不同佐剂对猪附红细胞体亚单位疫苗免疫效果的影响

为比较不同佐剂的猪附红细胞体亚单位疫苗的免疫效果,笔者通过试验提取猪附红细胞体抗原,并将其分别与白油佐剂、弗氏佐剂、明矾佐剂及铝胶佐剂混合,免疫小鼠。应用间接ELISA方法比较不同佐剂的猪附红细胞体亚单位疫苗的免疫效果。结果表明：白油佐剂组免疫效果最好,抗体水平在一周后有明显上升,其次为与弗氏佐剂组、铝胶佐剂组和明矾佐剂组。该试验为猪附红细胞体亚单位疫苗的研究提供了可靠的理论依据,为该病的防治奠定了一定的基础。     

不同佐剂对猪附红细胞体亚单位疫苗免疫效果的影响

为比较不同佐剂的猪附红细胞体亚单位疫苗的免疫效果,本试验提取猪附红细胞体抗原,并将其分别与白油佐剂、弗氏佐剂、明矾佐剂及铝胶佐剂混合,免疫小鼠。应用间接ELISA方法比较不同佐剂的猪附红细胞体亚单位疫苗的免疫效果。结果表明:白油佐剂组免疫效果最好,抗体水平在一周后有明显上升,其次为弗氏佐剂组、铝胶佐剂组和明矾佐剂组。该试验为猪附红细胞体亚单位疫苗的研究提供了可靠的理论依据,为该病的防治奠定了一定的基础。     

纳米磷酸钙作为猪瘟多肽疫苗佐剂的研究

为了探索纳米磷酸钙(NCaP)作为猪瘟多肽疫苗佐剂的可行性,本研究利用猪瘟多肽疫苗作为抗原,制备纳米磷酸钙吸附猪瘟多肽疫苗。将制备好的疫苗免疫兔后,分别检测兔抗体水平和攻毒后体温变化情况。检测结果显示,纳米磷酸钙对猪瘟多肽吸附率达70%;免疫后纳米磷酸钙佐剂组抗体水平低于弗氏佐剂,但与游离猪瘟多肽疫苗组相比,有较大提高;除弗氏佐剂组外,所有实验兔均有体温升高现象,纳米磷酸钙佐剂组体温升高的时间与空白组相比推迟16 h。结果表明,NCaP作为佐剂相对游离猪瘟多肽可提高其免疫效果,但其诱导的抗体水平低于弗氏佐剂组。     

虎源FPV主要抗原表位区基因的原核表达及免疫原性分析

为筛选出制备猫瘟热亚单位疫苗的最佳佐剂类型与首选抗原片段,本试验对虎源FPV-HLJ株VP2全长基因及其截短基因片段PAB进行原核表达,切胶法纯化的表达蛋白经Western blot分析后,分别与氢氧化铝胶佐剂及弗氏佐剂混合,免疫BALB/c小鼠,间接ELISA方法检测小鼠体液免疫水平,初步评价各疫苗的免疫效果。结果表明:VP2、PAB融合蛋白主要以包涵体形式表达,切胶法获得的高纯度表达蛋白均能与鼠抗虎源FPV阳性血清发生特异性反应。纯化蛋白各佐剂免疫组免疫小鼠后均引起较强的特异性抗体IgG反应,其中PAB蛋白各免疫佐剂组抗体水平明显高于VP2蛋白相应免疫佐剂组,且PAB蛋白+氢氧化铝胶组与PAB蛋白+弗氏佐剂组抗体水平无显著差异(P0.05),但与未加佐剂组差异显著(P0.01)。研究证实,PAB蛋白具有良好的免疫原性,可与氢氧化铝胶佐剂协同用于FPV亚单位疫苗的制备。     

大肠杆菌不同佐剂灭活苗血清抗体与卵黄抗体消长规律的研究

笔者用分离得到的猪源致病性大肠杆菌制成灭活疫苗免疫产蛋鸡,定期检测蛋鸡血清和卵黄的抗体效价。试验结果表明,免疫蛋鸡血清中和卵黄中抗体的消长规律相似,同一时期的血清抗体比卵黄抗体较高。同时比较3种不同佐剂疫苗产生的效果,结果表明,氟氏佐剂和氢氧化铝佐剂疫苗产生的抗体效价较白油佐剂疫苗产生的抗体效价高,但氟氏佐剂疫苗和白油佐剂疫苗较氢氧化铝佐剂疫苗抗体持续时间长。综合比较,氟氏佐剂较其他2种佐剂产生的抗体效价高、持续时间长。     

重组CRM197-4GnRH去势疫苗抗原分子的原核表达及免疫效果评价

通过大肠杆菌表达系统对CRM₁₉₇-4GnRH重组去势疫苗抗原进行表达,采用不同种类佐剂制备疫苗,研究其对大鼠的免疫去势作用,为获得1种免疫效果好、有效期长、生产成本低的促性腺激素释放激素(GnRH)免疫去势基因工程疫苗提供借鉴。采用白喉毒素无毒突变体(CRM₁₉₇)与4拷贝GnRH串联,将CRM₁₉₇-4GnRH基因与表达载体连接,并通过优化试验确定大肠杆菌的最优表达条件,用SDS-PAGE和Western blot鉴定目标蛋白的表达情况。将融合蛋白与不同佐剂乳化,制备成9种测试疫苗后分别免疫大鼠,测定血清中GnRH抗体滴度和睾酮浓度。结果显示,重组大肠杆菌的最优表达条件为诱导时间表达5 h,培养基为LB液体培养基,IPTG诱导浓度为0.5 mmol/L;通过SDS-PAGE和Western blot试验,表明目标融合蛋白已成功表达;动物试验结果表明,重组疫苗水包油包水佐剂组、重组疫苗油包水佐剂组、GnRH+Th抗原(辅助型T细胞抗原)表位(G1抗原)油包水佐剂组测试疫苗免疫大鼠较其他组GnRH抗体滴度显著升高、睾酮浓...     

乳酸杆菌DNA对鸡新城疫油苗和禽流感油苗免疫增强作用的研究

本试验提取乳酸杆菌L.caseiZhang的DNA作佐剂,按每只鸡100ug剂量添加到新城疫油苗及禽流感病毒H5亚型油苗中。新城疫和禽流感试验分别设空白对照组(Ⅰ组)、无佐剂疫苗组(Ⅱ组)、细菌核酸佐剂组(Ⅲ组)、弗氏完全佐剂组(Ⅳ组)、弗氏完全佐剂核酸免疫协同组(Ⅴ组)5组进行免疫,分别于15和28日龄首免,两周后二免。二免后每周采样用血凝及血凝抑制法检测血清抗体水平,并用MTT法检测脾脏T淋巴细胞增殖反应。结果显示,添加核酸组鸡HI抗体和T淋巴细胞增殖反应均极显著高于单独免疫疫苗组(P<0.01);而免疫组均极显著高于空白对照组(P<0.01);细菌核酸组HI抗体略高于弗氏完全佐剂组,但T淋巴细胞转化率极显著高于弗氏完全佐剂组(P<0.01);同时弗氏完全佐剂核酸免疫协同组与核酸组无显著性差异。由此得出,乳酸杆菌L.caseiZhangDNA对新城疫油苗和禽流感病毒H5亚型油苗具有免疫增强作用;乳酸杆菌L.caseiZhangDNA与弗氏完全佐剂无明显的免疫协同作用。     

适用于蛋鸡免疫的猪源ETEC菌毛蛋白佐剂的筛选

为筛选卵黄抗体的优良佐剂,以3株猪源致病性大肠杆菌(ETEC)菌毛蛋白为模式抗原,分别与6种佐剂制备多价菌毛蛋白疫苗,免疫海兰褐壳蛋鸡。结果显示,3次免疫后,所有试验组蛋鸡产蛋率均有不同程度下降,但SPA、ISCOMs、弗氏佐剂、蜂胶佐剂组的产蛋率下降程度较为轻微;对鸡体血清抗体和制备获得的卵黄抗体的效价检测结果均显示:SPA和ISCOMs组虽较SPB、SPC、弗氏佐剂组抗体水平略低,但持久度较好,且通过对蛋鸡外观形态和注射部位组织变化情况的观察显示,两者不会刺激鸡体热反应或炎症反应,无任何表观副作用。因此,更适合作为蛋鸡制备猪源ETEC卵黄抗体的优良佐剂。     

不同佐剂新城疫疫苗对鸡体液免疫的影响

佐剂是疫苗研究的重要组成部分.选择合适的佐剂不仅可以增强疫苗的免疫效果,而且可以提高机体的抵抗力.不同佐剂对体液免疫会产生不同的作用.本研究选用白油佐剂、弗氏佐剂和蜂胶佐荆与新城疫病毒抗原混合制成疫苗,免疫雏鸡,探讨不同佐剂对鸡体液免疫的影响.结果表明,三种佐剂疫苗均有保护力,试验鸡饲养至61日龄均未发生新城疫,其中弗氏佐剂的效果最好,蜂胶佐剂的效果次之,油佐剂组检测到雏鸡61日龄时仍未出现抗体高峰值.     

中药黄芪多糖的免疫佐剂作用

分别将黄芪多糖提取物、白油-吐温、氢氧化铝作为佐剂制备金黄色葡萄球菌灭活苗混合免疫家兔和奶牛,观察家兔免疫前后血清抗体滴度变化以及攻毒后白细胞数目变化和淋巴细胞百分比变化。对奶牛免疫后测其血清抗体和乳汁中体细胞数变化,结果注射疫苗后乳汁中体细胞数都有所下降,黄芪多糖组较其它组下降较快,幅度较大。黄芪多糖和抗原混合物免疫动物后未见不良反应,且抗原免疫血清抗体比氢氧化铝和白油-吐温佐剂组的抗体滴度增加快,幅度显著增高且抗体维持一个较高水平,而攻毒后家兔白细胞数和淋巴细胞数也较氢氧化铝和白油-吐温佐剂组增加较多。因此黄芪多糖是一种有效的佐剂。     

成年雄性小鼠经LHRH去势苗免疫后的性功能恢复

用LHRH去势苗免疫性成熟雄鼠，免疫期对小鼠的性行为、受孕情况、睾丸剖检和组织学变化以及精子数量和活动能力进行检查，以确定去势效果。待抗体水平自然下降后再对小鼠的上述指标进行检查，以判断性功能恢复情况。结果显示抗体下降后，小鼠可以重新获得生育能力，表明LHRH去势苗作用是可逆的。     

Effects of 17beta-estradiol and tamoxifen on the selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) mRNA expression in male reproductive organs of rats

The selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) is highly expressed in testes under gonadotropin control. The expression patterns of PHGPx mRNA by 17beta-estradiol (E2) as an estrogen and tamoxifen (Tam) as an estrogen antagonist were investigated in the reproductive organs of male rats. Twelve-week-old male Sprague-Dawley rats were subcutaneously injected with E2 (7.5 microg/kg/day) or Tam (5 mg/kg/day) for 1 week. The E2 treatment significantly increased the levels of PHGPx mRNA in both testes and prostates, whereas the Tam treatment significantly decreased the levels of PHGPx mRNA, compared to the vehicle control (p<0.01). The treatment with E2 or Tam slightly decreased the levels of PHGPx mRNA in epididymides. In histopathological examination, severe vacuolization and depletion of germ cells in the seminiferous tubules, cell debris in the tubular lumen, and mild proliferative changes in interstitial tissues were observed in the testes of Tam-treated rats, whereas only mild spermatogonial proliferation was observed in the seminiferous tubules of E2-treated rats. There were no typical histopathological changes in the epididymides of any of the laboratory rats but mild epithelial proliferation in the prostates of E2- and Tam-treated rats. These results suggest that PHGPx mRNA expression may be influenced by estrogen in the male reproductive organs.     

Effects of antiepileptic drugs carbamazepine and sodium valproate on fertility of male rats

The effects of carbamazepine and sodium valproate on fertility of male rats were studied. The tested drugs were given orally to male rats for 30 and 60 consecutive days. Mating performance, sex organs weights, semen quality, plasma concentrations of sexual hormones as well as histopathological findings were the criteria used to evaluate the reproductive efficiency of treated males. Oral administration of carbamazepine and sodium valproate for 30 and 60 consecutive days significantly decreased the testicular weight, sperm cell concentration, live sperms and percentage of progressively motile spermatozoa. Both drugs significantly increased the percentage of morphologically abnormal spermatozoa. A decrease in plasma testosterone, FSH and LH and an increase in prolactin levels were observed in the treated groups. Histopathological examination showed mild to moderate degenerative changes in the testes of the treated rats while the prostate glands and seminal vesicles appeared normal. A recovery period of 30 days was accompanied by marked changes in the tested parameters towards initial values.     

Comparison of alum-absorbed or non-alum-absorbed oil emulsion vaccines containing either pilate or non-pilate Bacteroides nodosus cells in inducing and maintaining resistance of sheep to experimental foot rot

Highly pilate (P) or non-pilate (NP) cells of Bacteroides nodosus were compounded into oil emulsion (O) either with or without prior absorption onto alum (A). The abilities of these four preparations (referred to as PAO, NPAO, PO and NPO vaccines) to stimulate antibody production and to protect sheep from foot rot were compared. Two injections of PAO vaccine protected sheep against homologous challenge 12 weeks after the second dose by PO, NPO and NPAO vaccines were less effective. Sheep were protected against homologous challenge for 14 weeks after a single dose of PAO vaccine and for 22 weeks after three doses; an ameliorative effect was still evident 40 weeks after the third dose. Protection against challenge with two heterologous strains was demonstrated at six weeks after three doses of vaccine. A numerical system of scoring the lesions also confirmed that foot rot in vaccinated sheep challenged outside the 'protective' period of the vaccine was somewhat less severe than in controls. PAO vaccine induced much higher and more persistent titres of agglutinins than the other vaccines tested. There was a relationship between agglutinin titres and resistance to homologous challenge.     

Evaluation of the efficacy of immunocastration vaccine composed of gonadotrophin-releasing hormone conjugated with Salmonella typhimurium flagellin in rats

Immunocastration is an alternative method to replace surgical castration that is commonly performed in domestic and pet animals. In this study, a new immunocastration vaccine was developed, and its efficacy was evaluated in male rats. Six tandem copies of gonadotrophin-releasing hormone (GnRH) peptide were genetically fused to Salmonella typhimurium flagellin fljB (STF2) that is a ligand of toll-like receptor 5 (TLR5). The recombinant STF2-GnRH protein expressed in Escherichia coli was used as the immunocastration vaccine. Sixteen male rats were equally assigned to four groups. Excluding the control rats, three groups were immunized with 100, 200 and 400 μg of the STF2-GnRH vaccine, respectively. All of the immunized rats developed significantly higher titres of antibodies to GnRH than the control rats. The size and weight of both testes and epididymides from the immunized rats were significantly smaller than those of the control rats. Testicular tissues in the immunized rats demonstrated atrophy of seminiferous tubules and decreased numbers of both spermatogonia and spermatocytes. These data indicate that the newly developed STF2-GnRH vaccine has a potent immunogenicity to GnRH and efficiently suppresses the development of testes in rats.     

Effect of vaccination against GnRH (Bopriva®) in the male pubertal calf

The objective of this experiment was to investigate the effectiveness of a newly developed bovine anti-GnRH vaccine (Bopriva?, Pfizer Animal Health, Australia). A total of 12 peripubertal bull calves aged between 6 and 8 months were used, 2 randomly selected animals served as controls. Animals were vaccinated twice at an interval of 4 weeks with 1ml of Bopriva? (400 μg GnRH-protein-conjugate) subcutaneously in the neck and observed for a total of 36 weeks. Scrotal circumference was measured every week and blood samples were also taken weekly for the determination of testosterone and GnRH antibodies. Three months after the second injection (booster), 5 animals were slaughtered and their testes histologically examined. GnRH antibody titers rapidly began to rise after the second vaccination and reached peak values 3 weeks later. Testosterone concentrations decreased to values below 0.5 ng/ml serum 1 week after the booster and remained at this low level for at least 10 weeks. The following increase of testosterone occurred individually within 11 and 23 weeks after the booster injection. Histological examination of testes in vaccinated animals showed an incomplete spermatogenesis with impaired or no production of spermatids and a reduced diameter of seminiferous tubules. From our results we conclude that in the peripubertal bull two injections with the new bovine anti-GnRH vaccine 4 weeks apart is effective in suppressing testicular growth and testosterone secretion during at least 10 weeks after the booster injection.     

Effects of experimental cryptorchidism on sperm motility and testicular endocrinology in adult male rats

The effect of induced cryptorchidism on testicular function and sperm motility was investigated. Bilateral cryptorchidism was created surgically in adult male rats (treated group), and sham-operated rats were used as a control group. Five rats from each group were sacrificed on days 1, 3, 5, and 7 after surgery. The percentage of motile spermatozoa began to decrease 1 day after the operation, followed by an abrupt decline 3 and 5 days later in cryptorchid rats. Furthermore, there were significant decreases in the other sperm motility parameters 5 days after inducement of cryptorchidism. In cryptorchid rats, plasma concentrations of LH, FSH, testosterone, and inhibin B were significantly lower than in the control group 1 day after the operation. Thereafter, plasma concentrations of LH, FSH, and testosterone gradually increased in the cryptorchid rats. On the other hand, plasma concentrations of inhibin B showed a further decline from day 3 after the operation onward. Concentrations of immunoreactive (ir)-inhibin, but not testosterone, in testicular interstitial fluid were remarkably increased until 3 days after surgery in the cryptorchid rats, and declined thereafter. Testicular response to human chorionic gonadotropin (hCG) for testosterone release was decreased in the cryptorchid rats compared with the control rats, indicating that heat stress to testes resulted in a reduction of the activity of Leydig cells and Sertoli cells. These results clearly indicate that heat stress to the testes resulted in a significant reduction of sperm activity within 3 days, and this was followed by changes in testicular endocrine function.     

Decrease of pituitary adenylate cyclase activating polypeptide and its type I receptor mRNAs in rat testes by ethanol exposure

The present study was designed to evaluate the effect of ethanol on pituitary adenylate cyclase activating polypeptide (PACAP) and its typ I (PAC1) receptor expression in adult rat testes. Ethanol (3 g/kg i.p., 15% v/v in saline) was administrated to adult male rats for 10 days. Using northern blot analysis, the present study showed the reduction of PACAP mRNA levels in rat testes by ethanol administration. Also, ethanol decreased the expression level of PAC1 receptor in testes. In particular, in situ hybridization clearly showed the decrease of PAC1 receptor mRNA expression in Leydig cells, which produce testosterone. Furthermore, the serum level of testosterone was significantly decreased in the ethanol-treated group. In conclusion, our findings suggest that the decrease of PACAP and PAC1 receptor expression in rat testes by ethanol exposure may partly contribute to the suppression of male reproductive activity.     

外生殖器发育不全公鹅睾丸组织学及血浆睾酮水平研究

本试验以天府肉鹅母系 (四川白鹅 ,WW)、天府肉鹅父系 (川朗鹅 ,WL)、朗德鹅 (LL)为试验材料 ,研究不同品种公鹅和外生殖器官发育不同 (发育正常 :WLn、LLn ,发育异常 :WLa、LLa)公鹅睾丸的形态学、组织学特性、超微结构及血浆睾酮水平。结果表明 :公鹅睾丸重量、体积、曲细精管直径、生殖上皮高度、生殖细胞总数的数密度、精原细胞的数密度、精母细胞的数密度、精细胞的数密度、精子的数密度、间质细胞的数密度 ,WW公鹅显著大于WLn和LLn ;WLn大于LLn ,但差异不显著 ;WLn、LLn显著大于WLa和LLa ;WLa与LLa间差异不显著。公鹅睾丸超微结构可观察到各级生殖细胞数量WW多于WLn和LLn ;WLn、LLn多于WLa和LLa ;血浆睾酮水平 ,WW公鹅显著高于WLn和LLn ;WLn、LLn显著高于WLa和LLa ;WLn显著高于LLn ,WLa显著高于LLa。睾丸重量、体积及定量组织学指标与阴茎发育情况、血浆睾酮水平高低有关 ,因此可以通过观测公鹅阴茎发育情况、血浆睾酮水平 ,来对种公鹅进行选择。     

对牦牛和普通牛种间杂种公牛精子生成水平的研究

本文对牦牛和普通牛种间杂种公牛睾丸组织及电刺激所采精液进行了观测。结果表明,种间杂种公牛随代数增加,睾丸逐渐恢复产生精子的机能。种间杂种3代公牛有的可产生形态正常的精子。种间杂种3代是可育或不育(不能产生精子)的临界代。