Assessment of gold nanoparticles as a size-dependent vaccine carrier for enhancing the antibody response against synthetic foot-and-mouth disease virus peptide

To assess the ability of gold nanoparticles (GNPs) to act as a size-dependent carrier, a synthetic peptide resembling foot-and-mouth disease virus (FMDV) protein was conjugated to GNPs ranging from 2 to 50 nm in diameter (2, 5, 8, 12, 17, 37, and 50 nm). An extra cysteine was added to the C-terminus of the FMDV peptide (pFMDV) to ensure maximal conjugation to the GNPs, which have a high affinity for sulfhydryl groups. The resultant pFMDV-GNP conjugates were then injected into BALB/c mice. Immunization with pFMDV-keyhole limpet hemocyanin (pFMDV-KLH) conjugate was also performed as a control. Blood was obtained from the mice after 4, 6, 8, and 10 weeks and antibody titers against both pFMDV and the carriers were measured. For the pFMDV-GNP immunization, specific antibodies against the synthetic peptide were detected in the sera of mice injected with 2, 5, 8, 12, and 17 nm pFMDV-GNP conjugates. Maximal antibody binding was noted for GNPs of diameter 8-17 nm. The pFMDV-GNPs induced a three-fold increase in the antibody response compared to the response to pFMDV-KLH. However, sera from either immunized mouse group did not exhibit an antibody response to GNPs, while the sera from pFMDV-KLH-immunized mice presented high levels of binding activity against KLH. Additionally, the uptake of pFMDV-GNP in the spleen was examined by inductively coupled plasma mass spectroscopy (ICP-MS) and transmission electron microscopy (TEM). The quantity of GNPs that accumulated in the spleen correlated to the magnitude of the immune response induced by pFMDV-GNP. In conclusion, we demonstrated the size-dependent immunogenic properties of pFMDV-GNP conjugates. Furthermore, we established that GNPs ranging from 8 to 17 nm in diameter may be ideal for eliciting a focused antibody response against a synthetic pFMDV peptide.

[1]  J. Slot,et al.  A new method of preparing gold probes for multiple-labeling cytochemistry. , 1985, European journal of cell biology.

[2]  Lawrence Tamarkin,et al.  Colloidal Gold: A Novel Nanoparticle Vector for Tumor Directed Drug Delivery , 2004, Drug delivery.

[3]  Dakrong Pissuwan,et al.  Therapeutic possibilities of plasmonically heated gold nanoparticles. , 2006, Trends in biotechnology.

[4]  M. Natan,et al.  Seeding of Colloidal Au Nanoparticle Solutions. 2. Improved Control of Particle Size and Shape , 2000 .

[5]  Warren C W Chan,et al.  Elucidating the mechanism of cellular uptake and removal of protein-coated gold nanoparticles of different sizes and shapes. , 2007, Nano letters.

[6]  Arezou A Ghazani,et al.  Assessing the effect of surface chemistry on gold nanorod uptake, toxicity, and gene expression in mammalian cells. , 2008, Small.

[7]  Keishiro Tomoda,et al.  Biodistribution of colloidal gold nanoparticles after intravenous administration: effect of particle size. , 2008, Colloids and surfaces. B, Biointerfaces.

[8]  R. May,et al.  Antibodies to Keyhole Limpet Hemocyanin Cross-React with an Epitope on the Polysaccharide Capsule of Cryptococcus neoformans and Other Carbohydrates: Implications for Vaccine Development 1 , 2003, The Journal of Immunology.

[9]  A. Gharavi,et al.  GDKV-induced antiphospholipid antibodies enhance thrombosis and activate endothelial cells in vivo and in vitro. , 1999, Journal of immunology.

[10]  E. Engvall,et al.  Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. , 1971, Immunochemistry.

[11]  M. Zwick,et al.  Filamentous phage as an immunogenic carrier to elicit focused antibody responses against a synthetic peptide. , 2006, Vaccine.

[12]  B. Baxt,et al.  Foot-and-Mouth Disease , 2004, Clinical Microbiology Reviews.

[13]  A. Tomii,et al.  Production of anti-platelet-activating factor antibodies by the use of colloidal gold as carrier. , 1991, Japanese journal of medical science & biology.

[14]  S. A. Staroverov,et al.  Obtainment of Polyclonal Antibodies to Clenbuterol with the Use of Colloidal Gold , 2007, Immunopharmacology and immunotoxicology.

[15]  J. Shiver,et al.  Preclinical study of influenza virus A M2 peptide conjugate vaccines in mice, ferrets, and rhesus monkeys. , 2004, Vaccine.

[16]  L. Pirofski,et al.  Immunogenicity and Efficacy of Cryptococcus neoformans Capsular Polysaccharide Glucuronoxylomannan Peptide Mimotope-Protein Conjugates in Human Immunoglobulin Transgenic Mice , 2004, Infection and Immunity.

[17]  J. Escribano,et al.  Induction of a virus-specific antibody response to foot and mouth disease virus using the structural protein VP1 expressed in transgenic potato plants. , 2001, Viral immunology.

[18]  Sai T Reddy,et al.  Exploiting lymphatic transport and complement activation in nanoparticle vaccines , 2007, Nature Biotechnology.

[19]  R. May,et al.  High Affinity Mimotope of the Polysaccharide Capsule of Cryptococcus neoformans Identified from an Evolutionary Phage Peptide Library1 , 2002, The Journal of Immunology.

[20]  K. Strohmaier,et al.  Location and characterization of the antigenic portion of the FMDV immunizing protein. , 1982, The Journal of general virology.

[21]  T. Tumpey,et al.  Matrix Protein 2 Vaccination and Protection against Influenza Viruses, Including Subtype H5N1 , 2007, Emerging infectious diseases.

[22]  D. Fuller,et al.  Powder and particle-mediated approaches for delivery of DNA and protein vaccines into the epidermis. , 2003, Comparative immunology, microbiology and infectious diseases.

[23]  Y. Hung,et al.  Assessment of the In Vivo Toxicity of Gold Nanoparticles , 2009, Nanoscale research letters.

[24]  Richard A. Houghten,et al.  Protection against foot-and-mouth disease by immunization with a chemically synthesized peptide predicted from the viral nucleotide sequence , 1982, Nature.

[25]  Markus Klinger,et al.  Generation of Peptide Mimics of the Epitope Recognized by Trastuzumab on the Oncogenic Protein Her-2/neu1 , 2004, The Journal of Immunology.

[26]  Katrin Schwarz,et al.  Nanoparticles target distinct dendritic cell populations according to their size , 2008, European journal of immunology.

[27]  Yingchun Hou,et al.  Development of Peptide Mimotopes of Lipooligosaccharide from Nontypeable Haemophilus influenzae as Vaccine Candidates , 2003, The Journal of Immunology.

[28]  Yu-Cheng Chang,et al.  Microwave Heating for the Preparation of Nanometer Gold Particles , 2003 .

[29]  Arezou A Ghazani,et al.  Determining the size and shape dependence of gold nanoparticle uptake into mammalian cells. , 2006, Nano letters.

[30]  J. Shieh,et al.  Enhancement of the immunity to foot-and-mouth disease virus by DNA priming and protein boosting immunization. , 2001, Vaccine.

[31]  Petra Krystek,et al.  Particle size-dependent organ distribution of gold nanoparticles after intravenous administration. , 2008, Biomaterials.

[32]  Leon Hirsch,et al.  Nanoshell-Enabled Photonics-Based Imaging and Therapy of Cancer , 2004, Technology in cancer research & treatment.

[33]  N. Beekman,et al.  Highly immunogenic and fully synthetic peptide-carrier constructs targetting GnRH. , 1999, Vaccine.