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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


Affiliations
1 School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
2 Department of Animal Genetics and Breeding, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
3 Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, Central Agricultural University , Mizoram, India
 

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.


Keywords

Adjuvants, Aseel, Lipopolysaccharide, Newcastle Disease, Vaccine.
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  • Janeway, C.A. Jr. and Medzhitov, R. (2002) Innate immune recognition. Annu. Rev. Immunol., 20: 197-216.
  • Watanabe, S. and Inoue, J. (2013) Intracellular delivery of lipopolysaccharide induces effective Th1-immune responses independent of IL-12. PLoS One, 8: e68671.
  • Khalifeh, M.S., Amawi, M.M., Abu-Basha, E.A. and Yonis, I.B. (2009) Assessment of humoral and cellular-mediated immune response in chickens treated with tilmicosin, florfenicol, or enrofloxacin at the time of Newcastle disease vaccination. Poultry Sci., 8: 2118-2124.
  • Beutler, B., Jiang, Z. and Georgel, P. (2006) Genetic analysis of host resistance: Toll-like receptor signaling and immunity at large. Annu. Rev. Immunol., 24: 353-389.
  • Livak, K.J. and Schmittgen, T.D. (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-[Delta][Delta]CT method. Methods, 25: 402-408.
  • OIE. (2015) Manual of Diagnostic Tests and Vaccines for Terrestrial Animals: Mammals, Birds and Bees, Biological Standards Commission. 7th ed. Available from: http:// www.oie.int/fileadmin/Home/eng/Health_standards/ tahm/2.03.14_NEWCASTLE_DIS.pdf. Last accessed on 06-11-2017.
  • Avery, S., Rothwell, L., Degen, W.G., Schijns, V.E., Young, J., Kaufman, J., Kaiser, P. (2004) Characterization of the first non-mammalian T2 cytokine gene cluster: The cluster contains functional single-copy genes for IL-3, IL-4, IL-13 and GMCSF, a gene for IL-5 which appears to be a pseudogene, and a gene encoding another cytokine-like transcript, KK34. J. Interferon Cytokine Res., 24: 600-610.
  • Rauw, F., Gardin, Y., Palya, V., Anbari, S., Lemaire, S., Boschmans, M., Van den Berg, T. and Lambrecht, B. (2010) Improved vaccination against Newcastle disease by an in-ovo recombinant HVT-ND combined with an adjuvanted live vaccine at day-old. Vaccine, 28: 823-833.
  • Zhang, L., Tian, X. and Zhou, F. (2007) Vaccination with Newcastle disease vaccine and CpG oligodeoxynucleotides induces specific immunity and protection against Newcastle disease virus in SPF chicken. Vet. Immunol. Immunopathol., 115: 216-222.
  • Taylor, J.M., Ziman, M.E., Huff, J.L., Moroski, N.M., Vajdy, M. and Solnick, J.V. (2006) Helicobacter pylori lipopolysaccharide promotes a Th1 type immune response in immunized mice. Vaccine, 24: 4987-4994.
  • Sachan, S., Ramakrishnan, S., Annamalai, A.K., Sharma, B.K., Malik, H., Saravanan, B.C., Jain, L., Saxena, M., Kumar, A. and Krishnaswamy, N. (2015) Adjuvant potential of resiquimod with inactivated Newcastle disease vaccine and its mechanism of action in chicken. Vaccine, 33: 4526-4532.
  • Greenbaum, D., Colangelo, C., Williams, K. and Gerstein, M. (2003) Comparing protein abundance and mRNA expression levels on a genomic scale. Genome Biol., 4: 117.
  • Arous, J.B., Deville, S., Pal, J.K., Baksi, S. and Bertrand, F. (2013) Reduction of Newcastle disease vaccine dose using a novel adjuvant for cellular immune response in poultry. Procedia. Vaccinol., 7: 28-33.
  • Taylor, J.M., Ziman, M.E., Canfield, D.R., Vajdy, M. and Solnick, J.V. (2008) Effects of a Th1 versus a Th2 biased immune response in protection against Helicobacter pylori challenge in mice. Microb. Pathog., 44: 20-27.
  • Levast, B., Awate, S., Babiuk, L., Mutwiri, G., Gerdts V. and Van den Hurk, S. (2014) Vaccine potentiation by combination adjuvants. Vaccines, 2: 297-322.
  • Kawai, T. and Akira, S. (2011) Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity, 34: 637-650.
  • Abu-Baker, F., Masoud, H. (2016), Synthesis, characterization, and immunological properties of LPS-based vaccines composed of O-polysaccharides conjugated with recombinant exoprotein a from Pseudomonas aeruginosa. Adv. Microbiol., 6: 332-342.

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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 Views: 174  |  PDF Views: 0

Authors

Rabia Bhardwaj
School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
Ramneek Verma
School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
Dipak Deka
School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
P. P. Dubey
Department of Animal Genetics and Breeding, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
J. S. Arora
School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
R. S. Sethi
School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India
T. C. Tolenkhomba
Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, Central Agricultural University , Mizoram, India
C. S. Mukhopadhyay
School of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana - 141 001, Punjab, India

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.


Keywords


Adjuvants, Aseel, Lipopolysaccharide, Newcastle Disease, Vaccine.

References