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Molecular conservation of MSP4 and MSP5 in Anaplasma marginale and A. centrale vaccine strain
T Molad1, K A Brayton, G H Palmer
1Division of Parasitology, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
Abstract:
Anaplasma centrale msp4 and msp5 genes were cloned and sequenced, and the recombinant proteins were expressed. The identity between Anaplasma marginale and A. centrale MSP4 was 83% in the nucleotide sequences and 91.7% in the encoded protein sequences. A. centrale msp5 nucleotide sequences shared 86.8% identity with A. marginale msp5, and there was 92.9% homology between A. centrale and A. marginale encoded amino acids of the MSP5 protein. Southern blots hybridized with probes derived from the msp4 and msp5 central regions indicate that msp4 and msp5 of A. centrale are encoded by single copy genes. Recombinant MSP4 and MSP5 fusion proteins reacted with anti-A. marginale monoclonal antibodies ANAR76A1 and ANAF16C, respectively, demonstrating the conservation of conformation-sensitive B-cell epitopes between A. centrale and A. marginale. These data demonstrate the structural and antigenic conservation of MSP4 and MSP5 in A. centrale and A. marginale. This conservation is consistent with the cross-protective immunity between A. marginale and A. centrale and supports the development of improved vaccines based upon common outer membrane proteins.
Insights
Anaplasma centrale and Anaplasma marginale share highly conserved outer membrane proteins MSP4 and MSP5. This structural and antigenic similarity explains their cross-protective immunity and aids vaccine development.
Area of Science:
- Veterinary Microbiology
- Molecular Biology
- Immunology
Background:
- Anaplasma marginale and Anaplasma centrale are tick-borne pathogens causing significant economic losses in cattle.
- Outer membrane proteins (OMPs) are crucial for Anaplasma pathogenesis and host immune responses.
- MSP4 and MSP5 are key OMPs involved in Anaplasma infections.
Purpose of the Study:
- To investigate the genetic and antigenic conservation of Anaplasma centrale msp4 and msp5 genes and their encoded proteins.
- To compare these genes and proteins with those of Anaplasma marginale.
- To assess the implications of this conservation for cross-protective immunity and vaccine development.
Main Methods:
- Cloning and sequencing of Anaplasma centrale msp4 and msp5 genes.
- Expression of recombinant MSP4 and MSP5 proteins.
- Nucleotide and amino acid sequence analysis and comparison with Anaplasma marginale.
- Southern blot hybridization to determine gene copy number.
- Immunological assays using monoclonal antibodies to detect conserved epitopes.
Main Results:
- High nucleotide (83-86.8%) and amino acid (91.7-92.9%) identity between A. centrale and A. marginale MSP4 and MSP5.
- Southern blots confirmed msp4 and msp5 as single-copy genes in A. centrale.
- Recombinant MSP4 and MSP5 proteins reacted with anti-A. marginale monoclonal antibodies, indicating conserved B-cell epitopes.
- Demonstrated structural and antigenic conservation of MSP4 and MSP5.
Conclusions:
- The MSP4 and MSP5 proteins of A. centrale and A. marginale are structurally and antigenically conserved.
- This conservation explains the observed cross-protective immunity between these two Anaplasma species.
- The findings support the development of improved vaccines targeting common outer membrane proteins for Anaplasma control.
