Intestinal and serum antibody response in gnotobiotic piglets to oral immunization with Escherichia coli

The local and systemic immune response to a formolized E. coli oral vaccine was investigated in 13 gnotobiotic piglets. Beginning at ten days of age animals received a daily dose of 10¹⁰ or 10¹¹ bacteria, on ten consecutive days. Intestinal loop tests with one animal of each group on day 26 showed protection which was more pronounced in the animal dosed 10¹⁰ bacteria compared with the other immunized piglet. Immunoglobulin class-specific antibodies to O and K antigens were determined by ELISA technique. In serum no IgG or IgA antibodies were found, whereas IgM-anti O149 antibodies in both immunized groups reached their highest level at day 4 of dosing and decreased thereafter. IgM-anti K88 antibodies were first detected at day 10 of dosing. Both immunized groups had comparable serum levels at days 20 and 30. Also in gut secretion the IgM antibody response was predominant, and higher levels were found in the 10¹⁰ group than in the 10¹¹ group. IgG and IgA antibody response were also detected in secretion.     

The immune response in a dog to Encephalitozoon cuniculi infection

The immune response to Encephalitozoon cuniculi infection in a dog was investigated by means of the indirect fluorescent antibody test, the leucocyte migration inhibition test and the radial immunodiffusion test for serum IgG and IgM levels. Specific antibodies were detected within 7 days of infection and they persisted for 370 days. A cell-mediated immune response was detected from Day 13 following infection until Day 97. Histopathological examination showed plasma cell infiltration of the kidneys, meninges, lung, bladder, smooth muscle and spleen.     

高致病性猪繁殖与呼吸综合征减毒疫苗在仔猪体内的抗体消长规律

本文比较了不同日龄和母源抗体水平的仔猪接种高致病性猪繁殖与呼吸综合征减毒疫苗（JXA1-R株）后的抗体平均值、抗体阳性率和变异系数等指标。结果表明,对于母源抗体水平和整齐度不高的猪群,15日龄和30日龄接种减毒疫苗的免疫效果相似;对于母源抗体水平和整齐度较高的猪群,采用30日龄进行减毒疫苗接种可以产生较好的效果。本研究结果为制定适合连云港地区的高致病性猪繁殖与呼吸综合征免疫程序提供了依据。     

伪狂犬病病毒囊膜蛋白ISCOMs免疫原性测定

应用伪狂犬病病毒囊膜蛋白与Quil A结合制备囊膜蛋白免疫刺激复合物(PRV-ISCOMs)，并通过ELISA、血清中和试验和淋巴细胞转化试验(Brdu-ELISA法)测定其诱导小鼠体液和细胞免疫应答水平。结果如下：1)小鼠分别接种3、7、10ug的PRV-ISCOMs后，于接种后PI7d血清中可测出ELISA IgG而抗体，随后抗体水平逐渐升高。间隔21d加强免疫后，FLISA IgG抗体水平进一步提高，且中和抗体效价均在1：22以上，而未加佐剂的囊膜蛋白对照组均无中和抗体；2)淋巴细胞转化试验初步结果表明PRV-ISCOMs能诱导小鼠的细胞免疫应答。3)免疫保护力测定显示免疫鼠能抵抗强毒攻击。上述结果表明PRV-ISCOMs具有良好的免疫原性，能诱导小鼠的体液免疫和细胞免疫应答。     

山羊实验感染绵羊进行性肺炎病毒的抗体应答反应

用琼脂凝胶免疫扩散试验（ＡＧＩＤＴ）和免疫印迹试验（ＩＢＡ）对山羊实验感染绵羊进行性肺炎病毒（ＯＰＰＶ）的抗体应答反应进行了研究。结果,用ＡＧＩＤＴ和ＩＢＡ都可在接毒山羊的血清中检测到ＯＰＰＶ的抗体。ＡＧＩＤＴ最早于接毒后１５ｄ检测到抗体;ＩＢＡ最早于接毒后４ｄ检测到抗ＯＰＰＶ的ｇｐ４４和ｐ２８的抗体,以后又陆续检测到抗ｐ９４、ｐ１４和ｇｐ１２５的抗体。由此看出,ＩＢＡ比ＡＧＩＤＴ更为敏感。本研究结果表明,ＯＰ－ＰＶ可在山羊体内诱生较强的体液免疫应答反应,因此用ＯＰＰＶ通过山羊体传代的方法可能会得到具有良好抗原性的ＯＰＰＶ毒株。     

Reconstitution of immunosuppression mice with mononuclear cells from donors sensitized to foot-and-mouth disease virus (FMDV)

Passive transfer experiments were performed to serve as a basis for analyzing the immune response of adult mice to FMDV infection. Animals were irradiated (750 rad: 1 lethal dose 50%) and reconstituted with allogeneic mononuclear cells from blood, spleen, thymus and peritoneal cavity from donors 2 and 8 days post-inoculation (p.i.). Donors were primed with 10 000 suckling mouse 50% lethal doses of FMDV strain O1 Campos. The following parameters were studied in recipient mice challenged with 10 000 suckling mouse 50% lethal doses of the same virus: (1) viremia; (b) FMDV neutralizing antibody titres; (c) sheep red blood cell (SRBC) hemagglutinating antibody titres. Viremia was substantially prolonged in irradiated control mice, which did not produce detectable antibodies to FMDV or SRBC. In contrast, the span of viremia was markedly shorter in animals reconstituted with cells obtained 8 days p.i. and its eclipse coincided with the onset of neutralizing antibody production. An equally efficient antibody response to the inoculation of SRBC was observed in these animals. No effect was detected after the transfer of cells obtained 2 days p.i. It is concluded that the humoral immune response plays a predominant role in the recovery from FMDV experimental infection in adult mice.     

Immunoreactive protein antigens of Mycoplasma pulmonis in experimentally infected rats

Mycoplasma pulmonis was cultured in modified Hayflick's medium, washed in 0.25 M NaCl, and solubilized by 2.5% sodium dodecyl sulfate. Protein antigens of M pulmonis separated by polyacrylamide-gel electrophoresis were blotted onto nitrocellulose strips. Specific-pathogen-free rats were inoculated intranasally with M pulmonis. The serum samples of these rats were obtained periodically and used to react with fractionated M pulmonis antigens which were fixed on the nitrocellulose strips. The antigen-antibody reactions were further recognized by 125I-labeled antiglobulin. Detection of immunoreactive antigens was obtained by autoradiography. Antibody response was not detected in serum obtained 7 days after rats were inoculated, and by 14 days, a slight response to several proteins was found. At 28 days after rats were inoculated, many immunoreactive antigens were detected. Generally, antibodies against antigens of moderate to low molecular weight appeared early in the infection, and antibodies against antigens of high molecular weight appeared late. Important immunoreactive antigens thus identified can readily be distinguished from more than 58 different M pulmonis antigens detectable by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. The humoral antibody response was measured by an enzyme-linked immunosorbent assay. The immunoglobulin G antibodies were initially detected at low level at 7 days after rats were inoculated. These humoral antibody responses reached maximum by 28 days. The increase in serum antibody titer corresponded with numbers of immunoreactive antigens detected by immunoradio-binding assay. The information gained by this investigation may improve our understanding of the antigenicity of M pulmonis and the immune response of rats exposed to M pulmonis.     

兔IL-6基因真核表达载体的构建及其对pcDNA-VP60DNA疫苗佐剂效应研究

为研究兔白介素6（interleukin6,IL-6）真核表达质粒（pcDNA-IL-6）对核酸疫苗免疫效果的影响,本文构建了真核表达质粒pcDNA-IL-6,并将其与核酸疫苗pcDNA-VP60联合免疫家兔,以pcDNA-VP60和质粒载体pcDNA3．1（＋）作对照,用血凝抑制试验检测试验兔体内特异性抗体水平。结果表明：真核重组质粒pcDNA-IL-6对重组质粒pcDNA-VP60均具有免疫增强作用,从免疫后7d到70d,pcDNA-VP60／pcDNA-IL6联合免疫组抗体水平均高于pcDNA-VP60免疫组,差异具有显著统计学意义。     

Characterization of the bovine secondary in vitro antibody response

In the present report, the bovine secondary in vitro antibody response to keyhole limpet hemocyanin is described. The induction of anti-KLH antibody was not dependent upon the presence of mitogen but was antigen specific (KLH vs ovalbumin). Furthermore, this response was dependent upon cell density (10(6) per well), antigen dose (1 to 10(-5) ug/culture) and time in culture (5 days). The antibody produced was specific for KLH as measured in several binding assays. An unresponsive state was detected with high concentrations of KLH (more than 10 ug per culture) which was not due to the formation of immune complexes but to the inactivation of B and/or T cells. The induction of the antibody response was dependent on the presence of macrophages (syngeneic or allogeneic) and their presence could not be replaced with 2-mercaptoethanol.     

小反刍兽疫疫苗临床免疫效果试验

为评估小反刍兽疫弱毒活疫苗(75/1株)的临床免疫效果,通过免疫试验、免疫场的跟踪检测以及荧光RT-PCR检测方法,对疫苗免疫安全性、免疫抗体持续期、抗体效价消长情况进行了临床免疫测试和评估。结果显示,疫苗免疫安全性良好,免疫后14d可以检测到疫苗核酸,免疫后7d开始产生特异性抗体,抗体持续时间达12个月以上。研究表明临床用小反刍兽疫疫苗具有良好的免疫原性,可产生良好的临床免疫效果。     

猪囊虫病基因工程疫苗的体液与细胞免疫反应

为了探讨以 Ｉ Ｓ Ｃ Ｏ Ｍ 作佐剂的猪囊虫病基因工程疫苗的免疫机理,对其诱导的体液免疫与细胞免疫反应进行了测定。用上述疫苗免疫 ９ 头试验猪,采用间接 Ｅ Ｌ Ｉ Ｓ Ａ 检测体液免疫反应及通过淋巴细胞转化试验、 Ａ Ｎ Ａ Ｅ染色试验、 Ｅ玫瑰花环形成试验等检测细胞免疫反应;用该疫苗和铝胶苗分别免疫昆明小鼠各 ２０ 只,分别检测体液免疫反应和 Ｔ 淋巴细胞抑制／杀伤亚群的动态变化。体液免疫的检测结果显示,免疫后 ７ 天即出现抗体,２１ 天后抗体全部转阳,持续的时间不少于 １９３ 天,效价明显高于铝胶苗;细胞免疫检测结果显示,免疫猪外周血 Ｔ淋巴细胞转化率、 Ａ Ｎ Ａ Ｅ＋ 细胞和粗粒型 Ａ Ｎ Ａ Ｅ＋ 细胞、 Ｅ Ｒ Ｆ Ｃ和 Ｅａ Ｒ Ｆ Ｃ细胞显著升高,免疫小鼠 Ｔ淋巴细胞抑制／杀伤亚群显著升高;与铝胶苗及对照组比较,差异极显著。以上结果表明猪囊虫病基因工程疫苗可同时激发动物的体液免疫和细胞免疫反应,增强了机体的免疫调节功能及杀伤性 Ｔ 淋巴细胞功能。     

鸡传染性法氏囊病CH80活疫苗免疫效果研究

试验用鸡传染性法氏囊病CH80活疫苗和某进口鸡传染性法氏囊活疫苗分别免疫18 000 AA肉鸡,于免疫前1d,免疫后14d,31d分别检测IBD抗体和新城疫HI抗体;免疫后4d观察法氏囊病理变化并测定囊重指数;出栏时(45日龄),计算成活率、体重、料肉比及生产指数等。结果表明,CH80活疫苗免疫后,抗体水平略高于对照组,并可一直维持到肉鸡出栏;对法氏囊的损伤也明显小于对照组;对新城疫HI抗体没有明显的影响;总体经济效益也略高于对照组。     

Identification of anti-idiotypic antibody in dogs

There is general agreement that idiotype/anti-idiotype (id/anti-id) networks are important mechanisms of immune regulation, based primarily on studies conducted using inbred laboratory animals. To determine if anti-idiotypic antibody (anti-id) could be induced during an immune response in outbred dogs, the dogs were immunized to the hapten-carrier combination dinitrophenol-ascaris (DNP-ASC) and subsequently immunized with autologous antibody in complete Freund's adjuvant (CFA). Auto-anti-id was detected in three of five dogs during the DNP-ASC response. A cross-reactive anti-id was detected in dogs immunized with autologous antibody when a mouse monoclonal antibody was used as the id. These experiments further suggest that the regulation of the immune response via network interaction, as first illucidated in inbred animals, may occur in dogs.     

Humoral immune response of water buffalo monitored with three different antigens of Toxocara vitulorum

Humoral immune response of water buffalo naturally infected with Toxocara vitulorum was monitored using three different antigens of this parasite in serum and colostrum of buffalo cows and calves. Soluble extract (Ex) and excretory/secretory (ES) larval antigens and perienteric fluid antigen (Pe) of adult T. vitulorum were used to measure the antibody levels by an indirect ELISA. Serum of 7-12 buffalo cows for the first 365 days and colostrum of the same number of buffalo cows for the first 60 days of parturition, and serum of 8-10 buffalo calves for the first 365 days after birth were assayed. The ELISA detected antibodies against all three T. vitulorum antigens in the colostrum and serum of 100% of buffalo cows and calves examined. The highest antibody levels against Ex, ES and Pe antigens were detected in the buffalo cow sera during the perinatal period and were maintained at high levels through 300 days after parturition. On the other hand, colostrum antibody concentrations of all three antigens were highest on the first day post-parturition, but decreased sharply during the first 15 days. Concomitantly to the monitoring of immune response, the parasitic status of the calves was also evaluated. In calves, antibodies passively acquired were at the highest concentrations 24 h after birth and remained at high levels until 45 days coincidentally with the peak of T. vitulorum infection. The rejection of the worms by the calves occurred simultaneously with the decline of antibody levels, which reached their lowest levels between 76 and 150 days. Thereafter, probably because of the presence of adults/larvae stimulation, the calves acquired active immunity and the antibodies started to increase slightly in the serum and plateaued between the days 211 and 365. All three antigens were detected by the serum antibodies of buffalo calves; however, the concentration of anti-Pe antibody was higher than anti-EX and anti-ES, particularly after 90 days of age. By conclusion, the buffalo cows develop immunity and keep high levels of antibodies against T. vitulorum-Ex, ES and Pe antigens and these antibodies are transferred to their calves through the colostrum. This passively acquired immunity does not protect the calves against the acquisition of the infection, but these antibodies, passively or actively acquired, may have an important role during worm rejection by the calves and prevention of intestinal reinfection.     

CDV免疫蛋鸡血清抗体与卵黄抗体的消长规律及其相关性分析

为了解产蛋鸡对犬瘟热病毒杭原的免疫反应及其血清抗体和卵黄抗体的消长规律及相关性,为制备卵黄抗体提供依据,将犬瘟热病毒接种Vero细胞和DF1细胞进行传代并测定其病毒滴度,选取细胞病变出现早,病毒滴度高的Vero细胞毒作为接种病毒抗原免疫蛋鸡。通过每10d进行蛋鸡免疫一次,四免后每30d加强免疫一次的方法进行免疫。免疫前和免疫后每10d采血分离血清和每5d收集鸡蛋提取卵黄抗体,用琼脂扩散法和间接ELISA法测定其抗体水平。结果显示,血清抗体比卵黄抗体的滴度高,两者间呈正相关性,卵黄抗体出现较血清抗体迟3～5d,卵黄抗体在三免后3～5d就已达到较高水平,此时即可开始收集鸡蛋制备卵黄抗体。     

H5N1亚型禽流感灭活疫苗免疫鸵鸟后抗体消长规律及免疫程序的研究

为了探讨鸵鸟对H5亚型禽流感灭活疫苗的免疫原性,做好鸵鸟禽流感的防控工作,采用哈尔滨兽医研究所研制的重组H5N1亚型禽流感疫苗,不同剂量免疫鸵鸟,用HI方法检测母源抗体和免疫抗体,根据母源抗体的衰减和免疫抗体的消长规律确定首免和再免日龄.结果表明,雏鸵鸟母源抗体能维持约3周～8周;8周龄时分组首免,C1组产生的免疫反应优于C2组,抗体峰值能达到7.50 log2,维持时间为9周左右;17周龄时C1组以3.0 mL/羽进行二免,2周后抗体达到7.40log2,3周～7周抗体维持在高峰值,最高达8.80log2,以后逐渐下降,有效抗体水平能维持至接种后25周左右.二免后25周进行三免,以5 mL/羽三免后2周抗体可达到9.80log2,2周～4周抗体维持在最高峰,以后缓慢下降,期间抗体时有起伏,但有效抗体水平约可维持1年时间.三免后45周内抗体水平合格率均在70%以上.根据抗体消长规律,初步推荐了鸵鸟的免疫程序.     

Intestinal antibody response after vaccination and infection with rotavirus of calves fed colostrum with or without rotavirus antibody

The intestinal and systemic antibody response of calves vaccinated and/or challenged with rotavirus was studied employing isotype-specific ELISAs for the detection of IgG1, IgG2, IgM and IgA antibodies to rotavirus. Monoclonal antibodies to bovine immunoglobulin isotypes of proven specificity were used as conjugated or catching antibody. Five days after oral inoculation (dpi) of a 5-day-old gnotobiotic calf with rotavirus, IgM rotavirus antibodies were excreted in faeces, followed 5 days later by IgA rotavirus antibodies. The increase in IgM rotavirus antibody titre coincided with the inability to detect further rotavirus excretion. Faeces IgM and IgA rotavirus antibody titres fell to low levels within 3 weeks post infection. IgG1 and IgG2 rotavirus antibodies were not detected in faecal samples. In serum, antibodies to rotavirus of all four isotypes were detected, starting with IgM at 5 dpi. Two SPF-calves, which were fed colostrum free of rotavirus antibodies, were vaccinated with a modified live rotavirus vaccine and challenged with virulent rotavirus 6 days later. Upon vaccination, the calves showed an antibody response similar to the response of the infected gnotobiotic calf. Intestinal IgM rotavirus antibodies were excreted before or on the day of challenge and appeared to be associated with protection against challenge infection with virulent virus and rotavirus-induced diarrhoea. In 3 control calves, which were challenged only, the antibody patterns also resembled that of the gnotobiotic calf and again the appearance of IgM rotavirus antibodies coincided with the end of the rotavirus detection period. Two other groups of 3 SPF-calves were treated similarly, but the calves were fed colostrum with rotavirus antibodies during the first 48 h of life. These calves excreted passively acquired IgG1 and IgG2 rotavirus antibodies in their faeces from 2 to 6 days after birth. After vaccination, no IgM or IgA antibody activity in serum or faeces was detectable. Upon challenge, all calves developed diarrhoea and excreted rotavirus. Seven to 10 days after challenge low levels of IgM rotavirus antibody were detected for a short period. These data indicate that the intestinal antibody response of young calves to an enteric viral infection is associated with the excretion of IgM antibodies, immediately followed by IgA antibodies. This response is absent or diminished in calves with passively acquired specific antibodies which may explain the failure to induce a protective intestinal immune response by oral vaccination with modified live rotavirus of calves fed colostrum containing rotavirus antibodies.     

Partial inhibition of the humoral immune response of pigs after early postnatal immunization

The ability of young pigs to be immunized during the postnatal period was studied. Eight groups of pigs born on the same day from 3 sows were injected with hen egg-white lysozyme in Freund's incomplete adjuvant at days 0, 1, 2, 3, 4, 5, 7, and 10 after birth. The serum antibody titers were determined each week. Results indicated that pigs injected within the first 3 days of life exhibited a delay of 10 days in the appearance of the humoral antibodies, compared with the antibody response observed in pigs injected later. Serum antibody titers were markedly lower in the early immunized pigs. The secondary immune response was similar in all pigs. This partial inhibition is not directly linked to the corticoids present in the serum at the immunization day. Possible reasons for this impairment of the humoral immune response were reviewed.     

The Response of Ponies to Myxovirus influenzae A-equi 2 I. Serum and Antibody Titres Following Exposure

The antibody response in serum and nasal secretions of groups of ponies vaccinated or infected with Myxovirus influenzae A-equi 2 was examined. Following infection by aerosol with live virus, a weak antibody response was recorded in both serum and secretions. Antibody levels were undetectable in secretions at 31 days after infection.

After primary intramuscular vaccination with killed virus, using sodium alginate as an adjuvant, antibody was detected only in the serum. However, following revaccination, a pronounced antibody response was demonstrated in both serum and secretions. Antibody was still detectable in all four ponies when tested 135 days later.

Only a serum antibody response was detected in ponies after primary intramuscular vaccination with a commercial vaccine. Upon revaccination nasal antibody occurred in all ponies but this only persisted for about 30 days.

Neither serum nor nasal antibody response occurred following intranasal vaccination and revaccination with a killed virus vaccine.