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1From the Center for Neurobiology and Vaccine Development, Ophthalmology Research, Department of Surgery, Cedars-Sinai Medical Center Burns and Allen Research Institute, Los Angeles, California; and the 2Department of Ophthalmology, University of California Los Angeles School of Medicine, Los Angeles, California.
PURPOSE. To compare the effectiveness of immunization with "naked" DNA corresponding to the genes encoding five HSV-1 glycoproteins, gB, gC, gD, gE, and gI (5gP DNA), with immunization with the five glycoproteins (5gP protein). Also, to compare immunization of 5gP protein in Montanide ISA 720 (SEPPIC, Paris, France), an adjuvant recently approved for use in humans, with immunization of 5gP protein in Freund’s adjuvant.
METHODS. BALB/c mice were vaccinated with 5gP DNA or 5gP protein emulsified in ISA 720 or Freund’s adjuvant. Neutralizing antibody titers were determined by plaque-reduction assays. IL-2, -4, and -12 and IFN-
levels were determined by ELISA after in vitro stimulation of spleen cells. After ocular challenge with 2 x 105 plaque-forming units [pfu] per eye of HSV-1 strain McKrae, virus replication in the eye, survival, blepharitis, corneal scarring, and latency were determined.
RESULTS. Neutralizing antibody titers (approximately 1:800–1:1200), corneal scarring (trace) and survival (100%) were similar for all vaccine groups, including 5gP DNA. Compared with the other vaccine groups, the 5gP DNA group had less ocular virus replication, as judged both by maximum virus titer and time of viral clearance. ISA 720 appeared more effective than Freund’s against ocular virus replication and eye disease. The 5gP DNA–vaccinated mice had less blepharitis and latency than any other group and had the highest levels of IL-12 and IFN-. All vaccine groups had similar levels of IL-2.
CONCLUSIONS. The 5gP DNA vaccine appeared to be more effective than the corresponding protein subunit vaccine, regardless of adjuvant. Emulsification of the 5gP protein in ISA 720 appeared to be more effective than emulsification in Freund’s adjuvant.
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