Mucosal immune responses elicited by wild-bird-origin avian paramyxovirus-type 1 isolates in chickens

Local immunity in the respiratory tract of the chicken. II. The secretory immune response to newcastle disease virus and the role of IgA

Objective: Isolation, serological and molecular identification and differentiation between velogenic and lentogenic of Newcastle disease virus (NDV) strains. Design: Descriptive study. Procedures: In this study, A total of 24 Pooled samples were collected from Dakahlia, Egypt, from 2017 to 2018 .The virus was isolated in 10 days old (ECEs) and serologically identified in third egg passage allantoic fluid by Haemagglutination (HA), Haemagglutination inhibition test (HIT) and agar gel precipitation test (AGPT). Molecular confirmation was done by reverse transcriptase-polymerase chain reaction (RT-PCR). Genetic diversity between velogenic and lentogenic NDV strains was done by RT-PCR amplification of 254bp F gene fragment from velogenic NDVs and sequence analysis of 362bp NDV F gene fragment. Results: Out of 24 tested samples, 20 samples were collected from allontoic fluid of 3rd passage positive by HA test, 18 samples by HIT, 16 samples by AGPT and 18 isolates were positive with RT-PCR amplification of 362bp F gene fragment. LaSota strain gave positive results in HA, HIT, AGPT and RT-PCR amplification of 362bp fragment. Interestingly, 254bp fragment failed to be amplified from LaSota strain. The phylogenic analysis revealed that our isolates were related to Egyptian velogenic genotype VII NDVs. The deduced amino acid sequence of the F protein cleavage site of our isolates was characteristic of velogenic NDVs (112R-R-Q-K-R↓F117) while LaSota strain had characteristic lentogenic NDVs cleavage site (112G-R-Q-G-R↓L117). Conclusion and clinical relevance: RT-PCR and sequence analysis in this study can be used for differentiation between velogenic and lentogenic NDV strains.

Selective isolation of avian influenza virus from cloacal samples containing mixed infection of avian influenza and Newcastle disease viruses

Four isolants of Newcastle disease virus (NDV) obtained from free-flying Canada geese in the Atlantic flyway were characterized and compared with the B1, LaSota, Roakin, and Texas-GB strains of NDV. The 4 isolants were identified as lentogenic strains on the basis of embryo mean death times of greater than 100 hours and relative lack of pathogenicity for 1-day-old and 3-week-old chickens. The hemagglutinin of 2 of the isolants was stable at 56 C for 15 minutes, 1 for 30 minutes, and the other for at least 2 hours. When held at 4 C for 24 hours, none of the 4 isolants could be eluted from chicken erythrocytes. All 4 of the isolants produced plaques in chicken embryo fibroblast cell cultures without additives, whereas the B1 and LaSota strains did not. The 4 lentogenic NDV isolants from Canada geese differed in many respects from currently used commerical vaccine strains (B1 and LaSota).

BackgroundInfectious diseases including Newcastle disease (ND) impair poultry productivity and represent a significant burden to sustainable agriculture in Nigeria. This study aimed to investigate the active circulation and seroprevalence of Newcastle disease virus (NDV) caused by virulent forms of avian paramyxovirus type-1 (APMV-1) in poultry at live bird markets (LBMs) across Nigeria.MethodsA cross-sectional study of 18 LBMs was conducted within three states in Nigeria (Kano, Oyo, and Abuja). Paired swab and tissue samples (n = 413) were collected from birds on FTA cards and tested for APMV-1 using real-time reverse transcription polymerase chain reaction (rRT-PCR). A subset of rRT-PCR-positive samples were selected for whole genome sequencing based on the originating species (chicken, duck, geese), date, and market location to provide a broad range of isolates for characterisation. Blood samples (n = 405) were also collected from birds and the seroprevalence of APMV-1 antibodies was measured using an enzyme-linked immunosorbent assay (ELISA).ResultsAPMV-1 RNA was detected in 21.5% (89/413) of samples from LBMs by rRT-PCR. At least one APMV-1 positive sample was detected in 55.6% (10/18) of LBMs. The largest proportion of APMV-1-positive markets was in Kano (83.3%, 5/6), whereas the lowest was in Oyo (16.7%, 1/6). Assessment of genetic data demonstrated that genotype XIV.2 APMV-1 was circulating within Nigeria with the viruses detected clustering closely with other Nigerian isolates described previously. The seroprevalence of APMV-1 in birds was 45.9% (186/405) and 94.4% (17/18) of LBMs had at least one APMV-1 seropositive sample (i.e., with at least one APMV-1-antibody-positive bird). The LBMs in Kano had the lowest seroprevalence (88.3%, 5/6).ConclusionsThis study demonstrated that APMV-1 continues to circulate in LBMs in Nigeria. LBM traders, poultry producers, and related industry and policy stakeholders should be aware of the occurrence of APMV-1 and how ND may negatively impact upon poultry production and the livelihoods of poultry farmers and LBM traders. Training initiatives aimed at improving the knowledge of APMV-1 infection and improvements in biosecurity practises and the role of disease mitigation through vaccination are required to reduce the impact of this threat to food security.

Newcastle disease virus (NDV) is a member of the genus Avulavirus of the Paramyxoviridae family of enveloped negative-stranded RNA viruses. The virus causes respiratory, neurological, or enteric disease in many species of birds, resulting in significant losses to the poultry industry worldwide. Strains of the virus are classified into three pathotypes based on the severity of disease in chickens. Avirulent strains that produce mild or asymptomatic infections are termed lentogenic, whereas virulent strains are termed velogenic. Strains of intermediate virulence are termed mesogenic. The envelope of NDV virions contains two types of glycoproteins, the hemagglutinin-neuraminidase (HN) and fusion (F) proteins. HN mediates three functions: 1) virus attachment to sialic acid-containing receptors; 2) neuraminidase activity that cleaves sialic acid from progeny virions to prevent self-aggregation; and, 3) complementation of the F protein in the promotion of fusion. Though it is widely accepted that cleavage of a fusion protein precursor is the primary determinant of NDV virulence, it is not the sole determinant. At least two other proteins, HN and the V protein, contribute to virulence. The V protein possesses interferon (IFN) antagonistic activity. The long-range goal of these studies is to understand the roles of HN and V in the differential virulence patterns exhibited by members of the NDV serotype. The first aim is to compare the IFN antagonistic activity of the V protein from a lentogenic and a mesogenic strain of the virus. The results of this study demonstrate that the V protein of the mesogenic strain Beaudette C (BC) exhibits greater IFN antagonistic activity than that of the lentogenic strain La Sota. Hence, the IFN antagonistic activities of the two V proteins correlate with their known virulence properties. Comparison of the C-terminal regions of La Sota and BC V proteins revealed four amino acid differences. The results demonstrate that the IFN antagonistic activity of La Sota V increases when any one of these residues is mutated to the corresponding residue in BC V. Conversely, the IFN antagonistic activity of BC V decreases when any one of these four residues is mutated to the corresponding residue in La Sota V. However, no single residue accounts for the difference in IFN antagonistic activity between the two V proteins. Also, analysis of La Sota V and BC V proteins with multiple mutations in these positions revealed that the four residues are collectively responsible for the difference in the IFN antagonistic activity of the two V proteins. Finally, characterization of chimeric La Sota/BC V proteins showed that the N-terminal region also contributes to the IFN antagonistic activity of V. Contrary to an earlier report, results described here demonstrate that the NDV V protein does not target STAT1 for degradation. However, both La Sota and BC V proteins target interferon regulatory factor (IRF)-7 for degradation and promote the conversion of full-length IRF-7 to a…

3100 Background: Newcastle disease virus (NDV), an avian paramyxovirus, is tumor selective and oncolytic by induction of apoptosis. Preclinical and clinical studies in patients with glioblastoma (GBM) using NDV demonstrated occasional clinical benefits with no major side effects. Limitations to the use of NDV as virotherapy of GBM is the inefficient delivery into cancer cells in the brain. Methods: Mesenchymal stromal cells (MSCs) can migrate towards cancer cells. We examined potential delivery of oncolytic effect of NDV (MTH-68/H) against glioma cell lines and glioma stem cells (GSCs) and the ability of MSCs to deliver NDV to glioma cells and GSCs in culture. Results: NDV induced a dose-dependent cell death in the glioma cells U87, A172 and U251 with maximal effects at 10 MOI. In contrast, we found only small level of apoptosis or changes in self-renewal in three GSCs infected with NDV. We found that MSCs derived from bone marrow, adipose tissue and cord were successfully infected by NDV and were able to deliver the virus to co-cultured glioma cells and GSCs. In addition, treatment of glioma cells and GSCs with culture supernatant of infected MSCs increase apoptosis of glioma cells as compared to the effect of direct infection of glioma cells. Moreover, the culture supernatants of the infected MSCs induced cell death in GSCs that were resistant to the oncolytic effect of NDV, suggesting that factor(s) secreted by the infected MSCs sensitized the glioma cells and GSCs to the cytotoxic effects of NDV. Using antibody array and ELISA we identified TRAIL as the factor secreted from infected MSCs. Indeed, treatment of infected glioma cells with TRAIL increased the cytotoxic effect of NDV and sensitized GSCs to the oncolytic effects of NDV. Conclusions: MSCs can be employed to deliver NDV to GBM. In addition, MSCs can also sensitize glioma cells and GSCs to oncolysis by NDV. Considering the resistance of GSCs to chemotherapy and radiation therapy, treatment of GBM with MSC-mediated targeted oncolytic NDV may provide a new clinical tool for treatment of GBM and eradication of GSCs.

Newcastle disease virus: virus replication in the Harderian gland stimulates lacrimal IgA; the yolk sac provides early lacrimal IgG

We compared detection of infectious bronchitis virus (IBV) by quantitative RT-PCR (qRT-PCR) in tears and trachea of IBV-infected chickens and found that quantitative detection of IBV RNA in tears is more sensitive than in tracheal homogenates. Furthermore, we demonstrated that IBV contained in chicken lachrymal fluid is infectious and that tears of IBV-infected chickens can be used to infect naive chickens. We compared the immune responses to IBV in the Harderian gland and cecal tonsils of immunocompetent chickens and chickens infected with chicken anemia virus (CAV) and/or infectious bursal disease virus (IBDV). Flow cytometry analyses of lymphocytes in Harderian glands and cecal tonsils indicated that the relative abundance of IgM+ B cells in the Harderian glands and cecal tonsils following exposure to IBV in combination with immunosuppressive viruses was reduced compared to chickens infected with IBV alone. CAV, but not IBDV, reduced the CD4+/CD8+ T cell ratios compared to chickens infected with IBV alone. Enzyme-linked immuno-spot forming assays on cells in the Harderian glands and cecal tonsils of IBV-infected chickens indicated that maximum IBV-specific IgA-secreting cell responses were reduced in chickens infected with CAV. IBDV co-infected chickens displayed a delayed IgA response to IBV. Thus immunosuppressive viruses reduced B cells and T helper cells in the Harderian glands and cecal tonsils in response to IBV, and slowed the kinetics and/or reduced the magnitude of the mucosal immune response against IBV. We have shown for the first time that CAV affects pathogen-specific B cell responses in a mucosal effector site.

Newcastle Disease Virus (NDV) has emerged as a promising oncolytic viral therapy for various human cancers; however, its effectiveness against cholangiocarcinoma (CCA) remains unexplored. This study presents the capability of the lentogenic LaSota strain of NDV to eliminate two CCA cell lines, KKU-055 and KKU-100, as well as the potential molecular mechanisms underlying this effect. Comprehensive transcriptome analysis revealed alterations in gene expression within several pathways in CCA cells following exposure to the LaSota strain NDV, including those involved in TNF-alpha signaling via NF-kB, interferon alpha response, apoptosis, and IL-6/JAK/STAT3 signaling pathways. We remarkably observed a contrasting alteration in the expression of CXCR4, GRAMD1B, IGFBP4, and TGM2 genes in KKU-055 and KKU-100 cells. In addition, gene network analysis highlighted CCNA2, CDK1, DDX58, DHX58, EXO1, GBP1, IFIH1, IFIT1, IFIT2, IFIT3, IRF7, ISIG15, MX1, OAS1, OAS2, PARP9, TOP2A and XAF1 as potential hub genes influencing the response of CCA cells to NDV LaSota strain. Our findings offer evidence supporting the promise of NDV-based therapies as potential strategies for eliminating CCA cells.

Background and Aim:Newcastle disease (ND) and avian influenza (AI) are two devastating diseases of poultry, which cause great economic losses to the poultry industry and disrupt food security in our country. The use of ND-AI inactive bivalent vaccine is very effective and economical to prevent and control ND and AI disease. Bivalent ND LaSota-AI H9N2 vaccine is not yet available in Indonesia. The inactivated vaccines used in poultry industry often require oil adjuvant to elicit a sufficient immune response. This study aimed to develop the bivalent inactive vaccines containing ND LaSota and AI H9N2 Sidrap isolate which are local isolates as poultry vaccine candidates, and formulated with two different commercial adjuvants, then compared.Materials and Methods:Two vaccines bivalent were prepared by emulsifying inactivated Newcastle disease virus (LaSota strain) and AI H9N2 Sidrap isolate viruses with Marcol white mineral oil and Montanide ISA70 adjuvants. Both of bivalent vaccines were tested for safety (physical and histopathological at the injection site) and efficacy in specific-pathogen-free chickens. Parameters used for the evaluation of the efficacy were immunogenicity by hemagglutination inhibition and protection percentage.Results:Both bivalent vaccines are safe to use. Post-vaccination (PV) immune response was observed using a hemagglutination inhibition test at 2, 3, 4, 5, 6, 7, and 8 weeks of PV. The bivalent vaccine B gives a better immune response to ND at 2, 3, and 4 weeks of PV (p<0.05) compared to the bivalent vaccine A, but in 5, 6, 7, and 8 weeks, the PV does not show differences in the immune response. The immune response to AI H9N2 showed differences at weeks 2 and 3 PV (p<0.05) with the bivalent vaccine B indicated higher immunity. A single immunization with both bivalent vaccines induces 100% protection in chickens that have been vaccinated against the deadly challenge with the virulent ND virus.Conclusion:Both of bivalent vaccines are safe to use and provide good efficacy against virulent ND viruses, but bivalent vaccine B (with Montanide ISA70 adjuvant) shows better immune response than bivalent vaccine A (Marcol white mineral oil adjuvant).

It was the aim of this study to determine the interaction between the Newcastle disease (ND) and infectious bursal disease (IBD) vaccines used to control these two important viral infections greatly affecting poultry industry worldwide. The commercially available vaccines in the Sudan were used. Haemagglutination inhibition (HI) and agar gel immunodiffusion (AGID) tests were employed to measure the Ab titres in chicks sera to ND and IBD respectively. Generally, IBD vaccine was reported to have adverse effect on the ND vaccine whereas the reverse was not true. The results obtained also revealed that better Ab response s against NDV were detected when ND vaccine was administered before IBD vaccine. The deleterious effect of IBD vaccine on Ab levels against NDV antigens was slightly (p< 0.05) low when IBD vaccine is administered at two weeks as compared to three weeks of chicken age. No variations in the Ab titres when chicks were boostered with ND vaccine containing LaSota or Komorov strain of the virus at 4 weeks were observed. However, slightly (p< 0.01) better Ab responses were noted for LaSota over Komorov strain. It was, therefore, concluded that vaccination of chicks with ND vaccine containing LaSota strain of the virus when they were 10 days followed by vaccination with IBD vaccine at two weeks and boostering with the same ND vaccine yielded better Ab responses but slightly lower protection levels.

Newcastle disease (ND) is one of the most important avian diseases with considerable threat to the productivity of poultry all over the world. The disease is associated with severe respiratory, gastrointestinal, and neurological lesions in chicken leading to high mortality and several other production related losses. The aetiology of the disease is an avian paramyxovirus type-1 or Newcastle disease virus (NDV), whose isolates are serologically grouped into a single serotype but genetically classified into a total of 19 genotypes, owing to the continuous emergence and evolution of the virus. In Nigeria, molecular characterization of NDV is generally very scanty and majorly focuses on the amplification of the partial F gene for genotype assignment. However, with the introduction of the most objective NDV genotyping criteria which utilize complete fusion protein coding sequences in phylogenetic taxonomy, the enormous genetic diversity of the virus in Nigeria became very conspicuous. In this review, we examine the current ecological distribution of various NDV genotypes in Nigeria based on the available complete fusion protein nucleotide sequences (1662 bp) in the NCBI database. We then discuss the challenges of ND control as a result of the wide genetic distance between the currently circulating NDV isolates and the commonest vaccines used to combat the disease in the country. Finally, we suggest future directions in the war against the economically devastating ND in Nigeria.

Turkeys given primary vaccination with an inactivated Newcastle (ND) vaccine failed to respond satisfactorily in the serumvirus neutralization test and challenge of immunity via the respiratory tract. Revaccination did not effect a satisfactory anamnestic response. LaSota strain Newcastle disease virus (NDV), however, applied by eyedrop or in drinking water, stimulated excellent immunity by the same criteria. Response was unsatisfactory when LaSota NDV was given intramuscularly. Bl-strain NDV given by eyedrop or drinking water induced good immunity but of lower magnitude than from the LaSota strain. Cell-culture ND vaccine resulted in relatively poor protection from infection of the respiratory tract, and the virus-neutralizing antibody titers were relatively low or marginal. The potential use of lentogenic NDV strains in turkey vaccination programs was discussed.

This research was carried out at the Department of Microbiology, University of Agricultur e Faisalabad, Pakistan in order to study humoral immune response to Newcastle disease vaccine (Lastoa strain) in broilers. For this purpose, 30-day-old broiler chicks were procured from Olympia chicks. The chicks were divided into three groups containing 10 chicks each. The HI-Ab-titre against ND virus in the control group showed an overall Ab-titre with GMT=3.99.The HI-Ab-response in broilers chicks with single dose of Lastoa vaccine ranged from 1:8 to 1:32.This group showed an overall HI-Ab- titer with GMT=13.931. While the HI-Ab titer against ND virus was found to be in the range of 1:16 to 1:128 in broilers chicks which were vaccinated with the 1 dose of ND virus (LaSota strain) at the age of 2 weeks followed by a booster dose with st the same strain at 3 weeks of age. The overall HI-Ab-titre against LaSota strain of ND virus came out to be GMT=55.692. During present investigation a significant increase in HI-Ab-titre (GMT) against ND virus was observed in broilers from single to boostered vaccination group.