Salivary gland delivery of pDNA-cationic lipoplexes elicits systemic immune responses

V. Sankar, L. Baccaglini, M. Sawdey, C. J. Wheeler, S. R. Pillemer, B. J. Baum, J. C. Atkinson

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


Objective: To test the ability of two cationic lipoplexes, Vaxfectin and GAP-DLRIE/DOPE, to facilitate transfection and elicit immune responses from plasmid DNAs (pDNAs) after retrograde instillation into salivary glands. Methods: Two pDNA expression vectors encoding either the influenza NP protein or human growth hormone (hGH) were complexed with the cationic lipid transfection reagents, GAP-DLRIE/DOPE or Vaxfectin, and delivered to the submandibular glands of rats. Samples from rats receiving the influenza NP protein pDNA and cationic lipoplexes were analyzed for anti-influenza NP-specific IgG1, IgG2a, and IgG2b in serum, and IgA in saliva, by an enzyme- linked immunosorbent assay (ELISA). Cytotoxic T-cell lymphocyte (CTL) assays were also performed. Transgene protein expression (hGH) was determined from extracts of submandibular glands of rats receiving hGH lipoplexes. Results: Serum antibodies (IgG) against the NP protein developed and were highest in all rats vaccinated with GAP-DLRIE/DOPE or Vaxfectin. The major serum IgG subclass stimulated by this route of immunization was IgG2b, followed by IgG2a. CTL assay results showed statistically significantly higher percentage killing in the Vaxfectin group than controls (P < 0.05). No rats developed IgA antibodies to NP protein in saliva. Animals receiving plasmid encoding hGH and either lipoplex expressed significantly higher amounts of hGH compared with those receiving the hGH plasmid and water. Although hGH expression was higher in the animals receiving pDNA/Vaxfectin (∼30-fold > pDNA/water), the difference with those receiving pDNA/GAP-DLRIE/DOPE (∼10-fold > pDNA/water) was not significant. Conclusions: Retrograde instillation of pDNA complexed with Vaxfectin into the salivary glands can stimulate cytotoxic and humoral responses to the influenza NP protein antigen. Optimization of the conditions required to stimulate humoral and secretory antibody formation may facilitate use of this tissue for specific clinical applications of pDNA immunization.

Original languageEnglish (US)
Pages (from-to)275-281
Number of pages7
JournalOral Diseases
Issue number6
StatePublished - Nov 1 2002


  • Cationic liposomes
  • Gene transfer
  • Plasmid DNA
  • Salivary
  • Vaccination

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Dentistry(all)


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