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Clinical and Vaccine Immunology, April 2008, p. 659-667, Vol. 15, No. 4
1071-412X/08/$08.00+0     doi:10.1128/CVI.00436-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Bovine Immune Response to Inoculation with Neospora caninum Surface Antigen SRS2 Lipopeptides Mimics Immune Response to Infection with Live Parasites

Timothy V. Baszler,^1* Varda Shkap,² Waithaka Mwangi,^1, Christopher J. Davies,¹ Bruce A. Mathison,¹ Monica Mazuz,² Dror Resnikov,³ Lea Fish,² Benjamin Leibovitch,² Lauren M. Staska,^1, and Igor Savitsky²

Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040,¹ Division of Parasitology, Kimron Veterinary Institute, Bet-Dagan, 50250 Israel,² Mutual Society for Insurance and Veterinary Services, Hahaklait, Israel³

Received 2 November 2007/ Returned for modification 19 December 2007/ Accepted 19 February 2008

Infection of cattle with Neospora caninum protozoa, the causative agent of bovine protozoal abortion, results in robust cellular and humoral immune responses, particularly CD4⁺ T-lymphocyte activation and gamma interferon (IFN-) secretion. In the present study, N. caninum SRS2 (NcSRS2) T-lymphocyte-epitope-bearing subunits were incorporated into DNA and peptide preparations to assess CD4⁺ cell proliferation and IFN- T-lymphocyte-secretion immune responses in cattle with predetermined major histocompatibility complex (MHC) genotypes. In order to optimize dendritic-cell processing, NcSRS2 DNA vaccine was delivered with granulocyte macrophage-colony-stimulating factor and Flt3 ligand adjuvant. The synthesized NcSRS2 peptides were coupled with a palmitic acid molecule (lipopeptide) and delivered with Freund's adjuvant. Cattle vaccinated with NcSRS2 DNA vaccine alone did not induce T-lymphocyte activation or IFN- secretion, whereas subsequent booster inoculation with NcSRS2-lipopeptides induced robust NcSRS2-specific immune responses. Compared to the response in control animals, NcSRS2-lipopeptide-immunized cattle had significantly increased NcSRS2-specific T-lymphocyte proliferation, numbers of IFN--secreting peripheral blood mononuclear cells, and immunoglobulin G1 (IgG1) and IgG2a antibody levels. The findings show that N. caninum NcSRS2 subunits bearing T-lymphocyte epitopes induced cell-mediated immune responses similar to the protective immune responses previously described against live parasite infection, namely T-lymphocyte activation and IFN- secretion. The findings support the investigation of NcSRS2 immunogens for protection against N. caninum-induced fetal infection and abortion in cattle.

Published ahead of print on 27 February 2008.

Present address: Department of Veterinary Pathobiology, Texas A&M University, College Station, TX 77843-4467.

Present address: Battelle Pacific Northwest National Laboratory, Richland, WA 99352.