Antiretroviral therapy immunologic non-response in a Brazilian population: association study using pharmaco- and immunogenetic markers

ABSTRACT Background: Antiretroviral therapy (ART) saved millions from HIV-1 infection and AIDS, but some patients do not experience adequate CD4+ T cells gain despite achieving viral suppression. The genetic component of this condition is not yet completely elucidated. Objective: To identify predictive genetic markers of immune response to ART. Methods: Case-control study. Out of 176 HIV-infected patients recruited in the city of Recife, Northeast Brazil, 67 patients with no immunologic response were the cases and the remaining 109 patients who responded were the controls. A set of 94 selected single nucleotide polymorphisms (SNPs) involved in antiretroviral drugs pharmacodynamic pathways and immune system homeostasis were genotyped, while the remaining 48 were ancestry informative markers (AIMs) for controlling for eventual hidden population structure. Results: Male patients were overrepresented in non-responder group (p = 0.01). Non-responders also started with lower absolute CD4+ T cell counts (p < 0.001). We found five SNPs significantly associated with the outcome, being three more frequent in non-responders than responders: rs2243250 (IL4) A allele (p = 0.04), rs1128503 (ABCB1) A allele (p = 0.03) and rs707265 (CYP2B6) A allele (p = 0.02), whereas the other two were less frequent in non-responders: rs2069762 (IL2) C allele (p = 0.004) and rs4646437 (CYP3A4) A allele (p = 0.04). Conclusion: Some significant univariate associations remained independently associated at multivariate survival analysis modeling, such as pre-treatment CD4+ T cells counts, IL2 and ABCB1 genotypes, and use of protease inhibitors, yielding a predictive model for the probability for immune response. More studies are needed to unravel the genetic basis of ART immunological non-response.

Síndrome de Kleine-Levin: hipótesis inmunogenética / Kleine-Levin syndrome: immunogenetic hypothesis

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Evaluation of the immunogenicity of Schistosoma mansoni egg surface

Abstract INTRODUCTION Immunogenicity of Schistosoma mansoni egg surface was examined to determine whether intact eggshells have lower antigenicity than ruptured eggs. METHODS: Swiss Webster mice were inoculated with intact or ultrasonicated S. mansoni eggs isolated from infected human feces. Mice were separated into four groups of six animals each and immunizations were performed approximately every 20 days during a 60-day period. Groups 1-4 were administered with saline solution, sonicated eggs with Freund's adjuvant, sonicated eggs without Freund's adjuvant, and intact eggs, respectively. IgG humoral immune response was assessed by ELISA using Soluble Egg Antigen produced from eggs isolated from the livers of infected mice. RESULTS Sonicated eggs co-administered with adjuvant induced the highest humoral response at 58 days, which was 11.9-fold (95% CI 6.2-17.5) greater than the response induced by saline solution. Sonicated eggs without adjuvant induced a 4.3-fold stronger response (95% CI 2.4-6.2) than normal saline. Intact eggs induced humoral response that was nominally twice stronger (95% CI 0.8-3.2) than that induced by normal saline but the effect did not reach statistical significance. CONCLUSIONS Soluble antigens are not abundant on the surface of S. mansoni eggs and/or are not secreted in sufficient quantities to induce a significant immune response to intact eggs. Assuming that isolation procedures had not damaged the eggs used for inoculation, our observations suggest that intact eggs either do not induce a significant immune response or, if they do, the mechanism involves insoluble antigens from the egg surface.

Resposta imunológica a antígenos de Hysterothylacium deardorffoverstreetorum de peixes teleósteos / Imunne response against Hysterothylacium deardorffoverestreetorum from teleost fish

Anisaquidose é uma doença provocada por parasitos da família Anisakidae e se caracteriza por manifestações gastrointestinais e alérgicas. O Anisakis simplex é o parasito mais patogênico ao homem e altamente alergênico. Porém, outros anisaquídeos também são danosos aos humanos, mas é desconhecida a imunogenicidade dessas larvas. O objetivo deste trabalho foi avaliar o potencial imunogênico do parasito Hysterothylacium deardorffoverestreetorum (HD) em modelo murino. Camundongos da linhagem BALB/c foram divididos em três grupos experimentais e receberam as preparações antigênicas obtidas de larvas de HD: extrato bruto de larvas (EBH), extrato secretado/ excretado de larvas (ESH) e extrato bruto de larvas após excreção/secreção (EEH). Amostras séricas foram obtidas em diferentes dias após imunização para determinação dos níveis de anticorpos específicos pelo ensaio imunoenzimático (ELISA). Os resultados demonstram aumento na produção de imunoglobulina (Ig) G após a segunda imunização, com aumento progressivo após a terceira imunização. Já em relação à IgE, a reatividade foi mais tardia, demonstrando aumento progressivo após a terceira imunização. Foi avaliada a imunidade celular por meio da intradermorreação, como resultado estatisticamente significativo em relação ao controle utilizado. Este experimento é a primeira descrição da potencialidade patogênica desse parasito em mamíferos e representa um avanço no diagnóstico da anisaquidose humana.(AU)

Anisaquidosis is a disease caused by parasites of Anisakidae family and is characterized by gastrointestinal and allergic reactions. The Anisakis simplex is a more pathogenic Anisakidae to humans and is highly allergenic. However, other species of this family also have characteristics that are harmful to humans, but little is known about the immunogenicity this parasites. The objective of this study was to experimentally assess the immunogenic potential of the parasite Hysterothylacium deardorffoverestreetorum (H.D) in mice. Mice of inbred BALB/c strain were divided into three groups and received three immunizations of the following antigenic preparations obtained from L3 larvae H.D: Crude larval extract of H.D (CEH) Extract secreted / excreted larvae H.D. (ESH) and crude extract of larvae after excretion / secretion (EEH). Serum samples were obtained on different days after immunization to determine the levels of circulating specific antibodies by enzyme-linked immunosorbent assay (ELISA). The results show increased production of immunoglobulin (Ig) G after the second immunization with a gradual increase after the third immunization. Regarding IgE reactivity, this occurred later, demonstrating a progressive increase only after the third immunization. Cellular immunity was evaluated by intradermal, and showed statistically significant result compared to the control used. This experiment is the first description of the pathogenic potential of this parasite in mammals and represents a breakthrough in the diagnosis of human Anisakidosis.(AU)

Enfoque inmunogenético de los fenotipos clínicos del asma / An immunogenetic approach to the clinical phenotypes of asthma

El asma bronquial es una enfermedad multifactorial compleja, caracterizada por una heterogeneidad genética que define diferentes fenotipos clínicos. Se realizó una revisión bibliográfica, empleando recursos disponibles en la red Infomed, con el objetivo de profundizar en la inmunogenética del asma, haciendo énfasis en los genes, cuyos productos tienen un papel activo en la inflamación de vías aérea y en la patogénesis de la enfermedad. Se reportan más de cien genes mayores y menores de susceptibilidad a esta enfermedad, la mayoría de estos genes codifican para proteínas, que juegan un papel clave dentro del sistema inmune o modulan la respuesta inflamatoria. Estos genes interactúan entre sí y con el ambiente y determinan el grado de complejidad de la respuesta del sistema inmune. La revisión permite un mejor entendimiento del asma, desde un enfoque inmunogenético, y una mejor comprensión de las variedades clínicas, el diagnóstico y propuestas terapéuticas para los pacientes asmáticos, acorde a cada fenotipo clínico(AU)

Bronchial asthma is a complex multifactorial disease characterized by a genetic heterogeneity that defines different clinical phenotypes. A review paper was carried out using the resources available in the Infomed network, with the aim of deepening the knowledge on asthma immunogenetics, emphasizing the genes which products have an active role in airway inflammation and in the pathogenesis of the disease. More than one hundred major or minor genes of susceptibility to this disease are reported. Most of these genes encode proteins that play a key role within the immune system or adjust the inflammatory response. These genes interact with each other and with the environment and determine the degree of complexity of the immune system response. The paper brings a better understanding of asthma, from an immunogenetic approach, and a better understanding of the clinical varieties, diagnosis and therapeutic proposals for asthmatic patients, according to each clinical phenotype(AU)

Detecção de proteínas imunorreativas de Rickettsia sp. cepa Mata Atlântica / Detection of immunoreactive proteins of Rickettsia sp. Atlantic Forest strain

A Febre Maculosa Brasileira (FMB) é uma doença infecciosa, transmitida por carrapatos ao homem. Uma nova riquetsiose humana foi descrita como causadora de Febre Maculosa no Estado de São Paulo, sendo denominada de Rickettsia sp. cepa Mata Atlântica. O presente trabalho teve como objetivo detectar e identificar proteínas com potencial de estimular o sistema imune de hospedeiro mamífero, desta nova cepa descrita. Para tanto, foi realizado a extração proteica total de Rickettsia sp. cepa Mata Atlântica. As proteínas extraídas foram fracionadas por eletroforese. As bandas proteicas foram transferidas para membranas de nitrocelulose por migração elétrica e submetidas à técnica de Western-blot, para detecção proteica. Ao todo sete proteínas imunorreativas foram detectadas. Duas proteínas apresentaram maior abundancia, com peso molecular, de 200 e 130 kDa respectivamente. Através da comparação de mapas proteômicos existentes e pelo peso molecular que estas proteínas apresentaram, sugere-se que as duas proteínas detectadas representem rOmpA (200 kDa) e rOmpB (130 kDa). As demais proteínas detectadas apresentaram menor ocorrência e peso molecular inferior a 78 kDa, podendo representar membros da família de antígenos de superfície celular (Sca - Surface cell antigen). As proteínas detectadas poderão servir como base de estudo na elaboração de métodos diagnósticos sensíveis e específicos, no desenvolvimento de vacinas, além de possibilitarem novos estudos para terapias mais eficazes.(AU)

Brazilian Spotted Fever (BSF) is an infectious disease transmitted by ticks to humans. A new human rickettsial infection was reported to cause spotted fever in the State of São Paulo and was named Rickettsia sp. Strain Atlantic Forest. This study aimed to detect and identify proteins with potential to stimulate the immune system of mammalian host of this new strain described. Therefore, we performed total protein extraction Rickettsia sp. Strain Atlantic Forest. The extracted proteins were fractionated by electrophoresis. The protein bands were transferred to nitrocellulose membrane by electrical migration and subjected to Western blot for protein detection. In all, seven immunoreactive proteins were detected. Two proteins showed higher abundance, with molecular weight of 200 and 130 kDa respectively. By comparing existing proteomic maps and the molecular weight of these proteins showed that, it is suggested that the two proteins detected representing rOmpA (200 kDa) and rOmpB (130 kDa). The other detected proteins had lower occurrence and molecular weight less than 78 kDa, which may represent members of the cell surface antigens Family (Sca - Surface cell antigen). The detected proteins may serve as a study based on the development of sensitive and specific diagnostic methods in the development of vaccines and they enable further studies to more effective therapies.(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.

Evaluación de la inmunogenicidad y la capacidad protectora homóloga de un candidato vacunal tetravalente de Leptospira, para uso veterinario / Evaluation of immunogenicity and protective homologous capacity of a tetravalent Leptospira vaccine candidate, for veterinary use, including serogroup Ballum

La inmunogenicidad es un parámetro importante en estudios de inmunógenos. En la leptospirosis la respuesta inmune humoral es vital para la resistencia a la infección, de ahí que este trabajo se haya propuesto evaluar la inmunogenicidad y la capacidad protectora homóloga de un candidato tetravalente que incluye al serogrupo Ballum en su formulación. Se trabajó con la cepa 245-12 clasificada como L. borgpetersenii serovar Ballum, aislada a partir de un caso confirmado de leptospirosis. Con esta cepa previamente caracterizada desde el punto de vista de su virulencia, se formularon 5 lotes de una preparación vacunal tetravalente, la cual contiene además las cepas Canicola, Icterohaemorrhagiae y Pomona contenidas en la vacuna Polivalente-Leptospira, de uso veterinario. Se evaluó la inmunogenicidad de esta preparación, mediante microaglutinación y la capacidad de protección homóloga en hámster sirio dorado, frente al reto con 100 DL50. Los animales se inmunizaron con dos dosis de la vacuna (0,1mL) con un intervalo de 15 días, el reto se llevó a cabo a los 14 días de concluido el esquema de inmunización. Los resultados obtenidos se compararon con los de la vacuna Polivalente-Leptospira. Todos los lotes de preparación tetravalente formulados cumplieron satisfactoriamente con los controles de calidad. Los lotes formulados en este estudio mostraron una significativa inmunogenicidad y capacidad de protección homologa frente al serogrupo Ballum, y lograron eliminar el estado de portador en los animales inmunizados. El presente trabajo, por primera vez, sienta las bases para formulaciones vacunales novedosas, en animales, conteniendo el serogrupo Ballum(AU)

Immunogenicity is an important parameter for studies of immunogens. The humoral immune response against leptospirosis is vital for resistance to the infection; hence this paper is proposed to evaluate the immunogenicity and protective homologous capacity of a tetravalent candidate that includes serogroup Ballum in its formulation. The strain 245-12 classified as L. borgpetersenii serovar Ballum was used in this paper, isolated from a confirmed case of leptospirosis. Five (5) batches of a tetravalent vaccine preparation were formulated with this strain previously characterized from the point of view of its virulence, which also contains the strains Canicola, Icterohaemorrhagiae and Pomona contained in the Polyvalent-Leptospira vaccine for veterinary use. The immunogenicity of this preparation was assessed by micro agglutination and homologous protection capacity in Golden Syrian Hamster before the challenge with 100 LD50. Animals were immunized with two doses of vaccine (0.1 mL) with an interval of 15 days; the challenge was carried out 14 days after the immunization schedule was ended. The results were compared with those obtained with the Polyvalent-Leptospira vaccine. All the batches of the tetravalent preparation formulated complied satisfactorily with the quality controls. The batches formulated in this study showed a significant immunogenicity and homologous protection capacity against serogroup Ballum and they were able to eliminate the carrier state in the immunized animals. This paper, for the first time sets the bases for novel vaccine formulations, in animals, containing the serogroup Ballum(AU)

Insights from animal models on the immunogenetics of leprosy: a review

A variety of host immunogenetic factors appear to influence both an individual's susceptibility to infection with Mycobacterium leprae and the pathologic course of the disease. Animal models can contribute to a better understanding of the role of immunogenetics in leprosy through comparative studies helping to confirm the significance of various identified traits and in deciphering the underlying mechanisms that may be involved in expression of different disease related phenotypes. Genetically engineered mice, with specific immune or biochemical pathway defects, are particularly useful for investigating granuloma formation and resistance to infection and are shedding new light on borderline areas of the leprosy spectrum which are clinically unstable and have a tendency toward immunological complications. Though armadillos are less developed in this regard, these animals are the only other natural hosts of M. leprae and they present a unique opportunity for comparative study of genetic markers and mechanisms associable with disease susceptibility or resistance, especially the neurological aspects of leprosy. In this paper, we review the recent contributions of genetically engineered mice and armadillos toward our understanding of the immunogenetics of leprosy.

Coping with genetic diversity: the contribution of pathogen and human genomics to modern vaccinology

Vaccine development faces major difficulties partly because of genetic variation in both infectious organisms and humans. This causes antigenic variation in infectious agents and a high interindividual variability in the human response to the vaccine. The exponential growth of genome sequence information has induced a shift from conventional culture-based to genome-based vaccinology, and allows the tackling of challenges in vaccine development due to pathogen genetic variability. Additionally, recent advances in immunogenetics and genomics should help in the understanding of the influence of genetic factors on the interindividual and interpopulation variations in immune responses to vaccines, and could be useful for developing new vaccine strategies. Accumulating results provide evidence for the existence of a number of genes involved in protective immune responses that are induced either by natural infections or vaccines. Variation in immune responses could be viewed as the result of a perturbation of gene networks; this should help in understanding how a particular polymorphism or a combination thereof could affect protective immune responses. Here we will present: i) the first genome-based vaccines that served as proof of concept, and that provided new critical insights into vaccine development strategies; ii) an overview of genetic predisposition in infectious diseases and genetic control in responses to vaccines; iii) population genetic differences that are a rationale behind group-targeted vaccines; iv) an outlook for genetic control in infectious diseases, with special emphasis on the concept of molecular networks that will provide a structure to the huge amount of genomic data.

Gene cloning and immunogenicity analysis of the structural proteins VP1-VP4 of enterovirus 71 / 南方医科大学学报

<p><b>OBJECTIVE</b>To clone the genes encoding the structural proteins VP1-VP4 of enterovirus 71 and investigate the immunogenicity of the expressed recombinant proteins.</p><p><b>METHODS</b>The VP1-VP4 cDNAs were amplified by RT-PCR from the extracted viral RNA and cloned into pMD19-T vector. The cloned VP1-VP4 genes were then inserted into the multi-cloning sites of plasmid pQE30a and expressed in E. coli M15 with IPTG induction. After washing with 8 mol/L urea and purification with Ni-affinity chromatography, the recombinant proteins obtained were tested for immunogenicity by Western blotting and ELISA using rabbit antisera against enterovirus 71 and Coxsackie Virus A16.</p><p><b>RESULTS</b>The recombinant VP1-VP4 proteins were highly expressed in E. coli M15 and the purified proteins could be specifically recognized by the rabbit sera against enterovirus 71.</p><p><b>CONCLUSION</b>The expressed enterovirus 71 structural proteins show good immunogenicity and can be used for developing enterovirus 71 vaccine and detection kits.</p>

Histocompatibility and Immunogenetics in Cord Blood Transplantation

This review of the immunogenetics of cord blood transplantation attempts to highlight the connections between classical studies and conclusions of the tissue transplantation field as a scholarly endeavor, exemplified by the work of Professor Hoecker, with the motivations and some recent and key results of clinical cord blood transplantation. The authors review the evolution of understanding of transplantation biology and find that the results of the application of cord blood stem cells to Transplantation Medicine are consistent with the careful experiments of the pioneers in the field, from the results of tumor and normal tissue transplants, histocompatibility immunogenetics, to cell and molecular biology. Recent results of the National Cord Blood Program of the New York Blood Center describe the functioning in cord blood transplantation of factors, well known in transplantation immunogenetics, like the Fl anti-parent effect and the tolerance-like status of donors produced by non-inherited maternal HLA antigens. Consideration of these factors in donor selection strategies can improve the prognosis of transplantation by characterizing "permissibility" in HLA-incompatible transplantation thereby increasing the probability of survival and reducing the likelihood of leukemic relapse.

Imunogenética das uveítes / Imunogenetics in the uveítes

Citocinas são moléculas envolvidas na comunicação intercelular nas respostas inflamatória e imune, desempenhando papel relevante nas uveítes. Polimorfismos dos genes responsáveis pela produção de determinadas citocinas têm sido relacionados com a ocorrência e a gravidade de algumas uveítes. Portanto, o presente trabalho tem como objetivo relatar essas possíveis associações, salientando o aspecto individual genético no prognóstico das uveítes.

Cytokines are molecules involved in intercellular communication in immune and inflammatory responses, playing an important role in uveitis. Genetic polymorphisms responsible for the production of certain cytokines have been associated with the occurrence and the severity of uveitis. Therefore, the present study has the purpose of describing these possible associations, pointing out the individual genetic background in the prognosis of uveitis.