Journal Article

Adjuvant potential of resiquimod with inactivated Newcastle disease vaccine and its mechanism of action in chicken

Saxena, Meeta, Annamalai, Arunsaravanakumar, Sachan, Swati, Sharma, Bal Krishan, Krishnaswamy, Narayanan, Ramakrishnan, Saravanan, Kumar, Ajay, Malik, Hina, Saravanan, B.C., Jain, Lata

2015

Overview

•Resiquimod (R-848) showed potential adjuvant capacity with Newcastle disease (ND) vaccine in SPF chicken.•R-848 enhanced antigen specific humoral as well as cellular immune responses.•R-848 up-regulated the expression of IFN-α, IFN-β, IFN-γ, IL-4, IL-1β, MHC-II and iNOS transcripts in the chicken spleen.•R-848 potentiated the protection capacity of inactivated ND vaccine against virulent ND virus challenge. Resiquimod (R-848), an imidazoquinoline compound, is a potent synthetic Toll-like receptor (TLR) 7 agonist. Although the solitary adjuvant potential of R-848 is well established in mammals, such reports are not available in avian species hitherto. Hence, the adjuvant potential of R-848 was tested in SPF chicken in this study. Two week old chicks were divided into four groups (10birds/group) viz., control (A), inactivated Newcastle disease virus (NDV) vaccine prepared from velogenic strain (B), commercial oil adjuvanted inactivated NDV vaccine prepared from lentogenic strain (C) and inactivated NDV vaccine prepared from velogenic strain with R-848 (D). Booster was given two weeks post primary vaccination. Humoral immune response was assessed by haemagglutination inhibition (HI) test and ELISA while the cellular immune response was quantified by lymphocyte transformation test (LTT) and flow cytometry post-vaccination. Entire experiment was repeated twice to check the reproducibility. Highest HI titre was observed in group D at post booster weeks 1 and 2 that corresponds to mean log2 HI titre of 6.4±0.16 and 6.8±0.13, respectively. The response was significantly higher than that of group B or C (P<0.01). LTT stimulation index (P≤0.01) as well as CD4+ and CD8+ cells in flow cytometry (P<0.05) were significantly high and maximum in group D. Group D conferred complete protection against virulent NDV challenge, while it was only 80% in group B and C. To understand the effects of R-848, the kinetics of immune response genes in spleen were analyzed using quantitative real-time PCR after R-848 administration (50μg/bird, i.m. route). Resiquimod significantly up-regulated the expression of IFN-α, IFN-β, IFN-γ, IL-1β, IL-4, iNOS and MHC-II genes (P<0.01). In conclusion, the study demonstrated the adjuvant potential of R-848 when co-administered with inactivated NDV vaccine in SPF chicken which is likely due to the up-regulation of immune response genes.

• Publisher: Elsevier Ltd,Elsevier Limited
• Subject: Adjuvant / adjuvants / Adjuvants, Immunologic - administration & dosage / agonists / Allergy and Immunology / Animal vaccines / Animals / Antibodies, Viral - blood / Avian orthoavulavirus 1 / Birds / Chicken / Chickens / chicks / Cytokines / Enzyme-Linked Immunosorbent Assay / Flow Cytometry / Gene Expression Profiling / gene expression regulation / Genes / hemagglutination / Hemagglutination Inhibition Tests / humoral immunity / Imidazoles - administration & dosage / Immune response / Immune response genes / inducible nitric oxide synthase / interferon-alpha / interferon-beta / interferon-gamma / interleukin-1beta / interleukin-4 / lymphocyte proliferation / Lymphocytes / Lymphocytes - immunology / mammals / mechanism of action / Newcastle disease / Newcastle Disease - prevention & control / Newcastle disease virus / oils / Poultry / quantitative polymerase chain reaction / Resiquimod / spleen / TLR agonist / Toll-like receptors / vaccination / Vaccines / Vaccines, Inactivated - administration & dosage / Viral Vaccines - administration & dosage / virulence