CIA免疫母鸡后子代雏鸡免疫器官T细胞变化的研究

鸡传染性贫血病（CIA）疫苗免疫母鸡后，对其子雏鸡免疫器官的T细胞变化进行了动态研究。结果发现，CIA疫苗免疫母鸡后，子代雏鸡免疫器官T细胞数量较未免疫对照子代雏鸡明显增加；强毒攻击子代雏鸡后，免疫攻毒组雏鸡免疫器官T细胞数量在13日龄内明显高于未免疫攻毒组雏鸡。表明CIA疫苗可使子代雏鸡免疫器官的细胞免疫功能增强，能抵卸强毒攻击。     

黄曲霉毒素B_1对雏鸡免疫器官影响的病理学观察

为探明黄曲霉毒素B1对雏鸡免疫器官组织学及超微结构的影响,将100只1日龄艾维茵健康公雏随机分为4组,分别喂以对照日粮和AFB1日粮(AFB1Ⅰ、Ⅱ、Ⅲ组日粮中AFB1添加量分别为0.15、0.3和0.6mg·kg-1),试验期21d。结果显示,AFB1Ⅱ组和Ⅲ组雏鸡的免疫器官脏器指数显著下降(P0.05)。AFB1组雏鸡的免疫器官组织学损伤表现:胸腺皮质区网状细胞周围见较多细胞核碎片;法氏囊淋巴滤泡内细胞核碎片增多,滤泡髓质区淋巴细胞减少;脾白髓区细胞核碎片增多。超微病理学观察,胸腺、法氏囊和脾内淋巴细胞线粒体肿胀,以染色质边移为特征的凋亡细胞数目增多。结果表明,摄食含0.15~0.6mg·kg-1 AFB1的日粮,可不同程度地抑制雏鸡免疫器官的发育,致免疫器官中的淋巴细胞数量减少、细胞核碎片增多。     

鸡传染性贫血病免疫母鸡后子代雏鸡免疫器官抗体生成细胞变化

鸡传染性贫血病疫苗免疫母鸡后，对其于代雏鸡免疫器官的抗体生成细胞变化进行了动态研究。结果发现，CIA疫苗免疫母鸡后，于代雏鸡免疫器官抗体生成细胞数量较未免疫对照于代雏鸡明显增加；强毒攻击于代雏鸡后，免疫攻毒组雏鸡免疫器官抗体生成细胞数量在27日龄内明显高于未免疫攻毒组雏鸡。表明CIA疫苗可使于代雏鸡免疫器官的体液免疫功能增强，能抵御强毒攻击。     

鸡贫血病—法氏囊病联合免疫母鸡后子代雏鸡免疫器官T细胞和抗体生成细胞数量的动态变化

对鸡传染性贫血病（CIA）－传染性法氏囊病（IBD）联合免疫母鸡后的子代雏鸡免疫器官的免疫学化变化进行了研究。结果发现,混合感染CIAV、IBDV雏鸡免疫器官T细胞和IgG,IgM,IgA抗体生成细胞数量在27日龄内明显未免疫对照组、联合免疫组和联合免疫攻毒组,表明感染CIAV、IBDV的雏鸡全身免疫功能显著下降,CAI－IBD联合免疫母鸡后,子代雏鸡T细胞胞和IgG,IgM,IgA抗体生成细胞数量在27日龄内,较未免疫对照组明显增加,表明CIA－IBD联合免疫母鸡可使子代雏鸡免疫器官的免疫功能增强,能抵御强毒攻击。     

马立克氏病胚胎免疫雏鸡体重和免疫器官增重变化

１８日龄鸡胚免疫接种火鸡疱疹病毒（ＨＶＴ）疫苗后,对出壳后雏鸡及其用马立克氏病（ＭＤ）病毒攻毒鸡体重与免疫器官增长效果的对比研究表明：相同饲养条件下胚胎免疫雏鸡的增长速度高于非免疫对照鸡,而且胚胎免疫组雏鸡免疫器官的生长发育普遍较相应的对照组雏鸡快。     

鸡马立克氏病胚胎免疫后淋巴器官免疫效果的观察

用火鸡疱疹病毒疫苗对孵化至１８日龄的鸡胚进行胚胎免疫注射，免疫后出壳雏鸡淋巴脏器的组织学和组织化学显示，１８日龄胚胎免疫鸡对淋巴器官的早期发育具有明显的免疫促进作用。     

实验性猪口蹄疫免疫病理形态学研究

选用健康仔猪13头,分成4组:健康对照组2头,健康攻毒组5头,免疫组和免疫攻毒级各3头。健康攻毒组经皮下注射猪口蹄疫病毒(FMDV);免疫组和免疫攻毒组均经皮下接种猪口蹄疫牛皮肤细胞弱毒苗(FMD-BC),观察1个月后,前者扑杀,后者再攻FMDV,1个月后扑杀。结果:健康攻毒组5头仔猪均出现口蹄疫(FMD)典型的临床症状和病现学变化,其淋巴结、脾脏不同程度的变性、坏死和出血,淋巴小结减少和缩小,淋巴细胞稀少,浆细胞呈坏死性变化,酯酶阳性(ANAE~ )细胞和次级淋巴小结均减少;免疫组未见任何FMD病理学变化,主要见全身各部位淋巴结及脾脏不同程度地增大,尤以肩前、髂下和脾门淋巴结更为显著,其淋巴小结显著增多、增大,淋巴细胞活化,大、中淋巴细胞增多,以过渡型和未成熟型浆细胞为主,ANAE~ 细胞和次级淋巴小结均增多;免疫攻毒组的变化与免疫组相似。     

HSP27在热应激致岭南黄鸡雏鸡免疫器官损伤中的作用

为了从热休克蛋白27(HSP27)基因角度揭示热应激致雏鸡免疫器官形态损伤的机制,试验将120只8日龄岭南黄鸡随机分为对照组和热应激组,并构建雏鸡热应激模型,采用组织切片和透射电镜方法对比分析其免疫器官在热应激条件下的显微结构和超微结构变化,并用Real-time PCR方法检测HSP27基因在免疫器官中mRNA的表达差异。结果表明:同日龄阶段比较,热应激组免疫器官发育不良,胸腺细胞数量减少,Hassall氏小体严重退化,胸腺细胞凋亡及坏死数量增多,染色质发生聚集,细胞器肿胀变性;脾脏组织疏松,脾小结生发中心不明显,动脉周围淋巴鞘变薄,淋巴细胞结构疏松,胞核内异染色质减少,线粒体数量增多、结构异常;法氏囊中淋巴滤泡数量减少,皮质和髓质分界模糊,淋巴细胞染色质聚集,胞浆中粗面内质网上核糖体脱落,线粒体肿胀空泡化。法氏囊中HSP27 mRNA相对表达量逐渐降低,在热应激第21天时相对表达量显著高于第28,35天;脾脏中HSP27 mRNA相对表达量逐渐升高,在第35天时相对表达量显著高于第21,28天;胸腺中HSP27mRNA相对表达量与日龄无明显关联,在第28天时相对表达量最低且显著低于第21天和第35天。说明热应激会造成岭南黄鸡雏鸡免疫器官损伤,而HSP27在这一过程中扮演着重要角色。     

铜中毒对雏鸡免疫功能影响的研究

180只1日龄艾维菌肉鸡健雏随机分为3组,分别喂以对照日粮(Cu 11.97 mg/kg)和铜中毒日粮(Cu 650 mg/kg,铜中毒Ⅰ组;Cu 850 mg/kg,铜中毒Ⅱ组)6周,观察铜中毒对免疫功能的影响.与对照组比较,两个铜中毒组雏鸡胸腺、脾脏和法氏囊的生长指数、淋巴细胞生长周期和外周血T淋巴细胞ANAE阳性率显著降低(P＜0.05或P＜0.01);病理组织学观察,淋巴免疫器官淋巴细胞显著减少并呈退性变.结果表明,铜中毒严重抑制淋巴免疫器官生长发育和外周血T淋巴细胞活化,并对淋巴免疫器官造成损伤,导致免疫功能下降.本中还就铜中毒对免疫功能的影响机理进行了讨论.     

青刺果种粕粉对鸡免疫器官发育的影响

本试验旨在研究青刺果种粕粉对鸡免疫器官发育的影响。将7日龄鸡分成试验组和对照组,每组设3个重复;对照组饲喂基础日粮,试验组饲粮分别添加10和15g/kg青刺果种粕粉,饲喂30d。第14、21和28日龄时分别取胸腺、法氏囊和脾脏称重并制作切片。结果表明,试验组鸡法氏囊、胸腺、脾脏脏器指数在各个日龄均显著或极显著高于对照组(P<0.05或P<0.01)。光镜下,14日龄时,试验组较对照组法氏囊皮质与髓质分界较清楚,淋巴滤泡形成,胸腺皮质不同程度增厚,脾脏形成动脉周围淋巴鞘;21日龄时,试验组较对照组法氏囊中淋巴细胞发育成熟,形态较小,淋巴滤泡直径增加,胸腺小体增多,皮质增厚,脾脏淋巴细胞致密;28日龄时,试验组与对照组比较,法氏囊淋巴滤泡较大,胸腺小体增大,可见朗罕氏细胞,脾小体和生发中心较发达。电镜下,试验组脾脏淋巴细胞形态均一,核仁增多,核染色质边移,随着日龄的增长,淋巴细胞活性增加。结论:青刺果种粕粉能促进鸡免疫器官的生长发育。     

鸡马立克氏病CVI988／Rispens疫苗免疫效力试验

应用荷兰农业部提供的鸡马立克氏病（ＭＤ）ＣＶＩ９８８／Ｒｉｓｐｅｎｓ Ⅰ型致弱种毒, 在农业部批准的符合ＧＭＰ 要求的生产车间研制出鸡马立克氏病ＣＶＩ９８８／Ｒｉｓｐｅｎｓ 疫苗。将按国际标准检验合格的三批疫苗及进口商品ＣＶＩ９８８／Ｒｉｓｐｅｎｓ 疫苗接种１ 日龄ＳＰＦ 雏鸡, 于７ 日龄经腹腔攻击鸡马立克氏病强毒（北京－ １ 株） 血毒, 全部鸡只隔离饲养观察至６０ 日龄并作全群剖检。经测定： 非免疫攻毒组１００％ 发病,健康对照组全部阴性, 三批国产ＣＶＩ９８８／Ｒｉｓｐｅｎｓ 疫苗保护指数分别为９０－０, ９０－０, ９３－３ , 进口商品苗保护率为９３－３ 。结果表明国产和进口ＣＶＩ９８８／Ｒｉｓｐｅｎｓ疫苗均能提供对ＭＤ 较高的免疫保护力, 国产疫苗的保护效果达到了国际同类产品的先进水平。     

鸡马立克氏病CVI988/Rispens疫苗免疫效力试验

本试验用北京市农林科学院畜牧兽医研究所制备的ＣＶＩ９８８／Ｒｉｓｐｅｎｓ疫苗和进口的ＣＶＩ９８８／Ｒｉｓｐｅｎｓ疫苗免疫１日龄来航鸡,７日龄以ＭＤＶ北京－１株血毒进行攻击,６０日龄全群剖检。经免疫效力试验两次测定,３批北京所制备的ＣＶＩ９８Ｒｉｓｐｅｎｓ疫苗产品保护指数分别为试验的９０．０、９０．０、９３．３和试验（２）的１００．０、１００．０、９４．５与进口商品ＣＶＩ／９８８Ｒｉｓｐｅｎｓ苗的保     

A comparative evaluation of the protective efficacy of rMd5deltaMeq and CVI988/ Rispens against a vv+ strain of Marek's disease virus infection in a series of recombinant congenic strains of White Leghorn chickens

Marek's disease (MD) is a lymphoproliferative disease of domestic chickens caused by a highly infectious, oncogenic alpha-herpesvirus known as Marek's disease virus (MDV). MD is presently controlled by vaccination. Current MD vaccines include attenuated serotype 1 strains (e.g., CVI988/Rispens), avirulent serotype 2 (SB-1), and serotype 3 (HVT) MDV strains. In addition, recombinant MDV strains have been developed as potential new and more efficient vaccines to sustain the success of MD control in poultry. One of the candidate recombinant MDV strains, named rMd5deltaMeq, was derived from Md5, a very virulent strain of MDV lacking the MDV oncogene Meq. Our earlier reports suggest that rMd5deltaMeq provided protection equally well or better than commonly used MD vaccines in experimental and commercial lines of chickens challenged with very virulent plus (vv+) strains of MDV. In this study, maternal antibody-positive (trial 1) and negative (trial 2) chickens from a series of relatively MD resistant lines were either vaccinated with the rMd5deltaMeq or CVI988/Rispens followed by infection of a vv+ strain of MDV, 648A, passage 10. This report presents experimental evidence that the rMd5deltaMeq protected significantly better than the CVI988/Rispens (P < 0.01) in the relatively resistant experimental lines of chickens challenged with the vv+ strain of MDV. Together with early reports, the rMd5deltaMeq appeared to provide better protection, comparing with the most efficacious commercially available vaccine, CVI988/Rispens, for control of MD in lines of chickens regardless of their genetic background.     

Field evaluations of safety and efficacy of an Australian Marek's disease vaccine

OBJECTIVE: To demonstrate the safety and efficacy of the Marek's Disease Virus-1 vaccine (strain BH 16) from field studies in comparison with the CVI 988 Rispens vaccine currently available in Australia. STUDY DESIGN: A small field trial was carried out on nine breeder flocks and a larger trial on 21 breeder flocks. All chickens were obtained from a commercial hatchery and each was vaccinated at hatch with cell-associated Herpes Virus of Turkeys vaccine. A group of chickens vaccinated with BH 16 vaccine was placed in one shed per property and the remainder were vaccinated with the Rispens vaccine and placed in the remaining sheds. At 25, 30, 35, and 40 weeks after hatch, the field veterinarian or farm manager examined all birds dying on two consecutive days in the designated placement sheds. RESULTS: In the small trial there was a significantly lower incidence of MD in birds vaccinated with the MDV-1 vaccine compared with the Rispens vaccine (P < 0.001). In a larger trial there was no difference in the incidence of MD between the treatment groups, due possibly to a lower rate of natural challenge. Egg production results and average weekly mortality results for both groups were similar. CONCLUSION: The present study describes an attenuated type 1 MD vaccine which is at least equivalent to a vaccine derived from the CVI 988 Rispens strain in terms of safety and efficacy when used in combination with HVT vaccine.     

表达H5亚型禽流感病毒HA基因的重组马立克氏病病毒的构建

采用聚合酶链反应或反转录聚合酶链反应扩增出H5亚型禽流感病毒(AIV)的HA基因、网状内皮增生症病毒的长末端重复序列(LTR)、马立克氏病病毒(MDV)Rispens CVI988毒株基因组的sorf 1和sorf 2序列、两端带loxp位点的lac/smGFP标志基因,构建含这些基因的转移载体质粒pMHA;以MDV Rispens CVI988毒株的基因组DNA和PMHA质粒DNA共转染鸡胚成纤维细胞(CEF),采用同源重组方法将LTR、lac/smGFP和HA基因插入到MDV基因组,获得重组病毒rMDV-HA/GFP;以cre介导的同源重组去除lac/smGFP标志基因,再转染CEF,获得仅带LTR启动子和HA基因的重组MDV疫苗毒株rMDV-HA.rMDV-HA仍保留了MDV RispensCVI988疫苗毒株的复制特点,并能稳定表达AIV的HA.     

马立克氏病病毒肿瘤Meq基因缺失株的保护性免疫作用

马立克氏病病毒（MDV）的致病性一直在不断增强中,甚至已出现了能抵抗CV1988/Rispens疫苗的特超强株。本实验室用BAC克隆技术构建了MDV中国野毒株的Meq基因缺失株,显示出在抗马立克氏病方面具有与美国Meq基因缺失株同样有效的保护性免疫效果,而且其自身没有明显的免疫抑制作用。对美国和中国的两个MDV的Meg基因缺失株的优缺点做了比较。     

鸡马立克病疫苗株CVI988/Rispens meq基因编辑及缺失毒株的构建与鉴定

meq是鸡马立克病病毒（MDV）最重要的致瘤基因，在马立克病（MD）肿瘤发生中发挥关键作用。同时，它在疫苗株和强毒株之间具有明显的序列差异性。本文利用CRISPR/Cas9基因编辑技术，以MDV疫苗株CVI988/Rispens meq基因为靶点，设计合成gRNA，克隆构建pX459-gRNA质粒，转染CEF并感染CVI988/Rispens，然后对meq基因编辑的病毒噬斑进行克隆纯化，经过PCR扩增、测序分析及IFA鉴定，成功构建1株meq基因编辑的缺失毒株CVI988Δmeq-C7，为后续筛选和鉴定抗MD疫苗株MEQ单抗及鉴别诊断研究奠定了基础。     

The efficacy of recombinant fowlpox vaccine protection against Marek's disease: its dependence on chicken line and B haplotype

Earlier studies have shown that the B haplotype has a significant influence on the protective efficacy of vaccines against Marek's disease (MD) and that the level of protection varies dependent on the serotype of MD virus (MDV) used in the vaccine. To determine if the protective glycoprotein gene gB is a basis for this association, we compared recombinant fowlpox virus (rFPV) containing a single gB gene from three serotypes of MDV. The rFPV were used to vaccinate 15.B congenic lines. Nonvaccinated chickens from all three haplotypes had 84%-97% MD after challenge. The rFPV containing gB1 provides better protection than rFPV containing gB2 or gB3 in all three B genotypes. Moreover, the gB proteins were critical, since the B*21/*21 chickens had better protection than chickens with B*13/*13 or B*5/*5 using rFPV with gB1, gB2, or gB3. A newly described combined rFPV/gB1gEgIUL32 + HVT vaccine was analyzed in chickens of lines 15 x 7 (B*2/*15) and N (B*21/*21) challenged with two vv+ strains of MDV. There were line differences in protection by the vaccines and line N had better protection with the rFPV/gB1gEgIUL32 + HVT vaccines (92%-100%) following either MDV challenge, but protection was significantly lower in 15 X 7 chickens (35%) when compared with the vaccine CVI988/Rispens (94%) and 301B1 + HVT (65%). Another experiment used four lines of chickens receiving the new rFPV + HVT vaccine or CVI988/Rispens and challenge with 648A MDV. The CVI 988/Rispens generally provided better protection in lines P and 15 X 7 and in one replicate with line TK. The combined rFPV/gB1gEgIUL32 + HVT vaccines protected line N chickens (90%) better than did CVI988/Rispens (73%). These data indicate that rFPV + HVT vaccines may provide protection against MD that is equivalent to or superior to CVI988/ Rispens in some chicken strains. It is not clear whether the rFPV/gB1gEgIUL32 + HVT vaccine will offer high levels of protection to commercial strains, but this vaccine, when used in line N chickens, may be a useful model to study interactions between vaccines and chicken genotypes and may thereby improve future MD vaccines.     

Differential quantification of cloned CVI988 vaccine strain and virulent RB-1B strain of Marek’s disease viruses in chicken tissues, using real-time PCR

The ‘gold standard’ vaccine against Marek’s disease in poultry is the CVI988/Rispens virus, which is not easily distinguishable, antigenically or genetically, from virulent Marek’s disease herpesvirus. Accurate differential measurement of the CVI988 vaccine and virulent viruses is important to investigate mechanisms of vaccinal protection. Minimal sequence differences between CVI988 and virulent MDV strains restrict the application of molecular diagnostic methods such as real-time PCR to distinguish between these viruses. The use of bacterial-artificial-chromosome (BAC) cloned CVI988 virus, which carries the BAC vector sequences in place of the U_s2 gene, allows its differential quantification from virulent strains using real-time PCR assays that target the BAC vector sequence and the U_S2 gene respectively. These novel assays allowed investigation of replication of both serotype-1 vaccine virus (cloned CVI988) and challenge virus (RB-1B strain) in tissues of individual chickens in an experimental vaccination-challenge model of Marek’s disease.