Targeting the Cryptococcus neoformans var. grubii cell wall using lectins: study of the carbohydrate-binding domain.
Cryptococcus neoformans var. grubii is considered to be the major cause of cryptococcosis in immunosuppressed patients. Understanding cell wall glycoproteins using lectins is of medical interest and can contribute to specific therapy. The aim of this study was to evaluate the carbohydrates on the cell wall of Cryptococcus neoformans var. grubii clinical isolates, using a fluorescein isothiocyanate-lectin binding protocol. Thirty yeast strains stocked in the culture collection were cultivated for 2 days at 30 °C with shaking. Cells were obtained by centrifugation, washed in phosphate-buffered saline, and a suspension of 107 cells/mL was obtained. To determine the binding profile of lectins, concanavalin A (Con A), wheat germ agglutinin (WGA), Ulex europaeus agglutinin I (UEA-I), and peanut agglutinin (PNA) conjugated to fluorescein were used. All the tested clinical isolates of Cryptococcus neoformans var. grubii were intensely stained by WGA, moderately stained by Con A, and weakly stained by PNA and UEA-I. Thus, Cryptococcus can be detected in clinical specimens such as blood and cerebrospinal fluid using the fluorescent lectin WGA, which may be considered as an option for detection in cases of suspected cryptococcosis with low laboratory sensitivity. Future applications may be developed using this basic tool.
Department of Mycology, Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves, s/n°-Cidade Universitária, Recife 50670-420, Brazil. pamella_ximenes@hotmail.com.
Pamella
P
de Brito Ximenes
Department of Biochemistry, Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves, s/n°-Cidade Universitária, Recife 50670-420, Brazil. ebeltrao@hotmail.com.
Eduardo Isidoro Carneiro
EI
Beltrão
Department of Pharmaceutical Sciences, Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves, s/n°-Cidade Universitária, Recife 50670-420, Brazil. daniellemacedo28@gmail.com.
Danielle Patrícia Cerqueira
DP
Macêdo
Department of Mycology, Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves, s/n°-Cidade Universitária, Recife 50670-420, Brazil. danielabuonafina@hotmail.com.
Maria Daniela Silva
MD
Buonafina
Department of Tropical Medicine, Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves, s/n°-Cidade Universitária, Recife 50670-420, Brazil. goncalves_reginaldo@hotmail.com.
Reginaldo Gonçalves
RG
de Lima-Neto
Department of Mycology, Universidade Federal de Pernambuco (UFPE), Av. Prof. Nelson Chaves, s/n°-Cidade Universitária, Recife 50670-420, Brazil. rejadel@yahoo.com.br.
Rejane Pereira
RP
Neves
D002241:Carbohydrates; D002473:Cell Wall / Q000378:metabolism*; D003208:Concanavalin A / Q000378:metabolism*; D003453:Cryptococcosis / Q000378:metabolism* / Q000382:microbiology / Q000473:pathology; D003455:Cryptococcus neoformans / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000378:metabolism*
D002241:Carbohydrates; D037102:Lectins; D003208:Concanavalin A