Neospora caninum: Antibodies directed against tachyzoite surface protein NcSRS2 inhibit parasite attachment and invasion of placental trophoblasts in vitro

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...

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Bibliographic Details
Published in:Experimental parasitology 2006-03, Vol.112 (3), p.172-178
Main Authors: Haldorson, Gary J., Stanton, James B., Mathison, Bruce A., Suarez, Carlos E., Baszler, Tim V.
Format: Article
Language:English
Subjects:
Adhesion/invasion assays
analysis of variance
Animals
ANOVA
antibodies
Antibodies, Monoclonal - immunology
antigen-antibody reactions
Antigens, Protozoan - immunology
Antigens, Surface - immunology
Biological and medical sciences
Blotting, Western - veterinary
cell adhesion
cell invasion
Cell Line
cell lines
Chlorocebus aethiops
Coccidiosis - immunology
Coccidiosis - transmission
Coccidiosis - veterinary
deoxyribonucleic acid
DMEM
DNA
Dulbecco’s modified Eagle’s medium
ELISA
enzyme-linked immunosorbant assay
Female
Fundamental and applied biological sciences. Psychology
Hanks’ balanced salt solution
HBSS
horseradish peroxidase
HRP
Immunohistochemistry - veterinary
Immunophenotyping - methods
Immunophenotyping - veterinary
Infectious Disease Transmission, Vertical - veterinary
Interferon Type I - genetics
Interferon Type I - immunology
Life cycle. Host-agent relationship. Pathogenesis
mAb
Mice
Mice, Inbred BALB C
Monoclonal antibodies
monoclonal antibody
NcSRS2
Neospora - immunology
Neospora - physiology
Neospora caninum
neosporosis
outer membrane proteins
PBS
PCR
phosphate buffered saline
placenta
Placenta - cytology
Placenta - parasitology
polymerase chain reaction
Pregnancy
Pregnancy Proteins - genetics
Pregnancy Proteins - immunology
Protozoa
Protozoan Proteins - immunology
Reverse Transcriptase Polymerase Chain Reaction - veterinary
Sheep
Simian virus 40
surface antigens
SV40
tachyzoites
Transfection - veterinary
transplacental transmission
trophoblast
Trophoblasts - immunology
Trophoblasts - parasitology
Vero Cells
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Summary: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-τ, 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.
ISSN:0014-4894
1090-2449
DOI:10.1016/j.exppara.2005.11.004