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Avian Marek's disease virus antibody ELISA kit Serum

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[#LSY-30016] Avian Marek's disease virus antibody ELISA kit Serum


LSY-30016 | Avian Marek's disease virus antibody ELISA kit Serum, 96 wells/kit
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(1) Inhibition of DNA-sensing pathway by Marek's disease virus VP23 protein through suppression of interferon regulatory factor 7 activation.[TOP]

Pubmed ID :30518647
Publication Date : //
The type I interferon (IFN) response is the first line of host innate immune defense against viral infection; however, viruses have developed multiple strategies to antagonize host IFN responses for efficient infection and replication. Here, we report that Marek's disease virus (MDV), an oncogenic herpesvirus, encodes VP23 protein as a novel immune modulator to block IFN-β activation induced by cyclic GMP-AMP synthase (cGAS) and stimulator of interferon genes (STING) in chicken fibroblasts and macrophages. VP23 overexpression markedly reduces viral DNA-triggered IFN-β production and promotes viral replication, while knockdown of VP23 during MDV infection enhances the IFN-β response and suppresses viral replication. VP23 selectively inhibits IFN regulatory factor 7 (IRF7) but not nuclear factor κB (NF-κB) activation. Furthermore, we found that VP23 interacts with IRF7 and blocks its binding to TANK-binding kinase 1 (TBK1), thereby inhibiting IRF7 phosphorylation and nuclear translocation, resulting in reduced IFN-β production. These findings expand our knowledge of DNA sensing in chickens and reveal a mechanism through which MDV antagonizes the host IFN response. Despite widespread vaccination, Marek's disease (MD) continues to pose major challenges for the poultry industry worldwide. MDV causes immunosuppression and deadly lymphomas in chickens, suggesting that this virus has developed a successful immune evasion strategy. However, little is known regarding the initiation and modulation of the host innate immune response during MDV infection. This study demonstrates that the cGAS-STING DNA-sensing pathway is critical for the induction of the IFN-β response against MDV infection in chicken fibroblasts and macrophages. An MDV protein, VP23, was found to efficiently inhibit the cGAS-STING pathway. VP23 selectively inhibits IRF7 but not NF-κB activation. VP23 interacts with IRF7 and blocks its binding to TBK1, thereby suppressing IRF7 activation and resulting in inhibition of the DNA-sensing pathway. These findings expand our knowledge of DNA sensing in chickens and reveal a mechanism through which MDV antagonizes the host IFN response.

Authors : Gao Li, Li Kai, Zhang Yu, Liu Yongzhen, Liu Changjun, Zhang Yanping, Gao Yulong, Qi Xiaole, Cui Hongyu, Wang Yongqiang, Wang Xiaomei,

(2) Suspension culture of Marek's disease virus and evaluation of its immunological effects.[TOP]

Pubmed ID :30518239
Publication Date : //
Marek's disease virus (MDV) is a cell-associated α-herpesvirus of chickens. It is difficult to perform its complete suspension culture. Therefore, MDV vaccines are currently produced using adherent primary chicken embryo fibroblasts and on a large scale is labor intensive and costly. In this study, the CVI988 strain of MDV was inoculated into chicken fibroblast cell line UMNSAH/DF-1 (DF-1) cultured by microcarrier suspension for proliferation experiment. Moreover, the effects of culture conditions, such as inoculation method, MOI, microcarrier concentration, and pH value, on the proliferation of MDV were investigated. The results demonstrated that the maximum viral load 64.76 ± 2.64 × 10 PFU/flask in a working volume of 100 mL could be obtained using synchronous cell seeding and inoculation method at an MOI of 0.02 and a microcarrier concentration of 5 g/L at pH 7.2. At the same time, the CVI988/DF-1 vaccines prepared by the microcarrier culture process and the traditional adherent cell culture process (CVI988/Rispense) were compared through animal experiments. We found a protective rate of 94.4% using the CVI988/DF-1 vaccine with specific pathogen free chickens that was equivalent to that of the commercial vaccine CVI988/Rispens (protection rate of 94.1%). In this study, the MDV CVI988/DF-1 vaccine prepared by the microcarrier suspension culture of DF-1 cells could provide effective immune protection for SPF chickens, providing a reference for the prevention and control of MD and further development of a large-scale bioreactor for producing the MD vaccine.

Authors : Wen Lianghai, Zhang Aiguo, Li Yanpeng, Lai Hanzhang, Li Huimin, Luo Qiong, Jin Shuangxing, Chen Ruiai,

(3) Establishment of xMAP for the simultaneous detection of antibodies to Newcastle disease virus and avian influenza virus.[TOP]

Pubmed ID :30476286
Publication Date : //
Using Luminex xMAP (x = analyte, MAP = multi-analyte profiling) technology, a serological method for the simultaneous detection of antibodies to Newcastle disease virus (NDV) and avian influenza virus (AIV) was established. Nano-magnetic beads coated with purified NDV protein and AIV nucleoprotein were incubated with serum samples. Using biotinylated rabbit anti-chicken IgY and streptavidin-R-phycoerythrin, the optical signals measured by a Luminex 200 detection system indicated the quantification of NDV or AIV antibodies in the serum. Specific pathogen-free (SPF) chicken serum was used as a negative control. The Luminex xMAP assay developed in this study demonstrated high specificity as there was no cross-reaction with antibodies to infectious laryngotracheitis virus, infectious bronchitis virus, infectious bursal disease virus, avian leukosis virus, and Marek's disease virus. The results from reproducibility experiments showed that intra-coefficients of variation were 3.36 and 9.23% and inter-coefficients of variation were 6.50 and 7.66% for NDV and AIV, respectively. The results also indicated that the Luminex xMAP assay was 16 times more sensitive for NDV antibody detection and 1,024 times more sensitive for AIV antibody detection compared to the enzyme-linked immunosorbent assay (ELISA). A total of 300 chicken serum samples were subjected to both Luminex xMAP assay and ELISA, showing the coincidence rates of 98.67 and 98% for NDV and AIV antibody detection, respectively. This study provides a new method for the simultaneous detection NDV and AIV antibodies in the serum with high specificity and sensitivity.

Authors : Wang Huanan, Cong Feng, Guan Jianchi, Xiao Li, Zhu Yujun, Lian Yuexiao, Huang Ren, Chen Meili, Guo Pengju,

(4) Transcripts of antibacterial peptides in chicken erythrocytes infected with Marek's disease virus.[TOP]

Pubmed ID :30463541
Publication Date : //
Chicken erythrocytes are involved in immunity through binding of toll-like receptors (TLRs) with their ligands to activate downstream signaling and lead to cytokine production in erythrocytes. Some avian β-defensins (AvBDs) are constitutively expressed in tissues and some others can be induced by various bacteria and viruses. However, the expression of AvBDs in erythrocytes has not yet been studied extensively.

Authors : Niu Sheng, Jahejo Ali Raza, Jia Fa-Jie, Li Xin, Ning Guan-Bao, Zhang Ding, Ma Hai-Li, Hao Wei-Fang, Gao Wen-Wei, Zhao Yu-Jun, Gao Shi-Min, Li Gui-Lan, Li Jian-Hui, Yan Fang, Gao Rong-Kun, Bi Yu-Hai, Han Ling-Xia, Gao George F, Tian Wen-Xia,

(5) A multiplex xTAG assay for the simultaneous detection of five chicken immunosuppressive viruses.[TOP]

Pubmed ID :30442149
Publication Date : //
Chicken anemia virus (CAV), avian reovirus (ARV), infectious bursal disease virus (IBDV), Marek's disease virus (MDV) and reticuloendotheliosis virus (REV) all cause immunosuppressive disease in birds through vertical or horizontal transmission. Mixed infections with these immunosuppressive pathogens lead to atypical clinical signs and obstruct accurate diagnoses and epidemiological investigations. Therefore, it is essential to develop a high-throughput assay for the simultaneous detection of these immunosuppressive viruses with high specificity and sensitivity. The aim of this study was to establish a novel method using a RT-PCR assay combined with fluorescence labeled polystyrene bead microarray (multiplex xTAG assay) to detect single or mixed viral infections.

Authors : Cong Feng, Zhu Yujun, Wang Jing, Lian Yuexiao, Liu Xiangnan, Xiao Li, Huang Ren, Zhang Yu, Chen Meili, Guo Pengju,

(6) A double recombinant herpes virus of turkeys for the protection of chickens against Newcastle, infectious laryngotracheitis and Marek's diseases.[TOP]

Pubmed ID :30404540
Publication Date : //
A double recombinant strain of herpes virus of turkeys (HVT) was constructed that contains the fusion (F) gene from Newcastle disease virus (NDV) and the gD plus gI genes from infectious laryngotracheitis virus (ILTV) inserted into a non-essential region of the HVT genome. Expression of the F protein was controlled by a human cytomegalovirus promoter, whereas expression of gD plus gI was driven by an ILTV promoter. The double recombinant vaccine virus (HVT-NDV-ILT) was fully stable genetically and phenotypically following extended passage in cell culture and infection of chickens. Safety of the vaccine virus was confirmed by overdose and backpassage studies in specific-pathogen-free chickens. Chickens vaccinated with a single dose of HVT-NDV-ILT administered by the in ovo route were highly protected from challenge with the velogenic NDV (GB Texas), ILTV (LT 96-3) and Marek's disease virus (GA 5) strains (97%, 94% and 97%, respectively). Similarly, chickens vaccinated with a single dose by subcutaneous (SC) route at 1 day of age were highly protected from challenge with the same three viruses (100%, 100%, and 88%, respectively). The protection level of a single dose given by in ovo or SC route against challenge with a virulent Marek's disease virus strain demonstrates that insertion of multiple genes from two different pathogens within the HVT genome had no adverse effect on the capacity of HVT to protect against Marek's disease. These results demonstrate that HVT-NDV-ILT is a safe and efficacious vaccine for simultaneous control of NDV, ILTV and Marek's diseases.

Authors : Gergen Linda, Cook Stephanie, Ledesma Brooke, Cress Wade, Higuchi Deb, Counts Diane, Cruz-Coy Julio, Crouch Colin, Davis Phillip, Tarpey Ian, Morsey M,

(7) Virus-Induced Immunosuppression in Chickens.[TOP]

Pubmed ID :30339511
Publication Date : //
A healthy immune system is a cornerstone for poultry production. Any factor diminishing the immune responses will affect production parameters and increase cost. There are numerous factors, infectious and noninfectious, causing immunosuppression (IS) in chickens. This paper reviews the three viral diseases that most commonly induce IS or subclinical IS in chickens: Marek's disease virus (MDV), chicken infectious anemia virus (CIAV), and infectious bursal disease virus (IBDV), as well as the interactions among them. MDV-induced IS (MDV-IS) affects both humoral and cellular immune responses. It is very complex, poorly understood, and in many cases underdiagnosed. Vaccination protects against some but not all aspects of MDV-IS. CIAV induces apoptosis of the hemocytoblasts resulting in anemia, hemorrhages, and increased susceptibility to bacterial infections. It also causes apoptosis of thymocytes and dividing T lymphocytes, affecting T helper functions, which are essential for antibody production and cytotoxic T lymphocyte (CTL) functions. Control of CIAV is based on vaccination of breeders and maternal antibodies (MAbs). However, subclinical IS can occur after MAbs wane. IBDV infection affects the innate immune responses during virus replication and humoral immune responses as a consequence of the destruction of B-cell populations. Vaccines with various levels of attenuation are used to control IBDV. Interactions with MAbs and residual virulence of the vaccines need to be considered when designing vaccination plans. The interaction between IBDV, CIAV, and MDV is critical although underestimated in many cases. A proper control of IBDV is a must to have proper humoral immune responses needed to control CIAV. Equally, long-term control of MDV is not possible if chickens are coinfected with CIAV, as CIAV jeopardizes CTL functions critical for MDV control.

Authors : Gimeno I M, Schat K A,

(8) Unraveling the role of B cells in the pathogenesis of an oncogenic avian herpesvirus.[TOP]

Pubmed ID :30337483
Publication Date : //
Marek's disease virus (MDV) is a highly oncogenic alphaherpesvirus that causes immunosuppression, paralysis, and deadly lymphomas in chickens. In infected animals, B cells are efficiently infected and are thought to amplify the virus and transfer it to T cells. MDV subsequently establishes latency in T cells and transforms CD4 T cells, resulting in fatal lymphomas. Despite many years of research, the exact role of the different B and T cell subsets in MDV pathogenesis remains poorly understood, mostly due to the lack of reverse genetics in chickens. Recently, Ig heavy chain J gene segment knockout (JH-KO) chickens lacking mature and peripheral B cells have been generated. To determine the role of these B cells in MDV pathogenesis, we infected JH-KO chickens with the very virulent MDV RB1B strain. Surprisingly, viral load in the blood of infected animals was not altered in the absence of B cells. More importantly, disease and tumor incidence in JH-KO chickens was comparable to wild-type animals, suggesting that both mature and peripheral B cells are dispensable for MDV pathogenesis. Intriguingly, MDV efficiently replicated in the bursa of Fabricius in JH-KO animals, while spread of the virus to the spleen and thymus was delayed. In the absence of B cells, MDV readily infected CD4 and CD8 T cells, allowing efficient virus replication in the lymphoid organs and transformation of T cells. Taken together, our data change the dogma of the central role of B cells, and thereby provide important insights into MDV pathogenesis.

Authors : Bertzbach Luca D, Laparidou Maria, Härtle Sonja, Etches Robert J, Kaspers Bernd, Schusser Benjamin, Kaufer Benedikt B,

(9) The emergence of the infection of subgroup J avian leucosis virus escalated the tumour incidence in commercial Yellow chickens in Southern China in recent years.[TOP]

Pubmed ID :30248239
Publication Date : //
A total of 81 clinical cases of suspected tumours were submitted to our laboratory from Yellow chicken farms in southern China during the years 2010 through 2017. The tumour-like tissue samples were closely examined for common oncogenic avian viruses in cell culture and further analysed using polymerase chain reaction (PCR). During 2010-2012, Marek's disease virus (MDV) mono-infection was found to be the dominant cause of the tumour incidences (52.4%, 11/21) followed by co-infection of MDV+ALVs (19.1%, 4/21). Starting from the year 2013 the mono-infection of avian leucosis virus subgroup J (ALV-J) became the dominant agent of the tumour cases (83.3%, 5/6). During the most recent four years (2014-2017), co-infections involving ALV-J and MDV or between ALV subgroups have increased (23.4% and 18.5%, respectively), but each of the co-infections was still slightly lower than the ALV-J mono-infection incidence (33.3%). In contrast to the dominant MDV mono-infection cases before 2013, more recently, the emerging ALV-J mono-infection and ALV-J co-infections were largely responsible for the occurrence of avian virus-induced tumour incidences in the commercial local Yellow breeds of chickens in southern China. These results indicate that eradication measures against ALV on all chicken farms, especially on farms with the Yellow chickens, ought to be enhanced to reverse this trend.

Authors : Li Haijuan, Wang Peikun, Lin Lulu, Shi Mengya, Gu Zhanming, Huang Teng, Mo Mei-Lan, Wei Tianchao, Zhang Huanmin, Wei Ping,

(10) Development of a sensitive and specific xMAP assay for detection of antibodies against infectious laryngotracheitis and bronchitis viruses.[TOP]

Pubmed ID :30241540
Publication Date : //
A serological method to simultaneously detect antibodies against infectious laryngotracheitis virus (ILTV) and infectious bronchitis virus (IBV) is imperative for the differential diagnosis and evaluation of antibodies titers after vaccination.

Authors : Wang Huanan, Cong Feng, Guan Jianchi, Xiao Li, Zhu Yujun, Lian Yuexiao, Huang Ren, Chen Meili, Guo Pengju,