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Neospora caninum: antibodies directed against tachyzoite surface protein NcSRS2 inhibit parasite attachment and invasion of placental trophoblasts in vitro
Authors:Haldorson Gary J  Stanton James B  Mathison Bruce A  Suarez Carlos E  Baszler Tim V
Affiliation:Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, WA 99164-7040, USA.
Abstract:Polyclonal and monoclonal antibodies to native Neospora caninum tachyzoite surface protein NcSRS2 were generated and tested in vitro for their ability to neutralize tachyzoite attachment to and invasion of host cells. Host cells included Vero cells and a newly cloned, immortalized ovine trophoblast cell line obtained from primary cultures of ovine placenta. The ovine trophoblasts had morphology consistent with fetal trophoblasts and expressed mRNA for interferon-tau, a marker for trophoblasts. Native NcSRS2 was used to immunize mice to obtain monospecific anti-NcSRS2 polyclonal serum and anti-NcSRS2 monoclonal antibodies. Compared to irrelevant antibodies, monospecific anti-NcSRS2 serum and two anti-NcSRS2 monoclonal antibodies, 100.2.4.4 and 119.4.9.10, significantly blocked invasion of tachyzoites into both trophoblasts and Vero cells. Parasite attachment, assessed by IFA, was significantly reduced by anti-NcSRS2 mAb 100.2.4.4 and monospecific serum. The findings provide rationale to investigate a role for antibodies to NcSRS2 in prevention of N. caninum transplacental transmission in vivo.
Keywords:Neospora caninum   Adhesion/invasion assays   Monoclonal antibodies   NcSRS2   DNA, deoxyribonucleic acid   mAb, monoclonal antibody   HBSS, Hanks’ balanced salt solution   DMEM, Dulbecco’s modified Eagle’s medium   SV40, Simian virus 40   PCR, polymerase chain reaction   ELISA, enzyme-linked immunosorbant assay   PBS, phosphate buffered saline   HRP, horseradish peroxidase   ANOVA, analysis of variance
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