ABSTRACT
BACKGROUND: The pathogenesis related protein PR10 (TcPR-10), obtained from the Theobroma cacao-Moniliophthora perniciosa interaction library, presents antifungal activity against M. perniciosa and acts in vitro as a ribonuclease. However, despite its biotechnological potential, the TcPR-10 has the P-loop motif similar to those of some allergenic proteins such as Bet v 1 (Betula verrucosa) and Pru av 1 (Prunus avium). The insertion of mutations in this motif can produce proteins with reduced allergenic power. The objective of the present work was to evaluate the allergenic potential of the wild type and mutant recombinant TcPR-10 using bioinformatics tools and immunological assays. METHODOLOGY/PRINCIPAL FINDINGS: Mutant substitutions (T10P, I30V, H45S) were inserted in the TcPR-10 gene by site-directed mutagenesis, cloned into pET28a and expressed in Escherichia coli BL21(DE3) cells. Changes in molecular surface caused by the mutant substitutions was evaluated by comparative protein modeling using the three-dimensional structure of the major cherry allergen, Pru av 1 as a template. The immunological assays were carried out in 8-12 week old female BALB/c mice. The mice were sensitized with the proteins (wild type and mutants) via subcutaneous and challenged intranasal for induction of allergic airway inflammation. CONCLUSIONS/SIGNIFICANCE: We showed that the wild TcPR-10 protein has allergenic potential, whereas the insertion of mutations produced proteins with reduced capacity of IgE production and cellular infiltration in the lungs. On the other hand, in vitro assays show that the TcPR-10 mutants still present antifungal and ribonuclease activity against M. perniciosa RNA. In conclusion, the mutant proteins present less allergenic potential than the wild TcPR-10, without the loss of interesting biotechnological properties.
Subject(s)
Allergens/immunology , Antigens, Plant/immunology , Cacao , Plant Proteins/immunology , Algorithms , Allergens/chemistry , Amino Acid Sequence , Animals , Antifungal Agents/pharmacology , Antigens, Plant/chemistry , Basidiomycota/cytology , Basidiomycota/drug effects , Bronchoalveolar Lavage , Cacao/chemistry , Cacao/immunology , Cell Count , Computational Biology , Databases, Protein , Female , Hydrophobic and Hydrophilic Interactions/drug effects , Immunoglobulin E/immunology , Lung/drug effects , Lung/immunology , Mice , Mice, Inbred BALB C , Microbial Viability/drug effects , Models, Molecular , Molecular Sequence Data , Mutant Proteins/chemistry , Mutant Proteins/immunology , Plant Proteins/chemistry , Plant Proteins/pharmacology , Ribonucleases/metabolism , Sequence Alignment , Structural Homology, ProteinABSTRACT
Cutaneous leishmaniasis affects millions of people around the world. Several species of Leishmania infect mouse strains, and murine models closely reproduce the cutaneous lesions caused by the parasite in humans. Mouse models have enabled studies on the pathogenesis and effector mechanisms of host resistance to infection. Here, we review the role of nitric oxide (NO), reactive oxygen species (ROS), and peroxynitrite (ONOO(-)) in the control of parasites by macrophages, which are both the host cells and the effector cells. We also discuss the role of neutrophil-derived oxygen and nitrogen reactive species during infection with Leishmania. We emphasize the role of these cells in the outcome of leishmaniasis early after infection, before the adaptive T(h)-cell immune response.
ABSTRACT
Os autores apresentam uma revisão da literatura sobre cervicovaginites causadas por Chlamydia trachomatis e discorrem sobre a epidemiologia, a teraia e o seguimento das pacientes. Atualmente mulheres que após o tratamento apresentam resultados positivos para C. trachomatis são consideradas como reinfectadas. Contudo, discutem estudos que mostram evidências de reemergência de infecções latentes persistentes em mulheres tratadas, ressaltando a importância do seguimento dessas paciente.
The authors present a literature review about cervicovaginitis caused by Chlamydia trachomatis and discuss the epidemiology, therapy and follow-up of the patients. Nowadays, women previously treated for chlamydial infection and presenting positive tests are considered reinfected. However, the authors discuss studies showing evidence of latent infection that persists after treatment, and therefore emphasize the importance of treatment follow-up.
Subject(s)
Female , Chlamydia trachomatis/pathogenicity , Chlamydia Infections/epidemiology , Chlamydia Infections/therapy , Chlamydia Infections/transmission , Brazil/epidemiology , Cross-Sectional Studies , Sexually Transmitted Diseases/transmission , Genital Diseases, Female , Prevalence , Risk Factors , Disease-Free SurvivalABSTRACT
BACKGROUND: There is no data concerning genotyping of Chlamydia trachomatis from Brazilian samples. GOAL: To characterize the genotype of C. trachomatis detected in women assisted at a STD public clinic and establish the prevalence of this infection in that population. STUDY DESIGN: Endocervical samples of a group of 100 women were tested for chlamydial infection with PCR directed to C. trachomatis cryptic plasmid. Genotyping of positive samples were done after omp1 amplification and sequencing. RESULTS: The overall prevalence of C. trachomatis infection was 19%, with the highest prevalence in women between 15 and 25 years old (68.4%). Four genotypes were found associated with endocervical infections: D, E, F, and K. Sequence analysis revealed a coinfection of genotypes D and E in 1 woman. CONCLUSIONS: To our knowledge this is the first study to characterize Brazilian C. trachomatis endocervical samples and Brazilian C. trachomatis genotype coinfection. Our results also emphasize the importance of routine diagnosis of C. trachomatis for the control of this STD.