Theileria equi claudin like apicomplexan microneme protein contains neutralization-sensitive epitopes and interacts with components of the equine erythrocyte membrane skeleton.
Abstract: Theileria equi is a widely distributed apicomplexan parasite that causes severe hemolytic anemia in equid species. There is currently no effective vaccine for control of the parasite and understanding the mechanism that T. equi utilizes to invade host cells may be crucial for vaccine development. Unlike most apicomplexan species studied to date, the role of micronemes in T. equi invasion of host cells is unknown. We therefore assessed the role of the T. equi claudin-like apicomplexan microneme protein (CLAMP) in the invasion of equine erythrocytes as a first step towards understanding the role of this organelle in the parasite. Our findings show that CLAMP is expressed in the merozoite and intra-erythrocytic developmental stages of T. equi and in vitro neutralization experiments suggest that the protein is involved in erythrocyte invasion. Proteomic analyses indicate that CLAMP interacts with the equine erythrocyte α-and β- spectrin chains in the initial stages of T. equi invasion and maintains these interactions while also associating with the anion-exchange protein, tropomyosin 3, band 4.1 and cytoplasmic actin 1 after invasion. Additionally, serological analyses show that T. equi-infected horses mount robust antibody responses against CLAMP indicating that the protein is immunogenic and therefore represents a potential vaccine candidate.
Publication Date: 2021-04-29 PubMed ID: 33927329PubMed Central: PMC8085155DOI: 10.1038/s41598-021-88902-4Google Scholar: Lookup
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- Journal Article
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Summary
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This research explores the role of claudin-like apicomplexan microneme protein (CLAMP), in the invasion mechanism of Theileria equi, a parasite causing severe anemia in equid species. The study provides evidence of CLAMP’s interaction with the membrane skeleton of equine erythrocytes and its potential viability as a vaccine candidate.
Overview of the Parasite and the Protein
- Theileria equi is a widely distributed parasite, responsible for severe anemia in equid species such as horses and donkeys. There’s currently no effective vaccine to control the parasite, hence the need for extensive study.
- The claudin-like apicomplexan microneme protein (CLAMP) under investigation in this study is an essential organ of the parasite suspected to be involved in the invasion process of host cells, particularly equine erythrocytes (red blood cells).
The Role of CLAMP in Invasion
- The researchers found that CLAMP is expressed during the merozoite and intra-erythrocytic stages of T. equi development— the stages within the life cycle of the parasite when it invades and develops within red blood cells.
- Neutralization experiments performed in vitro suggest that CLAMP plays a significant role in the invasion of erythrocytes by the parasite. These experiments involved crippling or “neutralizing” the function of the protein to determine if it would affect the parasite’s invasion capabilities.
CLAMP Interaction with Erythrocyte Components
- Proteomic analysis showed that during the initial stages of invasion, CLAMP interacts with α-and β-spectrin chains—an integral part of the equine erythrocyte membrane skeleton. This interaction is sustained, with CLAMP later associating with other proteins including the anion-exchange protein, tropomyosin 3, band 4.1, and cytoplasmic actin 1.
- This suggests that CLAMP may help T. equi anchor to or interact with these structures within the host cell to facilitate invasion and successful infection.
Potential for Vaccine Development
- The study also found that horses infected with T. equi produced strong antibody responses against CLAMP. Antibodies are our immune system’s natural defense against foreign invaders, and the response signifies that the protein is recognized as a threat by horses’ immune systems.
- This finding opens the potential for CLAMP to be a viable vaccine candidate, as it could induce a protective immune response in horses before they encounter the parasite, ultimately aiding in the control of this debilitating disease.
Cite This Article
APA
Onzere CK, Fry LM, Bishop RP, Silva MG, Bastos RG, Knowles DP, Suarez CE.
(2021).
Theileria equi claudin like apicomplexan microneme protein contains neutralization-sensitive epitopes and interacts with components of the equine erythrocyte membrane skeleton.
Sci Rep, 11(1), 9301.
https://doi.org/10.1038/s41598-021-88902-4 Publication
Researcher Affiliations
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA. cynthia.onzere@wsu.edu.
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA.
- Animal Disease Research Unit, Agricultural Research Service, US Department of Agriculture, P.O. Box 646630, Pullman, WA, 99164, USA.
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA.
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA.
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA.
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA.
- Department of Veterinary Microbiology and Pathology, Washington State University, P.O. Box 647040, Pullman, WA, 99164, USA. carlos.suarez@usda.gov.
- Animal Disease Research Unit, Agricultural Research Service, US Department of Agriculture, P.O. Box 646630, Pullman, WA, 99164, USA. carlos.suarez@usda.gov.
MeSH Terms
- Animals
- Antibodies, Protozoan / blood
- Antibodies, Protozoan / immunology
- Antigens, Protozoan / immunology
- Blood Proteins / metabolism
- Claudins
- Epitopes, B-Lymphocyte / immunology
- Erythrocyte Membrane / metabolism
- Erythrocytes / parasitology
- Horse Diseases / immunology
- Horse Diseases / parasitology
- Horses / blood
- Horses / parasitology
- Membrane Proteins / chemistry
- Membrane Proteins / genetics
- Membrane Proteins / immunology
- Membrane Proteins / metabolism
- Merozoites / genetics
- Merozoites / metabolism
- Neutralization Tests
- Protozoan Proteins / chemistry
- Protozoan Proteins / genetics
- Protozoan Proteins / immunology
- Protozoan Proteins / metabolism
- Theileria / growth & development
- Theileria / immunology
- Theileria / metabolism
- Theileria / pathogenicity
- Theileriasis / immunology
- Theileriasis / parasitology
Conflict of Interest Statement
The authors declare no competing interests.
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Citations
This article has been cited 3 times.- Loubens M, Marinach C, Paquereau CE, Hamada S, Hoareau-Coudert B, Akbar D, Franetich JF, Silvie O. The claudin-like apicomplexan microneme protein is required for gliding motility and infectivity of Plasmodium sporozoites. PLoS Pathog 2023 Mar;19(3):e1011261.
- Elsworth B, Keroack CD, Rezvani Y, Paul AS, Barazorda KA, Bauer NC, Tennessen JA, Sack SA, Moreira CK, Gubbels MJ, Meyers MJ, Zarringhalam K, Duraisingh MT. Babesia divergens host cell egress is mediated by essential and druggable kinases and proteases. Nat Microbiol 2026 Feb;11(2):492-506.
- Onzere CK, Bastos RG, Bishop RP, Suarez CE, Fry LM. Expression of IL-10 and TGF-β1 in horses experimentally infected with T. equi merozoites is associated with antibody production but not modulation of pro-inflammatory responses. Front Immunol 2024;15:1370255.
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