猪免疫口蹄疫灭活疫苗后外周血细胞亚群变化的研究

为了解动物在注射口蹄疫灭活疫苗后机体免疫状态的变化情况,本试验选取口蹄疫阴性猪进行疫苗免疫,利用流式细胞术检测方法来测定外周血中免疫细胞亚群的变化。结果显示,树突状细胞在免疫后能够快速产生应答,在24 h内其含量出现明显升高;B细胞和Th细胞含量在免疫后第10天都出现了明显升高,且两者之间的变化存在着高度相关性,而抗体水平的变化情况与这两种细胞含量的变化也高度相关;细胞毒性T细胞含量在免疫后也出现升高。这说明灭活疫苗免疫动物后不仅可以激活机体的体液免疫应答反应,产生高水平抗体,发挥免疫保护作用,更重要的是能够在短时间内激发机体的固有免疫应答,同时能够激活细胞免疫应答,全方位发挥免疫保护作用。     

鸡毒支原体F弱毒疫苗株细胞免疫特性研究

为了解鸡毒支原体（Mycoplasma gallisepticum,MG）F弱毒疫苗株的细胞免疫特性,分别以活菌浓度为109（A组）、106（B组）ccu/mL的MG F株及生理盐水（C组）点眼接种SPF鸡,采用流式细胞技术及T淋巴细胞增殖试验对免疫前后淋巴细胞亚类Th/T、Tc/T、Th/Tc及刺激指数（SI）的动态变化规律进行研究。结果显示免疫后A、B组的Th/T、Tc/T、SI明显升高,其中Th/T于d5、d7,Tc/T、SI于d7、d14,A组显著高于B组（P〈0.05）。研究结果表明,免疫MG F株可较好的提高鸡的细胞免疫,且MG F株活菌浓度与接种鸡外周血Th/T、Tc/T、SI呈正相关。     

Antibody and IFN-gamma responses induced by a recombinant canarypox vaccine and challenge infection with equine influenza virus

In horses, equine influenza virus (EIV) is a leading cause of respiratory disease. Conventional inactivated vaccines induce a short-lived immune response. By comparison, natural infection confers a long-term immunity to re-infection. An aim of new equine influenza vaccines is to more closely mimic natural infection in order to achieve a better quality of immunity. A new live recombinant vaccine derived from the canarypox virus vector and expressing haemagglutinin genes of EIV (subtype H3N8) has been developed. Stimulation of the immune system was studied after immunisation with this canarypox-based vaccine and challenge infection by exposure to a nebulised aerosol of EIV. The humoral immune response was evaluated by measuring serum antibody levels using the single radial haemolysis (SRH) assay. The cellular immune response was assessed by the measurement of interferon gamma (IFN-gamma) synthesis in peripheral blood mononuclear cells (PBMC). Clinical signs of the disease (temperature, coughing, nasal discharge, dyspnoea, depression and anorexia) and virus excretion were monitored after challenge infection. Clinical signs and virus shedding were significantly reduced in vaccinates compared with unvaccinated controls. EIV-specific immunity was stimulated by vaccination with a recombinant vaccine as serological responses were detected after immunisation. This study also provided the first evidence for increased IFN-gamma protein synthesis in vaccinated ponies following challenge infection with EIV compared with control ponies.     

Results of vaccination of Asian elephants (Elephas maximus) with monovalent inactivated rabies vaccine

OBJECTIVE: To evaluate the humoral immune response of Asian elephants to a primary IM vaccination with either 1 or 2 doses of a commercially available inactivated rabies virus vaccine and evaluate the anamnestic response to a 1-dose booster vaccination. ANIMALS: 16 captive Asian elephants. PROCEDURES: Elephants with no known prior rabies vaccinations were assigned into 2 treatment groups of 8 elephants; 1 group received 1 dose of vaccine, and the other group received 2 doses of vaccine 9 days apart. All elephants received one or two 4-mL IM injections of a monovalent inactivated rabies virus vaccine. Blood was collected prior to vaccination (day 0) and on days 9, 35, 112, and 344. All elephants received 1 booster dose of vaccine on day 344, and a final blood sample was taken 40 days later (day 384). Serum was tested for rabies virus-neutralizing antibodies by use of the rapid fluorescent focus inhibition test. RESULTS: All elephants were seronegative prior to vaccination. There were significant differences in the rabies geometric mean titers between the 2 elephant groups at days 35, 112, and 202. Both groups had a strong anamnestic response 40 days after the booster given at day 344. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed the ability of Asian elephants to develop a humoral immune response after vaccination with a commercially available monovalent inactivated rabies virus vaccine and the feasibility of instituting a rabies virus vaccination program for elephants that are in frequent contact with humans. A 2-dose series of rabies virus vaccine should provide an adequate antibody response in elephants, and annual boosters should maintain the antibody response in this species.     

ISCOM-matrix-based equine influenza (EIV) vaccine stimulates cell-mediated immunity in the horse

The humoral immune response induced by ISCOM-matrix (Immuno Stimulating COMplex-Matrix)-adjuvanted equine influenza virus (EIV) vaccine is well documented in horses. ISCOM-matrix adjuvanted vaccines against human influenza are strong inducers of cell-mediated immunity (CMI), including T cell proliferation and virus-specific cytotoxic T cell. In the horse, the CMI response to equine influenza vaccination is less well characterised. An ISCOM-based vaccine has been shown to induce interferon gamma (IFN-γ) synthesis, a CMI marker, in the horse, but this has not been shown for the ISCOM-matrix vaccine, which is a different formulation. The objective of this study was to measure EIV-specific IFN-γ synthesis after vaccination with an ISCOM-matrix-adjuvanted EIV vaccine. Equilis Prequenza is a commercialised inactivated EIV vaccine containing purified haemagglutinin (HA) and neuraminidase (NA) subunits adjuvanted with ISCOM-matrix. Six influenza-naïve Welsh mountain ponies were vaccinated twice with Equilis Prequenza at an interval of four weeks. Six control ponies received a placebo of physiological water. EIV-specific IFN-γ synthesis by peripheral blood lymphocytes and the antibody response to a panel of representative EIV isolates were measured prior to and after both injections. Immunisation with the ISCOM-matrix-based EIV vaccine stimulated significant EIV-specific IFN-γ synthesis and EIV-specific single radial haemolysis (SRH) antibody. In conclusion, EIV vaccine adjuvanted with ISCOM-matrix stimulates both antibody and a cellular immune response in the horse.     

Influence of the Th2 immune response established by Nippostrongylus brasiliensis infection on the protection offered by different vaccines against Chlamydophila abortus infection

Chlamydophila abortus is the aetiological agent of enzootic abortion in small ruminants in which it infects the placenta to cause abortion during the last trimester of gestation. In a mouse model, a Th1 immune response involving IFN-gamma production and CD8+ T cells is necessary for the infection to be resolved. The authors previously demonstrated that infection with Nippostrongylus brasiliensis, a rodent gastrointestinal nematode extensively used in experimental models to induce Th2 responses, alters the specific immune response against C. abortus infection, increasing bacterial multiplication in liver and reducing specific IFN-gamma production. The aim of the present work was to clarify whether a Th2 immune response has any influence on the success of vaccination using both inactivated and attenuated vaccines. The results showed that the Th2 response established prior to vaccination did not influence the induction of protection offered by the vaccines. However, the effectiveness of this protective response can be altered, depending on the adjuvant employed in the inactivated vaccines, when the Th2 response is established after vaccination, just before challenge with C. abortus.     

Effect of oral rotavirus/iscom vaccines on immune responses in gnotobiotic lambs.

A comparison of the effect on the immune responses in gnotobiotic lambs was made between an iscom vaccine prepared from recombinant rotavirus VP6 protein, an inactivated rotavirus/iscom-matrix vaccine and a vaccine comprising inactivated rotavirus alone. All three vaccines induced immunological priming and some degree of protection was observed after a single oral dose. However, different immune responses were induced in response to a virulent infection. The group vaccinated with the rotavirus/iscom-matrix vaccine showed a Th2-like response characterised by rotavirus-specific antibodies and a down-regulation of IFNgamma in jejunal Peyer's patches. Both Th1-like and Th2-like immune responses were induced in the group receiving the VP6 vaccine as seen by significantly increased expressions of IFNgamma and IL-6 in the jejunal Peyer's patch together with an increased percentage of CD8+ T cells in the intestine and rotavirus-specific antibodies at mucosal surfaces. Iscom vaccines given orally have the ability to induce both Th1-like and Th2-like immune responses in a ruminant model.     

Vaccination of Egyptian fruit bats (Rousettus aegyptiacus) with monovalent inactivated rabies vaccine.

Twenty-six captive, adult Egyptian fruit bats (Rousettus aegyptiacus) were tested for the presence of rabies virus neutralizing antibodies (RVNA) using a rapid fluorescent focus inhibition test before and after vaccination. The bats were randomly assigned into three treatment groups: group A (n = 10) bats each received one 0.1-ml dose of monovalent inactivated rabies vaccine, group B (n = 10) bats each received two 0.1-ml doses of vaccine given 30 days apart, and group C (n = 6) bats remained unvaccinated. Plasma was collected from all bats before vaccination and on days 14, 30, 60, and 360. All bats were seronegative before vaccination, and all unvaccinated animals remained negative throughout the study. Rabies virus neutralization titers remained above 0.5 IU/ml from day 30 through day 360 for both vaccinated groups. Group B had significantly higher titers on day 60. This study demonstrated a measurable humoral immune response after vaccination with an inactivated rabies vaccine, with two doses producing a higher level of RVNA. This study confirms the feasibility of a rabies vaccination program for Egyptian fruit bats.     

Enhancing immune responses to inactivated foot-and-mouth virus vaccine by a polysaccharide adjuvant of aqueous extracts from Artemisia rupestris L.

BackgroundNew-generation adjuvants for foot-and-mouth disease virus (FMDV) vaccines can improve the efficacy of existing vaccines. Chinese medicinal herb polysaccharide possesses better promoting effects.ObjectivesIn this study, the aqueous extract from Artemisia rupestris L. (AEAR), an immunoregulatory crude polysaccharide, was utilized as the adjuvant of inactivated FMDV vaccine to explore their immune regulation roles.MethodsThe mice in each group were subcutaneously injected with different vaccine formulations containing inactivated FMDV antigen adjuvanted with three doses (low, medium, and high) of AEAR or AEAR with ISA-206 adjuvant for 2 times respectively in 1 and 14 days. The variations of antibody level, lymphocyte count, and cytokine secretion in 14 to 42 days after first vaccination were monitored. Then cytotoxic T lymphocyte (CTL) response and antibody duration were measured after the second vaccination.ResultsAEAR significantly induced FMDV-specific antibody titers and lymphocyte activation. AEAR at a medium dose stimulated Th1/Th2-type response through interleukin-4 and interferon-γ secreted by CD4⁺ T cells. Effective T lymphocyte counts were significantly elevated by AEAR. Importantly, the efficient CTL response was remarkably provoked by AEAR. Furthermore, AEAR at a low dose and ISA-206 adjuvant also synergistically promoted immune responses more significantly in immunized mice than those injected with only ISA-206 adjuvant and the stable antibody duration without body weight loss was 6 months.ConclusionsThese findings suggested that AEAR had potential utility as a polysaccharide adjuvant for FMDV vaccines.     

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。     

Cytokine responses in camels (Camelus bactrianus) vaccinated with Brucella abortus strain 19 vaccine

In the present study, we determined the levels of cytokines produced by camel (Camelus bactrianus) peripheral blood mononuclear cells (PBMCs) in response to live attenuated Brucella abortus (B. abortus) S19 vaccine. Seven camels were vaccinated with commercial B. abortus S19 vaccine, and their cytokine responses were determined using a real-time PCR assay. Cytokine responses to B. abortus S19 were examined at 6 hr, 48 hr and 1, 2 and 3 weeks post-vaccination. Serological tests were performed to further confirm these immune responses. The results revealed that IFN-gamma and IL-6 were upregulated during the first week post-vaccination. Low level expressions of IL-1alpha, IL-1beta, TNFalpha and IL-10 and no expression of IL-2 and IL-4 were observed compared with the control camels. The findings showed that B. abortus stimulates cell-mediated immunity by directly activating camel Th1 cells to secrete IFN-gamma. This quantification of cytokine expression in camels is essential for understanding of Camelidae disease development and protective immune responses. This is the first report of in vivo camel cytokine quantification after vaccination.     

Antibody Responses Against Equine Influenza Virus Induced by Concurrent and by Consecutive Use of an Inactivated Equine Influenza Virus Vaccine and a Modified Live Equine Herpesvirus Type 1 Vaccine in Thoroughbred Racehorses

An inactivated equine influenza virus (EIV) vaccine and a live equine herpesvirus type 1 (EHV-1) vaccine are usually administered concurrently to Thoroughbred racehorses in Japan. The objective of this study was to evaluate whether concurrent administration of an inactivated EIV vaccine and a live EHV-1 vaccine in Thoroughbred racehorses influences the antibody response against EIV. We compared the antibody response against EIV in horses administered both vaccines on the same day (Group A; n = 27) and the response in horses administered an inactivated EIV vaccine first and then a live EHV-1 vaccine 1–2 weeks later (Group B; n = 20). In both groups, geometric mean hemagglutination inhibition (HI) titers against A/equine/Ibaraki/1/2007 and A/equine/Yokohama/aq13/2010 increased significantly after EIV vaccination. However, the percentage of horses that showed a twofold increase or greater in HI titers against A/equine/Yokohama/aq13/2010 was significantly higher in Group B (75%) than in Group A (37%; P = .02). These results suggest that the concurrent use of an inactivated EIV vaccine and a live EHV-1 vaccine reduced the immune response against EIV to some extent, and it would be better to use these vaccines consecutively, especially for naïve horses or horses whose vaccination history is incomplete.     

Role of polymorphonuclear neutrophils (PMNs) and NK cells in the protection conferred by different vaccines against Chlamydophila abortus infection

Ovine enzootic abortion (OEA) is caused by Chlamydophila abortus, an intracellular bacterium which acts by infecting the placenta, causing abortion in the last term of gestation. The main prevention strategy against OEA is the vaccination of flocks. An effective vaccine against C. abortus must induce a Th1-like specific immune response, which is characterized by the early production of IFN-gamma and the activation of CD8(+)T cells. Moreover, vaccine effectiveness could be modulated by the functioning of the innate immunity. The purpose of this study was to ascertain how polymorphonuclear neutrophils (PMNs) and NK cells might influence vaccine-induced protection. The live attenuated 1B vaccine and two inactivated experimental vaccines, adjuvated with aluminium hydroxide (AH) or QS-21 (QS), were used in PMN-depleted or NK cell-depleted mice. For PMN depletion, RB6-8C5 monoclonal antibody, which recognizes GR1(+) receptors (Robben, P.M., LaRegina, M., Kuziel, W.A., Sibley, L.D. 2005. Recruitment of Gr-1(+) monocytes is essential for control of acute toxoplasmosis. The Journal of Experimental Medicine 201, 1761-1769.) was used, while for NK cell-depletion the anti-asialo GM1 polyclonal antibody was used. The depletion of PMNs caused 100% mortality in non-vaccinated mice (NV) and 60% mortality in the AH-vaccinated mice by day 10 p.i., while both groups showed a significant increase in their bacterial burden in the liver by day 4 p.i. The depletion of NK cells caused mortality only in the NV group (50% by day 10 p.i.), although this group and the 1B vaccinated mice showed an increased bacterial burden in the liver at day 4 p.i. Our results suggest that the importance of PMNs in inactivated vaccines depends on the adjuvant chosen. The results also demonstrated that the importance of NK cells is greater in live vaccines than in inactivated vaccines.     

五加芪粉对鸡免疫新流二联灭活疫苗免疫效果的影响

研究了五加芪粉对鸡免疫新流二联灭活苗后免疫功能的影响。180只20日龄鸡随机分为6组,每组30只,分别为五加芪粉高、中、低剂量组、阳性药物对照组、免疫对照组、空白对照组。于免疫后第10、17、24、31和38天检测血清中新城疫抗体水平、禽流感抗体水平、IL-2含量,并于免疫后第38天检测免疫器官指数。结果表明,0.2、0.5 g/L的五加芪粉组与免疫对照组相比,显著或极显著的提高了新城疫(ND)和禽流感(AI)抗体水平(P0.05,P0.01)。不同剂量的五加芪粉组和紫锥菊末组与免疫对照组相比,显著或极显著的提高了法氏囊指数、脾脏指数和胸腺指数(P0.05,P0.01)。免疫给药后第31天,0.2、0.5、1 g/L的五加芪粉组、紫锥菊末组与免疫对照组相比,显著或极显著的提高了IL-2的水平(P0.05,P0.01),以上实验结果初步表明五加芪粉可以提高鸡免疫新流二联灭活疫苗免疫效果,增强机体的免疫力。     

Comparison between a Live and an Inactivated Vaccine against Newcastle Disease in Village Chickens. A Field Study in Northern Malawi

A total of 156 chickens in two villages in Malawi were marked and sampled. One hundred and fifteen of these were vaccinated against Newcastle disease immediately after blood sampling, using the V4 heat-resistant strain applied by eye-drop in one village and the inactivated Newcavac vaccine in the other village. A second blood sample was collected 4 weeks after vaccination. The samples were examined using an indirect ELISA test kit. The titre group median ranged from 2 to 3 before vaccination. Both vaccines led to a positive immune response. Newcavac induced higher and more homogeneous titres compared with the V4 vaccine. There was also an increase in the median of the control group where V4 live vaccine had been applied. The differences between the median titres induced by V4, Newcavac and controls were statistically significant.     

传染性法氏囊病疫苗免疫母鸡后子代雏鸡外周血液的B细胞和免疫球蛋白含量变化

应用免疫SPA菌体花环法和间接ELISE法对传染性法氏囊病（IBD）疫苗免疫母鸡后，其子代雏鸡外周血液B细胞数量和IgG、IgM、IgA含量进行了动态研究。结果发现：IBD疫苗免疫后，其子代雏鸡外周血液B细胞数量和IgG、IgM、IgA含量均不同程度地高于未免疫的相应对照雏鸡，表明IBD疫苗免疫母鸡后，其子代雏鸡外周血液的体液免疫功能明显增强。而传染性法氏囊病超强毒株攻击子代雏鸡后，未免疫子代雏鸡外周血液的上述各项指标均明显低于疫苗免疫的子代雏鸡，这与IBDV感染雏鸡后，其体液免疫中枢器官法氏囊严重损害，淋巴细胞变性坏死等有关，也是导致感染雏鸡免疫抑制的基础。     

Oral administration of Salmonella enterica serovar Typhimurium expressing swine interleukin-18 induces Th1-biased protective immunity against inactivated vaccine of pseudorabies virus

Enhancing and/or modulating innate and adaptive immunity by cytokines appears to be greatly useful to provide effective protective immunity against infectious diseases. However, an effective delivery system for mass administration in livestock industry is needed because of limitations such as cost, labor, time, and protein stability. Here the immunomodulatory functions of swine interleukine-18 (swIL-18), known as IFN-γ-inducing factor (IGIF), were evaluated in a vaccination model of pseudorabies virus (PrV) using attenuated Salmonella enterica serovar Typhimurium as the oral delivery system. The oral administration of S. enterica serovar Typhimurium expressing swIL-18 prior to vaccination with inactivated PrV vaccine induced enhanced levels of serum PrV-specific IgG and its IgG2 isotype, compared to administration of S. enterica serovar Typhimurium harboring the empty vector. Furthermore, S. enterica serovar Typhimurium expressing swIL-18 mounted Th1-biased cellular immune responses against PrV antigen, as evaluated by the production of IFN-γ and IL-4 from peripheral blood mononuclear cells of piglets. Subsequently, Th1-biased immunity induced by S. enterica serovar Typhimurium expressing swIL-18 showed rapid response and rendered piglets displayed more alleviated clinical signs following the virulent PrV challenge. Also, this alleviation of clinical signs was further confirmed by the reduction of nasal excretion of PrV after challenge. The present study demonstrates the extended use of immunomodulatory functions of swIL-18 orally delivered by attenuated S. enterica serovar Typhimurium.     

猪圆环病毒2型感染对伪狂犬疫苗免疫应答的影响

为明确PCV2感染对伪狂犬(PR)疫苗免疫应答的影响,本研究采用阻断ELISA方法对单独接种猪PR疫苗组(A组)及PCV2人工感染3周后接种猪PR疫苗组(PA组)不同时相血清中的猪PR病毒gB抗体进行检测;同时对不同时相前腔静脉血进行CD4+/CD8+流式细胞术及血常规分析。结果表明,在PCV2感染后2周至5周间,A组白细胞含量均高于PA组,随后PA组白细胞恢复至与A组略高的正常水平;在整个实验中,除接种猪PR疫苗后1周(WPI)和9周(WPI)外的所有时相PA组的淋巴细胞含量均略高于A组;PCV2感染后可使记忆/激活Th细胞数量略有升高,幼稚型Th细胞含量的下降;PCV2感染后2周～7周PA组Tc细胞均高于A组,在9WPIPA组Tc细胞数量显著下降(p0.05);除9WPI外,A组的S/N值均低于PA组。结果表明,PCV2感染看降低机体产生针对PRVgB特异性抗体水平,而且在一定程度上降低了幼稚型Th细胞及Tc细胞含量。     

Influence of the Th2 immune response established by Nippostrongylus brasiliensis infection on the protection offered by different vaccines against Chlamydophila abortus infection

Chlamydophila abortus is the aetiological agent of enzootic abortion in small ruminants in which it infects the placenta to cause abortion during the last trimester of gestation. In a mouse model, a Th1 immune response involving IFN-γ production and CD8⁺ T cells is necessary for the infection to be resolved. The authors previously demonstrated that infection with Nippostrongylus brasiliensis, a rodent gastrointestinal nematode extensively used in experimental models to induce Th2 responses, alters the specific immune response against C. abortus infection, increasing bacterial multiplication in liver and reducing specific IFN-γ production. The aim of the present work was to clarify whether a Th2 immune response has any influence on the success of vaccination using both inactivated and attenuated vaccines. The results showed that the Th2 response established prior to vaccination did not influence the induction of protection offered by the vaccines. However, the effectiveness of this protective response can be altered, depending on the adjuvant employed in the inactivated vaccines, when the Th2 response is established after vaccination, just before challenge with C. abortus.