Antigens of selected Acanthamoeba species detected with monoclonal antibodies

Turner, Marian L.; Cockerell, Emma J; Brereton, Helen M.; Badenoch, Paul R; Tea, Melinda N; Coster, Douglas J.; Williams, Keryn A.

doi:10.1016/j.ijpara.2005.03.015

Cited by 20 publications

(21 citation statements)

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“…These may be examined by staining with with calcofluor-white or immunoperoxidase [20] to aid in the detection of the trophozoites or cysts. Recent advances in immunohistological staining such as Acanthamoeba specific monoclonal antibodies reported by Turner et al [21] have added to the precision in which Acanthamoeba may be detected by traditional laboratory methods.…”

Section: Diagnosismentioning

confidence: 99%

Advances in the Diagnosis and Treatment of Acanthamoeba Keratitis

Clarke

Sinha

Parmar

et al. 2012

Journal of Ophthalmology

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Section: Diagnosismentioning

confidence: 99%

Advances in the Diagnosis and Treatment of Acanthamoeba Keratitis

Clarke

Sinha

Parmar

et al. 2012

Journal of Ophthalmology

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“…In a study performed on a single patient suffering from GAE, PCR was found to be the only technique to return a positive result for four CSF samples, dermal specimens, a bronchoalveolar lavage specimen, lung biopsy specimens, and brain biopsy specimens, while culture, microscopy, and immunofluorescence repeatedly returned negative results (594). Several monoclonal antibodies to surface antigens of Acanthamoeba cysts have also been developed, which could be useful for the development of new diagnostic tests in the future (563). Treatment.…”

Section: Free-living Amoebaementioning

confidence: 99%

Importance of Nonenteric Protozoan Infections in Immunocompromised People

et al. 2010

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SUMMARY There are many neglected nonenteric protozoa able to cause serious morbidity and mortality in humans, particularly in the developing world. Diseases caused by certain protozoa are often more severe in the presence of HIV. While information regarding neglected tropical diseases caused by trypanosomatids and Plasmodium is abundant, these protozoa are often not a first consideration in Western countries where they are not endemic. As such, diagnostics may not be available in these regions. Due to global travel and immigration, this has become an increasing problem. Inversely, in certain parts of the world (particularly sub-Saharan Africa), the HIV problem is so severe that diseases like microsporidiosis and toxoplasmosis are common. In Western countries, due to the availability of highly active antiretroviral therapy (HAART), these diseases are infrequently encountered. While free-living amoebae are rarely encountered in a clinical setting, when infections do occur, they are often fatal. Rapid diagnosis and treatment are essential to the survival of patients infected with these organisms. This paper reviews information on the diagnosis and treatment of nonenteric protozoal diseases in immunocompromised people, with a focus on patients infected with HIV. The nonenteric microsporidia, some trypanosomatids, Toxoplasma spp., Neospora spp., some free-living amoebae, Plasmodium spp., and Babesia spp. are discussed.

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“…Briefly, 5 × 10 5 log-phase trophozoites were allowed to adhere to 6-well plates in ATCC medium 712 for 30 min at 30°C. The adherent trophozoites were washed and replaced with 500 µl of non-nutrient medium (20 mM Tris-HCl (pH 8.8), 100 mM KCl, 8 mM MgSO 4 , 0.4 mM CaCl 2 and 1 mM NaHCO 3 ). In an Eppendorf tube, 4 µg of Midiprep (PureLink HiPure Midiprep Kit, Thermo Fisher Scientific) plasmid DNA was diluted to 100 µl with non-nutrient medium.…”

Section: Methodsmentioning

confidence: 99%

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

Samuelson

Magistrado-Coxen

Aqeel

et al. 2018

Preprint

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18Acanthamoeba castellanii, cause of keratitis and blindness, is an emerging pathogen because 19 of its association with contact lens use. The cyst wall contributes to pathogenesis as cysts are 20 resistant to sterilizing reagents in lens solutions and to antibiotics applied to the eye. We used 21 transmission electron microscopy, as well as structured illumination microscopy and probes for 22 cellulose and chitin, to show that purified cyst walls of A. castellanii retain an outer ectocyst 23 layer, an inner endocyst layer, and conical ostioles that connect the layers. Mass spectrometry 24 showed candidate cyst wall proteins are dominated by three families of lectins (named here 25 Luke, Leo, and Jonah), each of which binds to microcrystalline cellulose and to a lesser degree 26 chitin. A Jonah lectin, which has one choice-of-anchor A (CAA) domain, localizes to the 27 ectocyst layer of mature cyst walls. Luke lectins, which have two or three carbohydrate-binding 28 modules (CBM49), localize to the endocyst layer and ostioles. A Leo lectin, which has two 29 domains with eight Cys residues each (8-Cys), also localizes to the endocyst layer and 30 ostioles. In summary, the most abundant A. castellanii cyst wall proteins are three sets of 31 lectins, which have carbohydrate-binding modules that are conserved (CBM49s of Luke), 32 newly characterized (CAA of Jonah), or unique to Acanthamoebae (8-Cys of Leo). Despite 33 their lack of common ancestry, Luke and Leo lectins both localize to the endocyst layer and 34 ostioles, while the Jonah lectin localizes to the ectocyst layer. 35IMPORTANCE 36 Fifty years ago, investigators identified cellulose in the Acanthamoeba cyst wall, which has an 37 outer ectocyst layer, an inner endocyst layer, and conical ostioles that connect the layers. 38Here we show cyst walls also contain chitin and three large sets of cellulose-and chitin-binding 39 lectins, which have distinct localizations. The Acanthamoeba cyst wall, therefore, is more 40 complicated than cyst walls of Entamoeba and Giardia (causes of dysentery and diarrhea, 41 respectively), which have a single layer, a single glycopolymer, and small sets of one or two 42 lectins. In contrast, the Acanthamoeba cyst wall is far simpler than the walls of fungi and 43 plants, which have multiple layers, numerous glycopolymers, and hundreds of proteins. In 44 addition to providing a better understanding of the cell biology and biochemistry of the 45 Acanthamoeba cyst wall, these studies may lead to diagnostic antibodies that bind to cyst wall 46 proteins and/or therapeutics that target chitin. 47 100 file S1) (41). The GST-AcCBM49 expression construct was designed to replicate that used to 101 determine the carbohydrate binding properties of SlCBM49, which is a C-terminal 102 carbohydrate-binding module of the Solanum lycopersicum (tomato) cellulase SlGH9C (42). In 103 both intact cysts and purified cyst walls, GST-AcCBM49 predominantly labeled the ectocyst 104 layer, WGA highlighted the ostioles, and CFW stained the endocyst...

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Antigens of selected Acanthamoeba species detected with monoclonal antibodies

Cited by 20 publications

References 28 publications

Advances in the Diagnosis and Treatment of Acanthamoeba Keratitis

Advances in the Diagnosis and Treatment of Acanthamoeba Keratitis

Importance of Nonenteric Protozoan Infections in Immunocompromised People

The most abundant cyst wall proteins ofAcanthamoeba castellaniiare cellulose-binding lectins from three gene families that localize to distinct structures in cyst walls

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