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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2008.tde-25112008-134339
Document
Author
Full name
Priscilla Megumi Ueno
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2008
Supervisor
Committee
Timenetsky, Jorge (President)
Buim, Marcos Roberto
Cordova, Caio Mauricio Mendes de
Hirata, Mario Hiroyuki
Santelli, Glaucia Maria Machado
Title in Portuguese
Mycoplasma genitalium: curva de crescimento e interação com células humanas de cérvix (HeLa) e endometriais (EM42).
Keywords in Portuguese
Mycoplasma genitalium
Aderência
Curva de crescimento
Invasão citoplasmática
Invasão nuclear
Abstract in Portuguese
Mycoplasma (M.) genitalium é um importante agente de doença sexualmente transmissível sendo, responsável por uma série de desordens do trato urogenital humano. A aderência do micoplasma é um dos principais fatores de virulência na sua patogenicidade e conseqüente colonização nas células hospedeiras. Neste estudo, obte-se a curva de crescimento de duas cepas de M. genitalium (G37 e 1019V) utilizando-se da dosagem proteíca (BCA), densidade óptica (OD600) e PCR em tempo real. A cepa referencial G37 é de alta passagem e foi isolada de homens e a 1019V é de baixa passagem, sendo recentemente isolada de amostra clínica de cérvix humana. Utilizando-se da fase logarítimica obtida pela curva, comparou-se a dinâmica de interação destas cepas na célula epitelial de carcinoma de cérvix humana (HeLa) e na célula endometrial humana (EM42), em diferentes intervalos de tempo com auxílio de microscopia confocal. Apesar destas cepas divergirem na seqüência dos genes relacionados a aderência houve poucas variações entre as curvas de crescimento. A aderência e a invasão de M. genitalium nas células não fagocíticas confirmou os dados de literatura. Entretanto, após 30 minutos de contato com as células, detectou-se o antígeno de aderência de ambas as cepas na região intranuclear Este achado, indica uma nova característica desta espécie ainda não conhecida entre os molicutes.
Title in English
Mycoplasma genitalium: growth curve and interaction with HeLa cervical epithelioid cells and EM42 endometrial cells.
Keywords in English
Mycoplasma genitalium
Adhesion
Cytoplasmic invasion
Growth curve
Nuclear invasion
Abstract in English
Mycoplasma genitalium (Mg) is an important cause of sexual transmitted disease and has been implicated in a range of genital tract disorders.The adherence of mycoplasmas is a key virulence attribute, pathogenic features and consequences of host-cell colonization. Herein, we characterize growth properties of two Mg strains (G37 and 1019V) using BCA assay, OD600, CCU assay, real-time PCR. Based upon these strategies, we compared the behavior of similarly grown Mg variants coincubated with HeLa cervical epithelioid cells and EM42 endometrial cells over a dynamic time course.using laser scanning confocal microscopy. Mg G37 is a multiply passaged type strain isolated from a male while 1019V was recently isolated from human cervical samples and only minimally passaged. Both strains further diverge by sequence heterogeneities within adherence-related MG191 and MG192 genes. Despite these differences, our results identified only subtle variations in axenic growth for the two strains. Further and consistent with previous studies, a subset of adherent Mg organisms invaded host cells. However, intranuclear localization was observed, which occurred as early as 30 minutes after infection.
 
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Publishing Date
2009-05-11
 
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