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Balaji Narasimhan - One of the best experts on this subject based on the ideXlab platform.

  • evaluation of cpg odn adjuvanted Polyanhydride based intranasal influenza nanovaccine in pigs
    Veterinary Microbiology, 2019
    Co-Authors: Santosh Dhakal, Kathleen A Ross, Michael J Wannemuehler, Yashavanth Shaan Lakshmanappa, Shristi Ghimire, Sankar Renu, Bradley T Hogshead, Pauline Bernardo, Changwon Lee, Balaji Narasimhan
    Abstract:

    Abstract Influenza results in significant economic loss in the swine industry each year. A broadly protective swine influenza vaccine would have the dual benefit of protecting pigs from influenza A viruses (IAVs) and limiting their possible zoonotic transmission to humans. In this study, we developed Polyanhydride nanoparticles-based swine influenza vaccine (KAg + CpG-nanovaccine) co-encapsulating inacticated/killed soluble antigen (KAg) and Toll-like receptor (TLR)-9 agonist (CpG-ODN). The immunogenicity and protective efficacy of KAg + CpG-nanovaccine was compared with KAg vaccine containing five-times greater quantity of antigens following heterologous virus challenge. Prime-boost intranasally delivered KAg + CpG-nanovaccine induced significantly higher levels of cross-reactive antigen-specific IgA antibody responses in the nasal cavity, greater lymphoproliferative response in peripheral blood mononuclear cells (PBMCs), and higher IFN-γ secretion during antigen-induced recall responses of PBMCs and tracheobronchial lymph nodes cells compared to those immunized with KAg alone. Importantly, KAg + CpG-nanovaccine provided better protective efficacy through a significant reduction in influenza-induced fever, 16-fold reduction of nasal virus shedding and 80-fold reduction in lung virus titers compared to those immunized with soluble KAg. Our results indicated that CpG-ODN-adjuvanted Polyanhydride nanovaccine can induce higher mucosal antibody and cellular immune responses in pigs; and provide better protection as compared with intranasally delivered soluble KAg.

  • single dose of a Polyanhydride particle based vaccine generates potent antigen specific antitumor immune responses
    Journal of Pharmacology and Experimental Therapeutics, 2019
    Co-Authors: Emad I Wafa, Balaji Narasimhan, Kathleen A Ross, Sean M Geary, Jonathan T Goodman, Aliasger K Salem
    Abstract:

    Many factors affect vaccine efficacy. One of the most salient is the frequency and intervals of vaccine administration. In this study, we assessed the vaccine administration modality for a recently reported Polyanhydride-based vaccine formulation, shown to generate antitumor activity. Polyanhydride particles encapsulating ovalbumin (OVA) were prepared using a double-emulsion technique and subcutaneously delivered to mice either as a single-dose or as prime-boost vaccine regimens in which two different time intervals between prime and boost were assessed (7 or 21 days). This was followed by measurement of cellular and humoral immune responses, and subsequent challenge of the mice with a lethal dose of E.G7-OVA cells to evaluate tumor protection. Interestingly, a single dose of the Polyanhydride particle-based formulation induced sustained OVA-specific cellular immune responses just as effectively as the prime-boost regimens. In addition, mice receiving single-dose vaccine had similar levels of protection against tumor challenge compared with mice administered prime-boosts. In contrast, measurements of OVA-specific IgG antibody titers indicated that a booster dose was required to stimulate strong humoral immune responses, since it was observed that mice administered a prime-boost vaccine had significantly higher OVA-specific IgG1 serum titers than mice administered a single dose. These findings indicate that the requirement for a booster dose using these particles appears unnecessary for the generation of effective cellular immunity.

  • amphiphilic Polyanhydride based recombinant muc4β nanovaccine activates dendritic cells
    Genes & Cancer, 2019
    Co-Authors: Kasturi Banerjee, Balaji Narasimhan, Kathleen A Ross, Michael J Wannemuehler, Shailendra K Gautam, Prakash Kshirsagar, Gaelle Spagnol, Paul L Sorgen, Joyce C Solheim
    Abstract:

    Mucin 4 (MUC4) is a high molecular weight glycoprotein that is differentially overexpressed in pancreatic cancer (PC), functionally contributes to disease progression, and correlates with poor survival. Further, due to its aberrant glycosylation and extensive splicing, MUC4 is a potential target for cancer immunotherapy. Our previous studies have demonstrated the utility of amphiphilic Polyanhydride nanoparticles as a useful platform for the development of protein-based prophylactic and therapeutic vaccines. In the present study, we encapsulated purified recombinant human MUC4-beta (MUC4β) protein in Polyanhydride (20:80 CPTEG:CPH) nanoparticles (MUC4β-nanovaccine) and evaluated its ability to activate dendritic cells and induce adaptive immunity. Immature dendritic cells when pulsed with MUC4β-nanovaccine exhibited significant increase in the surface expressions of MHC I and MHC II and costimulatory molecules (CD80 and CD86), as well as, secretion of pro-inflammatory cytokines (IFN-γ, IL-6, and IL-12) as compared to cells exposed to MUC4β alone or MUC4β mixed with blank nanoparticles (MUC4β+NP). Following immunization, as compared to the other formulations, MUC4β-nanovaccine elicited higher IgG2b to IgG1 ratio of anti-MUC4β-antibodies suggesting a predominantly Th1-like class switching. Thus, our findings demonstrate MUC4β-nanovaccine as a novel platform for PC immunotherapy.

  • automated high throughput synthesis of protein loaded Polyanhydride nanoparticle libraries
    ACS Combinatorial Science, 2018
    Co-Authors: Jonathan T Goodman, Adam S Mullis, Lucas Dunshee, Akash Mitra, Balaji Narasimhan
    Abstract:

    The development of high-throughput techniques and combinatorial libraries can facilitate rapid synthesis and screening of biomaterial-based nanocarriers for drug and vaccine delivery. This study describes a high-throughput method using an automated robot for synthesizing Polyanhydride nanoparticles encapsulating proteins. Polyanhydrides are a class of safe and biodegradable polymers that have been widely used as drug and vaccine delivery vehicles. The robot contains a multiplexed homogenizer and has the capacity to handle parallel streams of monomer or polymer solutions to synthesize polymers and/or nanoparticles. Copolymer libraries were synthesized using the monomers sebacic acid, 1,6-bis(p-carboxyphenoxy)hexane, and 1,8-bis(p-carboxyphenoxy)-3,6-dioxactane and compared to conventionally synthesized copolymers. Nanoparticle libraries of varying copolymer compositions encapsulating the model antigen ovalbumin were synthesized using flash nanoprecipitation. The amount of the surfactant Span 80 was varied ...

  • efficacy of mucosal Polyanhydride nanovaccine against respiratory syncytial virus infection in the neonatal calf
    Scientific Reports, 2018
    Co-Authors: Jodi L Mcgill, Balaji Narasimhan, Shannon L. Haughney, Sean M. Kelly, Pankaj Kumar, Savannah Speckhart, Jamie Henningson, Randy E Sacco
    Abstract:

    Human respiratory syncytial virus (HRSV) is a leading cause of severe acute lower respiratory tract infection in infants and children worldwide. Bovine RSV (BRSV) is closely related to HRSV and a significant cause of morbidity in young cattle. BRSV infection in calves displays many similarities to RSV infection in humans, including similar age dependency and disease pathogenesis. Polyanhydride nanoparticle-based vaccines (i.e., nanovaccines) have shown promise as adjuvants and vaccine delivery vehicles due to their ability to promote enhanced immunogenicity through the route of administration, provide sustained antigen exposure, and induce both antibody- and cell-mediated immunity. Here, we developed a novel, mucosal nanovaccine that encapsulates the post-fusion F and G glycoproteins from BRSV into Polyanhydride nanoparticles and determined the efficacy of the vaccine against RSV infection using a neonatal calf model. Calves receiving the BRSV-F/G nanovaccine exhibited reduced pathology in the lungs, reduced viral burden, and decreased virus shedding compared to unvaccinated control calves, which correlated with BRSV-specific immune responses in the respiratory tract and peripheral blood. Our results indicate that the BRSV-F/G nanovaccine is highly immunogenic and, with optimization, has the potential to significantly reduce the disease burden associated with RSV infection in both humans and animals.

Michael J Wannemuehler - One of the best experts on this subject based on the ideXlab platform.

  • evaluation of cpg odn adjuvanted Polyanhydride based intranasal influenza nanovaccine in pigs
    Veterinary Microbiology, 2019
    Co-Authors: Santosh Dhakal, Kathleen A Ross, Michael J Wannemuehler, Yashavanth Shaan Lakshmanappa, Shristi Ghimire, Sankar Renu, Bradley T Hogshead, Pauline Bernardo, Changwon Lee, Balaji Narasimhan
    Abstract:

    Abstract Influenza results in significant economic loss in the swine industry each year. A broadly protective swine influenza vaccine would have the dual benefit of protecting pigs from influenza A viruses (IAVs) and limiting their possible zoonotic transmission to humans. In this study, we developed Polyanhydride nanoparticles-based swine influenza vaccine (KAg + CpG-nanovaccine) co-encapsulating inacticated/killed soluble antigen (KAg) and Toll-like receptor (TLR)-9 agonist (CpG-ODN). The immunogenicity and protective efficacy of KAg + CpG-nanovaccine was compared with KAg vaccine containing five-times greater quantity of antigens following heterologous virus challenge. Prime-boost intranasally delivered KAg + CpG-nanovaccine induced significantly higher levels of cross-reactive antigen-specific IgA antibody responses in the nasal cavity, greater lymphoproliferative response in peripheral blood mononuclear cells (PBMCs), and higher IFN-γ secretion during antigen-induced recall responses of PBMCs and tracheobronchial lymph nodes cells compared to those immunized with KAg alone. Importantly, KAg + CpG-nanovaccine provided better protective efficacy through a significant reduction in influenza-induced fever, 16-fold reduction of nasal virus shedding and 80-fold reduction in lung virus titers compared to those immunized with soluble KAg. Our results indicated that CpG-ODN-adjuvanted Polyanhydride nanovaccine can induce higher mucosal antibody and cellular immune responses in pigs; and provide better protection as compared with intranasally delivered soluble KAg.

  • amphiphilic Polyanhydride based recombinant muc4β nanovaccine activates dendritic cells
    Genes & Cancer, 2019
    Co-Authors: Kasturi Banerjee, Balaji Narasimhan, Kathleen A Ross, Michael J Wannemuehler, Shailendra K Gautam, Prakash Kshirsagar, Gaelle Spagnol, Paul L Sorgen, Joyce C Solheim
    Abstract:

    Mucin 4 (MUC4) is a high molecular weight glycoprotein that is differentially overexpressed in pancreatic cancer (PC), functionally contributes to disease progression, and correlates with poor survival. Further, due to its aberrant glycosylation and extensive splicing, MUC4 is a potential target for cancer immunotherapy. Our previous studies have demonstrated the utility of amphiphilic Polyanhydride nanoparticles as a useful platform for the development of protein-based prophylactic and therapeutic vaccines. In the present study, we encapsulated purified recombinant human MUC4-beta (MUC4β) protein in Polyanhydride (20:80 CPTEG:CPH) nanoparticles (MUC4β-nanovaccine) and evaluated its ability to activate dendritic cells and induce adaptive immunity. Immature dendritic cells when pulsed with MUC4β-nanovaccine exhibited significant increase in the surface expressions of MHC I and MHC II and costimulatory molecules (CD80 and CD86), as well as, secretion of pro-inflammatory cytokines (IFN-γ, IL-6, and IL-12) as compared to cells exposed to MUC4β alone or MUC4β mixed with blank nanoparticles (MUC4β+NP). Following immunization, as compared to the other formulations, MUC4β-nanovaccine elicited higher IgG2b to IgG1 ratio of anti-MUC4β-antibodies suggesting a predominantly Th1-like class switching. Thus, our findings demonstrate MUC4β-nanovaccine as a novel platform for PC immunotherapy.

  • Room Temperature Stable PspA-Based Nanovaccine Induces Protective Immunity
    Frontiers Media S.A., 2018
    Co-Authors: Danielle A. Wagner-muñiz, Balaji Narasimhan, Michael J Wannemuehler, Shannon L. Haughney, Sean M. Kelly
    Abstract:

    Streptococcus pneumoniae is a major causative agent of pneumonia, a debilitating disease particularly in young and elderly populations, and is the leading worldwide cause of death in children under the age of five. While there are existing vaccines against S. pneumoniae, none are protective across all serotypes. Pneumococcal surface protein A (PspA), a key virulence factor of S. pneumoniae, is an antigen that may be incorporated into future vaccines to address the immunological challenges presented by the diversity of capsular antigens. PspA has been shown to be immunogenic and capable of initiating a humoral immune response that is reactive across approximately 94% of pneumococcal strains. Biodegradable Polyanhydrides have been studied as a nanoparticle-based vaccine (i.e., nanovaccine) platform to stabilize labile proteins, to provide adjuvanticity, and enhance patient compliance by providing protective immunity in a single dose. In this study, we designed a room temperature stable PspA-based Polyanhydride nanovaccine that eliminated the need for a free protein component (i.e., 100% encapsulated within the nanoparticles). Mice were immunized once with the lead nanovaccine and upon challenge, presented significantly higher survival rates than animals immunized with soluble protein alone, even with a 25-fold reduction in protein dose. This lead nanovaccine formulation performed similarly to protein adjuvanted with Alum, however, with much less tissue reactogenicity at the site of immunization. By eliminating the free PspA from the nanovaccine formulation, the lead nanovaccine was efficacious after being stored dry for 60 days at room temperature, breaking the need for maintaining the cold chain. Altogether, this study demonstrated that a single dose PspA-based nanovaccine against S. pneumoniae induced protective immunity and provided thermal stability when stored at room temperature for at least 60 days

  • functionalization promotes pathogen mimicking characteristics of Polyanhydride nanoparticle adjuvants
    Journal of Biomedical Materials Research Part A, 2017
    Co-Authors: Yashdeep Phanse, Balaji Narasimhan, Brenda R Carrilloconde, Michael J Wannemuehler, Amanda E Ramertait, Scott R. Broderick, Krishna Rajan, Rajarshi Roychoudhury, Nicola L. B. Pohl, Bryan H Bellaire
    Abstract:

    Rational design of adjuvants and delivery systems will promote development of next-generation vaccines to control emerging and re-emerging diseases. To accomplish this, understanding the immune-enhancing properties of new adjuvants relative to those induced by natural infections can help with the development of pathogen-mimicking materials that will effectively initiate innate immune signaling cascades. In this work, the surfaces of Polyanhydride nanoparticles composed of sebacic acid (SA) and 1,6-bis(p-carboxyphenoxy) hexane were decorated with an ethylene diamine spacer partially modified with either a glycolic acid linker or an α-1,2-linked di-mannopyranoside (di-mannose) to confer "pathogen-like" properties and enhance adjuvanticity. Co-incubation of linker-modified nanoparticles with dendritic cells (DCs) elicited significant increases in surface expression of MHC I, MHC II, CD86, and CD40, and enhanced secretion of IL-6, IL-12p40, and TNF-α. An 800% increase in uptake of ethylene-diamine-spaced, linker and di-mannose functionalized Polyanhydride nanoparticles was also observed. Together, our data showed that linker-functionalized Polyanhydride nanoparticles demonstrate similar patterns of uptake, intracellular trafficking, particle persistence, and innate activation as did DCs exposed to Yersinia pestis or Escherichia coli. These results set the stage for rational selection of adjuvant chemistries to induce pathogen-mimicking immune responses. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2762-2771, 2017.

  • Polyanhydride nanovaccine against swine influenza virus in pigs
    Vaccine, 2017
    Co-Authors: Santosh Dhakal, Jonathan T Goodman, Kathryn Bondra, Yashavanth Shaan Lakshmanappa, Jagadish Hiremath, Duanliang Shyu, Kang Ouyang, Kyungil Kang, Steven Krakowka, Michael J Wannemuehler
    Abstract:

    We have recently demonstrated the effectiveness of an influenza A virus (IAV) subunit vaccine based on biodegradable Polyanhydride nanoparticles delivery in mice. In the present study, we evaluated the efficacy of ∼200nm Polyanhydride nanoparticles encapsulating inactivated swine influenza A virus (SwIAV) as a vaccine to induce protective immunity against a heterologous IAV challenge in pigs. Nursery pigs were vaccinated intranasally twice with inactivated SwIAV H1N2 (KAg) or Polyanhydride nanoparticle-encapsulated KAg (KAg nanovaccine), and efficacy was evaluated against a heterologous zoonotic virulent SwIAV H1N1 challenge. Pigs were monitored for fever daily. Local and systemic antibody responses, antigen-specific proliferation of peripheral blood mononuclear cells, gross and microscopic lung lesions, and virus load in the respiratory tract were compared among the groups of animals. Our pre-challenge results indicated that KAg nanovaccine induced virus-specific lymphocyte proliferation and increased the frequency of CD4+CD8αα+ T helper and CD8+ cytotoxic T cells in peripheral blood mononuclear cells. KAg nanovaccine-immunized pigs were protected from fever following SwIAV challenge. In addition, pigs immunized with the KAg nanovaccine presented with lower viral antigens in lung sections and had 6 to 8-fold reduction in nasal shedding of SwIAV four days post-challenge compared to control animals. Immunologically, increased IFN-γ secreting T lymphocyte populations against both the vaccine and challenge viruses were detected in KAg nanovaccine-immunized pigs compared to the animals immunized with KAg alone. However, in the KAg nanovaccine-immunized pigs, hemagglutination inhibition, IgG and IgA antibody responses, and virus neutralization titers were comparable to that in the animals immunized with KAg alone. Overall, our data indicated that intranasal delivery of Polyanhydride-based SwIAV nanovaccine augmented antigen-specific cellular immune response in pigs, with promise to induce cross-protective immunity.

Amanda E Ramertait - One of the best experts on this subject based on the ideXlab platform.

  • functionalization promotes pathogen mimicking characteristics of Polyanhydride nanoparticle adjuvants
    Journal of Biomedical Materials Research Part A, 2017
    Co-Authors: Yashdeep Phanse, Balaji Narasimhan, Brenda R Carrilloconde, Michael J Wannemuehler, Amanda E Ramertait, Scott R. Broderick, Krishna Rajan, Rajarshi Roychoudhury, Nicola L. B. Pohl, Bryan H Bellaire
    Abstract:

    Rational design of adjuvants and delivery systems will promote development of next-generation vaccines to control emerging and re-emerging diseases. To accomplish this, understanding the immune-enhancing properties of new adjuvants relative to those induced by natural infections can help with the development of pathogen-mimicking materials that will effectively initiate innate immune signaling cascades. In this work, the surfaces of Polyanhydride nanoparticles composed of sebacic acid (SA) and 1,6-bis(p-carboxyphenoxy) hexane were decorated with an ethylene diamine spacer partially modified with either a glycolic acid linker or an α-1,2-linked di-mannopyranoside (di-mannose) to confer "pathogen-like" properties and enhance adjuvanticity. Co-incubation of linker-modified nanoparticles with dendritic cells (DCs) elicited significant increases in surface expression of MHC I, MHC II, CD86, and CD40, and enhanced secretion of IL-6, IL-12p40, and TNF-α. An 800% increase in uptake of ethylene-diamine-spaced, linker and di-mannose functionalized Polyanhydride nanoparticles was also observed. Together, our data showed that linker-functionalized Polyanhydride nanoparticles demonstrate similar patterns of uptake, intracellular trafficking, particle persistence, and innate activation as did DCs exposed to Yersinia pestis or Escherichia coli. These results set the stage for rational selection of adjuvant chemistries to induce pathogen-mimicking immune responses. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2762-2771, 2017.

  • cellular internalization mechanisms of Polyanhydride particles implications for rational design of drug delivery vehicles
    Journal of Biomedical Nanotechnology, 2016
    Co-Authors: Balaji Narasimhan, Brenda R Carrilloconde, Michael J Wannemuehler, Amanda E Ramertait, Yashdeep Phanse, Paul Lueth, Bryan H Bellaire
    Abstract:

    Polyanhydride nanoparticles have emerged as a versatile delivery platform, due to their ability to encapsulate diverse drugs, immunogens, antibodies, and proteins. However, mechanistic studies on the effects of particle chemistry interactions with immune cells have yet to be described. Understanding the mechanism by which these particles are internalized by immune cells will enable rational selection of delivery vehicles for specific applications. In the present study, the internalization, mechanism(s) of uptake by monocytes, and intracellular fate of Polyanhydride nanoparticles were evaluated using copolymers based on 1,6-bis(p-carboxyphenoxy)hexane (CPH), sebacic acid (SA), and 1,8-bis(p-carboxyphenoxy)3,6-dioxaoctane (CPTEG). The results showed that 20:80 CPH:SA and 20:80 CPTEG:CPH nanoparticles were internalized to a greater extent by monocytes as compared to the 50:50 CPH:SA and 50:50 CPTEH:CPH nanoparticles. Further, cytochalasin-D treatment of cells inhibited uptake of all the particles, regardless of chemistry, indicating that actinmediated uptake is the primary mechanism of cellular entry for these particles. The insights gained from these studies were used to identify lead nanoparticle formulations to enhance treatment of intracellular bacterial infections. The use of doxycycline-loaded nanoparticles exhibited enhanced therapeutic efficacy compared to soluble drug in treating monocyte monolayers infected with the virulent intracellular pathogen Brucella abortus. Altogether, these studies demonstrate how rational design and selection of nanoscale delivery platforms can be used for a wide spectrum of biomedical applications.

  • sustained release and stabilization of therapeutic antibodies using amphiphilic Polyanhydride nanoparticles
    Chemical Engineering Science, 2015
    Co-Authors: Brenda R Carrilloconde, Michael J Wannemuehler, Amanda E Ramertait, Ross J Darling, Steven J Seiler, Balaji Narasimhan
    Abstract:

    Abstract Passive administration of antibodies (e.g., anti-serum or monoclonal antibodies) can be successfully used as treatments for infectious agents (e.g., human cytomegalovirus, HIV), chronic inflammation (e.g., anti-TNF), cancer (e.g., anti-HER2, anti-VEGF, anti-CD20), toxins (e.g., anti-ricin), and age-related diseases such as macular degeneration (e.g., anti-VEGF). As with the development of proteinaceous pharmaceuticals, one of the most challenging obstacles facing passive immunotherapies is the physical and the chemical instabilities of the antibodies, which invariably leads to loss of biological activity. In order to avoid these problems, appropriate delivery vehicles need to be designed that minimize the degradation, maximize the in vivo activity, and provide controlled release of the encapsulated biologically active protein. In this study, Polyanhydride nanoparticles were used for the delivery of stable and biologically active therapeutic antibodies. Tetanus antitoxin and anti-TNF-α monoclonal antibodies were encapsulated and released from Polyanhydride nanoparticles. The nanoparticles provided the ability to control antibody release kinetics and, additionally, preserved antibody functionality and bioactivity upon synthesis and release. In particular, amphiphilic Polyanhydride nanoparticles demonstrated the best combination of characteristics compatible for anti-serum or monoclonal antibody preservation and release, making them ideal candidates for use as a delivery system for therapeutic antibodies.

  • Polyanhydride nanovaccine platform enhances antigen specific cytotoxic t cell responses
    TECHNOLOGY, 2014
    Co-Authors: Lucas Huntimer, Balaji Narasimhan, Kathleen A Ross, Amanda E Ramertait, Ross J Darling, Nicole E Winterwood, Paola M Boggiatto, Michael J Wannemuehler
    Abstract:

    Polyanhydride nanoparticle-based vaccines (or nanovaccines) stabilize protein antigens, provide sustained antigen release leading to prolonged antigen presence, enhance activation of antigen presenting cells, and elicit protective immunity against respiratory infections upon challenge. However, induction of cell-mediated immunity when mice are immunized with Polyanhydride nanovaccines has not been evaluated. Using a transgenic ovalbumin-specific T cell adoptive transfer model, we report the induction of antigen-specific cytotoxic CD8+ T cells expressing an effector memory phenotype by seven days after immunization with nanovaccine formulations. Furthermore, mice immunized with Polyanhydride nanovaccines demonstrated enhanced recall responses after antigen re-exposure 35 days post-immunization indicating the activation and recruitment of antigen-specific memory CD8+ T cells to the site of antigen deposition.

  • functionalization of Polyanhydride microparticles with di mannose influences uptake by and intracellular fate within dendritic cells
    Acta Biomaterialia, 2013
    Co-Authors: Yashdeep Phanse, Balaji Narasimhan, Brenda R Carrilloconde, Michael J Wannemuehler, Amanda E Ramertait, Rajarshi Roychoudhury, Nicola L. B. Pohl, Bryan H Bellaire
    Abstract:

    Innovative vaccine delivery platforms can facilitate the development of effective single-dose treatment regimens to control emerging and re-emerging infectious diseases. Polyanhydride microparticles are promising vaccine delivery vehicles due to their ability to stably maintain antigens, provide tailored release kinetics and function as adjuvants. A major obstacle for the use of microparticle-based vaccines, however, is their limited uptake by dendritic cells (DCs). In this study, we functionalized the microparticle surface with di-mannose in order to target C-type lectin receptors (CLRs) on DCs. Polyanhydride particles based on sebacic acid (SA), 1,6-bis(p-carboxyphenoxy)hexane (CPH) and 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) were evaluated. Co-incubation of di-mannose-functionalized microparticles up-regulated the expression of CLRs on DCs. More importantly, di-mannose functionalization increased the uptake, as measured by the percentage of cells internalizing particles. The uptake of CPH:SA microparticles increased ∼20-fold, from 0.82% (non-functionalized) to 20.2%, and internalization of CPTEG:CPH microparticles increased ∼7-fold from 1.35% (non-functionalized) to 9.3% upon di-mannose functionalization. Both di-mannose-functionalized and non-functionalized particles trafficked to lysosomes. Together, these studies demonstrate that employing rational vaccine design principles, such as the targeting of CLRs on antigen-presenting cells, can enhance delivery of encapsulated antigens and potentially induce a more robust adaptive immune response.

Latrisha K. Petersen - One of the best experts on this subject based on the ideXlab platform.

  • © 2013 Petersen et al, publisher and licensee Dove Medical Press Ltd. This is an Open Access article
    2016
    Co-Authors: Latrisha K. Petersen, Balaji Narasimhan, Lucas Huntimer, Michael J Wannemuehler, Katharine Walz, Amanda Ramer-tait, Correspondence Balaji Narasimhan
    Abstract:

    which permits unrestricted noncommercial use, provided the original work is properly cited. International Journal of Nanomedicine 2013: 8 2213–2225 International Journal of Nanomedicine Combinatorial evaluation of in vivo distribution of Polyanhydride particle-based platforms for vaccine deliver

  • retention of structure antigenicity and biological function of pneumococcal surface protein a pspa released from Polyanhydride nanoparticles
    Acta Biomaterialia, 2013
    Co-Authors: Shannon L. Haughney, Latrisha K. Petersen, Michael J Wannemuehler, Amanda E Ramertait, Amy Danielle Schoofs, Janice D King, David E Briles, Balaji Narasimhan
    Abstract:

    Abstract Pneumococcal surface protein A (PspA) is a choline-binding protein which is a virulence factor found on the surface of all Streptococcus pneumoniae strains. Vaccination with PspA has been shown to be protective against a lethal challenge with S. pneumoniae, making it a promising immunogen for use in vaccines. Herein the design of a PspA-based subunit vaccine using Polyanhydride nanoparticles as a delivery platform is described. Nanoparticles based on sebacic acid (SA), 1,6-bis-(p-carboxyphenoxy)hexane (CPH) and 1,8-bis-(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG), specifically 50:50 CPTEG:CPH and 20:80 CPH:SA, were used to encapsulate and release PspA. The protein released from the nanoparticle formulations retained its primary and secondary structure as well as its antigenicity. The released PspA was also biologically functional based on its ability to bind to apolactoferrin and prevent its bactericidal activity against Escherichia coli. When the PspA nanoparticle formulations were administered subcutaneously to mice they elicited a high titer and high avidity anti-PspA antibody response. Together these studies provide a framework for the rational design of a vaccine against S. pneumoniae based on Polyanhydride nanoparticles.

  • combinatorial evaluation of in vivo distribution of Polyanhydride particle based platforms for vaccine delivery
    International Journal of Nanomedicine, 2013
    Co-Authors: Latrisha K. Petersen, Lucas Huntimer, Michael J Wannemuehler, Amanda E Ramertait, Katharine Walz, Balaji Narasimhan
    Abstract:

    Several challenges are associated with current vaccine strategies, including repeated immunizations, poor patient compliance, and limited approved routes for delivery, which may hinder induction of protective immunity. Thus, there is a need for new vaccine adjuvants capable of multi-route administration and prolonged antigen release at the site of administration by providing a depot within tissue. In this work, we designed a combinatorial platform to investigate the in vivo distribution, depot effect, and localized persistence of Polyanhydride nanoparticles as a function of nanoparticle chemistry and administration route. Our observations indicated that the route of administration differentially affected tissue residence times. All nanoparticles rapidly dispersed when delivered intranasally but provided a depot when administered parenterally. When amphiphilic and hydrophobic nanoparticles were administered intranasally, they persisted within lung tissue. These results provide insights into the chemistry- and route-dependent distribution and tissue-specific association of Polyanhydride nanoparticle-based vaccine adjuvants.

  • evaluation of biocompatibility and administration site reactogenicity of Polyanhydride particle based platform for vaccine delivery
    Advanced Healthcare Materials, 2013
    Co-Authors: Lucas Huntimer, Latrisha K. Petersen, Balaji Narasimhan, Kathleen A Ross, Chong Wang, Amanda E Ramertait, Katherine A Walz, Jesse M Hostetter, Michael J Wannemuehler
    Abstract:

    Efficacy, purity, safety, and potency are important attributes of vaccines. Polyanhydride particles represent a novel class of vaccine adjuvants and delivery platforms that have demonstrated the ability to enhance the stability of protein antigens as well as elicit protective immunity against bacterial pathogens. This work aims to elucidate the biocompatibility, inflammatory reactions, and particle effects on mice injected with a 5 mg dose of Polyanhydride nanoparticles via common parenteral routes (subcutaneous and intramuscular). Independent of polymer chemistry, nanoparticles more effectively disseminated away from the injection site as compared to microparticles, which exhibited a depot effect. Using fluorescent probes, the in vivo distribution of three formulations of nanoparticles, following subcutaneous administration, indicated migration away from the injection site. Less inflammation was observed at the injection sites of mice-administered nanoparticles as compared to Alum and incomplete Freund's adjuvant. Furthermore, histological evaluation revealed minimal adverse injection site reactions and minimal toxicological effects associated with the administration of nanoparticles at 30 days post-administration. Collectively, these results demonstrate that Polyanhydride nanoparticles do not induce inflammation as a cumulative effect of particle persistence or degradation and are, therefore, a viable candidate for a vaccine delivery platform.

  • evaluation of biocompatibility and administration site reactogenicity of Polyanhydride particle based platform for vaccine delivery
    Advanced Healthcare Materials, 2013
    Co-Authors: Lucas Huntimer, Latrisha K. Petersen, Balaji Narasimhan, Kathleen A Ross, Chong Wang, Amanda E Ramertait, Katherine A Walz, Jesse M Hostetter, Michael J Wannemuehler
    Abstract:

    Efficacy, purity, safety, and potency are important attributes of vaccines. Polyanhydride particles represent a novel class of vaccine adjuvants and delivery platforms that have demonstrated the ability to enhance the stability of protein antigens as well as elicit protective immunity against bacterial pathogens. This work aims to elucidate the biocompatibility, inflammatory reactions, and particle effects on mice injected with a 5 mg dose of Polyanhydride nanoparticles via common parenteral routes (subcutaneous and intramuscular). Independent of polymer chemistry, nanoparticles more effectively disseminated away from the injection site as compared to microparticles, which exhibited a depot effect. Using fluorescent probes, the in vivo distribution of three formulations of nanoparticles, following subcutaneous administration, indicated migration away from the injection site. Less inflammation was observed at the injection sites of mice-administered nanoparticles as compared to Alum and incomplete Freund's adjuvant. Furthermore, histological evaluation revealed minimal adverse injection site reactions and minimal toxicological effects associated with the administration of nanoparticles at 30 days post-administration. Collectively, these results demonstrate that Polyanhydride nanoparticles do not induce inflammation as a cumulative effect of particle persistence or degradation and are, therefore, a viable candidate for a vaccine delivery platform.

Kathleen A Ross - One of the best experts on this subject based on the ideXlab platform.

  • evaluation of cpg odn adjuvanted Polyanhydride based intranasal influenza nanovaccine in pigs
    Veterinary Microbiology, 2019
    Co-Authors: Santosh Dhakal, Kathleen A Ross, Michael J Wannemuehler, Yashavanth Shaan Lakshmanappa, Shristi Ghimire, Sankar Renu, Bradley T Hogshead, Pauline Bernardo, Changwon Lee, Balaji Narasimhan
    Abstract:

    Abstract Influenza results in significant economic loss in the swine industry each year. A broadly protective swine influenza vaccine would have the dual benefit of protecting pigs from influenza A viruses (IAVs) and limiting their possible zoonotic transmission to humans. In this study, we developed Polyanhydride nanoparticles-based swine influenza vaccine (KAg + CpG-nanovaccine) co-encapsulating inacticated/killed soluble antigen (KAg) and Toll-like receptor (TLR)-9 agonist (CpG-ODN). The immunogenicity and protective efficacy of KAg + CpG-nanovaccine was compared with KAg vaccine containing five-times greater quantity of antigens following heterologous virus challenge. Prime-boost intranasally delivered KAg + CpG-nanovaccine induced significantly higher levels of cross-reactive antigen-specific IgA antibody responses in the nasal cavity, greater lymphoproliferative response in peripheral blood mononuclear cells (PBMCs), and higher IFN-γ secretion during antigen-induced recall responses of PBMCs and tracheobronchial lymph nodes cells compared to those immunized with KAg alone. Importantly, KAg + CpG-nanovaccine provided better protective efficacy through a significant reduction in influenza-induced fever, 16-fold reduction of nasal virus shedding and 80-fold reduction in lung virus titers compared to those immunized with soluble KAg. Our results indicated that CpG-ODN-adjuvanted Polyanhydride nanovaccine can induce higher mucosal antibody and cellular immune responses in pigs; and provide better protection as compared with intranasally delivered soluble KAg.

  • single dose of a Polyanhydride particle based vaccine generates potent antigen specific antitumor immune responses
    Journal of Pharmacology and Experimental Therapeutics, 2019
    Co-Authors: Emad I Wafa, Balaji Narasimhan, Kathleen A Ross, Sean M Geary, Jonathan T Goodman, Aliasger K Salem
    Abstract:

    Many factors affect vaccine efficacy. One of the most salient is the frequency and intervals of vaccine administration. In this study, we assessed the vaccine administration modality for a recently reported Polyanhydride-based vaccine formulation, shown to generate antitumor activity. Polyanhydride particles encapsulating ovalbumin (OVA) were prepared using a double-emulsion technique and subcutaneously delivered to mice either as a single-dose or as prime-boost vaccine regimens in which two different time intervals between prime and boost were assessed (7 or 21 days). This was followed by measurement of cellular and humoral immune responses, and subsequent challenge of the mice with a lethal dose of E.G7-OVA cells to evaluate tumor protection. Interestingly, a single dose of the Polyanhydride particle-based formulation induced sustained OVA-specific cellular immune responses just as effectively as the prime-boost regimens. In addition, mice receiving single-dose vaccine had similar levels of protection against tumor challenge compared with mice administered prime-boosts. In contrast, measurements of OVA-specific IgG antibody titers indicated that a booster dose was required to stimulate strong humoral immune responses, since it was observed that mice administered a prime-boost vaccine had significantly higher OVA-specific IgG1 serum titers than mice administered a single dose. These findings indicate that the requirement for a booster dose using these particles appears unnecessary for the generation of effective cellular immunity.

  • amphiphilic Polyanhydride based recombinant muc4β nanovaccine activates dendritic cells
    Genes & Cancer, 2019
    Co-Authors: Kasturi Banerjee, Balaji Narasimhan, Kathleen A Ross, Michael J Wannemuehler, Shailendra K Gautam, Prakash Kshirsagar, Gaelle Spagnol, Paul L Sorgen, Joyce C Solheim
    Abstract:

    Mucin 4 (MUC4) is a high molecular weight glycoprotein that is differentially overexpressed in pancreatic cancer (PC), functionally contributes to disease progression, and correlates with poor survival. Further, due to its aberrant glycosylation and extensive splicing, MUC4 is a potential target for cancer immunotherapy. Our previous studies have demonstrated the utility of amphiphilic Polyanhydride nanoparticles as a useful platform for the development of protein-based prophylactic and therapeutic vaccines. In the present study, we encapsulated purified recombinant human MUC4-beta (MUC4β) protein in Polyanhydride (20:80 CPTEG:CPH) nanoparticles (MUC4β-nanovaccine) and evaluated its ability to activate dendritic cells and induce adaptive immunity. Immature dendritic cells when pulsed with MUC4β-nanovaccine exhibited significant increase in the surface expressions of MHC I and MHC II and costimulatory molecules (CD80 and CD86), as well as, secretion of pro-inflammatory cytokines (IFN-γ, IL-6, and IL-12) as compared to cells exposed to MUC4β alone or MUC4β mixed with blank nanoparticles (MUC4β+NP). Following immunization, as compared to the other formulations, MUC4β-nanovaccine elicited higher IgG2b to IgG1 ratio of anti-MUC4β-antibodies suggesting a predominantly Th1-like class switching. Thus, our findings demonstrate MUC4β-nanovaccine as a novel platform for PC immunotherapy.

  • ph responsive microencapsulation systems for the oral delivery of Polyanhydride nanoparticles
    Biomacromolecules, 2018
    Co-Authors: Lindsey A Sharpe, Balaji Narasimhan, Kathleen A Ross, Julia Vela E Ramirez, Olivia M. Haddadin, Nicholas A. Peppas
    Abstract:

    Multicompartmental polymer carriers, referred to as Polyanhydride-Releasing Oral MicroParticle Technology (PROMPT), were formed by a pH-triggered antisolvent precipitation technique. Polyanhydride nanoparticles were encapsulated into anionic pH-responsive microparticle gels, allowing for nanoparticle encapsulation in acidic conditions and subsequent release in neutral pH conditions. The effects of varying the nanoparticle composition and feed ratio on the encapsulation efficiency were evaluated. Nanoparticle encapsulation was confirmed by confocal microscopy and infrared spectroscopy. pH-triggered protein delivery from PROMPT was explored using ovalbumin (ova) as a model drug. PROMPT microgels released ova in a pH-controlled manner. Increasing the feed ratio of nanoparticles into the microgels increased the total amount of ova delivered, as well as decreased the observed burst release. The cytocompatibility of the polymer materials were assessed using cells representative of the GI tract. Overall, these r...

  • pH-Responsive Microencapsulation Systems for the Oral Delivery of Polyanhydride Nanoparticles
    2018
    Co-Authors: Lindsey A. Sharpe, Balaji Narasimhan, Kathleen A Ross, Julia Vela E Ramirez, Olivia M. Haddadin, Nicholas A. Peppas
    Abstract:

    Multicompartmental polymer carriers, referred to as Polyanhydride-Releasing Oral MicroParticle Technology (PROMPT), were formed by a pH-triggered antisolvent precipitation technique. Polyanhydride nanoparticles were encapsulated into anionic pH-responsive microparticle gels, allowing for nanoparticle encapsulation in acidic conditions and subsequent release in neutral pH conditions. The effects of varying the nanoparticle composition and feed ratio on the encapsulation efficiency were evaluated. Nanoparticle encapsulation was confirmed by confocal microscopy and infrared spectroscopy. pH-triggered protein delivery from PROMPT was explored using ovalbumin (ova) as a model drug. PROMPT microgels released ova in a pH-controlled manner. Increasing the feed ratio of nanoparticles into the microgels increased the total amount of ova delivered, as well as decreased the observed burst release. The cytocompatibility of the polymer materials were assessed using cells representative of the GI tract. Overall, these results suggest that pH-dependent microencapsulation is a viable platform to achieve targeted intestinal delivery of Polyanhydride nanoparticles and their payload(s)