Genome-wide DNA methylation profile in the peripheral blood of cocaine and crack dependents

Objective: Cocaine use disorders (CUDs) represent a major public health problem in many countries. To better understand the interaction between the environmental modulations and phenotype, the aim of the present study was to investigate the DNA methylation pattern of CUD patients, who had concomitant cocaine and crack dependence, and healthy controls. Methods: We studied DNA methylation profiles in the peripheral blood of 23 CUD patients and 24 healthy control subjects using the Illumina Infinium HumanMethylation450 BeadChip arrays. Results: Comparison between CUD patients and controls revealed 186 differentially methylated positions (DMPs; adjusted p-value [adjP] < 10-5) related to 152 genes, with a subset of CpGs confirmed by pyrosequencing. DNA methylation patterns discriminated CUD patients and control groups. A gene network approach showed that the EHMT1, EHMT2, MAPK1, MAPK3, MAP2K1, and HDAC5 genes, which are involved in transcription and chromatin regulation cellular signaling pathways, were also associated with cocaine dependence. Conclusion: The investigation of DNA methylation patterns may contribute to a better understanding of the biological mechanisms involved in CUD.

Polimorfismo de los antígenos leucocitarios humanos en receptores cubanos de trasplante relacionado de progenitores hematopoyéticas / Polymorphism of human leukocyte antigens in Cuban recipients of related hematopoietic stem cell transplants

Introducción: El trasplante relacionado de células progenitoras hematopoyéticas (TCPH) es una alternativa terapéutica curativa para los pacientes con ciertos tipos de hemopatías o de inmunodeficiencias, en la que se selecciona como donante a un familiar del receptor. Objetivo: Caracterizar el sistema de antígenos leucocitarios humanos (HLA) en receptores de TCPH relacionado. Métodos: Se realizó un estudio descriptivo y transversal en el departamento de Histocompatibilidad del Instituto de Hematología e Inmunología desde enero 2013 hasta diciembre de 2015. Se tipificaron 75 genes HLA mediante la técnica de reacción en cadena de la polimerasa con cebadores de secuencia específico, de baja resolución a 117 pacientes con criterio de TCPH. Para el análisis inmunogenético se empleó el programa Arlequín 3.5.2.2. Resultados: Fueron más frecuentes los genes HLA-A*02, HLA-B*35, HLA-DQB1*03, HLA-DRB1*03 y HLA-DRB1*04, los haplotipos de dos loci HLA-A*02 B*35, HLA-DQB1*03 DRB1*04 y el haplotipo extendido HLA-A*03 B*07 DQB1*06 DRB1*15. Conclusiones: Los genes del sistema HLA en pacientes cubanos candidatos a TCPH relacionado presentaron frecuencias similares a las descritas en poblaciones generales de Cuba y el mundo, aunque con características distintivas en algunos genes y haplotipos(AU)

Introduction: Related hematopoietic progenitor cell (TCPH) transplantation is a curative therapeutic alternative for patients with certain types of hemopathies or immunodeficiencies, in which a recipient family member is selected as a donor. Objective: To characterize the human leukocyte antigen (HLA) system in related TCPH receptors. Methods: A descriptive and cross-sectional study was conducted in the Histocompatibility department of the Institute of Hematology and Immunology from January 2013 to December 2015. 75 HLA genes were typed using the polymerase chain reaction technique with specific sequence primers, from Low resolution to 117 patients with TCPH criteria. For the immunogenetic analysis, the Harlequin 3.5.2.2 program was used. Results: The genes HLA-A * 02, HLA-B * 35, HLA-DQB1 * 03, HLA-DRB1 * 03 and HLA-DRB1 * 04, the haplotypes of two HLA-A * 02 B * 35 loci were more frequent , HLA-DQB1 * 03 DRB1 * 04 and the extended haplotype HLA-A * 03 B * 07 DQB1 * 06 DRB1 * 15. Conclusions: The genes of the HLA system in Cuban patients related to TCPH presented frequencies similar to those described in general populations of Cuba and the world, although with distinctive characteristics in some genes and haplotypes(AU)

Seguimiento inmunológico con anticuerpos anti-HLA por tecnología Luminex™, ¿se puede prevenir el rechazo agudo celular y humoral en trasplante de intestino? / Anti-HLA antibody immunological follow up by the Luminex TM. Is it possible to prevent the acute cell and humoral immune rejection in small bowel transplant? Report of one case

Introducción. El trasplante de intestino mejora la supervivencia de pacientes con falla intestinal secundaria al síndrome de intestino corto. Estos receptores tienen gran riesgo de rechazo agudo, por lo cual, de manera protocolaria y como método de referencia, se practican biopsias intestinales. En este reporte de caso se hizo el seguimiento inmunológico de anticuerpos anti-HLA por tecnología Luminex™ (LSA) de un paciente con trasplante de intestino más biopsias por protocolo para un diagnóstico temprano, y una adecuada correlación histológica. Presentación del caso. Se trata de un paciente de 20 años de edad con síndrome de intestino corto, que ingresó a la Fundación Valle del Lili (Cali, Colombia) y requirió un trasplante aislado de intestino. El seguimiento inmunológico se hizo con tecnología Luminex™ y biopsias intestinales mensuales. Según la tamización contemplada en el protocolo previo al trasplante, el paciente tuvo anticuerpos anti-HLA (PRA de clase I y II) negativos; y a los 11 meses después del trasplante, los anticuerpos anti-HLA de clase I y II fueron positivos. Con la prueba de LSA se detectó un anticuerpo específico contra donantes (Donor Specific Antibodies, DSA) y varios anticuerpos contra otros subtipos moleculares. Se tomó una biopsia que mostró un leve rechazo celular agudo y se inició tratamiento con plasmaféresis. Hasta 21 meses después del trasplante, el paciente no ha presentado rechazos clínicos y ha tenido una adecuada evolución clínica y paraclínica Conclusión. Este es el primer trasplante de intestino en nuestro centro, en el que se hace seguimiento inmunológico con tecnología Luminex™. Consideramos que la detección con DSA es un buen marcador de rechazo agudo humoral, que permitiría una aproximación diagnóstica y una intervención oportuna

Background: Small bowel transplant improves survival of the recipients that have intestinal failure secondary to short bowel syndrome. These patients have a high risk of acute rejection; for this reason bowel biopsies are performed as protocol and is the gold standard. Immunological follow-up of anti-HLA antibodies with Luminex® technology (LSA) was carried out in a patient with intestinal transplant and biopsies were performed to achieve an early diagnosis and a suitable histological correlation. Case report: A 20-year-old patient with short bowel syndrome secondary to extensive intestinal resection due to a complicated appendicitis underwent isolated bowel transplantation. The post-transplant immunological follow-up was performed with LSA and monthly intestinal biopsies. Antibodies with mean fluorescence intensity greater than 1500 were positive. During the pre-transplant protocol, the patient was screened for anti-HLA antibodies with negative results. Eleven months post-transplant, the screening test for anti-HLA Class I and II antibodies was positive; the specificity of the LSA test detected one specific donor antibody (DSA) and several antibodies against other molecular subtypes. The biopsy result was a mild acute cellular rejection and plasmapheresis therapy was started. The patient has not presented a clinical rejection, and at 21 months post-transplantation exhibits an adequate clinical and paraclinical evolution. Conclusions: This is the first small bowel transplant where immunological follow-up is done with LSA. We believe that the detection of DSA is a marker of acute humoral rejection that allows a diagnostic approach and a timely intervention

Trasplante renal con HLA idéntico de donante vivo y cadavérico: experiencia de la Fundación Valle de Lili, Cali, Colombia / Renal transplant with identical HLA with living and cadaver donor: Experience at Fundación Valle de Lili, Cali, Colombia

Introducción: En el trasplante renal con HLA idéntico los episodios de rechazo agudo son menores y tienen mejores tasas de supervivencia del injerto, comparado con los receptores con HLA no idéntico; a pesar de esto, persiste el dilema en cuanto al retiro o la disminución de la dosis de inmunosupresión. El objetivo de este trabajo es describir la experiencia de los trasplantes renales con HLA idéntico de donante vivo y cadavérico que se han realizado en la Fundación Valle del Lili desde 1995 hasta 2014. Material y métodos. De los 1.462 trasplantes renales realizados se incluyeron aquellos con HLA idéntico. Se hizo un análisis estadístico descriptivo para todas las variables consideradas y, para subgrupos seleccionados, el análisis de supervivencia y de rechazo agudo se hizo con el método de Kaplan-Meier. Para el análisis se usó Stata 12.0®. Resultados. Se practicaron 29 trasplantes renales con HLA idénticos. La mayoría fueron en hombres de raza mestiza y lo más frecuente fue una etiología desconocida de la enfermedad renal terminal. Dos pacientes presentaron rechazo agudo, y la supervivencia de los injertos a 1, 5, 10 y 15 años, fue de 100%, 93,7 %, 75 % y 75 %, respectivamente; la supervivencia de los pacientes a los 1, 5, 10 y 15 años, fue de 100%, 93,7 %, 84,3 % y 84,3 %, respectivamente. Conclusiones. Los receptores HLA idénticos poseen una supervivencia prolongada del injerto con menos tasas de rechazo agudo.

Introduction: Kidney transplantation is the treatment of choice for patients with end-stage renal disease (ESRD). Graft rejection is much lower in terms of acute rejection and improved graft survival in renal transplantation with HLA-identical compared to non-identical HLA receptors. The aim of this work is to describe the experience of HLA identical kidney transplantation from live and deceased donors that have been performed at Valle de Lili Foundation since 1995 to 2014. Material and methods. From the 1,462 kidney transplants performed those with HLA-identical were identified, a descriptive statistical analysis was performed for all variables considered in the analysis and for selected subgroups, the analysis of survival and acute rejection was made with the Kaplan-Meier method. Stata 12.0 was used for the analysis. Results: A total of 29 HLA-identical kidney transplants were performed. Most were men of mixed race; the main etiology of ESRD was unknown. Two patients had acute rejection and graft survival at five, ten and fifteen years was 93.7%, 75% and 75% respectively, patient survival at five, ten and fifteen years was 93.7%, 84.3% and 84.3% respectively. Conclusion: HLA-identical receptors have a prolonged survival of the graft with less acute rejection rates.

Monitoreo de anticuerpos anti-HLA en pacientes con insuficiencia renal crónica en lista de espera para trasplante / Anti-hla antibodies monitoring in patients with chronic re- nal failure on waiting list for renal transplant in Paraguay

INTRODUCCIÓN: El estudio de anticuerpos anti-HLA en el suero del paciente en lista de espera para trasplante renal es fundamental para optimizar la elección de un donante así como el esquema de inmunosupresión de inducción y mantenimiento acorde al riesgo inmunológico. Estos anticuerpos pueden Encontrarse de manera preexistente al trasplante como resultado de exposición del individuo a transfusiones sanguíneas, embarazos y trasplantes previos. El objetivo del estudio fue determinar la incidencia de inmunización frente a antígenos de HLA, los factores asociados y su impacto en pacientes en espera de un trasplante renal. MATERIAL Y MÉTODOS: En este estudio, observacional retrospectivo de corte trasversal, fueron incluidos 254 pacientes en lista de espera para trasplante renal que acudieron al Laboratorio Central de Salud Pública en el período comprendido entre julio de 2013 y julio de 2015. RESULTADOS: De los 254 pacientes estudiados, 30% presentaron anticuerpos anti-HLA. El evento sensibilizante más significativo fue la exposición a un trasplante previo, presentando anticuerpos anti-HLA el 84% de los candidatos a retrasplante (p<0,05). En segundo lugar se encontraron las mujeres multíparas, presentando un PRA (Panel Reactivo de Anticuerpos) positivo el 69% de ellas (p<0,05). Por último, el 24% de los pacientes poli-transfundidos presentaron anticuerpos anti HLA (p<0,05). CONCLUSIONES: En el trascurso de los dos años de estudio, 51 pacientes fueron trasplantados, de los cuales un solo paciente presentaba anticuerpos anti-HLA antes del trasplante. Esto indica claramente que la inmunización frente a antígenos de HLA representa una barrera para el acceso al trasplante

INTRODUCTION: Anti-HLA antibodies determination in the serum of patients on a waiting list for renal transplant is essential to optimize donor selection as well as for the induction and maintenance immunosuppression scheme, according to immunological risk. These antibodies could be present before transplantation as a result of being exposed to blood transfusions, pregnancies and previous transplants. The objective of the study was to determine immunization against HLA antigens, associated factors and their impact on the waiting list for a renal transplant. METHODS: In this observational retrospective cross sectional study, 254 patients on the waiting list for renal transplant were included. These patients attended the Public Health central laboratory between July 2013 and July 2015. RESULTS: 30% of the 254 studied patients presented anti-HLA antibodies. The most significant sensitizing event was the exposure to a previous transplant (p=<0.05). Multiparous women were in second place, 69% of them presenting positive PRA (panel reactive antibodies) (p=<0.05). Finally 24% of poly transfused patients presented anti-HLA antibodies (p=<0.05). CONCLUSIONS: During the 2 year of the study, 51 patients were transplanted, presenting only one of them anti-HLA antibodies before transplantation. This results clearly indicate that the immunization against HLA represents a barrier for transplantation access

Selection of Thymocytes Expressing Transgenic TCR Specific for a Minor Histocompatibility Antigen, H60

Minor histocompatibility antigens are MHC-bound peptides and contribute to the generation of allo-responses after allogeneic transplantation. H60 is a dominant minor H antigen that induces a strong CD8 T-cell response in MHC-matched allogeneic transplantation settings. Here, we report establishment of a TCR transgenic mouse line named J15, wherein T cells express TCRs specific for H60 in complex with H-2K(b), and different fates of the thymocytes expressing J15 TCRs in various thymic antigenic environments. Thymocytes expressing the J15 TCRs were positively selected and differentiated into CD8+ single positive (SP) cells in the thymus of C57BL/6 mice, wherein the cognate antigen H60 is not expressed. However, thymocytes were negatively selected in thymus tissue where H60 was transgenically expressed under the control of the actin promoter, with double-positive stages of cells being deleted. Despite the ability of the H60H peptide (LTFHYRNL) variant to induce cytotoxic activity from H60-specific CTL lines at ~50% of the activity induced by normal H60 peptides (LTFNYRNL), J15-expressing thymocytes were positively selected in the thymus where the variant H60H was transgenically expressed. These results demonstrate that a single amino-acid change in the H60 epitope peptide influences the fate of thymocytes expressing the cognate TCR.

Gestação: um desafio imunológico / Pregnancy: an immune challenge

Diversos estudos demonstram a importância de aspectos imunológicos na gestação. Durante a gestação ocorre a implantação do embrião no útero materno, onde irá se desenvolver até o final da gravidez. Dentre os aspectos imunes, pode-se citar a importância da modulação dos linfócitos T, das células natural killers (NK) e das diversas citocinas existentes no organismo materno. A tolerância materna ao feto parece ser mediada por hormônios maternos específicos e pela expressão do antígeno leucocitário humano G (HLA-G) característico na gravidez. Outros estudos sugerem que a rejeição fetal e complicações durante a gravidez podem ocorrer devido à presença de antígenos de histocompatibilidade menor (mHAg), adquiridos pela mãe a partir do compartilhamento sanguíneo com o feto, e devido à presença de anticorpos maternos contra o espermatozoide e contra o feto. O objetivo desta revisão é descrever os aspectos imunológicos que permitem a tolerância materna ao feto na gestação, assim como possíveis causas para a rejeição do embrião e complicações durante a gravidez.(

Several studies demonstrate the importance of immunological aspects of pregnancy. During pregnancy,the embryo is implanted in the womb, where it will develop until the end of pregnancy. Amongst the immune aspects, the importance of the modulation of T lymphocytes, natural killers (NK) cells and many cytokines in maternal organism can be mentioned. The maternal tolerance to the fetus appearsto be mediated by specific maternal hormones and by the expression of human leukocyte antigen G (HLA-G) - characteristic in pregnancy. Other studies suggest that fetal rejection and complications during pregnancy may occur because of the presence of minor histocompatibility antigens (mHAg), acquired by blood sharing of the mother with the fetus, and because of the presence of maternal antibodies against the sperm and against the fetus. The purpose of this review is to describe the immunological aspects that allow maternal tolerance to the fetus during pregnancy, as well as possible causes forrejection of the embryo and complications during pregnancy.

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).

Estudios de HLA en Colombia / HLA research in Colombia

Más de cincuenta años de historia de la medicina y las ciencias biológicas han sido testigos del desarrollo de los conceptos genéticos centrales a la respuesta inmune animal y humana. Los trabajos de George Snell y Jean Dausset, entre otros, identificaron los grupos de genes responsables del reconocimiento inmunológico de lo propio y lo ajeno, en el humano (sistema HLA) y en el ratón (sistema H2). De entonces a hoy se ha descrito la enorme diversidad de alelos y antígenos de histocompatibilidad, usando primero técnicas serológicas y más recientemente describiendo la diversidad de esos sistemas en términos moleculares. En Colombia iniciamos hace años el estudio de la variabilidad humana, utilizando tanto estos marcadores como otra serie de proteínas séricas y eritrocitarias. en 1989 se realizó un estudio de proteínas séricas (C3, BF, HP and TF) en varias poblaciones colombianas, donde se encontró una gran variación en las frecuencias génicas (Bernal y cols 1986). Posteriormente se realizaron estudios poblacionales de HLA (Bernal JE. 1991, Bernal 1995, Briceño I. 1996, Middleton 2000, Trachtenberg EA, 1996), en los que se describió la diversidad y la marcada restricción alélica en poblaciones amerindias en comparación con otros grupos poblacionales.

Suero de conejo con actividad de complemento para tipificación del complejo mayor de histocompatibilidad humano / Rabbit serum with complement activity to typify human major histocompatibility complex

En el ensayo de microlinfocitotoxicidad, el suero de conejo como fuente de complemento desempeña un rol esencial en la clasificación del complejo mayor de histocompatibilidad humano, tanto para la tipificación antigénica como en el cross-match linfocitario de la pareja donante-receptor antes de realizar los trasplantes. En este trabajo, se obtuvieron 3 lotes experimentales de dicho hemoderivado con óptimos indicadores de rendimiento, actividad biológica y estabilidad. El exanguíneo de los animales se realizó por la técnica de yugulación. El suero fue separado por centrifugación en condiciones ambientales y de temperatura controladas. Posteriormente, el producto se sometió a filtración esterilizante y se tomaron muestras para los ensayos de calidad. El estudio mostró que el producto conserva la actividad biológica al menos durante 18 meses y que los valores de citotoxicidad se encuentran dentro de los límites de aceptación para su empleo en los ensayos de histocompatibilidad pretrasplante

Microlinfocitotoxicity assay is one of the serological methods used to classify human major histocompatibility complex. In this technique the rabbit´s serum as complement source, plays an essential role in antigens determination and in lymphocytes cross match assay as necessary stage before the selection of the best donor-receiver pair to realize the organ transplant. Three experimental lots of normal rabbit serum were developed with excellent indicators of efficiency, biological activity and stability. Rabbit's blood was obtained through jugulating technique and serum was separated by centrifugation in controlled environment and temperature´s conditions. The sterilization process was performed by filtration and the samples were taken for quality´s control assays. Stability studies showed that the product kept the biological activity at least during 18 months after it was processed and cytotoxicity's values were adequate for its employment in histocompatibility pre-transplant assays

Recombination in human leukocyte antigen region in two Asian Indian families.

BACKGROUND: Recombination (crossing over) may generate novel haplotypes that can be beneficial to a population against recently introduced pathogens. It may lead to the generation of new alleles. SETTINGS AND DESIGN: A prospective study at a tertiary care centre. AIM: To report two rare cases of crossing over in HLA region. MATERIALS AND METHODS: Tissue-typing was done by sequence specific primers (SSP) for DR locus and by both SSP and serology for Class I which was reconfirmed on fresh samples. RESULTS: In one patient crossing over had taken place in the region of A locus resulting in inheritance of A*01 instead of expected A*11. In second family crossing over had taken place in region of DRB1 locus and the sibling inherited DRB1*08 instead of DRB1*10. CONCLUSIONS: Possibility of recombination must be considered when interpreting implausible tissue-typing results of families worked up for BMT.

Estudo da associação do HLA-B*57 com o controle da viremia em coorte de indivíduos recém infectados pelo HIV-1 / Association between HLA-B*57 and viremia control in recently HIV-1-infected subjects

INTRODUÇÃO: Após a infecção aguda pelo HIV-1, um grupo privilegiado de pessoas consegue controlar a replicação viral sem o uso de antirretrovirais para níveis de viremia abaixo dos limites de detecção pelos testes disponíveis. Alguns alelos HLA estão associados à menor replicação viral durante a infecção recente e menor progressão para doença causada pelo HIV-1, sendo o HLA-B*57 o que está mais associado a esse efeito protetor. OBJETIVO: O objetivo deste trabalho foi confirmar a associação do HLA-B*57 com o melhor controle da viremia em indivíduos com infecção recente pelo HIV-1. MÉTODOS: Foram analisadas amostras de 228 indivíduos de uma coorte prospectiva em acompanhamento na cidade de São Paulo, identificados com infecção recente pelo HIV-1, pelo algoritmo STARHS (serologic testing algorithm for recent human HIV seroconversion). Foram realizadas a contagem de linfócitos T CD4+ e CD8+, carga viral do HIV-1 e tipificação dos alelos HLA. RESULTADOS: Dos 208 indivíduos analisados para o locus B, 15 indivíduos (7,2%) expressam o alelo HLA-B*57. O alelo HLA-B*57 foi fortemente correlacionado com os indivíduos que apresentam parâmetros laboratoriais favoráveis. A presença do HLA-B*57 foi associada com maior contagem de linfócitos T CD4+ basal (p=0, 043) e menor carga viral basal (p=0, 001). Dos 15 indivíduos que expressam o HLA-B*57, oito (53,3%) apresentaram-se com a viremia menor que 400 cópias/ml na visita inicial (Grupo A) e sete (46,6%) apresentaram-se com viremia maior que 400 cópias/mL, todos eles na ausência de terapia antirretroviral. A contagem de linfócitos T CD4+, entretanto, não foi diferente entre os dois grupos. CONCLUSÃO: Concluindo, este estudo indica que indivíduos que expressam o alelo HLA-B*57, apresentam melhor resposta imunológica na infecção recente demonstrada por melhor padrão laboratorial na contagem de linfócitos T CD4+, mas que perfis diferentes de controle podem existir nesses indivíduos. A diferença entre o comportamento da...

INTRODUCTION: After HIV-1 infection, a privileged group of subjects control viral replication to low levels, without the use of antiretroviral drugs. Some HLA alleles are associated with this control and to slower progression to immunodeficiency, especially the HLA-B*57. AIM: The aim of this study was to confirm the association of HLA-B*57 with viral control in recently HIV-1-infected subjects. MATERIALS: A cohort of recently HIV-1-infected 228 subjects, prospectively followed in the City of São Paulo, were identified using STARHS (serologic testing algorithm for recent human HIV seroconversion). CD4+, CD8+ T cell counts, viral loads, and HLA typing were performed in the participants samples. RESULTS: HLA typing was performed in 208 out of 228 subjects. Of those, 15 (7.2%) were HLA-B*57. This genotype was strongly correlated with favorable laboratory outcomes. HLA-B*57 subjects presented higher CD4+ T cell counts (p=0.043) and lower viral loads at the baseline visit (p=0.001). Eight (53.3%) out of 15 HLA-B*57 subjects presented undetectable viral load at the baseline visit (Group A) and seven (46.6%)had detectable viremia (Group B). However, the CD4+ T cell counts were not different between the two groups. CONCLUSION: In conclusion, this study points to the protective association of HLA-B*57 allele with better laboratory outcomes in HIV-1 infection, demonstrated by better CD4+ T cell counts and lower viral loads. Nevertheless, different profiles may exist within this group of subjects. The diverse viral control in such subjects may help better understand the pathogenesis of HIV-1 infection.

Umbilical cord blood banking and transplantation: a short review

It is more than 20 years since the first cord blood transplant [CBT] was performed, following the realisation that cord blood [CB], which is normally wasted, is rich in progenitor cells and capable of rescuing haematopoiesis. Since then it has been appreciated that CB is rich in stem cells, and has many other features not the least of which is its ability to rescue the transplanted patient without a rigid need for full human lymphocyte antigen [HLA] compatibility. Also it is easily accessible, relatively free from infections and poses no medical risk to the donor. However, the quantity of the stem cells is rather small, thus predominantly restricting its use to children or adults requiring double units. In Oman, we have taken a keen interest in stem cell research and also CBT. We see such activities as an avenue for our patients, for whom a compatible bone marrow [BM] or a peripheral blood donor cannot be found, to have an alternative in the form of CBT. This has encouraged us to establish a national voluntary cord blood bank [CBB] which is a valuable option open to a selected group of patients, as compared to the controversial private CBB. This national CBB will have a better representation of HLA-types common in the region, an improvement on relying on banks in other countries. Considering the need for stem cell transplant/therapy in this country, it is only appropriate that this sort of bank is established to cater for some of these requirements

Polymeric Gene Delivery for Diabetic Treatment

Several polymers were used to delivery genes to diabetic animals. Polyaminobutyl glycolic acid was utilized to deliver IL-10 plasmid DNA to prevent autoimmune insulitis of non-obese diabetic (NOD) mouse. Polyethylene glycol grafted polylysine was combined with antisense glutamic acid decarboxylase (GAD) MRNA to represent GAD autoantigene expression. GLP1 and TSTA (SP-EX4) were delivered by bioreducible polymer to stop diabetic progression. Fas siRNA delivery was carried out to treat diabetic NOD mice animal.

Predictive Value of Donor Specific Antibody Measured by Luminex Single Antigen Assay for Antibody Mediated Rejection after Kidney Transplantation / 대한이식학회지

BACKGROUND: Luminex panel reactive antibody (PRA) is a method that is well known for its high sensitivity and specificity. By using a single antigen assay, the presence or absence of donor specific antibody (DSA) can be determined and its strength can be quantified in terms of the mean fluorescence intensity (MFI). In this study, we analyzed the correlation between the pre-transplant PRA and DSA measured by the Luminex method and the post-transplant clinical features after kidney transplantation. METHODS: A total of 123 pre-transplant sera samples from kidney transplanted patients were tested. Luminex-PRA identification tests were performed using a Luminex fluoroanalyzer and a LifeCodes class I, II ID Kits. Single antigen assay by the Luminex method was used for detecting DSA and its MFI. RESULTS: The positive Luminex-PRA group included more highly-sensitized patients such as women, patients with a previously positive lymphocyte cross match test and patients who were undergoing retransplantation. There was no correlation between the acute rejection rate and positive PRA on the Luminex-PRA. However, pretransplant DSA detected by the single antigen assay was significantly associated with episodes of antibody mediated rejection (P=0.047, OR=10.2), and DSA with higher MFI values (MFI> or =3,000) was associated with antibody mediated rejection (P=0.023). CONCLUSIONS: Although pre-transplant positive PRA was not correlated with acute rejection episodes, the DSA measured by the Luminex single antigen assay seems to have a predictive value for post-transplant antibody mediated rejection.

Characterization of CTL Clones Specific for Single Antigen, H60 Minor Histocompatibility Antigen

BACKGROUND: Disparities of Minor H antigens can induce graft rejection after MHC-matched transplantation. H60 has been characterized as a dominant antigen expressed on hematopoietic cells and considered to be an ideal model antigen for study on graft-versus-leukemia effect. METHODS: Splenocytes from C57BL/6 mice immunized with H60 congenic splenocytes were used for establishment of H60-specific CTL clones. Then the clones were characterized for proliferation capacity and cytotoxicity after stimulation with H60. Clone #14, #15, and #23 were tested for the TCR binding avidity to H60-peptide/H-2Kb and analyzed for TCR sequences. RESULTS: H60-specific CTL clones showed different levels of proliferation capacity and cytotoxic activity to H60-stimulation. Clones #14, #15, and #23 showed high proliferation activity, high cytotoxicity, and low activities on both aspects, respectively, and have TCRs with different binding avidities to H60-peptide/H-2Kb with t 1/2 values of 4.87, 6.92, and 13.03 minutes, respectively. The TCR usages were Valpha12D-3-01+Jalpha11-01 and Vbeta12-1-01+Dbeta1-01+J2-7-01 for clone #14, Valpha13D-1-02+Jalpha34-02 and Vbeta13-1-02+Dbeta2-01+Jbeta2-7-01 for clone #15, and Valpha16D+Jalpha45-01 and Vbeta12-1-01+Dbeta1-01+Jbeta2-5-01 for clone #23. CONCLUSION: The results will be useful for modeling GVL and generation TCR transgenic mouse.

HLA- promiscuous Th1-cell reactivity of MPT64[Rv1980c], a major secreted antigen of mycobacterium tuberculosis, in healthy subjects

To determine HLA-promiscuous Th1-cell reactivity of MPT64 [Rv1980c] in healthy humans. Peripheral blood mononuclear cells [PBMCs] were obtained from healthy subjects [n=61] and HLA typed genomically. PBMCs were tested for Th1-cell reactivity [antigen-induced proliferation and interferon-gamma secretion] with complex antigens [whole cells, culture filtrate and cell walls] and single secreted antigens [Ag85B, MPT64 and MPB70] of Mycobacterium tuberculosis. In addition, culture-filtrate-induced T-cell lines were established fromPBMCs of 14 donors and tested with the above antigens in Th1-cell assays. Furthermore, 3 T-cell lines were tested for cytotoxic activity against MPT64-pulsed monocytes/macrophages, and a T-cell line was analysed for HLA restriction in antigen presentation using anti-HLA class I and II monoclonal antibodies and HLA-DR-typed antigen-presenting cells. PBMCs showed strong Th1-cell reactivity with all of the complex mycobacterial antigens, whereas MPT64 induced moderate Th1-cell reactivity, which was comparable to the reactivity induced by other previously characterized secreted antigens of M.tuberculosis, i.e. Ag858 and MBP70. Furthermore, HLA heterogeneity of the responding donors suggested that MPT64 was presented to Th1 cells promiscuously. Testing of the T-cell lines confirmed that Th1 cells contributed to the promiscuous antigen-specific reactivity observed with PBMCs and exhibited cytotoxic activity against MPT64- pulsed monocytes/macrophages. In addition, a T-cell line investigated for HLA restriction analysis showed that MPT 64 was presented to T cells in association with HLA-DR molecules in a promiscuous manner. MPT64 is promiscuously recognized by human Th1 cells with cytotoxic activity and therefore deserved consideration as a candidate vaccine against tuberculosis in humans

Myasthenia gravis and psoriasis vulgaris

Myasthenia gravis is a rare autoimmune disorder in which antibodies form against acetylcholine nicotinic postsynaptic receptors at the myoneural junction. Psoriasis vulgaris is a chronic, recurring, and an inflammatory skin disease. Myasthenia gravis and psoriasis are both autoimmune diseases and correlated with specific human histocompatibility antigens. In this report, a 53-year-old woman who has myasthenia gravis accompanied with psoriasis vulgaris is presented. To conclude, this association is extremely rare and the pathogenetic etiology was thought to depend on a generalized immunological disturbance

MHC non -HLA gene polymorphisms in transplantation.

Despite many studies on non-HLA genetic polymorphism its role in transplantation is still not well understood. The NK cell receptor gene, MICA gene and Minor histocompatibility (mHag) system makes the puzzle still more intriguingly complex. Studies on cytokine gene polymorphism have enlightened some interesting associations such as the effect of donor IL-6 genotype on acute rejection in renal transplantation. In the bone marrow transplant where each polymorphism is taken as a risk factor for GVHD necessitates prospective testing of non-HLA gene polymorphism and hence, transplant outcome. Various typing methods are now available to identify the non-HLA genetic polymorphisms. A scenario can be envisaged where polymorphisms associated with transplant outcome are tested prior to transplantation at the same time as HLA typing.

Detection of heterogeneous type II collagen transplantation mediated cytotoxic response in vitro / 生物医学工程学杂志

To detect the cellular immunity state of New Zealand white rabbit immunized by pig type II collagen. The New Zealand white rabbit was immunized by type II collagen for sixty days. The plasma was collected at a regular interval and the anti-type II collagen antibodies were examined. At the sixtieth day, the peripheral circular lymphocytes and the lymphocytes separated from spleen cells of rabbit and lymph nodes were collected and were stimulated by type II collagen in vivo again. The regulation of reactive cellular proliferation caused by the stimulation was detected. The experiment samples were divided into two groups. The first group was the positive control group by adding different concentrations of PHA and the non-specific immunity was assayed. The different concentrations of type II collagen were added to the second group and the specific immunity was assayed. The lymphocytes of normal rabbits showed proliferation by PHA stimulation but no proliferation by the first stimulation of type II collagen. Obvious proliferation due to the stimulation of both PHA and type II collagen in the immunized rabbit were observed. It shows that certain concentration of heterogeneous collagen may cause an increase of anti-type II collagen antibody in immunized rabbit and may cause a proliferation of lymphocytes in rabbit spleen and peripheral blood. The heterogeneous type II collagen causes cellular immunity in vivo.