O probiótico Saccharomyces boulardii modula a resposta de IgG2a em camundongos expostos aos antígenos de Leishmania infantum / Probiotic Saccharomyces boulardii modulates IgG2a response in miceexposed to Leishmania infantum antigens

O objetivo do presente estudo foi avaliar os efeitos do probiótico Saccharomyces boulardii na modulação da resposta imune humoral de animais expostos a antígenos de Leishmania infantum. Para isso, 16 camundongos BALB/c foram imunizados com antígeno particulado de Leishmania infantum e divididos em dois grupos experimentais, um composto por animais suplementados e outro por animais não suplementados com o probiótico. Amostras de sangue dos animais foram colhidas semanalmente durante o período experimental e submetidas ao Ensaio da Imunoabsorbância Ligado à Enzima indireto para avaliação dos títulos de IgG totais e o perfil dos isotipos de IgG produzidos (IgG1 e IgG2a). A suplementação com o probiótico não exacerbou a produção de IgG total em comparação ao grupo controle, não havendo diferenças significativas entre os dois grupos. Porém, as soroconversões de IgG2a foram mais elevadas no grupo suplementado, no qual registrou-se um aumento de 1,46 vezes no final do experimento. Assim, a suplementação com S. boulardii foi capaz de modular a resposta de IgG2a/IgG1 nos animais expostos aos antígenos de Leishmania infantum.

The aim of the present study was to evaluate the effects of the Saccharomyces boulardii probiotic on the modulation of humoral immune response in animals exposed to Leishmania infantum antigens. For this, 16 BALB/c mice were immunized with Leishmania infantum particulate antigen and divided into two experimental groups, one consisting of supplemented animals and the other not probiotic supplemented animals. Blood samples from the animals were taken weekly during the experimental period and subjected to the Indirect Enzyme-Linked Immunosorbance Assay for evaluation of total IgG titers and the profile of the produced IgG isotypes (IgG1 and IgG2a). Probiotic supplementation did not exacerbate total IgG production compared to the control group, with no significant differences between the two groups. However, IgG2a seroconversions were higher in the supplemented group, which showed a 1.46-fold increase at the end of the experiment. Thus, supplementation with S. boulardii was able to modulate the IgG2a/IgG1 response in animals exposed to Leishmania infantum antigens.

Characteristics and Clinical Significance of de novo Donor-Specific Anti-HLA Antibodies after Kidney Transplantation

BACKGROUND: The association of de novo donor-specific anti-human leukocyte antigens (HLA) antibodies (DSA) and development of antibody-mediated rejection (AMR) in kidney transplant recipients (KTRs) is still undetermined. METHODS: We prospectively screened de novo DSA in 167 KTRs during 32 months after kidney transplantation (KT). Timing of DSA detection was at 3, 6, and 12 months post-transplant and annually thereafter and when clinically indicated. DSA levels were determined by Luminex assays and expressed as mean fluorescence intensity (MFI). We evaluated the incidence, characteristics of DSA, and association between DSA and tacrolimus trough levels or AMR. RESULTS: De novo DSA developed in 16 KTRs (9.6%) and acute AMR occurred more commonly in KTRs with de novo DSA compared to KTRs without de novo DSA (18.8% vs. 0%, P < 0.001). All de novo DSA were against class II antigens. The mean number of DSA was 1.8 ± 1.2 and the average MFI of DSA was 7,399 ± 5,470. Tacrolimus trough level during the first 0–2 months after KT was an independent predictor of DSA development (hazard ratio, 0.70; 95% confidence interval, 0.50–0.99; P = 0.043). No differences were found in the number of DSA, average MFI of DSA, and tacrolimus levels during the first year between de novo DSA-positive KTRs with AMR and those without AMR. CONCLUSION: The results of our study suggest that monitoring of DSA and maintaining proper tacrolimus levels are essential to prevent AMR during the initial period after KT.

Avaliação de antígeno leucocitário humano classe I e II em pacientes pediátricos com gastrite e úlcera duodenal ativas e Helicobacter pylori positivo / Evaluation of human leukocyte antigen class i and ii antigens in helicobacter pylori-positive pediatric patients with active gastritis and duodenal ulcer

ABSTRACT BACKGROUND: As being the first bacteria determined to be carcinogenic, Helicobacter pylori (H. pylori) is a pathogen localized in the stomach in more than half of the world population. Some earlier studies have found a relation between tissue histocompatibility antigens and gastric cancers depending on the regions. OBJECTIVE: The present study aimed to determine the distribution of human leukocyte antigen (HLA) class I and class II antigens in H. pylori-positive pediatric patients with active gastritis and duodenal ulcer, excluding cancer cases, in our center. METHODS: The study included 40 patients diagnosed with H. pylori-positive active gastritis and duodenal ulcer and 100 controls consisting of healthy donor candidates. The HLA class I and class II antigens were studied in the isolated DNA samples using the polymerase chain reaction sequence-specific oligonucleotide probes. RESULTS: The frequency of HLA-B*51 antigen was significantly higher in the patient group than in the control group (40% vs 17%; P=0.003). There was no difference between the two groups in terms of the frequencies of HLA-A, HLA-C, HLA-DR, and HLA-DQ antigens. CONCLUSION: It was determined that HLA-B*51 plays a critical role in H. pylori infection.

RESUMO CONTEXTO: Determinada como sendo a primeira bactéria cancerígena, o Helicobacter pylori (H. pylori) é um patógeno localizado no estômago em mais da metade da população mundial. Alguns estudos anteriores têm encontrado uma relação entre câncer gástrico e antígenos de histocompatibilidade de tecido dependendo das regiões. OBJETIVO: O presente estudo teve como objetivo determinar a distribuição em nosso centro do antígeno leucocitário humano (HLA) de classe I e antígenos classe II em pacientes pediátricos H. pylori-positivos com gastrite e úlcera duodenal ativas, excluindo casos de câncer. MÉTODOS: O estudo incluiu 40 pacientes H. pylori-positivos diagnosticados com gastrite e úlcera duodenal ativas e 100 controles consistindo de candidatos doadores saudáveis. Foram estudadas nas amostras de DNA isoladas o antígeno leucocitário humano classe I e antígenos classe II, utilizando-se as cadeias de sequência específica de polimerase do oligonucleotideo. RESULTADOS: A frequência do antígeno HLA - B * 51 foi significativamente maior no grupo de pacientes do que no grupo controle (40% vs 17%; P=0,003). Não houve diferença entre os dois grupos em termos das frequências dos antígenos HLA-A, HLA-DR, HLA-DQ e HLA-C. CONCLUSÃO: Determinou-se que o HLA - B * 51 desempenha um papel crítico na infecção pelo H. pylori.

Study of polymorphisms of HLA class Ⅰ (-A, -B, -C) and class Ⅱ (DRB1, DQA1, DQB1, DPA1, DPB1) genes among ethnic Hans from Southern China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the genetic polymorphisms of human leukocyte antigen (HLA)- A, B, C, DRB1, DQA1, DQB1, DPA1and DPB1among ethnic Hans from southern China.</p><p><b>METHODS</b>481 randomly selected individuals were genotyped using a polymerase chain reaction (PCR) sequence-based typing (SBT) method for the above genes. Their allele frequencies were determined by direct counting.</p><p><b>RESULTS</b>In total, 28 HLA-A, 57 HLA-B, 28 HLA-C, 40 HLA-DRB1, 18 HLA-DQA1, 17 HLA-DQB1, 6 HLA-DPA1and 21 HLA-DPB1alleles were identified. Among these, common alleles (with allelic frequencies > 0.05) included A*1101, A*2402, A*0207, A*3303, A*0201, B*40:01, B*46:01, B*58:01, B*13:01, B*15:02, C*01:02, C*07:02, C*03:04, C*03:02, C*08:01, C*03:03, C*04:01, DRB1*09:01, DRB1*15:01, DRB1*12:02, DRB1*08:03, DRB1*03:01, DRB1*04:05, DRB1*11:01, DQA1*01:02, DQA1*03:02, DQA1*03:03, DQA1*06:01, DQA1*01:03, DQA1*05:05, DQA1*01:04, DQA1*03:01, DQA1*05:01, DQB1*03:01, DQB1*03:03, DQB1*06:01, DQB1*05:02, DQB1*03:02, DQB1*02:01, DQB1*03:02, DQB1*06:02, DPA1*02:02, DPA1*01:03, DPA1*02:01, DPB1*05:01, DPB1*02:01, DPB1*13:01, DPB1*04:01and DPB1*02:02.For each of the locus, the overall frequencies of common alleles were 75.57%, 52.81%, 78.28%, 62.16%, 86.70%, 77.23%, 95.32% and 81.59%, respectively.</p><p><b>CONCLUSION</b>The allelic frequencies of the 8 selected HLA loci among ethnic Hans from southern China may served as a reference for anthropology, legal medicine, transplantation and disease association studies.</p>

Imunohematologia veterinária: antígenos eritrocitários caninos / Veterinary immunohematology: canine erythrocyte antigen / Inmunohematología veterinaria: antígenos eritrocitarios caninos

A imunohematologia veterinária vem ganhando interesse nos últimos anos devido a maior acessibilidade a tecnologias de detecção de antígenos e anticorpos, interesse dos donos e médicos veterinários em buscar uma melhor qualidade de vida para os animais e as necessidades de transfusões com o menor índice possível de reações indesejadas. Os cães possuem antígenos presentes na membrana de suas células vermelhas, podendo causar reações durante e após transfusões. Diferentemente de humanos e felinos, cães não possuem anticorpos naturais para os principais antígenos, a priori podendo ser transfundidos com qualquer tipo sanguíneo sem consequências posteriores, porém, se submetidos a uma segunda transfusão, sendo essa de um tipo sanguíneo incompatível e previamente sensibilizados, as chances de ocorrer reações transfusionais graves aumentam drasticamente, ocasionando danos ao animal, podendo levá-lo à morte. Por conta desses riscos se faz necessário uma maior atenção aos tipos sanguíneos desses animais onde 8 sistemas são reconhecidos internacionalmente classificados como sistema DEA, sendo eles DEA 1 e seus subtipos (DEA 1.1; DEA1.2; DEA 1.3); DEA 3; DEA 4; DEA 5; DEA 6; DEA 7 e DEA 8, e recentemente um novo sistema denominado Dal. Não há disponível ainda soros para os sistemas DEA 6 e DEA 8, tornando a pesquisa sobre esses antígenos dificultosa.(AU)

Veterinary immunohematology is gaining interest in recent years due to greater accessibility to antigen and antibody detection technologies, the interests of pet owners and veterinarians in seeking a better quality of life for animals, and requirement of transfusions with the lowest possible rate of collateral reactions. Dogs have antigens present in the membrane of their red blood cells that can cause reactions during and after transfusions. Unlike humans and cats, dogs do not have natural antibodies to the key antigens, and a priori they can be transfused with any type of blood without any further consequences. However, if they are ever subjected to a second transfusion, if using incompatible blood types and being previously sensitized, the likelihood of having serious transfusion reactions drastically increase, causing damage to the animal, which may even lead it to death. Due to those risks, greater attention is required to the blood type of those animals, which present 8 systems, internationally recognized and classified as the DEA system, namely DEA 1 and its subtypes (DEA 1.1; DEA 1.2; DEA 1.3); DEA 3; DEA 4; DEA 5; DEA 6; DEA 7 and DEA 8, and recently a new system referred to as Dal. No serum is yet available for DEA 6 and DEA 8 systems, hindering the research on those antigens.(AU)

La inmunohematología veterinaria ha ganado atención en los últimos años debido mayor accesibilidad a tecnologías de detección de antígenos y anticuerpos, interés de dueños y médicos veterinarios en buscar mejor calidad de vida para los animales y las necesidades de transfusiones con menor índice posible de reacciones indeseadas. Los perros poseen antígenos presentes en la membrana de sus células rojas, pudiendo causar reacciones durante y después de transfusiones. Diferentemente de humanos y felinos, perros no tienen anticuerpos naturales para los principales antígenos, a priori, pudiendo ser transfundidos con cualquier tipo de sangre sin consecuencias posteriores, todavía, si sometidos a una segunda transfusión, siendo esa de un tipo sanguíneo incompatible y previamente sensibilizados, la posibilidad de ocurrir reacciones transfusional grave aumenta drásticamente, ocasionando daños al animal, pudiendo llevarlo a la muerte. Por esos riesgos se hace necesario más atención a los tipos sanguíneos de esos animales, donde 8 sistemas son reconocidos internacionalmente y clasificados como sistema DEA, siendo ellos DEA 1 y sus subtipos (DEA 1.1; DEA 1.2; DEA 1.3); DEA 3; DEA 4; DEA 5; DEA 6; DEA 7 y DEA 8, y recién un nuevo sistema denominado Dal. No hay aún disponible sueros para los sistemas DEA 6 y DEA 8, haciendo dificultosa la investigación sobre esos antígenos.(AU)

The immunogenetics of multiple sclerosis. The frequency of HLA-alleles class 1 and 2 is lower in Southern Brazil than in the European population / Imunogenética na esclerose múltipla. A frequência dos alelos HLA classe 1 e 2 no sul do Brasil são menores que da população Europeia

ABSTRACT Objective To study the HLA of class 1and 2 in a multiple sclerosis (MS) population to verify the susceptibility for the disease in the Southern Brazil. Methods We analyzed patients with MS and controls, by direct sequencing of the genes related to HLA DRB1, DQB1, DPB1, A, B and C alleles with high resolution techniques. Results We found a lower frequency of all HLA alleles class 1 and 2 in MS and controls comparing to the European population. Several alleles had statistical correlation, but after Bonferroni correction, the only allele with significance was the HLA-DQB1*02:03, which has a positive association with MS. Conclusions Our data have different frequency of HLA-alleles than the previous published papers in the Southeast Brazil and European population, possible due to several ethnic backgrounds.

RESUMO Objetivo Estudo do HLA classes 1 e 2 em pacientes com esclerose múltipla (EM) a fim de verificar a susceptibilidade para a doença em uma população do Sul do Brasil. Métodos Foram analisados por sequenciamento direto de alta resolução os genes relacionados com os HLA DRB1, DQB1, DPB1, A, B e C em casos de EM comparados com uma população controle normal. Resultados Foi encontrado uma frequência menor dos alelos dos HLA classe 1 e 2 nos casos de EM e controles quando comparado com a população Europeia. Diversos alelos mostraram correlação estatística, mas depois da correção de Bonferroni, somente o alelo do HLA-DQB1*02:03 foi positivo para a EM. Conclusões Encontramos frequência diferente dos alelos do HLA relatados previamente nos Sudeste do Brasil e Europeus, possivelmente devido a origem étnica diferente da população estuda.

Lack of association between alopecia areata and HLA class I and II in a southeastern Brazilian population

Abstract: Background: Alopecia areata (AA) is a common disorder of unknown etiology that affects approximately 0.7% to 3.8% of patients among the general population. Currently, genetic and autoimmune factors are emphasized as etiopathogenic. Studies linking Human Leukocyte Antigens (HLA) to AA have suggested that immunogenetic factors may play a role in the disease's onset/development. Objectives: To investigate an association between AA and HLA class I/II in white Brazilians. Methods: Patients and control groups comprised 33 and 112 individuals, respectively. DNA extraction was performed by column method with BioPur kit. Allele's classification was undertaken using the PCR-SSO technique. HLA frequencies were obtained through direct counting and subjected to comparison by means of the chi-square test. Results: Most patients were aged over 16, with no familial history, and developed partial AA, with no recurrent episodes. Patients showed a higher frequency of HLA-B*40, HLA-B*45, HLA-B*53 and HLA-C*04 compared with controls, although P was not significant after Bonferroni correction. Regarding HLA class II, only HLA-DRB1*07 revealed statistical significance; nevertheless, it featured more prominently in controls than patients (P=0.04; Pc=0.52; OR=0.29; 95%; CI=0.07 to 1.25). P was not significant after Bonferroni correction. Conclusions: The development of AA does not seem to be associated with HLA in white Brazilians, nor with susceptibility or resistance. The studies were carried out in populations with little or no miscegenation, unlike the Brazilian population in general, which could explain the inconsistency found.

Immunologic properties of differentiated and undifferentiated mesenchymal stem cells derived from umbilical cord blood

The expression of immunogenic markers after differentiation of umbilical cord blood (UCB)-derived mesenchymal stem cells (MSC) has been poorly investigated and requires extensive in vitro and in vivo testing for clinical application. The expression of human leukocyte antigen (HLA) classes on UCB-derived MSC was tested by Fluorescence-activated cell sorting analysis and immunocytochemical staining. The undifferentiated MSC were moderately positive for HLA-ABC, but almost completely negative for HLA-DR. The MSC differentiated to chondrocytes expressed neither HLA-ABC nor HLA-DR. The proliferation of MSC was not significantly affected by the allogeneic lymphocytes stimulated with concanavalin A. The responder lymphocytes showed no significant decrease in proliferation in the presence of the MSC, but the apoptosis rate of the lymphocytes was increased in the presence of MSC. Taken together, these findings indicate that UCB-derived MSC differentiated to chondrocytes expressed less HLA class I and no class II antigens. The MSC showed an immunomodulatory effect on the proliferation and apoptosis of allogeneic lymphocytes. These data suggest that the differentiated and undifferentiated allogeneic MSC derived from umbilical cord blood can be a useful candidate for allogeneic cell therapy and transplantation without a major risk of rejection.

Expansion and function of MHC restricted killer T cells derived from umbilical cord blood / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to expand the cytotoxic T lymphocytes (CTL) through inducing the differentiation of umbilical blood monomuclear cells (UBMNC) by using various combination of cytokines, and to investigate the functions of expanded CTL.</p><p><b>METHODS</b>The MNC were isolated by ficoll density gradient centrifugation. Then, the PHA-P, IFN-γ combined with IL-2, IL-15 and other cytokines were used for induction and expansion of the cord blood-derived CTL. The biological function of CTL was examined by phenotype analysis, cytotoxic tests and real-time fluorescence quantitative PCR.</p><p><b>RESULTS</b>After expansion for 15 days, the cell number increased by 1522% ± 137%. The content of CD3(-)CD8(-) cells in uncultured cord blood MNC was 95%, and the CD3(+)CD8(+) CTL cells reached 82.77% in cultured cord blood MNC after expansion for 15 days. The expanded CTL cell showed the cytotoxic activity against K562 and HeLa cell line. The killing rate of MNC was 61.88 ± 1.08%. After expansion, the killing rate could reach to 90% with the average value of 90.33 ± 2.02%. The expanded CTL cells highly expressed some key cytokines, such as granzyme A, granzyme B, GM-CSF, granulysin, IFN-γ, TGF-β, TNF-α and perforin. Compared with the control group, the expression of IFN-γ and TGF-β significantly increased (P < 0.05), and the other factors dramatically increased (P < 0.01).</p><p><b>CONCLUSION</b>The cord blood-derived CTL can be expanded by different combinations of cytokines. These protocols may provide alternative choices for CTL cell expansion in tumor adoptive immunotherapy.</p>

Functional disruption of human leukocyte antigen II in human embryonic stem cell

BACKGROUND: Theoretically human embryonic stem cells (hESCs) have the capacity to self-renew and differentiate into all human cell types. Therefore, the greatest promise of hESCs-based therapy is to replace the damaged tissues of patients suffering from traumatic or degenerative diseases by the exact same type of cells derived from hESCs. Allo-graft immune rejection is one of the obstacles for hESCs-based clinical applications. Human leukocyte antigen (HLA) II leads to CD4+ T cells-mediated allograft rejection. Hence, we focus on optimizing hESCs for clinic application through gene modification. RESULTS: Transcription activator-like effector nucleases (TALENs) were used to target MHC class II transactivator (CIITA) in hESCs efficiently. CIITA(-/-)hESCs did not show any difference in the differentiation potential and self-renewal capacity. Dendritic cells (DCs) derived from CIITA(-/-)hESCs expressed CD83 and CD86 but without the constitutive HLA II. Fibroblasts derived from CIITA(-/-)hESCs were powerless in IFN-γ inducible expression of HLA II. CONCLUSION: We generated HLA II defected hESCs via deleting CIITA, a master regulator of constitutive and IFN-γ inducible expression of HLA II genes. CIITA(-/-)hESCs can differentiate into tissue cells with non-HLA II expression. It's promising that CIITA(-/-)hESCs-derived cells could be used in cell therapy (e.g., T cells and DCs) and escape the attack of receptors' CD4+ T cells, which are the main effector cells of cellular immunity in allograft.

Analyses of the TCR repertoire of MHC class II-restricted innate CD4+ T cells

Analysis of the T-cell receptor (TCR) repertoire of innate CD4+ T cells selected by major histocompatibility complex (MHC) class II-dependent thymocyte-thymocyte (T-T) interaction (T-T CD4+ T cells) is essential for predicting the characteristics of the antigens that bind to these T cells and for distinguishing T-T CD4+ T cells from other types of innate T cells. Using the TCRmini Tg mouse model, we show that the repertoire of TCRalpha chains in T-T CD4+ T cells was extremely diverse, in contrast to the repertoires previously described for other types of innate T cells. The TCRalpha chain sequences significantly overlapped between T-T CD4+ T cells and conventional CD4+ T cells in the thymus and spleen. However, the diversity of the TCRalpha repertoire of T-T CD4+ T cells seemed to be restricted compared with that of conventional CD4+ T cells. Interestingly, the frequency of the parental OT-II TCRalpha chains was significantly reduced in the process of T-T interaction. This diverse and shifted repertoire in T-T CD4+ T cells has biological relevance in terms of defense against diverse pathogens and a possible regulatory role during peripheral T-T interaction.

43 kDa and 66 kDa, two blood stage antigens induce immune response in Plasmodium berghei malaria.

The hunt for an effective vaccine against malaria still continues. Several new target antigens as candidates for vaccine design are being explored and tested for their efficacy. In the present study the sera from mice immunized with 24,000 × g fraction of Plasmodium berghei has been used to identify highly immunogenic blood stage antigens. The protective antibodies present in immune sera were covalently immobilized on CNBr activated sepharose 4B and used for affinity chromatography purification of antigens present in blood stages of P. berghei. Two polypeptides of 66 and 43 kDa molecular weights proved to be highly immunogenic. They exhibited a strong humoral immune response in mice as evident by high titres in ELISA and IFA. Protective immunity by these two antigens was apparent by in vivo and in vitro studies. These two proteins could further be analysed and used as antigens in malaria vaccine design.

Experimental study on H2-Ab1 gene expression in the nasal mucosa of mice with allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the level of H2-Ab1 in the nasal mucosa of mice with allergic rhinitis.@*METHOD@#Twenty-four female 129/sv mice were divided into 2 groups: ovalbumin (OVA) group and control. The allergic rhinitis models were induced by classical method with OVA. After the last challenge, the pathological differences between the two groups were surveyed. The levels of H2-AB1 were measured by ELISA and quantitative real time PCR.@*RESULT@#The expression of H2-Ab1 is higher in subjects with AR than that in controls (P < 0.05).@*CONCLUSION@#The levels of H2-Ab1 were highly increased in allergic rhinitis group, which might be associated with the pathogenesis of allergic rhinitis.

Hallazgo de patrones para péptidos-vacuna con capacidad de acople universal en moléculas de HLA-II / Identification of patterns for peptide-vaccines in universal capacity coupling HLA-II molecules / Identificagáo de padroes para peptídeos-vacina com capacidade de acoplamento universal em moléculas HLA-II

Partiendo del alineamiento múltiple de secuencias proteicas humanas obtenidas de las bases de datos del National Center of Biotechnology Information (NCBI) y su posterior análisis espacial tridimensional, se estableció la existencia de un patrón de acople universal para péptidos presentados por las moléculas de histocompatibilidad HLA-II (DR, DP y DQ), siendo una base para el diseño de vacunas proteicas. Estos patrones espaciales fueron claramente exhibidos por los residuos altamente conservados de los tres tipos de moléculas de HLA-II. La aplicación de este nuevo hallazgo permitió diseñar péptidos con mejores valores de acople péptido-HLA-II, que los generados por el péptido de acople universal conocido como CLIP (class Il-associated invariant chain peptide).

Starting from the multiple alignment of human protein sequences obtained from the NCBI database (National Center of Biotechnology Information) and subsequent three-dimensional spatial analysis, the existence of a pattern of universal coupling to peptides presented by MHC molecules HLA-II (DR, DP and DQ) was established, being a basis for the design of protein vaccines. These spatial patterns were clearly exhibited by highly conserved residues of the three kinds of HLA-II molecules. The application of this new finding made it possible to design peptides with better Peptide -HLA-II coupling values than those generated by the universal coupling peptide called CLIP (class II-associated invariant chain peptide).

FA partir do alinhamento múltiplo de sequéncias de proteínas humanas obtidas a partir das bases de dados do NCBI (National Center of Biotechnology Information) e análise espacial tridimensional subsequente, estabeleceu-se a existéncia de um padráo de acoplamento universal para peptídeos apresentados pelas moléculas de histocompatibilidade HLA-II (DR, DP e DQ), sendo uma base para o desenho de vacinas proteicas. Estes padroes espaciais foram claramente exibidos pelos residuos altamente conservados dos trés tipos de moléculas de HLA-II. A aplicagáo deste novo achado permitiu desenhar peptídeos com melhores valores de acoplamento peptídeo-HLA-II, do que aqueles gerados pelo peptídeo de acoplamento universal conhecido como CLIP (classe II-peptídeo associado a cadeia invariante).

ROS kinase fusions are not common in Chinese patients with cholangiocarcinoma / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expressions of different forms of ROS fusions in Chinese patients with cholangiocarcinoma (CCA).</p><p><b>METHODS</b>RT-PCR was employed to examine formalin-fixed and paraffin-embedded CCA samples from stage I-IV patients for detection of ROS fusions involving Fused in Glioblastoma (FIG), solute carrier protein (SLC34A2) and major histocompatibility complex class II invariant chain (CD74). Serpin peptidase inhibitor clade A member 1 (SERPINA1) was detected as the reference gene.</p><p><b>RESULTS</b>In all the 56 CCA samples, 80.4% (45/56) were positive for SERPINA1 expression as evaluable samples. Of these evaluable samples, none expressed the ROS fusions.</p><p><b>CONCLUSION</b>ROS fusions are not common in Chinese CCA patients.</p>

Identification of differentially expressed genes in gauze-exposed omentum of dogs using differential display RT-PCR

Molecular mechanisms governing peritonitis caused by the presence of aseptic gauze have remained unclear. To identify the genes involved, sterile gauze-exposed omentum was collected at 0, 6, 12, 24, and 48 h intervals, and analyzed by differential display RT(reverse transcription)-PCR. Among over 1,200 bands, 230 bands were found differentially expressed. These bands represented the fragment sizes of approximately 200 to 1,500 bp. The eight fragments were expressed differentially in the treatment group but not in the control. The sequences of two bands were similar to those of genes associated with the inflammatory process and a band was related to repair and regeneration process. Another one was related with spermatogonia and the rest four were unknown. Additionally, amplicons corresponding to the full-length sequences of two inflammatory gene fragments were synthesized by rapid amplification of cDNA end PCR. One showed 99% similarity to the major histocompatibility complex class II dog leukocyte antigen-DR beta chain and the other was canis familiaris proteasome beta type 3. Results of the present study suggested that sterile gauze induced the differential expression of genes in the omentum involved in inflammation and healing process.

HLA among Brazilian patients with spontaneous chronic urticaria and positive autologous serum skin test / HLA em brasileiros com urticária crônica espontânea e teste cutâneo do soro autólogo positivo

BACKGROUND: An association between class I and II alleles of the major histocompatibility complex and idiopathic chronic urticaria has previously been observed in different populations, but there are still no studies on Brazilian populations in this regard. OBJECTIVE: The involvement of the major histocompatibility complex classes I and II (loci A, B and DR) in Brazilian patients with idiopathic chronic urticaria and a positive autologous serum skin test was investigated and compared with a healthy population group. METHODS: DNA was extracted from the blood of 42 patients with idiopathic chronic urticaria and major histocompatibility complex classes I and II alleles were determined using the polymerase chain reaction and a laboratory test for oligonucleotide hybridization using a single-filament probe. The frequencies of these alleles in patients with chronic urticaria were compared with the frequencies in 1000 genetically unrelated voluntary blood donors from the same region of Brazil. The diagnosis of idiopathic chronic urticaria was based on the patients' clinical history and routine laboratory tests. Only the patients with positive autologous serum skin test were selected. The allele distribution resulted from the patient and control groups were analyzed using odds ratios and 95% confidence intervals. RESULTS: No statistically significant differences were found between the positive autologous serum skin test patients with chronic urticaria and the control group. CONCLUSIONS: We found that in this population group, there was no specific association between the HLA alleles studied and chronic urticaria. We believe that further population studies are needed in order to investigate the possible existence of this association.

FUNDAMENTOS: A associação entre os alelos do MHC classe I e II e a urticária crônica idiopática tem sido previamente constatada em diferentes populações, sendo que na população brasileira ainda não existem estudos a este respeito. OBJETIVOS: Foi estudado o envolvimento do MHC classe I e II (locci A, B e DR) em pacientes brasileiros com urticária crônica idiopática e teste cutâneo do soro autólogo positivo, comparando-se com um grupo populacional saudável. MÉTODOS: O DNA foi extraído do sangue de 42 pacientes com urticária crônica idiopática e o MHC classe I e II determinado por reação em cadeia da polimerase e teste laboratorial de hibridização de oligonucleotídeo com sonda de filamento único. A freqüência destes alelos em pacientes com urticária crônica idiopática foi comparada com a de 1000 doadores de sangue voluntários e geneticamente não relacionados, da mesma região do Brasil. O diagnóstico de urticária crônica idiopática foi baseado na história clínica do paciente e exames laboratoriais de rotina; foram selecionados apenas os pacientes com teste cutâneo do soro autólogo positivo. O resultado da distribuição alélica entre o grupo de pacientes e o grupo controle foi analisado através do odds rate com o cálculo do intervalo de confiança de 95% (95% IC). RESULTADOS: Não foram encontradas diferenças com significância estatística entre os pacientes com urticária crônica teste cutâneo do soro autólogo positivos e o grupo controle. CONCLUSÕES: Verificamos que neste grupo populacional estudado não houve associação específica entre os alelos HLA estudados e a urticária crônica; acreditamos na necessidade de outros estudos populacionais, para podermos verificar a possível existência desta associação.

S100, CD68, and MHC class II molecule expression in cervical high- and low-grade HPV-induced lesions / Expressão de S100, CD68 e moléculas de MHC classe II em lesões cervicais de alto e baixo grau induzidas por HPV

INTRODUCTION: Some human papillomavirus (HPV) types are involved in malignant processes in the cervical epithelium, with 99 percent of cases attributed to oncogenic HPV infection. This study aimed to detect S100, CD68, and major histocompatibility complex class II (MHC-II) molecules in cervical uterine epithelial samples in patients with high- and low-grade lesions induced by HPV. METHODS: Fifty-eight samples from patients who were confirmed positive or negative for high-risk oncogenic HPV DNA, had histopathological diagnosis of cervical intraepithelial neoplasia (CIN) of grades I, II, or III, or were negative for intraepithelial lesion or malignancy were subjected to immunohistochemistry reaction to S100 protein, CD68, and MHC-II (HLA-DR alpha chain). RESULTS: The presence of MHC-II predominated in samples exhibiting histopathological alterations (p < 0.05). S100 detection was more numerous in carcinoma samples (CIN III) (75 percent). Presence of this protein correlated significantly (p < 0.05) with histopathological findings and viral load. CONCLUSIONS: A small expression of CD68 was observed, which may be explained by the observation in our study having been made on random microscopic fields and not on specific areas. The findings, such as the presence of S100 protein and MHC-II expression in samples with histological alterations, could suggest that the immune system fails to control HPV replication at the early stages of infection. Further studies with larger prospective data are necessary to confirm this result.

INTRODUÇÃO: Alguns tipos de papilomavirus humano (HPV) estão envolvidos em processos malignos no epitélio cervical, com 99 por cento dos casos atribuídos à infecção por HPV oncogênico. O objetivo deste estudo foi detectar a proteína S100, CD68 e moléculas de MHC-II (complexo principal de histocompatibilidade classe II) em amostras de epitélio cervical uterino, de pacientes com lesões de alto e baixo grau induzidas pelo HPV. MÉTODOS: Cinquenta e oito amostras de pacientes positivos ou negativos, confirmados, para DNA de HPV de alto ou baixo risco oncogênico, e que tiveram diagnóstico histopatológico de neoplasia intraepithelial cervical (NIC) de graus I, II ou III ou foram negativas para lesão intraepithelial e malignidade (NILM), foram submetidas à reação de imunohistoquímica (IHQ) para proteína S100, CD68 e MHC-II (HLA-DR cadeia alfa). RESULTADOS: A presença da molécula MHC-II predominou em amostras exibindo alterações histopatológicas (p < 0,05). A detecção de S100+ foi mais numerosa em amostras com carcinoma (NIC III) (75 por cento). A presença dessa proteína correlacionou-se significantemente (p < 0,05) com achados histopatológicos e a carga viral. CONCLUSÕES: Pequena expressão CD68+ foi observada, uma possível explicação seria que em nosso estudo as observações foram feitas em campo microscópicos aleatórios e não em áreas específicas. Os achados como a presença de S100 e a expressão de MHC-II, em amostras com alterações histológicas, podem sugerir que o sistema imune falha em controlar a replicação do HPV nas fases iniciais da infecção. Maiores estudos, com mais dados prospectivos, são necessários para confirmar esses resultados.

Avaliação da hemopoese e da resposta imune inata mediada por macrófagos em camundongos submetidos à recuperação nutricional após desnutrição protéica / Evaluation of hematopoiesis and innate immune response mediated by macrophages in mice submitted to nutritional recovery after protein malnutrition

A desnutrição protéico-energética (DPE) afeta mais de 1 bilhão de pessoas no mundo, principalmente crianças, idosos e pacientes hospitalizados. Ela provoca alterações metabólicas e hormonais, além de afetar o tecido hemopoético. O comprometimento da hemopoese provoca anemia e leucopenia, modificando a resposta imune inata e adquirida do organismo. Dessa forma, é comum a associação entre desnutrição e infecção, levando ao comprometimento do tratamento e aumento da morbidade e mortalidade de indivíduos hospitalizados. Após a recuperação nutricional, é relatada a reversão das alterações bioquímicas e hormonais, bem como das alterações na hemopoese e na resposta imune. Porém, muitos resultados são controversos, existindo dúvidas quanto à reversibilidade das alterações. Assim, nos propusemos a avaliar os efeitos da recuperação nutricional nos parâmetros bioquímicos, hormonais, hematológicos e imunológicos em modelo murino de desnutrição. Os animais desnutridos apresentaram perda de peso significativa, redução de proteínas totais, albumina, glicose, insulina e IGF-1, bem como aumento de glutamina plasmática, glutamina sintetase muscular e corticosterona. Houve redução dos parâmetros hepáticos e musculares, bem como alteração na sensibilidade à insulina, evidenciada pelos testes de OGTT e ITT. Todas as alterações descritas caracterizam o quadro de desnutrição. Após a recuperação nutricional, alguns parâmetros foram normalizados, mas as concentrações de glicose, insulina e IGF-1 permaneceram reduzidas. Da mesma forma, as alterações na concentração de DNA hepático e na sensibilidade à insulina permaneceram nos animais renutridos. A pancitopenia periférica e hipocelularidade da medula óssea e do baço observadas nos animais desnutridos foram revertidas após a renutrição. A avaliação de macrófagos peritoniais mostrou reversão parcial do comprometimento da capacidade e adesão e espraiamento, bem como da atividade fungicida nos animais renutridos. A produção de peróxido de hidrogênio continuou baixa após a recuperação nutricional, enquanto a produção de óxido nítrico voltou a aumentar. O comprometimento da produção de citocinas pró-inflamatórias decorrente da desnutrição não foi completamente revertido, visto que, em camundongos Swiss Webster, somente a produção de TNF-α retornou ao normal, enquanto em camundongos C56BL/6J a produção de nenhuma citocina foi restabelecida. A avaliação da via de sinalização do fator de transcrição NFkB mostrou alteração na expressão de MyD88, TRAF-6, IkKß e IkBα em animais desnutridos. Após a recuperação nutricional, algumas dessas proteínas não retornaram ao normal. Os animais desnutridos também apresentaram comprometimento da ativação de NFkB, que não foi normalizada após a recuperação nutricional. Sendo assim, é possível afirmar que o retorno a uma dieta normoprotéica não é suficiente para reverter todas as alterações causadas pela desnutrição

Protein-energy malnutrition (PEM) affects more than 1 billion people worldwide, mainly children, elderly and hospitalized patients. It causes metabolic and hormonal changes, besides affecting hematopoietic tissue. Impaired hematopoiesis causes anemia and leukopenia, modifying innate and acquired immune response of the organism. Thus, it is common the association between malnutrition and infection, leading to impaired treatment and increasing morbidity and mortality in hospitalized patients. After nutritional recovery, it is reported reversal of biochemical and hormonal changes, as well as, reversal of changes in hematopoiesis and immune response. However, many results are controversial, and there are doubts about the reversibility of the changes. Thus, we proposed to evaluate the effects of nutritional recovery biochemical, hormonal, haematological and immunological parameters in a murine model of malnutrition. The malnourished animals showed significant weight loss, reduction in total protein, albumin, glucose, insulin and IGF-1, as well as increased plasma glutamine, corticosterone and muscle glutamine synthetase. There was a reduction in muscle and liver parameters as well as change in insulin sensitivity, evidenced by the tests of OGTT and ITT. All modifications described characterize the malnutrition. After nutritional recovery, there was normalization of some parameters, but the concentrations of glucose, insulin and IGF-1 remained low. Likewise, changes in hepatic DNA concentration and insulin sensitivity remained in renourished animals. Peripheral pancytopenia and hypocellularity in bone marrow and spleen observed in malnourished animals were reversed after refeeding. The evaluation of peritoneal macrophages showed partial reversal of impairment of adhesion and spreading ability, as well as fungicidal activity in animals renourished. The hydrogen peroxide production remained low after nutritional recovery, while nitric oxide production increased again. Impaired production of proinflammatory cytokines due to malnutrition was not completely reversed, whereas in Swiss Webster mice, only the production of TNF-α returned to normal, whereas in C56BL/6J mice no cytokine production was restored. The assessment of the signalling pathway of transcription factor NFkB showed alterations in the expression of MyD88, TRAF-6 IkKß and IkBα in malnourished animals. After nutritional recovery, some of these proteins didn't return to normal. Malnourished animals also showed impaired activation of NFkB, which wasn't normalized after nutritional recovery. Therefore, it is possible to say that the return to a normal diet is not enough to reverse all the changes caused by malnutrition