Virulence factors and genetic variability of vaginal Candida albicans isolates from HIV-infected women in the post-highly active antiretroviral era

Vulvovaginal candidiasis (VVC) in HIV-infected (HIV+) women is a serious public health problem. However, little is known about the virulence mechanisms of vaginal Candida albicans from HIV+ women in the post-highly active antiretroviral therapy (HAART) era. Here, we report a comparative analysis of the expression of key virulence factors and genetic variability of 26 vaginal C. albicans strains isolated from HIV+ women undergoing HAART and 18 from HIV-uninfected (HIV-) women. In general, we observed that C. albicans from HIV+ women receiving HAART showed lower expression of virulence factors compared with C. albicans from HIV- women, except for the proteinase activity which is highly expressed. The results in HIV-women further suggest that virulence factors appear to be expressed in response to the yeast stress, in the presence of an adequate immune response. Furthermore, the RAPD results showed a high heterogeneity among isolates from both groups of women. These findings in HIV+ women using HAART will help to improve the monitoring of vaginal yeast infections and the quality of life of patients.

Vulvovaginal candidiasis.; Candida albicans; HIV; Virulence; Genetic variability

Authorship

Pâmela Cristina Mastellaro Delvas Zanni

Universidade Estadual de Maringá, Departamento de Análises Clínicas e Biomedicina, Laboratório de Micologia Médica, Maringá, Paraná, BrazilUniversidade Estadual de MaringáBrazilMaringá, Paraná, BrazilUniversidade Estadual de Maringá, Departamento de Análises Clínicas e Biomedicina, Laboratório de Micologia Médica, Maringá, Paraná, Brazil

Universidade Estadual de Maringá, Departamento de Análises Clínicas e Biomedicina, Laboratório de Citologia Clínica, Maringá, Paraná, BrazilUniversidade Estadual de MaringáBrazilMaringá, Paraná, BrazilUniversidade Estadual de Maringá, Departamento de Análises Clínicas e Biomedicina, Laboratório de Citologia Clínica, Maringá, Paraná, Brazil

Patrícia de Souza Bonfim-Mendonça

Melyssa Negri

Sandra Sayuri Nakamura

Lucélia Donatti

Universidade Federal do Paraná, Departamento de Biologia Celular, Curitiba, Paraná, BrazilUniversidade Federal do ParanáBrazilCuritiba, Paraná, BrazilUniversidade Federal do Paraná, Departamento de Biologia Celular, Curitiba, Paraná, Brazil

Terezinha Inez Estivalet Svidzinski

Márcia Edilaine Lopes Consolaro

Correspondence to: Marcia Edilaine Lopes Consolaro. Universidade Estadual de Maringá, Departamento de Análises Clínicas e Biomedicina, Laboratório de Citologia Clínica, Av. Colombo, 5790, Zona 7, CEP 87020-900, Maringá, PR, Brazil. Tel: +55 44 3011-4795, Fax: +55 44 3011-4860. E-mail: melconsolaro@gmail.com

CONFLICT OF INTERESTS

The authors report no conflicts of interest. The authors alone are responsible for the content and the writing of the paper.

SCIMAGO INSTITUTIONS RANKINGS

Figures

Figures (6)

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Figure 1
Proteinase activity (106 cells/ mL) of vaginal Candida albicans isolated from HIV positive (HIV+) and negative (HIV-) women overall and in clinical groups: A) Proteinase activity in HIV+ and HIV- groups; B) Proteinase activity in HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (- three different experiments with samples tested in triplicate), *p<0.05 significant difference between HIV+ and HIV groups. ##p<0.05 significant difference between HIV+INF and HIV-INF

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Figure 2
Cell surface hydrophobicity (CSH) of vaginal Candida albicans isolated from HIV positive (HIV+) and negative (HIV-) women overall and in clinical groups: A) CSH (%) in HIV+ and HIV- groups; B) CSH (%) HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (three different experiments with samples tested in triplicate).

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Figure 3
Adhesion and biofilm formation capacity (absorbance/cm2) of vaginal Candida albicans isolated from HIV positive (HIV+) and negative (HIV-) women overall and in clinical groups: A) Adhesion capacity in HIV+ and HIV- groups; B) Adhesion capacity in HIV+ and HIV- groups with colonization (COL) and infection (INF); C) Biofilm formation in HIV+ and HIV- groups; D) Biofilm formation in HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (three different experiments with samples tested in triplicate), *p<0.05 significant difference between HIV+ and HIV- groups. #p<0.05 significant difference between HIV+COL and HIV-COL. ##p<0.05 significant difference between HIV+INF and HIV-INF.

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Figure 4
Scanning electron photomicrographs showing vaginal Candida albicans from the HIV+ and HIV- groups adhered to the microplate surface after 2 h of contact: A) and B) HIV+ group adhesion showing yeasts in the blastoconidia form attached to the surface, isolated or in small clusters; C) and D) HIV- group adhesion showing several yeasts in the blastoconidia form attached to the surface, isolated or in large clusters. In A) and C) initial extracellular matrix material (red arrow) observed close to the blastoconidia or with microorganisms embedded. In B) and D) budding yeasts (blue arrows). Scale bars: A) and C), 10 µm; B) and D), 5 µm.

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Figure 5
Germ tube (GT) formation capacity (%) and length (µm) of vaginal Candida albicans isolated from HIV+ and HIV- women overall and in clinical groups: A) GT (%) in HIV+ and HIV- groups; B) GT in HIV+ and HIV- groups with colonization (COL) and infection (INF); C) GT length in HIV+ and HIV- groups; D) HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (three different experiments with samples tested in triplicate). *p<0.05 significant difference between HIV + and HIV- groups.

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Figure 6
Dendrogram generated by RAPD analysis of vaginal Candida albicans isolated from HIV + and HIV- women using the UPGMA (unweighted pair-group method with arithmetical average) grouping, with SAB calculated by the coefficient of Dice: A) and C) Yeast dendrograms using the OPA-18 primer; B) and D) Yeast dendrograms were constructed using the OPE-18 primer results. The numbers indicated on the right side of dendrograms represent the registration of each yeast in the culture collection of the Medical Mycology Laboratory of the State University of Paraná, Maringá (UEM), Brazil.

Figure 1 Proteinase activity (106 cells/ mL) of vaginal Candida albicans isolated from HIV positive (HIV+) and negative (HIV-) women overall and in clinical groups: A) Proteinase activity in HIV+ and HIV- groups; B) Proteinase activity in HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (- three different experiments with samples tested in triplicate), *p<0.05 significant difference between HIV+ and HIV groups. ##p<0.05 significant difference between HIV+INF and HIV-INF

Figure 2 Cell surface hydrophobicity (CSH) of vaginal Candida albicans isolated from HIV positive (HIV+) and negative (HIV-) women overall and in clinical groups: A) CSH (%) in HIV+ and HIV- groups; B) CSH (%) HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (three different experiments with samples tested in triplicate).

Figure 3 Adhesion and biofilm formation capacity (absorbance/cm2) of vaginal Candida albicans isolated from HIV positive (HIV+) and negative (HIV-) women overall and in clinical groups: A) Adhesion capacity in HIV+ and HIV- groups; B) Adhesion capacity in HIV+ and HIV- groups with colonization (COL) and infection (INF); C) Biofilm formation in HIV+ and HIV- groups; D) Biofilm formation in HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (three different experiments with samples tested in triplicate), *p<0.05 significant difference between HIV+ and HIV- groups. #p<0.05 significant difference between HIV+COL and HIV-COL. ##p<0.05 significant difference between HIV+INF and HIV-INF.

Figure 4 Scanning electron photomicrographs showing vaginal Candida albicans from the HIV+ and HIV- groups adhered to the microplate surface after 2 h of contact: A) and B) HIV+ group adhesion showing yeasts in the blastoconidia form attached to the surface, isolated or in small clusters; C) and D) HIV- group adhesion showing several yeasts in the blastoconidia form attached to the surface, isolated or in large clusters. In A) and C) initial extracellular matrix material (red arrow) observed close to the blastoconidia or with microorganisms embedded. In B) and D) budding yeasts (blue arrows). Scale bars: A) and C), 10 µm; B) and D), 5 µm.

Figure 5 Germ tube (GT) formation capacity (%) and length (µm) of vaginal Candida albicans isolated from HIV+ and HIV- women overall and in clinical groups: A) GT (%) in HIV+ and HIV- groups; B) GT in HIV+ and HIV- groups with colonization (COL) and infection (INF); C) GT length in HIV+ and HIV- groups; D) HIV+ and HIV- groups with colonization (COL) and infection (INF). Each bar represents the mean ± SD of the results from all isolates in each group (three different experiments with samples tested in triplicate). *p<0.05 significant difference between HIV + and HIV- groups.

Figure 6 Dendrogram generated by RAPD analysis of vaginal Candida albicans isolated from HIV + and HIV- women using the UPGMA (unweighted pair-group method with arithmetical average) grouping, with SAB calculated by the coefficient of Dice: A) and C) Yeast dendrograms using the OPA-18 primer; B) and D) Yeast dendrograms were constructed using the OPE-18 primer results. The numbers indicated on the right side of dendrograms represent the registration of each yeast in the culture collection of the Medical Mycology Laboratory of the State University of Paraná, Maringá (UEM), Brazil.

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Virulence factors and genetic variability of vaginal Candida albicans isolates from HIV-infected women in the post-highly active antiretroviral era

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[1] Correspondence to: Marcia Edilaine Lopes Consolaro. Universidade Estadual de Maringá, Departamento de Análises Clínicas e Biomedicina, Laboratório de Citologia Clínica, Av. Colombo, 5790, Zona 7, CEP 87020-900, Maringá, PR, Brazil. Tel: +55 44 3011-4795, Fax: +55 44 3011-4860. E-mail: melconsolaro@gmail.com