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1.
目的研究表达HPV—16结构蛋白L1和12的重组痘苗病毒(rVVL1L2)的免疫效果。方法以重组痘苗病毒rVVL1L2免疫C57BIJ6小鼠,用酶联免疫(ELISA)和酶联免疫斑点(ELISPOT)方法检测重组痘苗病毒诱发小鼠产生的体液免疫和细胞免疫应答水平;利用C3肿瘤细胞在C57BIJ6小鼠中的成瘤模型,观察重组痘苗病毒在抗肿瘤移植实验和肿瘤生长抑制实验中对小鼠的免疫保护效果。结果重组痘苗病毒rVVL1L2免疫的小鼠,可以检测到针对L1和12特异的抗体、L1165-175肽特异性的、分泌IFN-的T细胞;同时可以观察到免疫后的C57BI/6小鼠,可以有效预防HPV.16相关肿瘤细胞(1.5×10^5C3细胞)的攻击;对已产生的肿瘤,可以延缓肿瘤细胞的生长速度。结论重组痘苗病毒rVVL1L2可以有效诱发小鼠产生体液免疫和细胞免疫应答,为研究预防和治疗HPV-16感染的疫苗提供了实验资料。  相似文献   

2.
目的构建共表达人乳头瘤病毒16型(HPV16)L1、L2、E6、E7蛋白的非复制型重组痘苗病毒人用疫苗株。方法以痘苗病毒为载体、利用同源重组技术筛选共表达HPV16L1、L2、E6、E7蛋白的重组痘苗病毒并对其进行鉴定。结果该病毒在CEF细胞上连续传至第15代,经斑点杂交结果表明重组病毒基因组中有L1、L2、E6、E7基因插入;经WesternBlot检测,重组病毒能稳定表达HPV16L1、L2、E6、E7蛋白。结论非复制型重组痘苗病毒NTVJE6E7CKL1L2可作为预防和治疗HPV16相关肿瘤及其癌前病变候选疫苗。  相似文献   

3.
目的:构建用于子宫颈癌治疗的HPV16型E6和E7重组痘苗病毒实验性疫苗株,并对其抗肿瘤免疫效果进行初步评价。方法:以痘苗病毒为载体、利用同源重组技术构建共表达HPV16 E6和E7基因的重组痘苗病毒。该病毒免疫C57BL/6小鼠后,检测其免疫原性和抗移植瘤生长情况。结果:PCR结果显示,重组病毒VmE6E7的TK基因内插入了分别由痘苗病毒早晚期启动子H6和7.5K表达的ME6和ME7-1基因。动物实验结果表明,rVmE6E7在C57BL/6小鼠体内可诱发E6和E7特异性抗体产生,被免疫小鼠能够抵抗HPV16 E6E7转化的同系肿瘤细胞的攻击。结论:获得1株用于宫颈癌治疗的HPV16型实验疫苗株,为进一步研制人用HPV16型疫苗株奠定了基础。  相似文献   

4.
目的:评价表达HPV16 L1、L2E7的非复制重组痘苗病毒疫苗的抗肿瘤免疫反应.方法:采用肿瘤预防、肿瘤治疗和肿瘤切除等方法,观察疫苗NTVJL1/L2E7的抗肿瘤效果,用酶联免疫斑点(ELISPOT)和CTL检测该疫苗在小鼠体内诱发的细胞免疫应答.结果:在肿瘤预防和肿瘤治疗试验中,疫苗可以分别使60%和50%的小鼠免受HPV16阳性的治疗细胞攻击,肿瘤切除试验中,疫苗可以使70%的小鼠免于肿瘤复发,与对照组之间的差异具有显著性.ELISPOT和CTL均检测出强的特异性细胞免疫水平.结论:NTVJL1/L2E7能够在小鼠体内诱发出理想的抗肿瘤免疫反应,可以作为防治宫颈癌的候选疫苗.  相似文献   

5.
目的 构建表达HPV18E7E6融合蛋白的重组痘苗病毒,并对E7E6蛋白的免疫原性进行研究.方法 将去除了转化活性的HPV18E6、E7基因融合,插入痘苗病毒重组质粒,通过同源重组构建表达HPV18E7E6的重组痘苗病毒,观察其免疫效果.结果 构建了表达E7E6融合蛋白的重组痘苗病毒,PCR鉴定及测序表明融合基因序列与设计相符,正确插入到痘苗病毒TK区域;Western-Blot检测表明该重组病毒能表达HPV18E7E6融合蛋白.免疫后的小鼠可产生E6、E7特异性抗体,但ELISPOT没检测到E7肽库刺激小鼠脾细胞产生分泌IFN-丫的阳性反应.结论 构建了一株表达HPV18E7E6融合蛋白的重组痘苗病毒,可以有效诱发小鼠产生针对E6、E7的体液免疫,但不能诱发产生相应的细胞免疫,为进一步研究不同动物模型中HPV18E6E7的细胞免疫特点提供了实验基础.  相似文献   

6.
目的 原核表达人乳头瘤病毒(HPV)6型L2AN360E7E6融合蛋白并对其免疫效果进行初步评价.方法 用重叠PCR将HPV6b 12(1~360 bp)、E7、E6三个基因片段融合,原核表达HPV6bL2△N360E7E6融合蛋白,蛋白纯化后与Al(OH)3、CpG佐剂配伍肌内注射免疫C57BL/6小鼠,使用IFN-γ ELISPOT与ELISA分别对其细胞免疫和体液免疫效果进行评价.结果 蛋白+CpG佐剂组与其他免疫组相比,针对E7与E6均有明显较强的细胞免疫反应;各免疫组均能检测到高滴度的抗L2的抗体,但各组之间无明显差异.结论 利用pQE30原核表达系统成功克隆、表达和纯化了HPV6bL2△N360E7E6融合蛋白,且该蛋白与合适佐剂配伍能在C57BL/6小鼠体内诱发强的细胞免疫和体液免疫反应,为该蛋白的后期研究奠定了基础.  相似文献   

7.
目的 选出适合于治疗性疫苗研制的HPV16E7突变基因。方法 对表达野生型和突变型E7蛋白的重组痘苗病毒所诱发的细胞免疫反应和抗肿瘤活性进行比较研究。结果 表达突变型ME7-1(24G26G)的重组痘苗病毒VmE7-1与表达野生型E7的VwE7相同,可诱发特异性抗体和CTL的产生,明显推迟成瘤时间并且保护部分小鼠抵抗肿瘤细胞的攻击;而表达突变型ME7-2(24G26G91G)的重组痘苗病毒VmE7-2免疫小鼠后难以有效的激发细胞免疫反应,在抗肿瘤移植实验中也不具明显的免疫保护作用。结论 E7突变基因ME7-1可作为候选基因用于HPV16治疗性疫苗的研制。  相似文献   

8.
目的 筛选人乳头瘤病毒18型E6、E7蛋白在小鼠中的T细胞表位.方法 以重组痘苗病毒rVVJ18 E7、E6分别免疫C57BL/6和BALB/c小鼠,利用覆盖E6和E7蛋白全长序列的肽库或截短的多肽,用酶联免疫斑点方法 (ELISPOT)和细胞内因子染色检测其所诱发的细胞免疫反应.结果 重组痘苗病毒rVVJ18 E7、E6免疫的两种品系小鼠均可检测到E6肽库刺激产生的特异性细胞免疫反应,经筛选确定E667-75(KCIDFYSRI)为C57BL/6小鼠、E660-68(IPHAAGHKC)为BALB/c小鼠识别的CD8+的T细胞表位.两种小鼠中均未检测到E7蛋白诱发的细胞免疫反应.结论 筛选到分别被BALB/c和C57BL/6小鼠识别的两条针对E6蛋白的不同T细胞表位,为今后评价HPV18疫苗中E6蛋白的细胞免疫效果提供了实验依据.  相似文献   

9.
目的 构建人乳头瘤病毒16型(HPV16)L1-ETc嵌合基因并表达融合蛋白,以期获得防治HPV16感染及相关肿瘤的疫苗.方法 以HPV16型的中国人野毒株为模板,利用PCR克隆技术制备HPV16 L1-E7c嵌合基因,并转入原核表达载体pET28a( ),获得pET28a( )-L1-E7c表达质粒.以Western blot方法鉴定融合蛋白与HPV16 L1和E7抗体的特异性结合.应用蛋白纯化仪纯化HPV16 L1-E7c融合蛋白,经过复性后,电镜观察病毒样颗粒(cVLP)的形态结构.纯化的蛋白免疫动物,测定疫苗抗肿瘤的细胞免疫及抗体产生情况.结果 经过序列分析和酶切鉴定表明成功构建了HPV16L1-E7c嵌合基因,并在大肠杆菌中可高效表达L1-E7c融合蛋白.此融合蛋白具有HPV16 L1和E7的抗原性,纯化的蛋白复性后可形成嵌合病毒样颗粒(cVLP),纯化的蛋白免疫动物后,产生特异性细胞和体液免疫反应,具有抗肿瘤活性.结论 构建的嵌合基因HPV16L1-E7c可有效表达HPV16 L1-E7c重组蛋白并形成cVLP.此蛋白在动物实验中具有疫苗的免疫保护作用,为HPV16疫苗的研究奠定了实验基础.  相似文献   

10.
目的构建能表达L1E7融合蛋白的原核表达菌株,纯化蛋白,并观察其免疫效果。方法用PCR方法分别扩增出C末端部分缺失的HPV16L1基因和HPV16E7编码基因N端部分序列。将上述基因连接,构建融合基因L1ΔCE7N并将其插到原核表达载体pGEX-2T中进行融合蛋白表达纯化,然后观察其免疫效果。结果L1ΔCE7N融合基因测序结果表明,序列与设计相符,读码框架正确。将其插入原核表达质粒在大肠埃希菌中获得高效表达;经Wester-Blot鉴定在相对分子质量约85×103处有特异性表达带,与预期相符。用亲和层析和分子筛可纯化L1ΔCE7N融合蛋白,将其免疫C57BL/6小鼠,结果表明融合蛋白能诱发高滴度L1、E7抗体,并能保护小鼠免受TC-1肿瘤细胞的攻击。结论本实验在原核系统中高效表达并纯化了L1ΔCE7N融合蛋白,该蛋白可作为预防和治疗HPV16感染以及相关肿瘤的候选疫苗株。为研制HPV16预防治疗性疫苗探索一条经济、易普及的途径。  相似文献   

11.
Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc2 (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.  相似文献   

12.
HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.  相似文献   

13.
Renal dysplasia and asplenia in two sibs   总被引:2,自引:0,他引:2  
A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.  相似文献   

14.
Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.  相似文献   

15.
Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.  相似文献   

16.
Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996  相似文献   

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18.
Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999  相似文献   

19.
A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.  相似文献   

20.
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