脂酰肌醇3激酶（PI3K）/丝氨酸-苏氨酸激酶（Akt）信号通路在信号转导的调控中扮演着重要角色,能调节细胞增殖、凋亡、代谢、运动、血管生成等生物过程。与其他信号通路相比,PI3K/Akt信号通路的组成部分更庞大,在肿瘤中更多见。目前已证实多种肿瘤中存在PI3K/Akt信号通路的超活化,对肿瘤细胞的存活、生长、运动、血管生成和代谢意义重大。因此,抑制PI3K和与通路相关的成分可能会使肿瘤生长受抑,使患者预后改善。PI3K/Akt信号通路抑制剂包括针对单一成分的抑制剂和双重抑制剂。目前大量的PI3K抑制剂已在临床前期研究中取得良好结果,有些已经在血液恶性肿瘤和实体肿瘤中进行了临床试验。在此综述中,我们简单的总结了PI3K-AKt通路的研究成果,讨论了PI3K抑制剂从临床前研究到临床研究的发展前景。     

白藜芦醇通过PI3K/Akt通路及线粒体途径抑制软骨肉瘤

 目的：探讨白藜芦醇抑制软骨肉瘤的机制及对线粒体途径和PI3K/Akt通路的影响。方法：SW1353软骨肉瘤细胞培养至对数生长期后设对照组和白藜芦醇处理组,药物处理组用25、50和100 μmol/L白藜芦醇处理24 h、48 h或72 h。采用CCK8法检测SW1353软骨肉瘤细胞的活力,Hoechst 33258荧光染色观察细胞凋亡,Western blotting检测activated caspase-3、Bcl-2、Bax、Akt和p-Akt蛋白在细胞中表达情况,细胞划痕实验观察细胞迁移情况。结果：白藜芦醇处理后细胞活力下降,呈时间-剂量依赖性（P<0.01）。Hoechst  33258染色检测可见药物处理组有明显的细胞凋亡核象。Western blotting检测显示药物处理组activated caspase-3和Bax蛋白表达上调,Bcl-2蛋白和p-Akt蛋白表达下调,总Akt改变不显著。细胞划痕试验显示,白藜芦醇能显著抑制SW1353细胞的迁移。结论：白藜芦醇能够诱导软骨肉瘤凋亡,部分是通过线粒体途径及PI3K/Akt信号通路发挥作用。     

ROCK通过抑制PI3K/Akt通路促进高糖诱导的心肌细胞凋亡

目的:探讨Rho相关卷曲螺旋激酶(ROCK)是否通过调控PI3K/Akt信号通路参与高糖诱导的原代心肌细胞凋亡。方法:培养原代Wistar乳鼠心肌细胞,用α-横纹肌肌动蛋白(α-SCA)免疫组化法进行鉴定;建立心肌细胞高糖模型,采用5.5、33和40 mmol/L葡萄糖作用于细胞48 h。采用MTT法检测细胞活力;RT-qPCR检测各组心肌细胞中ROCK1和ROCK2的表达水平;流式细胞术检测各组心肌细胞的凋亡水平;Western blot检测ROCK1、ROCK2、cleaved caspase-3、Bcl-2、PI3K、Akt和p-Akt的蛋白水平;为了证实ROCKs对PI3K/Akt信号通路的调控作用,将细胞分为对照组(5.5 mmol/L葡萄糖处理细胞)、高糖组(33 mmol/L葡萄糖处理细胞)和高糖加Y27632(ROCK抑制剂)组,Western blot检测ROCK1、ROCK2、PI3K、Akt和p-Akt的蛋白水平。结果:高糖培养48 h后,33和40 mmol/L葡萄糖组细胞相对活力分别为(79.71±2.43)%和(68.41±7.49)%,与对照组相比显著降低...     

米诺环素通过调控PI3K/Akt通路抑制硝普钠诱导的PC12细胞凋亡

 目的： 探讨PI3K/Akt信号通路在米诺环素（minocycline,MC）抑制硝普钠(sodium nitoprusside,SNP)诱导的PC12细胞凋亡中的作用。方法：将体外培养的PC12细胞分为4组：空白对照组、SNP组、MC+SNP组和PI3K抑制剂LY294002+ MC+SNP组。用四甲基偶氮唑盐（MTT）法检测细胞活力,流式细胞术检测细胞凋亡;Western blotting检测不同时点（0.5、1、2、3 h）各处理组PI3K/Akt通路蛋白p-Akt和Akt的表达。结果：SNP处理PC12细胞24 h能抑制细胞生长,加入10 μmol/L MC预处理30 min可明显提高细胞活力,降低细胞凋亡率（P<0.05）,抑制SNP诱导的PC12细胞凋亡。MC组的p-Akt表达高于其它组,而加入LY294002后可阻断MC的上述效应。结论：MC可通过调控PI3K/Akt通路抑制SNP诱导的PC12细胞凋亡。     

PI3K/Akt途径在Aβ诱导细胞凋亡过程中的作用

目的：探讨凝聚态β-淀粉样蛋白25-35片段（Aβ25-35）诱导细胞凋亡过程中PI3K/Akt转导通路的作用。方法: MTT法检测Aβ25-35作用不同时间后PC12细胞活力的变化;Annexin V-PI双染检测Aβ25-35作用不同时间后PC12细胞的凋亡情况;Western blotting检测Aβ25-35作用不同时间后p-AktSer473、p-GSK-3βSer9的水平变化。结果: 20μmol/L Aβ25-35作用不同时间（30 min、1 h、3 h、6 h、12 h、24 h、48 h）后,MTT结果显示细胞活力均明显下降（P<0.05）;Annexin V-PI结果显示,Aβ25-35作用后均可引起细胞凋亡（P<0.05）;Western blotting结果表明,Aβ25-35作用于细胞后,p-AktSer473和p-GSK-3βSer9的表达均出现迅速的下调（P<0.05）,在作用3 h时两者的表达出现迅速的上升,在作用6 h时又出现表达的下调,两者在该时间段的变化趋势相符。p-AktSer473在Aβ25-35作用于细胞12 h后出现逐渐回升趋势,但在48 h时仍没有达到正常水平;而p-GSK-3βSer9的表达在Aβ25-35作用于细胞12 h后出现迅速的升高（P<0.05）,而后逐渐恢复到正常水平。结论: PI3K/Akt信号通路可能参与了Aβ诱导的细胞损伤,本研究为AD的发病机制及选择合适的药物作用时点提供了新思路。     

紫草素通过PI3K/Akt通路抑制氧糖剥夺诱导的大鼠原代皮层神经元凋亡

目的:探讨紫草素对氧糖剥夺(OGD)损伤模型中大鼠原代皮层神经元的作用及机制。方法:用不同浓度(0. 02、0. 2、2和20μmol/L)紫草素对大鼠原代皮层神经元经进行预处理,再经OGD损伤处理,用乳酸脱氢酶(LDH)释放法和荧光素二乙酸酯/碘化丙啶(FDA/PI)双染法分别检测神经元活性和凋亡情况,选择最适紫草素浓度。然后,在加入紫草素之前提前加入LY294002(PI3K/Akt信号通路抑制剂,1μmol/L),用Wesern blot法检测神经元p-Akt(Ser473)水平变化,用LDH法和FDA/PI双染法检测神经元活性和凋亡率变化。结果:0. 2、2及20μmol/L的紫草素可显著提高神经元存活率(P 0. 05),同时还可使神经元内p-Akt(Ser473)水平显著升高(P 0. 05); LY294002可显著阻断紫草素对神经元p-Akt(Ser473)水平和凋亡率的影响(P 0. 05)。结论:紫草素可通过激活PI3K/Akt通路来减少OGD诱导的大鼠原代皮层神经元凋亡。     

槲皮苷通过抑制PI3K/AKT信号通路诱导胃癌SGC7901细胞凋亡

目的:探讨槲皮苷是否通过抑制PI3K/AKT信号通路诱导人胃癌SGC7901细胞凋亡。方法:选取SGC7901细胞作为研究对象,采用MTT法检测槲皮苷对SGC7901细胞的毒性作用并测定IC50值。实验分为对照组(不加药处理)、槲皮苷组(采用200μmol/L槲皮苷处理)、PI3K/AKT通路激动剂胰岛素样生长因子1(IGF-1)组(采用100μg/L IGF-1处理)和槲皮苷+IGF-1组(采用200μmol/L槲皮苷+100μg/L IGF-1共处理)。处理48 h后,采用流式细胞术检测细胞凋亡,Western blot法检测cleaved caspase-3、p-AKT(Ser473)、AKT、p-PI3K(Tyr508)和PI3K的蛋白水平。结果:从100μmol/L开始,随着槲皮苷处理浓度的逐渐升高,SGC7901细胞活力显著降低(P 0. 05),槲皮苷作用48 h的IC50值为275. 40μmol/L。200μmol/L槲皮苷作用SGC7901细胞48 h后,与对照组比较,细胞凋亡率和cleaved caspase-3蛋白水平显著上升(P 0. 05),p-AKT和p-PI3K蛋白水平显著降低(P 0. 05),然而IGF-1与槲皮苷共同作用时,IGF-1可逆转槲皮苷对SGC7901细胞的作用效果。结论:槲皮苷能够诱导胃癌SGC7901细胞凋亡,其作用机制可能与抑制PI3K/AKT信号通路的激活有关。     

PI3K/Akt/mTOR的信号传导通路在恶性肿瘤中的研究进展

真核生物对诸如生长因子受体、激素、细胞因子等刺激产生应答,激活细胞内不同信号传导通路,其中PI3K／Akt／mTOR是与细胞增殖和细胞凋亡关系最密切的信号传导通路之一。随着研究的不断深入,该通路在多种常见肿瘤如乳腺癌、结直肠癌、前列腺癌、卵巢癌、肝癌、肺癌及淋巴瘤等的发生发展中凸现出日益重要的作用,并为肿瘤的治疗提供了新的靶点。     

Adipophilin通过PI3K/Akt通路促进细胞内脂质蓄积

目的:探讨adipophilin促进细胞内脂质蓄积的可能机制,为动脉粥样硬化的防治提供参考。方法:分别通过q PCR、Western blot和油红O染色观察氧化型低密度脂蛋白(ox LDL)处理RAW264.7细胞不同时间后,细胞内Akt、p-Akt和adipophilin的蛋白水平及脂质蓄积情况;并检测PI3K/Akt抑制剂LY294002处理后,上述指标的变化;HEK293细胞过表达adipophilin后,检测Akt的活性;并用免疫共沉淀实验检测adipophilin与Akt之间的相互作用。结果:ox LDL处理细胞后,随着时间的延长,脂滴增多,Akt被活化,adipophilin表达增加,而LY294002处理则可抑制上述变化;过表达adipophilin后,p-Akt水平增高,但adipophilin与Akt之间未见直接相互作用。结论:Adipophilin可通过PI3K/Akt途径促进细胞内脂质蓄积,但可能不是通过直接相互关系发挥作用的。     

沙眼衣原体质粒蛋白pORF5生物学特性初步研究

目的 分析沙眼衣原体(Chlamydia trachomatis,Ct)隐蔽性质粒编码的质粒蛋白pORF5在感染细胞中的定位并初步探讨其生物学特征.方法 PCR扩增pORF5质粒蛋白编码基因,构建pORF5原核表达重组体并诱导表达融合蛋白;融合蛋白经Glutathione Sepharose亲和层析纯化后免疫小鼠,制备单克隆抗体和多克隆抗体,间接免疫荧光技术及免疫印迹鉴定抗体的特异性;分析pORF5质粒蛋白在感染细胞中的分布及表达模式特征.采用ELISA分析pORF5质粒蛋白在自然感染状态下的表达情况及免疫原性.结果 pORF5主要分布于宿主细胞质,但也少量分布在原体(EB)和网状体(RB)上,在细胞质中的分布模式与衣原体分泌蛋白酶样活性因子(CPAF)基本相似,激光共聚焦结果显示两者并不重叠;pORF5能与Ct生殖道感染患者血清发生强烈的免疫应答.结论 pORF5为衣原体分泌蛋白,并具有很强的免疫原性;在自然感染状态下,pORF5质粒蛋白基因被激活产生内源性靶蛋白.

Abstract：

Objective To localize and characterize the plasmid protein pORF5 in the Chlamydia trachomatis(Ct) infected cells. Methods The open reading frame encoding for pORF5 protein from the Ct plasmid was amplified and cloned into the pGEX-6p vector. The recombinant plasmid pGEX-pORF5 was transformed into XL1-blue E. coli to express fusion protein with the glutathione-s-transferase (GST). After purified with Glutathione Sepharose 4B beads, the pORF5 fusion protein was used to immunize mice to make monoclonal and polyclonal antibody. The antibodies were used to localize the endogenous pORF5 protein and detect the expression pattern in Chlamydia-infected cells using an indirect immunofluorescence assay (IFA). At the same time, ELISA was used to determine whether pORF5 plasmid protein was expressed and immunogenic during Ct infection in humans. Results pORF5 was detected a dominant signal in the cytosol of the Chlamydia-infected cells with a pattern similar to that of anti-CPAF. pORF5 also appeared in the RBs and EBs in small quantity. Athough pattern was similarly, pORF5 did not overlap with CPAF. pORF5 protein was strongly recognized antiserum in an ELISA. Conclusion The pORF5 plasmid protein was identified as a secreted protein with good immunogenicity, pORF5 gene was to express the endogenous target protein during human infection.     

17β-雌二醇通过PI3K-Akt信号通路抑制丙泊酚诱导皮层神经元凋亡

 目的: 探讨17β-雌二醇对丙泊酚诱导原代培养皮层神经元凋亡的影响及机制。方法: 原代培养7 d的大鼠皮层神经元,给予不同浓度丙泊酚和或17β-雌二醇处理12 h,用MTT法检测神经元存活率变化,Hoechst 33258核染色法检测神经元调亡,Western blot法测定神经元p-Akt蛋白的变化。结果: 与溶剂对照组比较,丙泊酚呈剂量依赖性降低神经元存活率(P<0.05);与丙泊酚组比较,17β-雌二醇呈剂量依赖性提高神经元存活率(P<0.05),PI3K/Akt激动剂IGF组神经元存活率显著增加(P<0.01)。与溶剂对照组比较,丙泊酚组神经元凋亡率显著增加(P<0.01),丙泊酚+17β-雌二醇组神经元凋亡率较丙泊酚组显著下降(P<0.01),LY294002预处理可阻断17β-雌二醇的作用,使细胞凋亡率增加(P<0.01)。与溶剂对照组比较,丙泊酚呈剂量依赖性降低神经元p-Akt蛋白水平(P<0.05);与丙泊酚组比较,17β-雌二醇呈剂量依赖性提高神经元p-Akt蛋白水平(P<0.05);与丙泊酚+17β-雌二醇组比较,LY294002预处理组p-Akt蛋白水平显著下降(P<0.01)。结论: 17β-雌二醇可抑制丙泊酚诱导的原代培养皮层神经元凋亡,其机制可能与激活PI3K-Akt信号通路有关。     

沙眼衣原体pORF5 质粒蛋白免疫优势片段的筛选与鉴定

目的:鉴定沙眼衣原体pORF5质粒蛋白的免疫原性,并进一步筛选和确定pORF5质粒蛋白免疫优势片段。方法:以沙眼衣原体D血清型DNA为模板,设计pORF5基因全长和9个不同片段特异引物进行PCR扩增,PCR产物经Bam HⅠ、NotⅠ双酶切后插入经同样双酶切的原核表达载体pGEX-6p中,构建pORF5质粒蛋白不同长度片段的原核表达重组体,重组体经PCR和测序鉴定后,转化XL1 Blue大肠杆菌表达10种不同长度的GST融合蛋白;ELISA方法检测pORF5质粒蛋白的免疫原性,Western blot鉴定pORF5质粒蛋白的免疫反应性;ELISA测定10个不同片段与沙眼衣原体生殖道感染患者血清、鼠免疫血清以及抗pORF5单克隆抗体的免疫反应性,鉴定pORF5质粒蛋白免疫优势片段。结果:pORF5质粒蛋白免疫原性强,能刺激机体产生高效价抗体;破坏pORF5质粒蛋白天然空间结构,其免疫反应性基本消失;在ELISA反应中,N端缺失66氨基酸的F6片段的免疫反应强度与pORF5全长基本相似,F2与F3出现较弱的免疫反应,其余片段免疫反应消失。结论:pORF5质粒蛋白为构象依赖性免疫优势抗原,其免疫优势表位和构象表位位于C端,本研究为进一步探讨pORF5质粒蛋白的生物学功能和疫苗的研制提供实验依据。     

沙眼衣原体pORF5质粒蛋白单克隆抗体2H4的纯化及其免疫学特性研究

目的 纯化抗沙眼衣原体pORF5单克隆抗体2H4并鉴定其免疫学特性.方法 大量培养pORF5单克隆抗体阳性杂交瘤细胞株并收集培养上清,采用G蛋白免疫亲和层析法纯化2H4单克隆抗体;酶联免疫吸附试验(ELISA)测定2H4效价及抗体亚类;Western blot鉴定其特异性;免疫荧光试验(IFA)检测2H4单克隆抗体的衣原体种属特异性.结果 纯化后2H4抗体的纯度高达93％;效价为1:1024,免疫球蛋白类型为IgG2a;2H4抗体不仅能特异性识别pORF5融合蛋白,而且能特异性识别Ct血清型A、D、L2、鼠农原体(MoPn)、鹦鹉热嗜农原体(6BC)质粒所编码的内源性pORF5蛋白,但不识别衣原体其他质粒蛋白和肺炎嗜衣原体(Cpn).结论 获得了高纯度的能特异识别pORF5质粒蛋白的单克隆抗体,为进一步研究pORF5蛋白结构和功能以及沙眼衣原体诊断试剂盒的研制奠定了良好的基础.     

Nicotine stimulated bone marrow-derived dendritic cells could augment HBV specific CTL priming by activating PI3K-Akt pathway

Our previous studies have revealed that nicotine-treated immature dendritic cells (imDCs) have anti-tumor effects in murine lymphoma models. The present study is to explore HBV-specific CTL priming and its cytolytic activities of nicotine-treated murine DCs, the mechanism of α7 nicotinic acetylcholine receptor (nAChR) up-regulation by nicotine and the efficiency of nicotine with other cytokines. To address these hypotheses, bone marrow-derived imDCs were stimulated by nicotine and expression of α7 nAChR was firstly determined by flow cytometry and Western blot. Then, DCs-dependent HBV-specific T cell proliferation and IL-12 secretion were secondly determined by BrdU cell proliferation assay and ELISA, respectively. The HBV-specific CTL priming and its activities were further explored by intraperitoneal transfer of nicotine treated imDCs. The mechanism of nicotine up-regulating α7 nAChR was finally explored by Western blot. The results showed that: first, the maximal activation of PI3K and Akt was reached at 30 and 60-120 min respectively after nicotine stimulation. Nicotine up-regulated the expression of α7 nAChR by activating PI3K-Akt pathway in murine DCs; secondly, nicotine stimulation could enhance DCs' ability of HBV-specific T cell proliferation and IL-12 secretion; thirdly, adoptive transfer of nicotine stimulated DCs could induce HBV specific CTL priming in vivo and those CTL had cytolytic activities; fourthly, nicotine had equal efficiencies to 2 ng/ml IFN-γ in DCs-mediated T cell proliferation. All these data presented here indicated that nicotine treated imDCs might be considered as a potential candidate for HBV immunotherapy.     

IL-12 inhibits glucocorticoid-induced T cell apoptosis by inducing GMEB1 and activating PI3K/Akt pathway

Interleukin (IL)-12 is an important pro-inflammatory cytokine that has been shown to play a role in T cell survival, at least in part by activating the PI3K/Akt pathway. Glucocorticoid modulatory element binding protein (GMEB)1 and 2 are closely related proteins that modify the glucocorticoid receptor binding locus and thus modulate glucocorticoid-mediated gene induction effects, including apoptosis. GMEB1 associates with caspases and prevents apoptosis of cells in the nervous system. We have observed, in preliminary studies, that IL-12 up-regulates GMEB mRNA in human T cells, and postulated that this may contribute to the anti-apoptotic effect of IL-12 on T cells, in particular with regard to glucocorticoid induced apoptosis. Here, we confirm that IL-12 rescue of dexamethasone induced T cell apoptosis involves the PI3K/Akt pathway and that IL-12 induces GMEB1 and GMEB2. A siRNA knockdown of GMEB1 reverses the protective effect of IL-12 on dexamethasone induced T cell apoptosis. Thus, IL-12 protects T cells from glucocorticoid induced apoptosis via PI3K/Akt pathway and via induction of GMEB1, which is likely to reduce transactivation of the glucocorticoid receptor and induction of apoptotic genes. As glucocorticoid induced apoptosis occurs both in physiological and pathological/therapeutic situations, and IL-12 is actively involved in a variety of inflammatory and immune responses, the ability of IL-12 to inhibit steroid responses and increase T cell survival through GMEB1 has wide ranging implications. Manipulating GMEB may be used therapeutically to enhance the resistance or the sensitivity to steroids.     

The PI3K-Akt pathway promotes microtubule stabilization in migrating fibroblasts

Directed cell migration is controlled by extracellular cues such as growth factors/chemokines and extracellular matrix. In a migrating cell, a subset of microtubules becomes stabilized, and this stabilization is implicated in the establishment and maintenance of cell polarity. It is still not fully understood, however, how extracellular cues regulate the dynamics of microtubules. Here we show that the PI3K-Akt signaling pathway plays a pivotal role in growth factor regulation of microtubule stability. Treatment of NIH 3T3 fibroblasts with platelet-derived growth factor (PDGF) increases the amount of stabilized microtubules, and this increase is abrogated by the addition of a PI3K inhibitor or by expression of a dominant-negative form of Akt (DN-Akt), but not by the addition of a MEK inhibitor. Expression of an active form of Akt slightly increases the bulk amount of stabilized microtubules. Stabilization of microtubules induced in edge cells in the wounded monolayer culture is also attenuated by the PI3K inhibitor treatment or by expression of DN-Akt. Given that Akt is activated at the leading edge of a migrating cell and plays an essential role in directed cell migration, these results reveal a novel mechanism linking extracellular cues to directed cell migration, namely Akt regulation of microtubule stability.     

质粒蛋白pORF5在沙眼衣原体感染细胞中的定位及免疫原性研究(英文)

目的分析沙眼衣原体隐蔽性质粒编码的质粒蛋白pORF5在感染细胞中的定位并初步探讨免疫原性特征。方法 PCR扩增pORF5质粒蛋白编码基因,构建原核表达重组体pGEX-6p/pORF5,重组体转化大肠杆菌XL1-blue中诱导表达融合蛋白;融合蛋白经Glutathione Sepharose亲和层析纯化后免疫小鼠,制备单克隆抗体和多克隆抗体,间接免疫荧光技术及免疫印迹鉴定抗体的特异性,并分析pORF5质粒蛋白在感染细胞中的分布特征。采用ELISA方法分析pORF5质粒蛋白的免疫原性。结果 pORF5原核表达重组体成功构建,融合蛋白在大肠杆菌中可溶性表达;pORF5主要分布于宿主细胞胞浆,但也少量分布在EB、RB上;pORF5能与衣原体患者血清及鼠免疫血清发生强烈地免疫反应。结论 pORF5为衣原体分泌蛋白,并具有很强的免疫原性。