Supercritical fluid extraction provides an enhancement to the immune response for orally-delivered hepatitis B surface antigen.
The hepatitis B virus continues to be a major pathogen worldwide despite the availability of an effective parenteral vaccine for over 20 years. Orally-delivered subunit vaccines produced in maize may help to alleviate the disease burden by providing a low-cost, heat-stable alternative to the parenteral vaccine. Oral subunit vaccination has been an elusive goal due to the large amounts of antigen required to induce an immunologic response when administered through the digestive tract. Here we show that high levels of HBsAg were obtained in maize grain, the grain was formed into edible wafers, and wafers were fed to mice at a concentration of approximately 300 μg/g. When these wafers were made with supercritical fluid extraction (SFE)-treated maize material, robust IgG and IgA responses in sera were observed that were comparable to the injected commercial vaccine (Recombivax(®)). In addition, all mice administered SFE wafers showed high secretory IgA titers in fecal material whereas Recombivax(®) treated mice showed no detectable titer. Increased salivary IgA titers were also detected in SFE-fed mice but not in Recombivax(®) treated mice. Wafers made from hexane-treated or full fat maize material induced immunologic responses, but fecal titers were attenuated relative to those produced by SFE-treated wafers. These responses demonstrate the feasibility of using a two-dose oral vaccine booster in the absence of an adjuvant to induce immunologic responses in both sera and at mucosal surfaces, and highlight the potential limitations of using an exclusively parenteral dosing regime.
Applied Biotechnology Institute, Cal Poly Tech Park, San Luis Obispo, CA 93407, USA.
Celine A
CA
Hayden
Applied Biotechnology Institute, Cal Poly Tech Park, San Luis Obispo, CA 93407, USA.
Emily M
EM
Smith
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843, USA.
Debra D
DD
Turner
Applied Biotechnology Institute, Cal Poly Tech Park, San Luis Obispo, CA 93407, USA.
Todd K
TK
Keener
Horticulture and Crop Science Department, California Polytechnic State University, San Luis Obispo, CA 93407, USA.
Jeffrey C
JC
Wong
Department of Statistics, California Polytechnic State University, San Luis Obispo, CA 93407, USA.
John H
JH
Walker
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, TX 77843, USA.
Ian R
IR
Tizard
Dairy Product Technology Center, California Polytechnic State University, San Luis Obispo, CA 93407, USA.
Rafael
R
Jimenez-Flores
Applied Biotechnology Institute, Cal Poly Tech Park, San Luis Obispo, CA 93407, USA. Electronic address: jhoward@appliedbiotech.org.
John A
JA
Howard
D000284:Administration, Oral; D000818:Animals; D025924:Chromatography, Supercritical Fluid; D006508:Hepatitis Antibodies / Q000097:blood; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis / Q000276:immunology*; D017325:Hepatitis B Vaccines / Q000276:immunology*; D018928:Immunity, Mucosal; D007070:Immunoglobulin A / Q000097:blood; D007071:Immunoglobulin A, Secretory / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D030821:Plants, Genetically Modified / Q000378:metabolism*; D022223:Vaccines, Subunit / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology; D003313:Zea mays / Q000235:genetics / Q000378:metabolism
D006508:Hepatitis Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D007070:Immunoglobulin A; D007071:Immunoglobulin A, Secretory; D007074:Immunoglobulin G; C075655:Recombivax HB; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic
Bioencapsulation; HBsAg; Hepatitis B; Immunogenicity; Mucosal; Oral vaccine; Plant vaccine; Supercritical fluid extraction