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Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken.

ABSTRACT: Background and Aim:Newcastle disease (ND) is considered one of the most important poultry diseases with chicken morbidity and mortality rates up to 100%. Current vaccination programs allow the use of live attenuated vaccines in the field to protect against the disease, which alone is inefficient and requires repeat booster doses. Toll-like receptor agonists (e.g., lipopolysaccharide [LPS]) as adjuvants are the ones, most extensively studied and have shown to be very promising in delivering a robust balanced immune response. In the present study, we have evaluated the potential of LPS to elicit a strong immune response with respect to the elicitation of both Th1 (cell-mediated) and Th2 (humoral) immune arms. Materials and Methods:A total of 72 apparently healthy 1-day-old indigenous unvaccinated chicks were randomly divided into six experimental Groups A to F (n=12). At 8-week of age chicks in Group A, C, and E were vaccinated with live attenuated La Sota strain ND vaccine along with LPS, bovine serum albumin, and normal saline solution, respectively, and those in Group B, D, and E were kept separately without vaccination. Sampling was done on days 0, 1, 3, 7, 14, 21, 35, and 60 after vaccination. After vaccination and respective adjuvant application, Th1 and Th2 cytokine expression were measured in mRNA of both blood and tissue samples. Results:The results were validated by, hemagglutination inhibition and enzyme-linked immunosorbent assay tests, to check for the humoral as well as cell-mediated immune response in blood serum levels. The results showed an increase in mRNA expression of the Th1 biased cytokines in Group A (LPS+NDV) as compared to the control groups. Similar mRNA expression pattern was seen in blood as well as tissue samples. Validation of results also indicates an increase in Cell-mediated Immunity as well as a humoral immune response in Group A (LPS+NDV). Conclusion:The results of the study provided enough evidence to consider LPS as a potential vaccine adjuvants candidate against ND in chicken.

SUBMITTER: Bhardwaj R

PROVIDER: S-EPMC5960781 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken.

Bhardwaj Rabia R Verma Ramneek R Deka Dipak D Dubey P P PP Arora J S JS Sethi R S RS Tolenkhomba T C TC Mukhopadhyay C S CS

Veterinary world 20180409 4

<h4>Background and aim</h4>Newcastle disease (ND) is considered one of the most important poultry diseases with chicken morbidity and mortality rates up to 100%. Current vaccination programs allow the use of live attenuated vaccines in the field to protect against the disease, which alone is inefficient and requires repeat booster doses. Toll-like receptor agonists (e.g., lipopolysaccharide [LPS]) as adjuvants are the ones, most extensively studied and have shown to be very promising in deliveri ...[more]

PMID: 29805207

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Project description:Newcastle disease virus (NDV) belongs to genus Avulavirus and family Paramyxoviridae. There are thirteen serotypes named APMV-I (Avian Paramyxovirus-I) to APMV-13 of which NDV has been designated as APMV-1. The disease has been reported worldwide affecting both domestic and wild avian species. Morbidity and mortality rates up to 100% have been reported in cases of unvaccinated flocks. Stringent vaccination schedule is practiced in endemic/disease prone areas in order to prevent the disease. Despite this, NDV outbreaks have been reported even in cases of vaccinated populations. In this study we describe detailed pathological and molecular investigation that were undertaken in an organized poultry farm from Bareilly region, Uttar Pradesh, India, involving layer flocks which succumbed to ND outbreak in spite of following strict vaccination protocol. The mortality rate ranged from 76.80 to 84.41% in different flocks with an average mortality of 79.50%. Necropsied birds had gross lesions suggestive of viscerotropic ND including petechial hemorrhages on the proventricular tips, intestinal lumen with necrotic areas covered with hemorrhages, hemorrhagic cecal tonsils, para-tracheal edema and mottling of spleen. The characteristic histopathological lesions were mainly seen in the blood vessels and lymphoid tissues. Vascular changes characterized by congestion, edema, and hemorrhage were found in majority of the organs. Lymphocytolysis in spleen and cecal tonsils was evident. Immunohistochemical studies revealed positive signals mostly in macrophage and lymphocytes. PCR assay was done to confirm the NDV genome, which revealed an amplicon size of 356 bp. The phylogenetic analysis revealed the resemblance of the present isolate (ADS01) with class II genotype NDV XIIIA. The isolate belonged to velogenic NDV as the Minimum Lethal Dose (MLD) and Mean Death Time (MDT) for the present isolate were 10-8 and 41 h, respectively. Thus this study clearly demonstrates that in spite of strict vaccination regime and biosecurity procedures, ND continues to be rampant. Hence it is important to effectively administer the present vaccine in addition to strains matching to the field isolates to provide longer and optimal protection against spreading of virus by means of reducing the extent of viral shedding.

Project description:BackgroundExposure to heat stress suppresses poultry immune responses, which can increase susceptibility to infectious diseases and, thereby, intensify the negative effects of heat on poultry welfare and performance. Identifying genes and pathways that are affected by high temperatures, especially heat-induced changes in immune responses, could provide targets to improve disease resistance in chickens. This study utilized RNA-sequencing (RNA-seq) to investigate transcriptome responses in the bursa of Fabricius, a primary immune tissue, after exposure to acute heat stress and/or subcutaneous immune stimulation with lipopolysaccharide (LPS) in a 2 × 2 factorial design: Thermoneutral + Saline, Heat + Saline, Thermoneutral + LPS and Heat + LPS. All treatments were investigated in two chicken lines: a relatively heat- and disease-resistant Fayoumi line and a more susceptible broiler line.ResultsDifferential expression analysis determined that Heat + Saline had limited impact on gene expression (N = 1 or 63 genes) in broiler or Fayoumi bursa. However, Thermoneutral + LPS and Heat + LPS generated many expression changes in Fayoumi bursa (N = 368 and 804 genes). Thermoneutral + LPS was predicted to increase immune-related cell signaling and cell migration, while Heat + LPS would activate mortality-related functions and decrease expression in WNT signaling pathways. Further inter-treatment comparisons in the Fayoumi line revealed that heat stress prevented many of the expression changes caused by LPS. Although fewer significant expression changes were observed in the broiler bursa after exposure to Thermoneutral + LPS (N = 59 genes) or to Heat + LPS (N = 146 genes), both treatments were predicted to increase cell migration. Direct comparison between lines (broiler to Fayoumi) confirmed that each line had distinct responses to treatment.ConclusionsTranscriptome analysis identified genes and pathways involved in bursal responses to heat stress and LPS and elucidated that these effects were greatest in the combined treatment. The interaction between heat and LPS was line dependent, with suppressive expression changes primarily in the Fayoumi line. Potential target genes, especially those involved in cell migration and immune signaling, can inform future research on heat stress in poultry and could prove useful for improving disease resistance.

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Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken.

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Validation of immunomodulatory effects of lipopolysaccharide through expression profiling of Th1 and Th2 biased genes in Newcastle disease virus vaccinated indigenous chicken.

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