TY - JOUR
T1 - Antigens of selected Acanthamoeba species detected with monoclonal antibodies
AU - Turner, Marian L.
AU - Cockerell, Emma J.
AU - Brereton, Helen M.
AU - Badenoch, Paul R.
AU - Tea, Melinda
AU - Coster, Douglas J.
AU - Williams, Keryn A.
N1 - Funding Information:
This work was supported by the National Health and Medical Research Council of Australia, the Ophthalmic Research Institute of Australia, and the Flinders Medical Centre Foundation. The authors thank Bret Robinson from the Australian Water Quality Centre, Bolivar, Australia for providing reference strains of Acanthamoeba , Graham Nye and Alex Szabo from Gribbles Pathology, Wayville, Australia, for providing sections of human cornea, and Kirsty Marshall for expert technical assistance.
PY - 2005/8
Y1 - 2005/8
N2 - Acanthamoeba species are ubiquitous soil and freshwater protozoa that have been associated with infections of the human brain, skin, lungs and eyes. Our aim was to develop specific antibodies to aid in rapid and specific diagnosis of clinically important isolates. Mice were variously immunised with live mixtures of Acanthamoeba castellanii strain 112 (AC112) trophozoites and cysts, or with sonicated, formalin-fixed or heat-treated trophozoites, or with a trophozoite membrane preparation. Eight hybridoma cell lines secreting monoclonal antibodies reactive with A. castellanii epitopes were generated. Seven of the new antibodies (designated AMEC1-3 and MTAC1-4) were isotyped as IgMκ and one (MTAC5) as IgG1κ. All of the novel antibodies bound to AC112 cysts, and MTAC4 and MTAC5 also bound to trophozoites as measured by flow cytometry on unfixed cells. Single chain antibody fragments that retained parental antibody binding characteristics were engineered from three of the hybridomas (AMEC1, MTAC3 and MTAC4). Four monoclonal antibodies (AMEC1, AMEC3, MTAC1, MTAC3) bound reliably to unfixed cysts of clinical isolates of A. castellanii (two strains) and Acanthamoeba polyphaga (two strains), belonging to Pussard-Pons morphological group II, and to Acanthamoeba lenticulata and Acanthamoeba culbertsoni, belonging to Pussard-Pons morphological group III. None of the antibodies bound to cysts or trophozoites of the environmental group I species, Acanthamoeba tubiashi. Antibodies AMEC1, MTAC3, MTAC4 and MTAC5 reacted with buffered formalin-fixed AC112 by immunohistochemistry, and also stained Acanthamoeba in sections of infected rat cornea and buffered formalin-fixed, paraffin-embedded infected human cornea. These antibodies may be useful in diagnosing pathogenic Acanthamoeba species in clinical specimens, provided that cysts are present.
AB - Acanthamoeba species are ubiquitous soil and freshwater protozoa that have been associated with infections of the human brain, skin, lungs and eyes. Our aim was to develop specific antibodies to aid in rapid and specific diagnosis of clinically important isolates. Mice were variously immunised with live mixtures of Acanthamoeba castellanii strain 112 (AC112) trophozoites and cysts, or with sonicated, formalin-fixed or heat-treated trophozoites, or with a trophozoite membrane preparation. Eight hybridoma cell lines secreting monoclonal antibodies reactive with A. castellanii epitopes were generated. Seven of the new antibodies (designated AMEC1-3 and MTAC1-4) were isotyped as IgMκ and one (MTAC5) as IgG1κ. All of the novel antibodies bound to AC112 cysts, and MTAC4 and MTAC5 also bound to trophozoites as measured by flow cytometry on unfixed cells. Single chain antibody fragments that retained parental antibody binding characteristics were engineered from three of the hybridomas (AMEC1, MTAC3 and MTAC4). Four monoclonal antibodies (AMEC1, AMEC3, MTAC1, MTAC3) bound reliably to unfixed cysts of clinical isolates of A. castellanii (two strains) and Acanthamoeba polyphaga (two strains), belonging to Pussard-Pons morphological group II, and to Acanthamoeba lenticulata and Acanthamoeba culbertsoni, belonging to Pussard-Pons morphological group III. None of the antibodies bound to cysts or trophozoites of the environmental group I species, Acanthamoeba tubiashi. Antibodies AMEC1, MTAC3, MTAC4 and MTAC5 reacted with buffered formalin-fixed AC112 by immunohistochemistry, and also stained Acanthamoeba in sections of infected rat cornea and buffered formalin-fixed, paraffin-embedded infected human cornea. These antibodies may be useful in diagnosing pathogenic Acanthamoeba species in clinical specimens, provided that cysts are present.
KW - Acanthamoeba
KW - Cornea
KW - Cyst
KW - Monoclonal antibody
KW - Trophozoite
UR - http://www.scopus.com/inward/record.url?scp=23044507324&partnerID=8YFLogxK
U2 - 10.1016/j.ijpara.2005.03.015
DO - 10.1016/j.ijpara.2005.03.015
M3 - Article
C2 - 15964573
AN - SCOPUS:23044507324
SN - 0020-7519
VL - 35
SP - 981
EP - 990
JO - International Journal for Parasitology
JF - International Journal for Parasitology
IS - 9
ER -