鸡眼区相关淋巴组织ND疫苗点眼免疫应答 Ⅱ.局部T淋巴细胞亚群的变化

本研究采用流式细胞仪技术对鸡眼区相关淋巴组织在鸡新城疫疫苗点眼免疫后的局部 Ｔ 淋巴细胞亚群的变化进行了研究。研究发现， Ｌａｓｏｔａ 系 Ｎ Ｄ 疫苗点眼免疫后眼区相关淋巴组织对 Ｎ Ｄ 疫苗可 产生良好的细胞免疫应答，免疫组 Ｃ Ｄ＋４ Ｔ及 Ｃ Ｄ＋８ Ｔ 淋巴细胞数量于点眼免疫后迅速升高，其百分比含量明显高于非免疫组。     

影响鸡免疫应答因素分析

鸡的免疫类型分非特异性免疫和特异性免疫.特异性免疫又分为细胞免疫和体液免疫.细胞免疫是指来自于骨髓中的淋巴细胞在胸腺依赖区中被诱导分化成熟为T细胞,在受到抗原或有丝分裂源的刺激后,分化增殖转化为致敏淋巴细胞所表现出来特异性免疫.这类免疫应答不能通过血清传递,只能通过致敏淋巴细胞所传递.体液免疫是指来自于骨髓的淋巴细胞在法氏囊依赖区诱导分化为成熟的B细胞.在抗原物质的刺激下产生抗体以及血液性抗体与相应抗原接触后引起一系列抗原抗体反应的免疫应答方式[1].分析和掌握影响鸡免疫应答因素,对于制定适合现地养禽场的免疫程序和选择合理疫苗,保证该养禽场生物安全生产具有重要意义.     

MTT法评价猪口蹄疫合成肽疫苗的细胞免疫应答

为评价猪口蹄疫合成肽疫苗免疫猪后的细胞免疫应答,用含有E、F两种多肽抗原的猪口蹄疫合成肽疫苗免疫猪,二免后两周采血分离猪外周血淋巴细胞,再用E、F两种合成肽及二者混合物对淋巴细胞刺激培养48 h,用MTT法检测特异性T淋巴细胞增殖反应。结果显示,在抗原E、F混合物浓度为50μg/mL时,抗原对免疫组淋巴结细胞的刺激增殖作用显著高于未免疫对照组（P〈0.01）。表明,该猪口蹄疫合成肽疫苗免疫猪能有效引起特异性T淋巴细胞免疫应答。     

五种旋毛虫抗原对猪的免疫保护作用研究

本实验研究了旋毛虫肌幼虫可溶性粗抗原、排泄分泌抗原（ＥＳ）、表面抗原（ＳＡ）及成虫ＥＳ、ＳＡ５种抗原对猪的免疫保护作用。结果５种抗原对猪均具有一定程度的免疫原性，可诱导猪体产生对攻击感染的抵抗力（减虫率），其中肌幼虫粗抗原为５５．２０％；肌幼虫ＥＳ为４２．５６％，肌幼虫ＳＡ为７２．２１％；成虫ＥＳ为３２．９２％；成虫ＳＡ为４２．１７％。免疫５种抗原后用肌幼虫“Ｂ”抗原、新生幼虫可溶性抗原及成虫可溶性抗原进行ＥＬＩＳＡ检测，均可测出血清抗体应答反应，其中以相应抗原测出的抗体应答较强烈。免疫５种抗原后猪外周血液中Ｂ淋巴细胞减少，Ｔｈ及Ｔｓ增加，Ｔｈ／Ｔｓ比值降低，呈暂时的细胞免疫抑制现象。     

CpG序列对副伤寒疫苗免疫小鼠应答反应的影响

研究了CpG序列作为分子免疫增强剂，对常规副伤寒疫苗免疫小鼠后，小鼠体液和细胞免疫反应的变化情况。试验结果显示，CpG序列作为免疫增强剂，可以使免疫小鼠的总IgG含量、血清特异性抗体效价、淋巴细胞IL—2的诱生活性和脾淋巴细胞的增殖反应有明显的提高。结果表明，CpG序列能显著增强免疫动物对常规疫苗的免疫应答反应水平，可以作为提高疫苗免疫效果的有效途径。     

影响马立克氏病毒感染的因素及疫苗免疫策略

马立克氏病毒是高度细胞结合性、嗜淋巴组织的α疱疹病毒,其致病过程包括淋巴细胞的溶细胞感染和潜伏感染以及易感鸡体内CD 4 T细胞的致瘤性转化。宿主的遗传抗性、细胞免疫、体液免疫以及相关细胞因子和淋巴细胞等对M DV的感染过程有重要影响。针对M D肿瘤抗原的靶向免疫应答的保护性抗原分子和宿主细胞因子疫苗是今后疫苗研究的方向。     

鸡转移因子的制和协及其对鸡免疫应答影响的研究

将新研制的鸡脾转移因子（TransferFactor,TF)与鸡新城疫、法氏减蛋综合征疫苗同时应用后，以淋巴细胞转化试验监测细胞免疫应答，血凝抑制试验（HI）监测体液免疫应答，结果表明，应用TF试验组的细胞应答能力和血清抗体效价与对照组比较，差异显著，说明TF与疫苗同时应用时具有协同作用，可显著地提高机体的细胞免疫和体液免疫功能，增强疫苗的免疫效果。     

鸡IFN-γ重组鸡痘病毒对鸡传染性腔上囊炎MB43弱毒疫苗免疫鸡免疫应答的影响

将重组鸡痘病毒（rFPV—IFN-γ）与IBDMB43弱毒疫苗联合接种SPF鸡，研究重组鸡IFN-γ对IBD疫苗免疫效果的影响。结果显示，rFPV—IFN-γ和MB43弱毒疫苗联合免疫组（MB43＋rFPV-IFN-）＂）及rlFN-γ和MB43弱毒疫苗联合免疫组（MB43＋rIFN-γ）在免疫后第2周即可检出ELISA抗体，比MB43疫苗单独免疫组早1周。用IBDVGx株攻击后，MB43＋rFPV-IFN-γ组和MB43＋rIFN-γ组免疫鸡的脾只有个别淋巴细胞核浓缩、核崩解，少数滤泡萎缩变性坏死；胸腺损伤程度轻于MB43疫苗单独免疫组和非免疫对照组。免疫后1周，3个免疫组外周血CD8^＋T淋巴细胞含量均显著高于非免疫对照组，CD4^＋T淋巴细胞含量变化不明显；攻毒后第2周，3个疫苗免疫组CD4^＋、CD8^＋T淋巴细胞含量均显著升高，各组之间CD4^＋、CD8^＋T淋巴细胞含量无明显差异。证实，rFPV—IFN-γ和rIFN-γ可加强疫苗的细胞免疫和体液免疫应答。     

动物抗独特型疫苗的研究进展

抗独特型疫苗（anti-idiotype vaccine）是免疫调节网络学说发展到新阶段的产物。抗独特型抗体可以模拟抗原,刺激机体产生与抗原特异性抗体具有同等免疫效应的抗体,由此制成的疫苗称为抗独特型疫苗,又称内影像疫苗。抗独特型疫苗不仅能诱导体液免疫应答,亦能诱导细胞免疫应答。近年来,国内外对有关独特型疫苗的报道越来越多。抗独特型疫苗可分为以下几类。     

免疫佐剂的研究进展

佐剂（A山uvants）是先于抗原或与抗原同时注射于动物体内，能非特异性地改变或增强机体对该抗原的特异性免疫应答的物质，亦称免疫增强剂。[1]佐剂可选择性的改变免疫应答类型，产生体液和／或细胞免疫。改变体液抗体种类，IgG亚类及抗体的亲和性。佐剂可改变抗原的构型，使疫苗诱导T－辅助细胞和细胞毒T淋巴细胞（CHL）反应。佐剂可改变兔疫反应为MHCⅠ型或MHCⅡ型、MHCⅠ型反应通常是在细胞内寄生的病原体如病毒感染时产生，诱导CTL，而蛋白质或多肽抗原则不能诱导。佐剂如免疫刺激复合物，QS21与蛋白质、多肽或灭活病毒结合可…     

Immune responses of breeding chickens to trivalent oil emulsion vaccines: responses to Newcastle disease and infectious bursal disease

Bivalent Newcastle disease (ND)/infectious bursal disease (IBD) and trivalent ND/IBD/infectious bronchitis (IB) inactivated oil emulsion vaccines were prepared in the laboratory and evaluated under field conditions. Broiler breeder parent chickens previously vaccinated with live vaccines were inoculated with commercial monovalent ND and experimental bivalent or trivalent oil emulsion vaccines. The commercial vaccine induced a higher initial ND haemagglutination inhibition (HI) response than the experimental vaccines but, by 34 weeks after vaccination, the mean ND HI levels were not significantly different in any of the three flocks. All three vaccines provided sufficient ND immunity to protect against the clinical disease and egg production losses. The IBD responses of both flocks vaccinated with oil emulsion vaccine were similar to each other and only slightly lower than those flocks vaccinated with monovalent IBD oil emulsion vaccine in earlier experiments. Six weeks after vaccination, sufficient immunity was transferred to protect all the progeny against IBD challenge up to 33 days of age and some of them up to 45 days of age. Thirty-four weeks after vaccination of the parents with oil emulsion vaccine, the progeny were totally immune up to 27 days of age and some of them were immune until 37 days. Application of oil emulsion vaccines in bivalent or trivalent form did not impair the responses of the chickens to the monovalent components.     

几种法氏囊活疫苗对鸡法氏囊损伤及免疫抑制作用的比较

探讨了鸡传染性法氏囊病（ＩＢＤ）活疫苗对鸡新城疫活苗免疫的影响。将四种ＩＢＤ活苗分别接种３０日龄ＳＰＦ公雏,４天后再免疫鸡新城疫ＬａＳｏｔａ系活疫苗,同时法氏囊损伤情况。试验结果表明,所选ＩＢＤ活疫苗对雏鸡法氏囊均有不同程度地损伤,并且诱导产生ＮＤ－ＨＩ抗体的时间被推迟。     

4株鸡IBD活疫苗对鸡ND免疫抗体的影响

４株不同毒力鸡传染性法氏囊病（ＩＢＤ）活疫苗,分别接种３０ｄＳＰＦ公维,４ｄ后再接种鸡新城（ＮＤ）Ｌａｓｏｔａ系活疫苗,同时每组捕杀５只,观察法氏囊损伤情况,另４组用同１天的ＳＰＦ公无接种ＮＤ－Ｌａｓｏｔｑ系疫苗４天后再接种不同毒力的ＩＢＤ疫苗。试验结果表明,４株ＩＢＤ活疫苗对雏鸡法氏囊不同程度损伤,并且对ＮＤ免疫抗体产生有明显抑制作用。     

Efficacy of avridine as an adjuvant for Newcastle disease virus antigen in chickens

Avridine, a lipoidal amine with interferon-inducing and adjuvant properties, was an effective adjuvant for Newcastle disease antigen (NDA) in chickens. Eleven vaccine lots were evaluated: 2 commercial water-in-oil vaccines, 4 experimental oil emulsion vaccines, 4 avridine-containing vaccines, and a control lot of nonadjuvanted antigen. Avridine significantly enhanced the immunologic responses of chickens against NDA. Chickens vaccinated with the avridine-containing vaccines had significantly higher antibody titers (hemagglutination inhibition) than did chickens vaccinated with the commercial vaccines. Experimental oil emulsion vaccines prepared from the same antigens as avridine-adjuvanted vaccines induced higher hemagglutination inhibition antibody titers after primary but not after booster vaccination. Use of avridine as an adjuvant for NDA in vaccines for chickens induced immunologic protection rates similar to those induced by oil emulsion vaccines, without causing the reactogenic and tissue residue problems associated with the use of oil vaccines in chickens.     

不同法氏囊病活疫苗对新城疫免疫效果影响的评估

为了评估不同毒力鸡传染性法氏囊病（Infectious Bursal Disease,IBD）活疫苗是否会对鸡的免疫应答产生影响,本实验选用A、B、C三种商品化IBD活疫苗免疫商品肉鸡,以鸡新城疫（Newcastle Disease,ND）疫苗抗体的消长规律为模型,评估3种I BD疫苗对ND抗体产生的影响.免疫后ND抗体检测结果显示,三种疫苗对ND抗体产生的影响差异不明显。结果提示,临床上选用IBD疫苗不必过于关注其毒力。     

Enhancement of mucosal immune responses by intranasal co-delivery of Newcastle disease vaccine plus CpG oligonucleotide in SPF chickens in vivo

Immunostimulatory CpG oligodeoxynucleotides (ODN) have been tested as immunoadjuvants for various vaccines in mice and human. Findings from previous reports suggest that CpG ODN can be used to enhance magnitude and balance of an immune response while reducing undesirable side effects of commercial vaccine, when delivered by parenteral route. Recently, it has been showed that CpG ODN is a promising mucosal adjuvant in mice, but data on mucosal immune responses induced by CpG ODN in other animals, especially in chickens, are scarce. Herein, we evaluated intranasal (IN) delivery of CpG ODN with newcastle disease (ND) vaccine (NDV) to determine its potential as a mucosal adjuvant to a commercial vaccine. CpG ODN augmented systemic (IgG in serum, T cell proliferation) and mucosal (IgA in intestinal washings and feces) immune responses against antigen. CpG ODN stimulated effectively both systemic and mucosal immune responses when delivered intranasally. Results from this study indicate that stimulatory CpG ODN is a potential effective mucosal adjuvant for the NDV in SPF chickens and may be applicable to husbandry animals.     

Enhancement of mucosal immune responses by intranasal co-delivery of Newcastle disease vaccine plus CpG oligonucleotide in SPF chickens in vivo

Immunostimulatory CpG oligodeoxynucleotides (ODN) have been tested as immunoadjuvants for various vaccines in mice and human. Findings from previous reports suggest that CpG ODN can be used to enhance magnitude and balance of an immune response while reducing undesirable side effects of commercial vaccine, when delivered by parenteral route. Recently, it has been showed that CpG ODN is a promising mucosal adjuvant in mice, but data on mucosal immune responses induced by CpG ODN in other animals, especially in chickens, are scarce. Herein, we evaluated intranasal (IN) delivery of CpG ODN with newcastle disease (ND) vaccine (NDV) to determine its potential as a mucosal adjuvant to a commercial vaccine. CpG ODN augmented systemic (IgG in serum, T cell proliferation) and mucosal (IgA in intestinal washings and feces) immune responses against antigen. CpG ODN stimulated effectively both systemic and mucosal immune responses when delivered intranasally. Results from this study indicate that stimulatory CpG ODN is a potential effective mucosal adjuvant for the NDV in SPF chickens and may be applicable to husbandry animals.     

鸡传染性法氏囊病活疫苗（B87株 CA株 CF株）安全性及免疫效果评价

通过观察疫苗免疫后鸡群的生长情况和临床症状,检测免疫后不同时期鸡群的ELISA抗体水平和攻毒保护率以及对鸡新城疫（ND）疫苗免疫后不同时间鸡群的HI抗体水平,评价和检测鸡传染性法氏囊病活疫苗（B87株 CA株 CF株）在石家庄地区临床应用的安全性、免疫效果、免疫持续期以及是否会产生免疫抑制。结果表明,试验疫苗接种蛋鸡和肉鸡后,均没有观察到因疫苗引起的不良反应,对蛋鸡和肉鸡均安全。疫苗接种蛋鸡后14 d、28 d、42 d、60 d和90 d,接种肉鸡后14 d、21 d、36 d攻毒保护率均达80%以上;商品蛋鸡接种疫苗后90 d其ELISA抗体水平仍高达6500以上。ND HI抗体检测结果表明,试验疫苗接种后对ND疫苗的免疫应答无显著影响,不引起免疫抑制。结论：试验疫苗安全、有效,质量稳定,适用于预防鸡传染性法氏囊病。     

Vaccination with Newcastle disease vaccine and CpG oligodeoxynucleotides induces specific immunity and protection against Newcastle disease virus in SPF chicken

Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) have been proven to be immunoprotective in mouse models. However, little work has been conducted on in vivo immune responses in chicken with CpG ODN. The objective of this study was to investigate the immunoadjuvant effects of CpG ODN to Newcastle disease (ND) vaccine and its protective effects against ND virus in SPF chicken. In this report, the titre of serum IgG to ND vaccine and the proliferation of lymphocytes were monitored in SPF chickens. The results demonstrated that the above-mentioned immune responses were significantly stronger in chickens that received CpG ODN than in the birds that received only ND vaccine. Furthermore, ND vaccine plus CpG ODN protected SPF chicken from challenge with an otherwise lethal dose of ND virus. These data suggest that CpG ODN holds considerable promise as an adjuvant for future vaccines against ND virus.     

IBD疫苗免疫SPF雏鸡局部黏膜组织的免疫学变化

应用组织化学染色法和免疫酶组织化学法,研究IBD疫苗免疫SPF雏鸡后,雏鸡哈德氏腺和盲肠扁桃体的T细胞及IgA、IgM和IgG抗体生成细胞数量的动态变化。结果发现,免疫雏鸡上述局部免疫组织的各项免疫指标较对照雏鸡均不同程度地增高,表明疫苗免疫能显著提高雏鸡眼部、消化道局部黏膜免疫功能;IBD强毒攻击后,对照雏鸡上述指标明显低于疫苗免疫雏鸡,且发生典型IBD全部死亡,而疫苗免疫雏鸡免疫保护率达60%。