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1.
Appl Microbiol Biotechnol ; 93(5): 2099-108, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21894481

RESUMO

Accurate diagnosis is critical for effective treatment of the invasive infection by Candida albicans. Here, we investigated whether a (99m) technetium (Tc)-labeled Fab' fragment of the monoclonal antibody specific for the C. albicans germ tube could specifically identify an invasive C. albicans infection. The germ tube of C. albicans was used as an immunogen to obtain monoclonal antibodies and the Fab' fragment of MAb03.2 C1-C2 with highest affinity and specificity was labeled with (99m)Tc. In vitro binding assays showed that the labeled Fab' preferentially bound to the germ tubes of C. albicans (4.23 ± 0.17 × 10(2) Bq per 1 × 10(7) cells). These values were significantly higher than those for blastospores of C. albicans, blastospores of heat-killed C. albicans, Aspergillus fumigatus, Staphylococcus aureus, and Escherichia coli (P < 0.05). By using in vivo biodistribution and planar imaging with single photon emission computed tomography, we demonstrated a significant specific accumulation of radioactivity in C. albicans-infected tissues. In summary, (99m)Tc-MAb03.2 C1-C2 Fab' is able to specifically accumulate in C. albicans-infected tissues, but not in tissue infected with A. fumigatus or bacteria or in a sterile inflammation. This study provides a new and specific radiopharmaceutical for the diagnosis of invasive C. albicans infections.


Assuntos
Anticorpos Antifúngicos , Anticorpos Monoclonais , Candida albicans/imunologia , Candida albicans/patogenicidade , Candidíase/diagnóstico , Candidíase/microbiologia , Fragmentos Fab das Imunoglobulinas , Sensibilidade e Especificidade , Coloração e Rotulagem/métodos , Tecnécio/metabolismo
2.
Indian J Biochem Biophys ; 49(1): 25-35, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22435141

RESUMO

Psoriasis vulgaris (PV) is a common autoimmune disease that involves the dysfunction of CD4+CD25+ regulatory T cells. FOXP3 is a key transcription factor in the development and function of CD4+CD25+ regulatory T cells. Previous studies have demonstrated a genetic association between the FOXP3 gene and some autoimmune diseases. To elucidate the association between the FOXP3 gene and the risk of PV, 408 patients diagnosed with PV and 363 age and sex-matched healthy controls from a cohort of the Chinese majority Han population were recruited. Four single nucleotide polymorphisms (rs2232365, rs3761547, rs3761548 and rs3761549) of the FOXP3 gene were analyzed using the polymerase chain reaction and ligase detection reaction. The major allele of three single nucleotide polymorphisms (SNPs - rs2232365 A, rs3761547 A and rs3761549 C) were associated with an increased risk of PV in a clinical subgroup of female patients, who were less than 40 yrs of age, had a family history of the disease and did not have disease complications (p < 0.05 for all parameters). The haplotype was structured between rs3761547 and rs3761549. An increased risk of PV was observed in haplotype A/A-T/T (p = 0.0055; adjusted OR = 3.188; 95% CI = 0.4354-23.34) and A/G-C/C (p = 0.0082; adjusted OR = 1.288; 95% CI = 0.1529-10.85) between rs3761547 and rs3761549. A synergistic effect was found among the three SNPs. Subjects with the rs2232365AA- rs3761547 AG + GG genotype were more susceptible to PV (p = 0.0393; OR = 2.90; 95% CI = 1.05-7.97). No correlation was found between rs3761548 and the onset of PV. Therefore, the FOXP3 polymorphisms appear to contribute to the risk of psoriasis among the Chinese majority Han population. These findings may aid in our understanding of the pathogenesis of psoriasis.


Assuntos
Fatores de Transcrição Forkhead/genética , Haplótipos/genética , Inteínas/genética , Polimorfismo de Nucleotídeo Único/genética , Psoríase/genética , Adulto , Povo Asiático/genética , China/epidemiologia , Estudos de Coortes , Feminino , Fatores de Transcrição Forkhead/metabolismo , Predisposição Genética para Doença , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Psoríase/epidemiologia , Risco
3.
Exp Dermatol ; 20(11): 945-7, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21801235

RESUMO

Biological agents are becoming increasingly popular for therapeutic applications in epidermal diseases. Ethosomes facilitate the transdermal/topical delivery of biological macromolecules. The mouse epidermal growth factor (mEGF) was selected as the model biological agent. The aim of this experiment was to determine the penetration pathways and biological functions of the mEGF ethosomal delivery system after its topical application. The mEGF ethosomal delivery system was topically applied on the dorsal skin of C57BL/6 mice at different time points. Freshly excised skin samples were obtained by skin biopsies and shock-frozen, and immunofluorescence was performed. The results showed that penetration of mEGF ethosomes was mainly through the pilosebaceous unit and partly through the intercellular domain. Biological agents encapsulated in the ethosomal delivery system could reach each site of the pilosebaceous unit. We also found that mEGF ethosomes had caused successful transition of the hair follicles from the telogen to the anagen phase of the hair cycle.


Assuntos
Fator de Crescimento Epidérmico/administração & dosagem , Fator de Crescimento Epidérmico/farmacocinética , Pele/efeitos dos fármacos , Pele/metabolismo , Administração Tópica , Animais , Sistemas de Liberação de Medicamentos , Folículo Piloso/efeitos dos fármacos , Folículo Piloso/crescimento & desenvolvimento , Folículo Piloso/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Permeabilidade , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/farmacocinética
4.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 23(4): 350-2, 2007 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-17428393

RESUMO

AIM: To prepare and identify the monoclonal antibody(mAb) specific for the germ tube of Candida albicans. METHODS: mAb were prepared via hybridoma technique. Indirect immunofluorescence (IIF) was used to analyze the activity and specificity of the mAb. The protection of the mAb was analyzed via the inhibition of the mAb to the germ tube formation and the adhesion of C.albicans to the epithelial and endothelial cells. The frozen hybridoma cells were cultured to analyze the ability of the secretion of the mAb. RESULTS: mAb03.2C1-C2 belongs to IgG1 subclass. Relative molecular mass (M(r)) of antigens recognized by the mAb was 156, 48, 22 and 12. The target antigen could be detected 30 minutes after the germ tube formation. The epitope might lie in the N-carbohydrate chain. mAb03.2C1-C2 was shown by IIF test to be specific to the surface of the germ tube or mycelium phase of C.albicans. In the protective mAb screening experiment, it was found that mAb3.2C1-C2 could inhibit C.albicans germ tube formation and the adhension of C.albicans to the epithelial and endothelial cells. CONCLUSION: In vitro, mAb03.2C1-C2 could inhibit the formation of germ tube of C.albicans, which could depress the invasiveness of C.albicans. This anti-C.albicans cell wall surface-specific mAb may be a good candidate for use in tests for the rapid differentiation of the two closely related species C.albicans and C.dubliniensis.


Assuntos
Anticorpos Monoclonais/imunologia , Candida albicans/imunologia , Animais , Western Blotting , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Células Endoteliais/microbiologia , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/microbiologia , Hibridomas/imunologia , Camundongos
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