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Identification of antigenic proteins from Neospora caninum recognized by bovine immunoglobulins M, E, A and G using
Yong-seung Shin1, Eung-goo Lee, Gee-wook Shin
1Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju, Republic of Korea.
Insights
This study identified key Neospora caninum antigenic proteins recognized by bovine antibodies. These immunodominant proteins, including HSP70 and actin, are crucial for developing new diagnostic markers and vaccines against Neospora caninum infections.
Area of Science:
- Veterinary Parasitology
- Immunology
- Proteomics
Background:
- Neospora caninum is a significant protozoan parasite causing abortion in cattle.
- Understanding the antigenic proteins of N. caninum is crucial for developing effective control strategies.
Purpose of the Study:
- To investigate the antigenic proteins of Neospora caninum (N. caninum) recognized by different classes of bovine immunoglobulins (IgM, IgE, IgA, IgG).
- To identify immunodominant proteins that could serve as targets for diagnostic markers and vaccines.
Main Methods:
- Immunoproteomics approach using two-dimensional gel electrophoresis and Western blotting with bovine immunoglobulins.
- Identification of antigenic proteins using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS).
Main Results:
- A total of 132, 84, 4, and 40 antigenic protein spots were identified against bovine IgM, IgE, IgA, and IgG, respectively.
- Several proteins, including HSP70, actin, NTPase, HSP60, pyruvate kinase, enolase, and GRA2, were identified as immunodominant.
- Identified antigenic proteins are involved in apicomplexan invasion, proliferation, and egress.
Conclusions:
- HSP70, actin, NTPase, HSP60, pyruvate kinase, enolase, putative ribosomal protein S2, NCDG-1, and GRA2 are identified as immunodominant antigens of N. caninum.
- These immunodominant proteins hold potential for the development of novel diagnostic tools and vaccines for bovine neosporosis.
Abstract:
Antigenic proteins of Neospora caninum (N. caninum) against bovine immunoglobulins M, E, A, and G were investigated by using immunoproteomics. Proteins of N. caninum (KBA-2) tachyzoite lysates separated by two-dimensional gel electrophoresis were transferred to polyvinylidene difluoride (PVDF) membranes, probed with different bovine immunoglobulin class and classified. Antigenic spots recognized were also identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis. 132, 84, 4, and 40 antigenic protein spots were recognized on N. caninum immunoblot profiles against bovine IgM, IgE, IgA, and IgG, respectively. Of these protein spots, the antigenic proteins recognized by either IgM, IgE, and IgG, or IgM and IgG were HSP70, pyruvate kinase, actin, NCDG-1, tubulin alpha-chain, and putative ribosomal protein S2. On the other hand, IgM, IgE, and IgA reacted with NTPase, HSP60, tubulin beta-chain, putative protein disulfide isomerase, enolase, lactate dehydrogenase, serine-threonine phosphatase, 14-3-3 protein homologue, and GRA2 protein. Most of the antigenic proteins identified were associated with the process of invasion, proliferation, and egression of apicomplexans. In our study, HSP70, actin, NTPase, HSP60, pyruvate kinase, enolase, putative ribosomal protein S2, NCDG-1, and GRA2 proteins were found to be immunodominant proteins, which may contribute to the development of diagnostic markers and vaccine.
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