Intranasal Delivery of Inactivated Influenza Virus and Poly(I:C) Adsorbed Corn-Based Nanoparticle Vaccine Elicited Robust Antigen-Specific Cell-Mediated Immune Responses in Maternal Antibody Positive Nursery Pigs

Abstract

We designed the killed swine influenza A virus (SwIAV) H1N2 antigen (KAg) with polyriboinosinic:polyribocytidylic acid [(Poly(I:C)] adsorbed corn-derived Nano-11 particle based nanovaccine called Nano-11-KAg+Poly(I:C), and evaluated its immune correlates in maternally derived antibody (MDA)-positive pigs against a heterologous H1N1 SwIAV infection. Immunologically, in tracheobronchial lymph nodes (TBLN) detected enhanced H1N2-specific cytotoxic T-lymphocytes (CTLs) in Nano-11-KAg+Poly(I:C) vaccinates, and in commercial vaccinates detected CTLs with mainly IL-17A+ and early effector phenotypes specific to both H1N2 and H1N1 SwAIV. In commercial vaccinates, activated H1N2- and H1N1-specific IFNγ+&TNFα+, IL-17A+ and central memory T-helper/Memory cells, and in Nano-11-KAg+Poly(I:C) vaccinates H1N2-specific central memory, IFNγ+ and IFNγ+&TNFα+, and H1N1-specific IL-17A+ T-helper/Memory cells were observed. Systemically, Nano-11-KAg+Poly(I:C) vaccine augmented H1N2-specific IFNγ+ CTLs and H1N1-specific IFNγ+ T-helper/Memory cells, and commercial vaccine boosted H1N2- specific early effector CTLs and H1N1-specific IFNγ+&TNFα+ CTLs, as well as H1N2- and H1N1-specific T-helper/Memory cells with central memory, IFNγ+&TNFα+, and IL-17A+ phenotypes. Remarkably, commercial vaccine induced an increase in H1N1-specific T-helper cells in TBLN and naive T-helper cells in both TBLN and peripheral blood mononuclear cells (PBMCs), while H1N1- and H1N2-specific only T-helper cells were augmented in Nano-11-KAg+Poly(I:C) vaccinates in both TBLN and PBMCs. Furthermore, the Nano-11-KAg+Poly(I:C) vaccine stimulated robust cross-reactive IgG and secretory IgA (SIgA) responses in lungs, while the commercial vaccine elicited high levels of serum and lung IgG and serum hemagglutination inhibition (HI) titers. In conclusion, despite vast genetic difference (77% in HA gene identity) between the vaccine H1N2 and H1N1 challenge viruses in Nano-11-KAg+Poly(I:C) vaccinates, compared to over 95% identity between H1N1 of commercial vaccine and challenge viruses, the virus load and macroscopic lesions in the lungs of both types of vaccinates were comparable, but the Nano-11-KAg+Poly(I:C) vaccine cleared the virus from the nasal passage better. These data suggested the important role played by Nano-11 and Poly(I:C) in the induction of polyfunctional, cross-protective cell-mediated immunity against SwIAV in MDA-positive pigs.

Lipid Nanoparticle-Mediated Lymphatic Delivery of Immunostimulatory Nucleic Acids

“…However, the high molecular weight and highly negative charge of PIC hinder its penetration of the plasma membrane. Previously, PIC has been delivered using various cationic materials such as polysaccharide [22], polymer [23], and lipid [24] nanoparticles. In these studies, cationic net charges per se have not shown to induce the non-specific uptake of PIC to the lymph nodes, indicating the need for specific ligands in the design of delivery systems.…”

Gut Microbiota of Obese Children Influences Inflammatory Mucosal Immune Pathways in the Respiratory Tract to Influenza Virus Infection: Optimization of an Ideal Duration of Microbial Colonization in a Gnotobiotic Pig Model

“…The isolated PBMCs and MNCs of TBLN and MLN were immunostained for T-helper/memory cells (CD3 + CD4 + CD8α + β-), cytotoxic T lymphocytes (CTLs) (CD3+CD4-CD8α + β + ), and myeloid cells (CD172 + ) using specific immune markers as described previously ( 31 , 76 , 79 ). The immunostaining procedure and gating pattern are described in our earlier studies ( 31 , 76 , 79 ) and the same was followed for this study.…”

“…The isolated PBMCs and MNCs of TBLN and MLN were immunostained for T-helper/memory cells (CD3 + CD4 + CD8α + β-), cytotoxic T lymphocytes (CTLs) (CD3+CD4-CD8α + β + ), and myeloid cells (CD172 + ) using specific immune markers as described previously ( 31 , 76 , 79 ). The immunostaining procedure and gating pattern are described in our earlier studies ( 31 , 76 , 79 ) and the same was followed for this study. The immunostained cells were fixed, and 100,000 events were acquired by using the BD Aria II flow cytometer (BD Biosciences, CA) and the data were analyzed using FlowJo software (Tree Star, Palo Alto, CA).…”

Supplementation of H7N9 Virus-Like Particle Vaccine With Recombinant Epitope Antigen Confers Full Protection Against Antigenically Divergent H7N9 Virus in Chickens

“…These findings indicate that fusion protein of the recombined influenza conserved epitopes can act as an adjuvant to enhance the crossprotection of influenza VLP vaccine against drifted influenza virus. Poly(I:C), a toll-like receptor (TLR)-3 ligand, is a potent adjuvant, intranasal delivery of influenza vaccine with Poly(I:C) elicited robust antigen-specific cell-mediated immune responses ( 27 , 28 ). Poly(I:C) has been identified to induce strong Th1 immune responses.…”