Influence of KIR genes and their HLA ligands in susceptibility to dengue in a population from southern Brazil.

Killer cell immunoglobulin-like receptors (KIR) form a group of regulatory molecules that specifically recognise human leukocyte antigen (HLA) class I molecules, modulating the cytolytic activity of natural killer cells. The purpose of this study was to investigate the influence of KIR genes and their class I HLA ligands in susceptibility to dengue fever in a population from southern Brazil through a case-control study. One hundred four subjects with confirmed diagnoses of dengue participated in this study, along with a control group of 172 individuals from the same geographic area. HLA and KIR genotyping was performed by polymerase chain reaction with sequence-specific oligonucleotide probes (PCR-SSOP) and with sequence-specific primer (PCR-SSP) techniques, respectively. Data analysis showed significant differences for the KIR2DS1 (54.8% vs 40.7%, P = 0.03), KIR2DS5 (50.0% vs 36.0%, P = 0.03) and KIR2DL5 (76.0% vs 56.4%, P = 0.001) genes. With regard to KIR-ligand pairs, positive associations with dengue were observed in KIR3DS1-Bw4 (45.2% vs 29.7%, P = 0.01), KIR3DL1-Bw4 (80.7% vs 65.1%, P < 0.001), KIR2DL1-C2 (75.0% vs 62.2%, P = 0.03) and KIR2DS1-C2 (40.4% vs 25.6%, P = 0.01) interactions, and a negative association in KIR2DL3-C1/C1 (18.2% vs 33.1%, P = 0.01). Furthermore, the analysis of KIR haplogroups showed a possible protective factor against dengue fever in individuals with the AA genotype. Taken together, these results suggest the existence of genetic predisposition to dengue fever in the population from southern Brazil.

Protective effect of HLA-DRB1 11 and predisposition of HLA-C 04 in the development of severe liver damage in Brazilian patients with chronic hepatitis C virus infection.

The objective of this study was to investigate human leucocyte antigen (HLA) genes in patients chronically infected with hepatitis C virus (HCV) and to analyse the possible role of these genes in the progression of chronic hepatitis C. One hundred and forty-five (145) Brazilian patients infected only with HCV genotype 1 were evaluated. HLA class I (A, B, C) and class II (DRB1, DQA1, DQB1) typing were carried out by PCR-SSO, through Luminex technology. Associations were found with protection against development of liver damage by both DRB1 11 (5.0% versus 18.2%, P=0.0016, OR=0.23, CI 95% = 0.09-0.58; Pc=0.0208) and DRB1 11-DQA1 05-DQB1 03 haplotype (4.2% versus 15.3%, P=0.0032; OR = 0.24, CI 95% = 0.08-0.64). Liver damage was associated with HLA-C 04 in patients with <20 years of infection (38.4% versus 9.1%, P = 0.002, OR = 6.25, CI 95%=1.97-19.7; Pc=0.0238). It is concluded that HLA alleles can influence the development of liver damage in HCV type-1 chronically infected Brazilian patients.

Influence of HLA alleles in response to treatment with pegylated interferon-alpha and ribavirin in patients with chronic hepatitis C.

The objective of this study was to analyse the possible role of HLA polymorphism of chronically infected hepatitis C virus patients in the response outcome to treatment with pegylated interferon-alpha plus ribavirin. To that end, 144 Brazilian patients infected only with genotype 1 of the virus were treated with pegylated interferon-alpha at 1.5 µg kg(-1) in conjunction with ribavirin (1000 mg if patient weight was <75 kg and 1250 mg if >75 kg) for 48 weeks. The patients did not have concomitant HBV or HIV infections or liver disease, did not undergo previous antiviral treatment, and were followed up for 24 weeks after the end of treatment to assure they presented a sustained virological response. Patients were classified according to response to treatment in responsive (SVR), nonresponsive (NRS) and relapsers (REL). HLA class I and class II typing were carried out through PCR-SSO using Luminex technology. A statistically higher frequency of DRB1*11 patients was observed in the SVR group (39.6% vs. 14.3%P = 0.0012; Pc = 0.0156; OR = 3.94; 95% CI = 1.8-8.8). HLA-DQB1*03 patients were also more frequent in the SVR group, but the P value lost significance after Bonferroni correction (62.3% vs. 41.7%P = 0.024; Pc = 0.14, OR = 2.3; 95% CI = 1.14-4.60). HLA class II antigens can positively influence the response to treatment with pegylated interferon-alpha and ribavirin.

Radiotherapy effect on frequency of Candida spp. and on virulence of C. albicans isolated from the oral cavity of head and neck cancer patients

The aim of this study was to investigate the diversity and prevalence of yeasts, and the virulence of C.albicans found in the oral cavity during the course of ionizing radiation treatment of patients with head and neck tumor (HNTP). Samples from 21 HNTP and 24 healthy controls were isolated and identified. C. albicans isolated from two patients during radiotherapy were analyzed for virulence factors. Radiotherapy induced a higher level of both yeast colonization (81% vs 33%) and non-albicans Candida (NAC) colonization (52.4% vs 4.0%) in HNTP than the control group. Patients were colonized by 5 different NAC species: C. glabrata, C. tropicalis, C. parapsilosis, C. krusei and C. kefir. On the other hand, C. albicans colonization was similar in patients and controls (6/21, 28.6% vs 7/24, 29.2%, respectively). Also, of the 11 patients assessed before and during radiotherapy, 5 (45.5%) were colonized before the start of treatment and another 5 (45.5%) during treatment. All of the latter were colonized by NAC species alone. Moreover, we observed a significant and continuous enhancement of C. albicans virulence as the radiotherapy progressed, in the two patients involved in this test. Thus, it is concluded that radiotherapy is an important predisposing factor for the oral candidiasis, including NAC species. Also, it may facilitate the development of more virulent C. albicans strains.

O objetivo deste estudo foi avaliar a diversidade e a prevalência de Cândida, bem como a virulência de Cândida albicans, isoladas da cavidade bucal no decurso de tratamento por radiações ionizantes de pacientes acometidos por tumores de cabeça e pescoço (PTCP). Amostras de 21 pacientes e 24 controles foram analisadas. C. albicans isoladas de dois pacientes ao longo do tratamento radioterápico foram avaliadas para fatores de virulência. A radioterapia induziu um grande aumento da colonização de Cândida como um todo (81% vs 33%) e Cândida não albicans (CNA) em particular (52.4% vs 4.0%) em PTCP quando comparado com controles não irradiados. Cinco espécies diferentes de CNA foram encontradas nos pacientes: C. glabrata, C. tropicalis, C. parapsilosis, C. krusei and C. kefir. Por outro lado, a colonização por C. albicans nestes pacientes e controles foi similar (6/21, 28.6% vs 7/24, 29.2%, respectivamente). Além disso, dos 11 pacientes que foram avaliados antes e durante o tratamento radioterápico, 5 pacientes (45,5%) foram colonizados antes do início da radioterapia e outros 5 (45,5%) durante o tratamento radioterápico. Destes últimos, todos foram colonizados apenas com espécies CNA. Observou-se, ainda, um aumento contínuo e significante da virulência de C. albicans com o progresso da radioterapia nos dois pacientes estudados. Conclui-se que o tratamento radioterápico é um importante fator de desenvolvimento de candidíase oral, incluindo candidíase por espécies não albicans, em pacientes portadores de tumor de cabeça e pescoço. A radioterapia pode, ainda, facilitar o desenvolvimento de cepas mais virulentas de C.albicans.

Surface modification of polystyrene and poly(ethylene terephtalate) by grafting poly(N-isopropylacrylamide).

In this work, poly(N-isopropylacrylamide) (PNIPAAm) was incorporated into previously oxidized PS and PET surfaces by grafting using two photo-initiation pathways. The incorporation of PNIPAAm was observed by drop water contact angle measurements, dyeing with Methylene Blue and AFM images analysis of the virgin and modified polymers. It was verified that the grafting process depends on the chemical surface environment. The grafted surfaces are hydrophilic below 32 degrees C and hydrophobic above this temperature. The transition is due to the incorporated PNIPAAm. This characteristic gives to the grafted materials potential to be applied as biomaterials.

Association of HLA-DR7 with rheumatic fever in the Brazilian population.

OBJECTIVE: Rheumatic fever (RF) is a multisystem inflammatory disease that develops as a sequel of untreated throat infection by the group A beta-hemolytic streptococcus. As HLA antigens are known to be important in controlling immunological responsiveness, studies have investigated HLA antigen association with RF. Studies with Caucasians, Black Americans, and Indians showed associations with HLA-DR4, DR2, and DR3, respectively. One study on a Brazilian population suggested an association with HLA-DR7 and HLA-DR53. We investigated the association between RF and antigens HLA-DR7 and DR53 in the white Brazilian population. METHODS: Thirty-five patients and 209 healthy individuals living in the northern region of the state of Parana, Brazil, were used as test and control groups, respectively. Classical statistical methods were used to compare HLA frequencies between these groups. Results. Data confirmed positive association with HLA-DR7 (46.7 vs. 25.7%; p = 0.015), but not with HLA-DR53 (54.3 vs. 44.5%; p = 0.28). The relative risk and etiologic fractions were 2.4 and 0.27%, respectively. CONCLUSION: Positive association between HLA-DR7 specificity and RF was observed in the white Brazilian population by 2 independent studies, supporting the hypothesis of the involvement of genetic factors in susceptibility of rheumatic fever.

Identification of two types of autoreactive T lymphocyte clones cultured from cardiac allograft-infiltrating cells incubated with recombinant mycobacterial heat shock protein 71.

Recent studies in several laboratories have advanced the concept that during cellular rejection, the allograft undergoes a stress response which regulates the expression of stress proteins (or heat shock proteins, hsp) and triggers the recruitment and activation of hsp-reactive lymphocytes. In a rat model of heterotopic heart transplants we have found that allograft-infiltrating lymphocytes respond to recombinant mycobacterial hsp and irradiated syngeneic spleen cells as a source of self-APC (antigen-presenting cells). This report describes T cell clones generated by culturing ACI into Lewis rat cardiac allograft-derived lymphocytes with mycobacterial hsp71, syngeneic spleen cells and IL-2 (interleukin-2). Two groups of self-APC-reactive T cell clones have been distinguished, all of them are CD3+, CD4+, CD8-. One group is referred to as hsp71-dependent, autoreactive T cells because these clones respond to self-APC but only in the presence of hsp71. No reactivity is seen with mycobacterial hsp65 or when hsp71 is tested with allo-PC from ACI donors or third-party APC from Brown Norway (BN) rats. Treatment of hsp71 with trypsin, polymyxin B or ATP-agarose chromatography abrogates the hsp71 effect thus indicating that structurally intact hsp71 must interact with self-APC which then activate hsp71-dependent, autoreactive T cells. The second group of clones reacts to self-APC and while their response does not require the presence of hsp71, their proliferation is often augmented by hsp71 but not by hsp65. These hsp71-independent, autoreactive clones do not respond to allo-APC from ACI donors or third-party APC from BN rats. Polymyxin or trypsin treatment had no significant effect on their proliferative responses. The data with the anti-TCR-alpha beta monoclonal antibody R73 offer additional evidence for two functionally different types of self-APC reactive CD4 cells infiltrating the allograft. R73 inhibits the proliferation of self-APC induced responses of hsp-71-independent clones as well as the allo-APC induced responses of alloreactive T cell clones. In contrast, this antibody augments the responses of hsp71-dependent T cells. Moreover, these clones can also proliferate in response to self-APC when hsp71 is substituted by R73. The hsp71-dependency of self-APC reactive T cell reactivity represents a previously unrecognized mechanism of cellular immunity to allografts. This mechanism might be related to the peptide binding properties of hsp71 and the ability of stress proteins to function as molecular chaperones in antigen processing.

Association of viral hepatitis A with HLA-A9.

HLA-A and B phenotyping was carried out, during a viral hepatitis A (VHA) epidemic in Brazil, on 47 unrelated patients with VHA and 53 unrelated healthy subjects of the same age, sex, social status, and ethnic origin. An increased frequency of HLA-A9 (44.7% vs. 15.1%) and a decrease of that of HLA-A3 (2.1% vs. 22.6%) were observed when compared with controls. After correction for the number of antigens tested, only the positive association with HLA-A9 was observed. It was found that the relative risk for an HLA-A9 carrier to develop VHA was 4.5.