TGF-ß1 polymorphism in American tegumentary leishmaniasis in a Southern Brazilian population.

INTRODUCTION: Genetic polymorphisms define the cytokine production leading to susceptibility or resistance to diseases. We studied the cytokine polymorphism in the development of tegumentary leishmaniasis (TL). METHODS: Genotyping of TNF-α, TGF-ß1, IFN-γ, IL-6, and IL-10 were performed by polymerase chain reaction assay. RESULTS: G and C alleles of TGF- ß1 (codon 25) were the most common in controls and patients, respectively. G/G was the most frequent genotype in controls, and G/C and C/C in patients. CONCLUSIONS: G/G genotype of codon 25 in TGF-ß1 appeared to confer resistance, and G/C and C/C genotypes, susceptibility to TL in this population.

TGF-ß1 polymorphism in American tegumentary leishmaniasis in a Southern Brazilian population

Abstract INTRODUCTION: Genetic polymorphisms define the cytokine production leading to susceptibility or resistance to diseases. We studied the cytokine polymorphism in the development of tegumentary leishmaniasis (TL). METHODS: Genotyping of TNF-α, TGF-β1, IFN-γ, IL-6, and IL-10 were performed by polymerase chain reaction assay. RESULTS: G and C alleles of TGF- β1 (codon 25) were the most common in controls and patients, respectively. G/G was the most frequent genotype in controls, and G/C and C/C in patients. CONCLUSIONS: G/G genotype of codon 25 in TGF-β1 appeared to confer resistance, and G/C and C/C genotypes, susceptibility to TL in this population.

Association between KIR genes and dust mite sensitization in a Brazilian population.

BACKGROUND: Killer cell immunoglobulin-like receptors (KIRs), found on the surface of natural killer (NK) cells, play a key role in controlling the innate response. Such response depends on a series of cellular interactions between these receptors and HLA activating/inhibiting ligands. Atopic diseases have been associated with genes that regulate cytokine production and HLA genes, which may either protect or predispose to hypersensitivity. OBJECTIVE: To verify an association study of KIR genes with sensitization to the following mites: Dermatophagoides farinae, Dermatophagoides pteronyssinus, and Blomia tropicalis. METHODS: A total of 341 children aged up to 14 years, were classified as mite-sensitive or mite-insensitive after undergoing a skin prick test for immediate allergic reactions. The presence/absence of KIR genes and their human leukocyte antigen (HLA) ligands was determined by polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO) with the commercial kit LabType™ using Luminex™. RESULTS: The frequencies of KIR genes and their respective class I HLA ligands and the frequency of haplotypes were performed in sensitive and insensitive individuals, and no significant differences were found. CONCLUSION: Our results suggest no influence of KIR genes on resistance/susceptibility to sensitization to dust mites.

MICA diversity and linkage disequilibrium with HLA-B alleles in renal-transplant candidates in southern Brazil.

The major histocompatibility complex (MHC) class I chain-related gene A (MICA) is located centromerically to the human leukocyte antigen (HLA)-B. The short distance between these loci in the MHC indicates the presence of linkage disequilibrium (LD). Similarly to the HLA, the MICA is highly polymorphic, and this polymorphism has not been well documented in different populations. In this study, we estimated the allelic frequencies of MICA and the linkage disequilibrium with HLA-B alleles in 346 renal-transplant candidates in southern Brazil. MICA and HLA were typed using the polymerase chain reaction-sequence-specific primer method (PCR-SSO), combined with the Luminex technology. A total of 19 MICA allele groups were identified. The most frequent allele groups were MICA*008 (21.6%), MICA*002 (17.0%) and MICA*004 (14.8%). The most common haplotypes were MICA*009-B*51 (7.8%), MICA*004-B*44 (6.06%) and MICA*002-B*35 (5.63%). As expected from the proximity of the MICA and HLA-B loci, most haplotypes showed strong LD. Renal patients and healthy subjects in the same region of Brazil showed statistically significant differences in their MICA polymorphisms. The MICA*027 allele group was more frequent in renal patients (Pc = 0.018, OR: 3.421, 95% CI: 1.516-7.722), while the MICA*019 allele group was more frequent in healthy subjects (Pc = 0.001, OR: 0.027, 95% CI: 0.002-0.469). This study provided information on the distribution of MICA polymorphisms and linkage disequilibrium with HLA-B alleles in Brazilian renal-transplant candidates. This information should help to determine the mechanisms of susceptibility to different diseases in patients with chronic kidney disease, and to elucidate the mechanisms involved in allograft rejection associated with MICA polymorphisms in a Brazilian population.

Cytokine Profile in Patients with Progressive Multiple Sclerosis and Its Association with Disease Progression and Disability.

Inflammation is the driving force for brain injury in patients with multiple sclerosis (MS). The objective of the present study is to delineate the serum cytokine profile in patients with progressive MS in a Southern Brazilian population compared with healthy controls and patients with relapsing-remitting MS (RRMS) and its associations with disease progression and disability. We included 32 patients with progressive MS, 126 with RRMS, and 40 healthy controls. The patients were evaluated using the Expanded Disability Status Scale (EDSS) and magnetic resonance imaging (MRI) with gadolinium. Serum interleukin (IL)-1ß, IL-6, IL-12, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, IL-10, IL-4, and IL-17 levels were assessed using an enzyme-linked immunosorbent assay. IL-1ß, IL-6, TNF-α, IFN-γ, IL-17, IL-4, and IL-10 levels were higher in progressive MS than in controls. Increased IL-1ß and IFN-γ and decreased IL-12 and IL-4 levels were found in progressive MS compared with RRMS. Patients with progressive MS with disease progression presented higher TNF-α, IFN-γ, and IL-10 levels than those without disease progression. Patients with progressive MS with disease progression showed a higher frequency of positive gadolinium-enhanced lesions in MRI; higher TNF-α, IFN-γ, and IL-17 levels; and decreased IL-12 levels compared with RRMS patients with progression. There was a significant inverse correlation between IL-10 levels and EDSS score in patients with progressive MS. The results underscore the complex cytokine network imbalance exhibited by progressive MS patients and show the important involvement of TNF-α, IFN-γ, and IL-17 in the pathophysiology and progression of the disease. Moreover, serum IL-10 levels were inversely associated with disability in patients with progressive MS.

Association between HLA genes and dust mite sensitivity in a Brazilian population.

BACKGROUND: Type I hypersensitivity, also known as IgE-mediated allergy, is a complex, multifactorial condition whose onset and severity are influenced by both genetic and environmental factors. Mite allergens stimulate the production of humoral response (IgE), especially in children, which is closely involved in atopic asthma and rhinitis. OBJECTIVE: This study aimed to investigate the association between HLA class I (-A, -B, and -C), and HLA class II (-DRB1) genes in individuals sensitive to dust mites (Dermatophagoides farinae, Dermatophagoides pteronyssinus, or Blomia tropicalis) and mite-insensitive controls. METHODS: 396 participants were grouped as mite-sensitive and mite-insensitive according to immediate hypersensitivity as determined by skin-prick tests, and to HLA genotyping by polymerase chain reaction-sequence specific oligonucleotide (PCR-SSO). RESULTS: After chi-square heterogeneity testing no significant differences were observed in HLA-A, B, and C genes, except for the HLA-DRB1 locus, which, showed a negative association for DRB1∗04, between mite-sensitive and mite-insensitive individuals. In high resolution, DRB1∗04:11 allele was significantly different from all other results (P=0.0042, OR=0.26, and 95%CI=0.09-0.70). The analysis stratified by etiologic agent confirmed these associations. CONCLUSION: Our results suggest a possible association between HLA-DRB1 genes and hypersensitivity to dust mites.

Immune-Inflammatory and Oxidative and Nitrosative Stress Biomarkers of Depression Symptoms in Subjects with Multiple Sclerosis: Increased Peripheral Inflammation but Less Acute Neuroinflammation.

There is evidence that activated immune-inflammatory and oxidative and nitrosative stress (IO&NS) pathways play a role in the pathophysiology of multiple sclerosis (MS) and depression. This study examines serum levels of interleukin (IL)-1ß, IL-4, IL-6, and IL-10; peroxides (LOOH); nitric oxide metabolites (NOx); albumin; ferritin; C-reactive protein (CRP); and tumor necrosis factor (TNF)-ß NcoI polymorphism (rs909253) and gadolinium-enhanced magnetic resonance imaging (MRI) scan in MS patients with (n = 42) and without (n = 108) depression and normal controls (n = 249). Depression is scored using the depressive subscale of the Hospital Anxiety and Depression Scale (HADS). The extent of neurological disability is measured using the Expanded Disability Status Scale (EDSS) at the same time of the abovementioned measurements and 5 years earlier. Disease progression is assessed as actual EDSS-EDSS 5 years earlier. Three variables discriminate MS patients with depression from those without depression, i.e., increased IL-6 and lower IL-4 and albumin. Binary logistic regression showed that MS with depression (versus no depression) was characterized by more gastrointestinal symptoms and disease progression, higher serum IL-6, and lower albumin levels. In subjects with MS, the HADS score was significantly predicted by three EDSS symptoms, i.e., pyramidal, gastrointestinal, and visual symptoms. Fifty-eight percent of the variance in the HADS score was predicted by gastrointestinal symptoms, visual symptoms, the TNFB1/B2 genotype, and contrast enhancement (both inversely associated). There were no significant associations between depression in MS and type of MS, duration of illness, age, sex, nicotine dependence, and body mass index. MS with depression is associated with signs of peripheral inflammation, more disability, disease progression, gastrointestinal and visual symptoms, but less contrast enhancement as compared to MS without depression. It is concluded that depression is part of the neurological symptoms of MS and that its expression is primed by peripheral inflammation while acute neuroinflammation and the TNFB1/B2 genotype may be protective.

HLA-A, HLA-B, and HLA-DRB1 Allele and Haplotype Frequencies in Renal Transplant Candidates in a Population in Southern Brazil.

BACKGROUND: Very few studies have examined the diversity of human leukocyte antigens (HLA) in the Brazilian renal transplant candidates. METHODS: The frequencies of the HLA-A, HLA-B, and HLA-DRB1 alleles, haplotypes and phenotypes were studied in 522 patients with chronic renal failure, renal transplant candidates, registered at the Transplant Centers in north/northwestern Paraná State, southern Brazil. Patients were classified according to the ethnic group (319 whites [Caucasians], 134 mestizos [mixed race descendants of Europeans, Africans, and Amerindians; browns or "pardos"] and 69 blacks). The HLA typing was performed by the polymerase chain reaction sequence-specific oligonucleotide method (PCR-SSO), combined with Luminex technology. RESULTS: In the analysis of the total samples, 20 HLA-A, 32 HLA-B, and 13 HLA-DRB1 allele groups were identified. The most frequent allele groups for each HLA locus were HLA-A*02 (25.4%), HLA-B*44 (10.9%), and HLA-DRB1*13 (13.9%). The most frequent haplotypes were HLA-A*01-B*08-DRB1*03 (2.3%), A*02-B*44-DRB1*07 (1.2%), and A*03-B*07-DRB1*11 (1.0%). Significant differences (P < 0.05) were observed in the HLA-A*68, B*08, and B*58 allele frequencies among ethnic groups. CONCLUSIONS: This study provides the first data on the HLA-A, HLA-B, and HLA-DRB1 allele, phenotype and haplotype frequencies of renal transplant candidates in a population in southern Brazil.

CCR2-V64I genetic polymorphism: a possible involvement in HER2+ breast cancer.

Many tumor cells express chemokines and chemokine receptors, and these molecules can affect both tumor progression and anti-tumor immune response. Genetic polymorphisms of some chemokine receptors were found to be closely related to malignant tumors, especially in metastasis process, including breast cancer (BC). Considering this, it was investigated a possible role for CCR2-V64I (C-C chemokine receptor 2) and CCR5-Δ32 (C-C chemokine receptor 5) genetic variants in BC context. Patients were divided into subgroups according to immunohistochemical profile of estrogen (ER) and progesterone (PR) receptors and the human epidermal growth factor receptor 2 (HER2) overexpression. No significant associations were found in relation to susceptibility (CCR2-V64I: OR 1.32; 95 % CI 0.57-3.06; CCR5-∆32: OR 1.04; 95 % CI 0.60-1.81), clinical outcome (tumor size, lymph nodes commitment and/or distant metastasis, TNM staging and nuclear grade) or therapeutic response (recurrence and survival). However, it was found a significant correlation between CCR2-V64I allelic variant and HER2 immunohistochemical positive samples (p = 0.026). All in all, we demonstrate, for the first time, a positive correlation between CCR2 receptor gene polymorphism and a subgroup of BC related to poor prognosis, which deserves further investigation in larger samples for validation.

Prevalence of Torque teno virus in healthy donors of Paraná State, southern Brazil

OBJECTIVE: To determine the prevalence of the Torque teno virus in healthy donors in the northern and northwestern regions of the state of Paraná, southern Brazil.METHODS: The Torque teno virus was detected by a nested polymerase chain reaction using a set of oligoprimers for the N22 region.RESULTS: The prevalence of the virus was 69% in 551 healthy blood donors in southern Brazil. There was no statistically significant difference between the presence of the virus and the variables gender, ethnicity and marital status. There was significant difference in the prevalence of the virus regarding the age of the donors (p-value = 0.024) with a higher incidence (74.7%) in 18- to 24-year-old donors.CONCLUSION: A high prevalence of Torque teno virus was observed in the population studied. Further studies are needed to elucidate the routes of contamination and the clinical implications of the virus in the healthy population.

Prevalence of Torque teno virus in healthy donors of Paraná State, southern Brazil.

OBJECTIVE: To determine the prevalence of the Torque teno virus in healthy donors in the northern and northwestern regions of the state of Paraná, southern Brazil. METHODS: The Torque teno virus was detected by a nested polymerase chain reaction using a set of oligoprimers for the N22 region. RESULTS: The prevalence of the virus was 69% in 551 healthy blood donors in southern Brazil. There was no statistically significant difference between the presence of the virus and the variables gender, ethnicity and marital status. There was significant difference in the prevalence of the virus regarding the age of the donors (p-value=0.024) with a higher incidence (74.7%) in 18- to 24-year-old donors. CONCLUSION: A high prevalence of Torque teno virus was observed in the population studied. Further studies are needed to elucidate the routes of contamination and the clinical implications of the virus in the healthy population.

Torque teno virus among dialysis and renal-transplant patients.

Patients who undergo dialysis treatment or a renal transplant have a high risk of blood-borne viral infections, including the Torque teno virus (TTV). This study identified the presence of TTV and its genome groups in blood samples from 118 patients in dialysis and 50 renal-transplant recipients. The research was conducted in a hospital in the city of Maringá, state of Paraná. The viral DNA, obtained from whole blood, was identified by using two nested Polymerase Chain Reactions (PCR). The frequencies of TTV were 17% and 36% in dialysis patients using the methodology proposed by Nishizawa et al . (1997) and Devalle and Niel (2004) , respectively, and 10% and 54% among renal-transplant patients. There was no statistically significant association between the frequency of the pathogen and the variables: gender, time in dialysis, time since transplant, blood transfusions, and the concomitant presence of hepatitis B, for either the dialysis patients or the renal-transplant recipients. Among dialysis patients and renal-transplant recipients, genogroup 5 was predominant (48% and 66% respectively), followed by genogroup 4 (37% and 48%) and genogroup 1 (23% and 25%). Genogroup 2 was present in both groups of patients. Some patients had several genogroups, but 46% of the dialysis patients and 51% of the renal-transplant recipients had only a single genogroup. This study showed a high prevalence of TTV in dialysis patients and renal-transplant recipients.

Analysis of platelet eluate for the elucidation of sensitization to HLA in kidney transplant candidate / Análise do eluato de plaqueta para a elucidação da sensibilização ao HLA em um candidato ao transplante de rim

While a 42-year-old male patient was being prepared for deceased-donor renal transplantation, anti-HLA-A2 antibodies were detected in the serum by enzyme-linked immunosorbent assay (ELISA) method. The patient denied any transfusion history and previous transplant. Crossmatch by complement dependent cytotoxicity (CDC) and CDC with anti -human globulin (CDC-AHG) proved negative with a four-cell panel with positive typing for HLA-A2. Adsorption of antibodies with platelets and analysis of eluate were suggested to elucidate discrepancies in results by ELISA and by CDC-AHG. ELISA showed that adsorbed serum with platelets did not reveal antibodies for HLA-A2 specificity and suggested that they were removed by their specific binding with HLA-A2 antigens on the platelet surface. Eluate analysis by ELISA showed antibodies for HLA-A2 specificity. No antibodies for HLA-A2 specificity in the non-adsorbed serum were detected by CDC-AHG method. Revision of patient's data showed that a previous transfusion had occurred, which may have been the source of HLA sensitization. The suggested method may be a contribution towards the evaluation of sensitivity between CDC-AHG and ELISA methods for characterizing antibodies in the patient's serum.

Enquanto um paciente do sexo masculino de 42 anos de idade estava sendo preparado para o transplante renal de doador falecido, anticorpos anti-HLA-A2 foram detectados no soro pelo método de ensaio imunoenzimático (ELISA). O paciente negava história de transfusão e transplante anterior. Prova-cruzada por citotoxicidade dependente de complemento (CDC) e CDC com antiglobulina humana (CDC-AGH) foram negativos com um painel de quatro células com tipagem positiva para HLA-A2. O método de adsorção de anticorpos com plaquetas e análise do eluato foi sugerido para explicar as discrepâncias dos resultados de ELISA e CDC-AGH. O método de ELISA mostrou que o soro adsorvido com plaquetas não revelou anticorpos para especificidade HLA-A2, sugerindo que eles foram removidos por meio de sua ligação específica com os antígenos HLA-A2 na superfície das plaquetas. A análise do eluato por ELISA mostrou anticorpos para especificidade HLA-A2. Nenhum anticorpo para especificidade HLA-A2 foi detectada no soro não adsorvido pelo método de CDC-AGH. Revisão dos dados do paciente mostrou que houve transfusão anterior, podendo ter sido a fonte de sensibilização HLA. O método sugerido é uma contribuição para avaliação da sensibilidade entre os métodos de CDC-AGH e ELISA em caracterizar anticorpos no soro do paciente.

Torque teno virus among dialysis and renal-transplant patients

Patients who undergo dialysis treatment or a renal transplant have a high risk of blood-borne viral infections, including the Torque teno virus (TTV). This study identified the presence of TTV and its genome groups in blood samples from 118 patients in dialysis and 50 renal-transplant recipients. The research was conducted in a hospital in the city of Maringá, state of Paraná. The viral DNA, obtained from whole blood, was identified by using two nested Polymerase Chain Reactions (PCR). The frequencies of TTV were 17% and 36% in dialysis patients using the methodology proposed by Nishizawa et al. (1997) and Devalle and Niel (2004), respectively, and 10% and 54% among renal-transplant patients. There was no statistically significant association between the frequency of the pathogen and the variables: gender, time in dialysis, time since transplant, blood transfusions, and the concomitant presence of hepatitis B, for either the dialysis patients or the renal-transplant recipients. Among dialysis patients and renal-transplant recipients, genogroup 5 was predominant (48% and 66% respectively), followed by genogroup 4 (37% and 48%) and genogroup 1 (23% and 25%). Genogroup 2 was present in both groups of patients. Some patients had several genogroups, but 46% of the dialysis patients and 51% of the renal-transplant recipients had only a single genogroup. This study showed a high prevalence of TTV in dialysis patients and renal-transplant recipients.

Human leukocyte antigen (HLA)-G and cervical cancer immunoediting: a candidate molecule for therapeutic intervention and prognostic biomarker?

While persistent infection with oncogenic types of human Papillomavirus (HPV) is required for cervical epithelial cell transformation and cervical carcinogenesis, HPV infection alone is not sufficient to induce tumorigenesis. Only a minor fraction of HPV infections produce high-grade lesions and cervical cancer, suggesting complex host-virus interactions. Based on its pronounced immunoinhibitory properties, human leukocyte antigen (HLA)-G has been proposed as a possible prognostic biomarker and therapeutic target relevant in a wide variety of cancers and viral infections, but to date remains underexplored in cervical cancer. Given the possible influence of HLA-G on the clinical course of HPV infection, cervical lesions and cancer progression, a better understanding of HLA-G involvement in cervical carcinogenesis might contribute to two aspects of fundamental importance: 1. Characterization of a novel diagnostic/prognostic biomarker to identify cervical cancer and to monitor disease stage, critical for patient screening; 2. Identification of HLA-G-driven immune mechanisms involved in lesion development and cancer progression, leading to the development of strategies for modulating HLA-G expression for treatment purposes. Thus, this systematic review explores the potential involvement of HLA-G protein expression and polymorphisms in cervical carcinogenesis.

Protein Expression and Codon 72 Polymorphism of TP53 Gene in Triple Negative Breast Cancer

A subgroup of tumor that has received attention is triple-negative breast cancer (TNBC), which presents phenotype of negative estrogen receptor, negative progesterone receptor and has no overexpression of HER2. TP53 acts as a tumor suppressor limiting the proliferation of damaged cells. A polymorphic site (rs1042522) of TP53 encodes either an arginine or a proline amino acid, but its biological significance remains unclear. This study aimed to investigate this variant and its expression in search for a possible involvement in TNBC susceptibility and clinical outcome. Genetic polymorphism was evaluated in 50 patients and 115 controls by PCR based methodology and immunohistochemistry was done with monoclonal antibody. Case-control study showed no positive or negative association (OR= 0.95; CI95%= 0.48-1.89). Comparison of genotypes and clinical outcome showed no significant results. Despite most of patients presented p53 positive staining by immunohistochemistry, there was no significant association in relation to prognostic parameters. Results demonstrated a lack of association between codon 72 polymorphism, susceptibility and prognosis of TNBC. Immunohistochemistry analysis should be done more carefully, since most of the patients had the somatic mutation of p53, which could be an indicator of prognostic value in TNBC.

Soroprevalência do Citomegalovirus Humano em amostras de pacientes dialíticos e transplantados renais / Seroprevalence for Human Cytomegalovirus in samples from dialysis and kidney transplanted patients

Objetivou-se investigar a soroprevalência do Citomegalovírus Humano em dialíticos e transplantados renais e a associação com a hemotransfusão e o sexo. Trata-se de estudo transversal, realizado nos anos de 2011 e 2012, no município de Maringá, PA, Brasil. A população estudada foi composta por 203 pacientes dialíticos e 53 transplantados renais e a identificação viral foi realizada por meio de técnica sorológica. Encontrou-se 96% (195) de soropositividade para o anti-HCMV-IgG em dialíticos e 100% nos transplantados; 5% (10) de soropositividade para o anti-HCMV-IgM em dialíticos e 37,7% (20) em transplantados. Realizando o teste exato de Fischer, não houve associação significante entre a soropositividade do anti-HCMV com a hemotransfusão e o gênero. A grande parcela de infectados sugere a importância do cuidado de enfermagem para evitar a infecção cruzada durante os procedimentos de rotina...

The aim of this study was at investigating the seroprevalence of Human Cytomegalovirus in dialysis and kidney transplantedpatients and the association with hemodialysis and sex. It is a cross sectional study made in 2011 and 2012 in the countyof Maringá, PR, Brazil. The studied population consisted of 203 dialysis patients and 53 kidney transplanted patients andthe viral identification was made through serological technique. The study found 96% (195) seropositivity for anti-HCMVIgG in dialysis patients and 100% in the transplanted ones; 5% (10) seropositivity for anti-HCMV-IgM in dialysis patientsand 37.7% (20) in transplanted ones. Performing Fisher’s exact test, there was no significant association between theseroprevalence of anti-HCMV with hemodialysis and gender. A large number of infected patients suggest the importance ofnursing care to avoid cross-infection during routine procedures...

El objetivo fue investigar la seroprevalencia de Citomegalovirus Humano en dialíticos y trasplantados renales y la asociacióncon la transfusión de sangre y el sexo. Estudio transversal, llevado a cabo de 2011 a 2012, en Maringá, PR, Brasil. La poblaciónfue compuesta de 203 pacientes dialíticos y 53 trasplantados renales y la identificación viral se realizó mediante la técnicaserológica. Se encontró 96% (195) de seropositividad para anti-HCMV-IgG en dialíticos y 100% en los trasplantados; 5%(10) de seropositividad para anti-HCMV-IgM en dialíticos y 37,7% (20) en trasplantados. Realizándose la prueba exacta deFisher, no hubo asociación significativa entre la seropositividad del anti-HCMV con la transfusión de sangre y género. A lagran parte de los infectados se señala la importancia de la atención de enfermería para prevenir la infección cruzada durantelos procedimientos de rutina...

Tumor necrosis factor beta NcoI polymorphism is associated with inflammatory and metabolic markers in multiple sclerosis patients.

To evaluate the association between the tumor necrosis factor beta (TNF-ß) NcoI polymorphism and inflammatory and metabolic markers in patients with multiple sclerosis (MS) patients and the association of these markers with disease disability, a 782 base-pair fragment of the TNF-ß gene was amplified from genomic DNA and digested with the NcoI restriction enzyme. The serum levels of numerous cytokines (IL-1ß, IL-12, IL-6, TNF-α, IFN-γ, IL-4, IL-10, and IL-17) serum lipid levels, plasma insulin levels, and the Homeostasis Model Assessment-Insulin Resistance (HOMA-IR) levels were evaluated in 123 female and 43 male patients with MS. Females carrying the TNFB2/B2 genotype presented with decreased IL-4 and IL-10 levels and increased TNF-α, glucose, insulin, and HOMA-IR levels; moreover, there were positive correlations between EDSS and glucose and between EDSS and HOMA-IR in these females. Males carrying the TNFB2/B2 genotype exhibited increased levels of TNF-α, IFN-γ, and IL-17 (p=0.0326) and decreased levels of IL-4, IL-10, insulin, and HOMA-IR; there was a positive correlation between EDSS and TNF-α levels. The TNFB2/B2 genotype of TNF-ß NcoI polymorphism was associated with increased inflammatory and metabolic markers and this association was different according to sex of MS patients.

Evaluation of the humoral immune response to human leukocyte antigens in Brazilian renal transplant candidates.

Pre-transplant sensitization to human leukocyte antigens (HLA) is a risk factor for graft failure. Studies of the immunological profile related to anti-HLA antibodies in Brazilian renal transplant candidates are few. In this study, we evaluated the humoral immune response to HLA antigens in 269 renal transplant candidates, in Paraná State, Brazil. The HLA typing was performed by the polymerase chain reaction sequence-specific oligonucleotide method (PCR-SSO) combined with Luminex technology, using an SSO-LABType commercial kit (One Lambda, Inc., Canoga Park, CA, USA). The percentages of panel-reactive antibodies (PRA) and the specificity of anti-HLA antibodies were determined using the LS1PRA and LS2PRA commercial kits (One Lambda, Inc.). The PRA-positive group consisted of 182 (67.7%) patients, and the PRA-negative group of 87 (32.3%) patients. The two groups differed significantly only with respect to gender. Females were the most sensitized. Among the 182 patients with PRA- positive, 62 (34.1%) were positive for class I and negative for class II, 39 (21.4%) were negative for class I and positive for class II, and 81 (44.5%) were positive for both classes I and II. The HLA-A*02, A*24, A*01, B*44, B*35, B*15, DRB1*11, DRB1*04 and DRB1*03 allele groups were the most frequent. The specificities of anti-HLA antibodies were more frequent: A34, B57, Cw15, Cw16, DR51, DQ8 and DP14. This study documented the profile of anti-HLA antibodies in patients with chronic renal failure who were on waiting lists for an organ in Paraná, and found high sensitization to HLA antigens in the samples.

Discrepancies in HLA typing: use of different methods to confirm doubtful or inconclusive results / Discrepâncias na tipagem HLA: diferentes métodos para confirmar resultados duvidosos ou inconclusivos

The major histocompatibility complex (MHC) is a set of genes found on the short arm of chromosome 6. MHC molecules in human beings are known as human leukocyte antigens (HLA). HLA polymorphism can be determined by serological and molecular typing methods, which may yield discordant results. The present analysis performed HLA typing of samples with discordant results by PCR-SSP and PCR-SSO, so that typing discrepancies could be clarified. The cross-sectional study analyzed 33 samples from individuals included in an HLA-disease association study. Discrepant alleles were observed in 6 of 33 samples. Discordant samples were retyped using One Lambda Micro SSP™, Dynal RELI™ SSO and Luminex™ SSO assays for HLA class I (HLA-A, HLA-B) and class II (HLA-DRB1) molecules. The three methods produced concordant results after HLA retyping. Human error occurred in interpreting the initial results, which led to discrepancies in the results obtained. The participation of experienced professionals and the availability of at least two different methods to confirm doubtful or inconclusive results are mandatory for effective HLA typing.

O complexo principal de histocompatibilidade (MHC) é um conjunto de genes encontrados no braço curto do cromossomo 6. Em humanos, as moléculas de MHC são conhecidas como antígenos leucocitários humanos (HLA). Polimorfismo HLA pode ser determinado por métodos de tipagem sorológica e molecular que são susceptíveis de produzir resultados discordantes. Este estudo teve como objetivo realizar a tipagem HLA de amostras com resultados discordantes por PCR-SSP e-SSO e para esclarecer discrepâncias de digitação. Este estudo transversal analisou 33 amostras de indivíduos incluídos em um estudo de associação HLA-doença. Alelos discrepantes foram observados em seis das 33 amostras. Amostras discordantes foram retyped usando One Lambda Micro SSP™, Dynal RELI™ SSO Luminex e ensaios ™ SSO para HLA de classe I (HLA-A, HLA-B) e classe II (HLA-DRB1) moléculas. Todos os três métodos apresentaram resultados concordantes após HLA redigitação. Houve erro humano na interpretação dos resultados iniciais o que levou a uma discrepância entre os resultados obtidos. Concluiu-se que a participação de profissionais experientes e com a disponibilidade de pelo menos dois métodos diferentes para confirmar os resultados duvidosos ou inconclusivos são essenciais para a tipagem de HLA eficaz.