Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.
Abstract
Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.
참고문헌 (26)
- Michael A. Page, "Acanthamoeba keratitis: a 12-year experience covering a wide spectrum of presentations, diagnoses, and outcomes", Journal of Ophthalmology, 2013, pp.1~6 (2013), Hindawi Limited[DOI]
- Arshi Singh, "Acanthamoeba Keratitis Versus Mixed Acanthamoeba and Bacterial Keratitis: Comparison of Clinical and Microbiological Profiles", Cornea, 39(9), pp.1112~1116 (2020), Ovid Technologies (Wolters Kluwer Health)[DOI]
- Anita Raghavan, "The Acanthamoeba–Fungal Keratitis Study", American Journal of Ophthalmology, 201, pp.31~36 (2019), Elsevier BV[DOI]
- Nicholas Fanselow, "Acanthamoeba Keratitis, Pathology, Diagnosis and Treatment", Pathogens, 10(3), pp.323~ (2021), MDPI AG[DOI]
- Nóra Szentmáry, "Acanthamoeba keratitis – Clinical signs, differential diagnosis and treatment", Journal of Current Ophthalmology, 31(1), pp.16~23 (2019), Medknow[DOI]
- Sharma S, "Evaluation of immunoperoxidase staining technique in the diagnosis of Acanthamoeba keratitis", Indian J Ophthalmol, 49, pp.181~186 (2001)
- Marian L. Turner, "Antigens of selected Acanthamoeba species detected with monoclonal antibodies", International Journal for Parasitology, 35(9), pp.981~990 (2005), Elsevier BV[DOI]
- Michele Martha Weber-Lima, "Acanthamoeba spp. monoclonal antibody against a CPA2 transporter: a promising molecular tool for acanthamoebiasis diagnosis and encystment study", Parasitology, 147(14), pp.1678~1688 (2020), Cambridge University Press (CUP)[DOI]
- A-Young Kang, "Production of a monoclonal antibody against a mannose-binding protein of Acanthamoeba culbertsoni and its localization", Experimental Parasitology, 192, pp.19~24 (2018), Elsevier BV[DOI]
- So-Min Park, "Production of a polyclonal antibody against inosine-uridine preferring nucleoside hydrolase of Acanthamoeba castellanii and its access to diagnosis of Acanthamoeba keratitis", PLOS ONE, 15(9), pp.e0239867~ (2020), Public Library of Science (PLoS)[DOI]
- Hae-Ahm Lee, "Chorismate mutase peptide antibody enables specific detection of Acanthamoeba", PLOS ONE, 16(4), pp.e0250342~ (2021), Public Library of Science (PLoS)[DOI]
- Jeffrey Field, "Purification of a <i>RAS<", i>-Responsive Adenylyl Cyclase Complex from <i>Saccharomyces cerevisiae<, Molecular and Cellular Biology, pp.1988, 8(5)[DOI]
- Noegel AA, "Assessing the role of the ASP56", CAP homologue of Dictyostelium discoideum and the requirements for subcellular localization, 1999, pp.112, 3195~3203
- G. V. Kakurina, "Adenylyl cyclase-associated protein 1: Structure, regulation, and participation in cellular processes", Biochemistry (Moscow), 83(1), pp.45~53 (2018), Pleiades Publishing Ltd[DOI]
- Andrew V. Hubberstey, "Cyclase‐associated proteins: CAPacity for linking signal transduction and actin polymerization", The FASEB Journal, 16(6), pp.487~499 (2002), Wiley[DOI]
- Haitao Zhang, "Mammalian Adenylyl Cyclase-associated Protein 1 (CAP1) Regulates Cofilin Function, the Actin Cytoskeleton, and Cell Adhesion", Journal of Biological Chemistry, 288(29), pp.20966~20977 (2013), Elsevier BV[DOI]
- Li Li, "CAP2 is a Valuable Biomarker for Diagnosis and Prognostic in Patients with Gastric Cancer", Pathology & Oncology Research, 26(1), pp.273~279 (2018), Springer Science and Business Media LLC[DOI]
- Lihua Xu, "Expression status of cyclase-associated protein 2 as a prognostic marker for human breast cancer", Oncology Reports, 36(4), pp.1981~1988 (2016), Spandidos Publications[DOI]
- Jia Fu, "Increased Expression of CAP2 Indicates Poor Prognosis in Hepatocellular Carcinoma", Translational Oncology, 8(5), pp.400~406 (2015), Elsevier BV[DOI]
- 문은경, "Comparison of Proteins Secreted into Extracellular Space of Pathogenic and Non-pathogenic Acanthamoeba castellanii", Parasites, Hosts and Diseases, 56(6), pp.553~558 (2018), 대한기생충학ㆍ열대의학회
- Daniel W. Clarke, "The pathophysiology of Acanthamoeba keratitis", Trends in Parasitology, 22(4), pp.175~180 (2006), Elsevier BV[DOI]
- Nicholas J. R. Maycock, "Update on Acanthamoeba Keratitis", Cornea, 35(5), pp.713~720 (2016), Ovid Technologies (Wolters Kluwer Health)[DOI]
- S. Sharma, "Patient characteristics, diagnosis, and treatment of non-contact lens related Acanthamoeba keratitis", British Journal of Ophthalmology, 84(10), pp.1103~1108 (2000), BMJ[DOI]
- Hassan Alizadeh, "Tear IgA and Serum IgG Antibodies Against Acanthamoeba in Patients With Acanthamoeba Keratitis", Cornea, 20(6), pp.622~627 (2001), Ovid Technologies (Wolters Kluwer Health)[DOI]
- Walochnik J., "Anti- Acanthamoeba IgG, IgM, and IgA immunoreactivities in correlation to strain pathogenicity", Parasitology Research, 87(8), pp.651~656 (2001), Springer Science and Business Media LLC[DOI]
- Elisa Alvarez-Curto, "cAMP production by adenylyl cyclase G induces prespore differentiation in<i>Dictyostelium<", i>slugs, 2007, pp.134(5), 959~966[DOI]