Aya Sadahiro

T-Cell Recognition of Paracoccidioides brasiliensis gp43-Derived Peptides in Patients with Paracoccidioidomycosis and Healthy Individuals

ABSTRACT Vaccines with synthetic peptides induce the immune response to epitopes that bind to several HLA alleles. By using a TEPITOPE algorithm, we selected and analyzed the T-cell responses of peripheral blood mononuclear cells from 29 paracoccidioidomycosis (PCM) patients to peptides of the immunodominant gp43 antigen of Paracoccidioides brasiliensis, the causative agent of PCM.

Association between the PTPN22 1858C/T gene polymorphism and tuberculosis resistance

Previous studies identified the functional polymorphism 1858C/T in the gene PTPN22 in association with several autoimmune diseases and with resistance to tuberculosis (TB). This study is the first to investigate the association between pulmonary TB and the PTPN22 1858C/T polymorphism in the Brazilian Amazon. We conducted a case-control study involving a group of 413 individuals, comprised of 208TB carriers and 205 controls. No significant association between the PTPN22 1858T allele frequency in controls (2.4%) and TB carriers (2.7%, p=0.982, odds ratio (OR)=0.89, 95% confidence interval=0.37-2.13) was identified in the Brazilian Amazon population. An additional evaluation by meta-analysis, however, suggested a protective role of the T allele in relation to TB (pooled OR=0.44, p=0.011). These results suggest that the PTPN22 1858T allele serves as a protective genetic factor for TB in those individuals who carry this minor allele.

The Robust and Modulated Biomarker Network Elicited by the Plasmodium vivax Infection Is Mainly Mediated by the IL-6/IL-10 Axis and Is Associated with the Parasite Load

Background. Recent studies have shown that the inflammatory process, including the biomarker production, and the intense activation of innate immune responses are greater in the malaria caused by Plasmodium vivax than other species. Here, we examined the levels of serum biomarkers and their interaction during acute malaria. Material and Methods. Blood samples were collected from P. vivax-infected patients at admission and from healthy donors. Levels of serum biomarkers were measured by Cytometric Bead Assay or ELISA. Results. P. vivax infection triggered the production of both inflammatory and regulatory biomarkers. Levels of IL-6, CXCL-8, IFN-γ, IL-5, and IL-10 were higher in P. vivax-infected patients than in healthy donors. On the other hand, malaria patients produced lower levels of TNF-α, IL-12p70, and IL-2 than healthy individuals. While the levels of IL-10 and IL-6 were found independent on the number of malaria episodes, higher levels of these cytokines were seen in patients with higher parasite load. Conclusion. A mixed pattern of proinflammatory and regulatory biomarkers is produced in P. vivax malaria. Analysis of biomarker network suggests that IL-10 and IL-6 are a robust axis in malaria patients and that this interaction seems to be associated with the parasite load.

HLA in Brazilian Ashkenazic Jews with chronic dermatophytosis caused by Trichophyton rubrum

The frequency of HLA (Human Leucocyte Antigens) was analyzed in 25 non-consanguineous Brazilian Ashkenazic Jews, resident in the city of Sao Paulo, Brazil, suffering from chronic dermatophytosis caused by T. rubrum, and in 25 non-infected individuals belonging to the same ethnic group. Statistically significant values (p<0.05) were observed for HLA-B14 associated with resistance to chronic dermatophytosis and HLA-DQB1*06 (p=0.05) possibly related to susceptibility. These findings suggest that genes on the chromosome 6, in the region of the major histocompatibility complex, may influence the development of chronic dermatophytosis.

PCR in the diagnosis of cutaneous tuberculosis

Visando melhorar o diagnostico laboratorial da Tuberculose Cutânea, foi realizado o estudo da aplicacao da tecnica de PCR em amostras de tecidos cutâneos macerados, descontaminados (com H2SO4 4% para eliminacao da microbiota normal), neutralizados (com NaOH 4%) e armazenadas a -20oC. Das 37 amostras submetidas ao estudo, 16,22% apresentavam baciloscopias positivas para bacilos alcool-acidos resistentes (metodo concentrado) e em 43,24% houve o isolamento do Mycobacterium tuberculosis em meio de cultivo Lowenstein-Jensen. Utilizando-se de primers para a regiao 16S rDNA do M. tuberculosis, o DNA micobacteriano foi detectado em 24,32% das biopsias. A sensibilidade e especificidade da PCR foram 43,7% e 90,4%, respectivamente. Devido a baixa sensibilidade e resultados divergentes entre as tecnicas bacteriologicas e PCR (para a sequencia 16S rDNA), as amostras foram repetidas em um novo PCR com primers para a regiao IS6110. Tanto a sensibilidade como a especificidade da PCR com primers para IS6110 alcancaram 100% em relacao ao cultivo. Os resultados confirmam a eficacia da PCR utilizando primers para a sequencia IS6110 e oferecem a possibilidade da tecnica ser aplicada em amostras congeladas enviadas por servicos que nao identificam o M. tuberculosis por tecnicas de biologia molecular.

The influence of a TNF gene polymorphism on the severity of rheumatoid arthritis in the Brazilian Amazon.

PURPOSE The aim of this study was to investigate the influence of the TNF -308 G/A polymorphism in the promoter region of the tumor necrosis factor-α gene on the susceptibility and severity of rheumatoid arthritis (RA) in individuals from the Brazilian Amazon. METHODS A total of 323 individuals-192 healthy controls without arthritis and 131 individuals suffering from arthritis-were genotyped for this polymorphism using a methodology based on PCR-RFLP. RESULTS The frequency of the A allele (TNF2) in rheumatoid arthritis sufferers was not significantly higher than in the controls (p=0.926; OR=0.97; confidence interval 0.54-1.76). However, using a logistic regression model, when the patients were stratified according to whether the manifestations were preponderantly articular or systemic, there was a strong association between the TNF2 allele and systemic arthritis (p=0.001; OR=5.89; confidence interval=1.98-17.5) as well as the use of anti-TNF immunotherapy (p=0.023; OR=1.10; confidence interval=1.00-1.14). The main factors that were found to influence the risk of extra-articular disease were age greater than or equal to 60 years (p=0.008; OR=4.06; confidence interval=1.45-11.38), disease duration greater than 10 years (p=0.031; OR=3.10; confidence interval=1.11-8.63) and positive rheumatoid factor (p=0.035; OR=2.07; confidence interval=1.05-4.09). CONCLUSIONS These results suggest that the TNF2 allele is associated with the more serious forms of the disease in individuals from the Brazilian Amazon but not with a risk for developing RA.

Interaction between smoking and HLA-DRB1*04 gene is associated with a high cardiovascular risk in Brazilian Amazon patients with rheumatoid arthritis.

Background Rheumatoid Arthritis (RA) is an autoimmune disease characterized by chronic inflammation of the joints that affects approximately 1% of the population worldwide. The HLA-DRB1 gene locus plays a major role in genetic susceptibility to RA, a condition that has been associated with a high cardiovascular morbidity and mortality in many studies. Methodology/Principal Findings The aim of this work was to investigate which types of HLA class II genes are associated with RA in patients from the Brazilian Amazon and their influence on high cardiovascular risk status in this population. For this purpose, a case-control study was carried out with a total of 350 non-Indian individuals made up of a cohort of 132 consecutive RA sufferers and 218 healthy controls. A χ2 test showed that HLADRB1*04 (p<0.0016; OR = 1.89; 95% CI = 1.29–2.79) and HLADRB1*10 (p = 0.0377; OR = 3.81; 95% CI = 1.16–12.50) are the major HLA genes associated with susceptibility to RA. A logistic regression model also showed that the interaction between HLADRB1*04 (p = 0.027; OR = 6.02; 95% CI = 1.21–29.7), age (p = 0.0001; OR = 1.26; 95% CI = 1.13–1.39) and smoking (p = 0.0001; OR = 23.6; 95% CI = 4.25–32.1) is associated with a probability of a high cardiovascular risk status at an early age. Conclusions/Significance The results of this study show for the first time that HLA class II type is associated with RA in Brazilian Amazon populations and that a specific interaction between the HLA-DRB1*04 gene and smoking is associated with a high cardiovascular risk status, as initially reported in the European population. This study therefore contributes to an understanding of gene-environment interactions in RA patients.

Generic human leukocyte antigen class II (DRB1 and DQB1) alleles in patients with paracoccidioidomycosis

Human leukocyte antigen (HLA) class II alleles are involved in antigen processing and in the presentation of antigens to T lymphocytes. Few studies have investigated HLA genes in paracoccidioidomycosis. In the present investigation, we analyzed the distribution of the HLA class II alleles DRB1 and DQB1 in 45 healthy volunteers and in 80 patients with paracoccidioidomycosis. The patients presented with various clinical forms of the disease, and allele distribution was evaluated individually in each presentation type. In patients with the unifocal chronic form of the disease, a mild clinical presentation in which lesions are restricted or localized, the HLA allele most commonly seen was DRB1*11 (p<0.039). This suggests that the participation of HLA antigens may influence the outcome of the host-parasite interaction in paracoccidioidomycosis, regulating the immune response to Paracoccidioides brasiliensis antigens.

Kinetics of IFN-gamma, TNF-alpha, IL-10 and IL-4 production by mononuclear cells stimulated with gp43 peptides, in patients cured of paracoccidioidomycosis

We analyzed the kinetics of cytokine production by mononuclear cells from 17 patients who had been treated for paracoccidioidomycosis, using the stimulus of gp43 peptide groups (43 kDa glycoprotein of Paracoccidioides brasiliensis) at 0.1 and 1 microM, gp43 (1 microg/ml) and crude Paracoccidioides brasiliensis antigen (PbAg; 75 microg/ml). IFN-gamma production was a maximum at 144 hours in relation to the G2 and G8 peptide groups at 1 microM and was greatest at 144 hours when stimulated by gp43 and by PbAg. The maximum TNF-alpha production was at 144 hours for the G2 group (0.1 microM) and for gp43. IL-10 production was highest after 48 and 72 hours for G7 and G6 at 1 microM, respectively. We also suggest the best time for analysis of IL4 production. These results may contribute towards future studies with gp43 peptides and encourage further investigations with the aim of understanding the influence of these peptides on the production of inflammatory and regulatory cytokines.

Alleles of HLA-DRB1*04 Associated with Pulmonary Tuberculosis in Amazon Brazilian Population.

Immunogenetic host factors are associated with susceptibility or protection to tuberculosis (TB). Strong associations of HLA class II genes with TB are reported. We analyzed the HLA-DRB1*04 alleles to identify subtypes associated with pulmonary TB and their interaction with risk factors such as alcohol, smoking, and gender in 316 pulmonary TB patients and 306 healthy individuals from the Brazilian Amazon. The HLA-DRB1*04 was prevalent in patients with pulmonary TB (p<0.0001; OR = 2.94; 95% CI = 2.12 to 4.08). Direct nucleotide sequencing of DRB1 exon 2 identified nine subtypes of HLA-DRB1*04. The subtype HLA-DRB1*04:11:01 (p = 0.0019; OR = 2.23; 95% CI = 1.34 to 3.70) was associated with susceptibility to pulmonary TB while DRB1*04:07:01 (p<0.0001; OR = 0.02; 95% CI = 0.001 to 0.33) to protection. Notably, the interaction between alcohol and HLA-DRB1*04:11:01 increased the risk for developing pulmonary TB (p = 0.0001; OR = 51.3; 95% CI = 6.81 to 386). Multibacillary pulmonary TB, the clinical presentation of disease transmission, was strongly associated with interaction to alcohol (p = 0.0026; OR = 11.1; 95% CI = 3.99 to 30.9), HLA-DRB1*04:11:01 (p = 0.0442; OR = 2.01; 95% CI = 1.03 to 3.93) and DRB1*04:92 (p = 0.0112; OR = 8.62; 95% CI = 1.63 to 45.5). These results show that HLA-DRB1*04 are associated with pulmonary TB. Interestingly, three subtypes, DRB1*04:07:01, DRB1*04:11:01 and DRB1*04:92 of the HLA-DRB1*04 could be potential immunogenetic markers that may help to explain mechanisms involved in disease development.