Antigenic Liposomes for Generation of Disease-specific Antibodies

Kyle J. Bednar; Lakeya Hardy; Johanna Smeekens; Dharmendra Raghuwanshi; Shiteng Duan; Mike D. Kulis; Matthew S. Macauley

doi:10.3791/58285

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Method Article

Antigenic Liposomes for Generation of Disease-specific Antibodies

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DOI:

10.3791/58285

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October 25th, 2018

Kyle J. Bednar^*¹ , Lakeya Hardy^*²^,³ , Johanna Smeekens^*³^,⁴ , Dharmendra Raghuwanshi⁵ , Shiteng Duan⁶ , Mike D. Kulis³^,⁴ , Matthew S. Macauley⁵^,⁷

¹Janssen R&D, ²Department of Microbiology and Immunology, University of North Carolina, ³UNC Food Allergy Initiative, University of North Carolina, ⁴Department of Pediatrics, University of North Carolina, ⁵Department of Chemistry, University of Alberta, ⁶Department of Molecular Medicine, Scripps Research Institute, ⁷Department of Medical Microbiology and Immunology, University of Alberta

^* These authors contributed equally

Summary

Described is the preparation of antigenic liposomal nanoparticles and their use in stimulating B-cell activation in vitro and in vivo. Consistent and robust antibody responses led to the development of a new peanut allergy model. The protocol for generating antigenic liposomes can be extended to different antigens and immunization models.

Abstract

Antibody responses provide critical protective immunity to a wide array of pathogens. There remains a high interest in generating robust antibodies for vaccination as well as understand how pathogenic antibody responses develop in allergies and autoimmune disease. Generating robust antigen-specific antibody responses is not always trivial. In mouse models, it often requires multiple rounds of immunizations with adjuvant that leads to a great deal of variability in the levels of induced antibodies. One example is in mouse models of peanut allergies where more robust and reproducible models that minimize mouse numbers and the use of adjuvant would be beneficial. Presented here is a highly reproducible mouse model of peanut allergy anaphylaxis. This new model relies on two key factors: (1) antigen-specific splenocytes are adoptively transferred from a peanut-sensitized mouse into a naïve recipient mouse, normalizing the number of antigen-specific memory B- and T-cells across a large number of mice; and (2) recipient mice are subsequently boosted with a strong multivalent immunogen in the form of liposomal nanoparticles displaying the major peanut allergen (Ara h 2). The major advantage of this model is its reproducibility, which ultimately lowers the number of animals used in each study, while minimizing the number of animals receiving multiple injections of adjuvant. The modular assembly of these immunogenic liposomes provides relatively facile adaptability to other allergic or autoimmune models that involve pathogenic antibodies.

Introduction

Food allergy affects 8% of children in the United States, and has increased in prevalence over the past decade¹. Allergy to peanut affects 1% of children and is not typically outgrown². Although several promising clinical trials are underway for the treatment of food allergy, including oral immunotherapy (OIT), sublingual immunotherapy (SLIT), and epicutaneous immunotherapy (EPIT), there are currently no FDA-approved treatment strategies for desensitizing peanut-allergic individuals³^,⁴^,⁵^,⁶^....