Relación entre la compatibilidad del HLA y la pérdida del injerto en trasplante renal de donante cadavérico: Un análisis por propensity score matching en Colombia / Relationship between HLA compatibility and graft loss in renal transplant from a deceased donor: An analysis by propensity score matching in Colombia

Introducción. En Colombia, solo un 24 % de los pacientes en lista recibieron un trasplante renal, la mayoría de donante cadavérico. Para la asignación de órganos se considera el HLA A-B-DR, pero la evidencia reciente sugiere que el HLA A-B no está asociado con los desenlaces del trasplante. El objetivo de este estudio fue evaluar la relevancia del HLA A-B-DR en la sobrevida del injerto de los receptores de trasplante renal. Métodos. Estudio de cohorte retrospectivo que incluyó pacientes trasplantados renales con donante cadavérico en Colombiana de Trasplantes, desde 2008 a 2023. Se aplicó un propensity score matching (PSM) para ajustar las covariables en grupos de comparación por compatibilidad y se evaluó la relación del HLA A-B-DR con la sobrevida del injerto renal por medio de la prueba de log rank y la regresión de Cox. Resultados. Se identificaron 1337 pacientes transplantados renales, de los cuales fueron mujeres un 38,7 %, con mediana de edad de 47 años y de índice de masa corporal de 23,8 kg/m2. Tras ajustar por PSM las covariables para los grupos de comparación, la compatibilidad del HLA A-B no se relacionó significativamente con la pérdida del injerto, con HR de 0,99 (IC95% 0,71-1,37) para HLA A y 0,75 (IC95% 0,55-1,02) para HLA B. Solo la compatibilidad por HLA DR fue significativa para pérdida del injerto con un HR de 0,67 (IC95% 0,46-0,98). Conclusión. Este estudio sugiere que la compatibilidad del HLA A-B no influye significativamente en la pérdida del injerto, mientras que la compatibilidad del HLA DR sí mejora la sobrevida del injerto en trasplante renal con donante cadavérico

Introduction. In Colombia, only 24% of patients on the waiting list received a renal transplant, most of them from cadaveric donors. HLA A-B-DR is considered for organ allocation, but recent evidence suggests that HLA A-B is not associated with transplant outcomes. The objective of this study was to evaluate the relevance of HLA A-B-DR on graft survival in kidney transplant recipients. Methods. Retrospective cohort study that included kidney transplant recipients with a cadaveric donor in Colombiana de Trasplantes from 2008 to 2023. A propensity score matching (PSM) was applied to adjust the covariates in comparison groups for compatibility, and the relationship of HLA A-B-DR with kidney graft survival was evaluated using the log rank test and Cox regression. Results. A total of 1337 kidney transplant patients were identified; of those, 38.7% were female, with median age of 47 years, and BMI 23.8 kg/m2. After adjusting the covariates with PSM for the comparison groups, HLA A-B matching was not significantly related to graft loss, with HR of 0.99 (95% CI 0.71-1.37) and 0.75 (95% CI 0.55-1.02), respectively. Only HLA DR matching was significant for graft loss with an HR of 0.67 (95% CI 0.46-0.98). Conclusions. This study suggests that HLA A-B matching does not significantly influence graft loss, whereas HLA DR matching does improve graft survival in renal transplantation with a cadaveric donor.

Current status of research on the genetic susceptibility of primary biliary cholangitis / 临床肝胆病杂志

Primary biliary cholangitis （PBC） is a liver autoimmune disease with a strong genetic tendency characterized by the degeneration and necrosis of bile duct epithelial cells， and it is often observed in middle-aged and elderly women. With the continuous development of genome-wide association studies， the genetic susceptibility of PBC has attracted more and more attention. This article elaborates on the research advances in the genetic susceptibility genes closely associated with PBC， in order to provide effective targets for the treatment of PBC.

Desensitization using IVIG alone for living-donor kidney transplant: impact on donor-specific antibodies / Dessensibilização usando somente IgIV para transplante renal com doador vivo: impacto nos anticorpos específicos contra o doador

Abstract Introduction: Sensitization to human leukocyte antigen is a barrier to. Few data have been published on desensitization using polyvalent human intravenous immunoglobulin (IVIG) alone. Methods: We retrospectively reviewed the of 45 patients with a positive complement-dependent cytotoxicity crossmatch (CDCXM) or flow cytometry crossmatch (FCXM) against living donors from January 2003 to December 2014. Of these, 12 were excluded. Patients received monthly IVIG infusions (2 g/kg) only until they had a negative T-cell and B-cell FCXM. Results: During the 33 patients, 22 (66.7%) underwent living donor kidney transplantation, 7 (21.2%) received a deceased donor graft, and 4 (12.1%) did not undergo transplantation. The median class I and II panel reactive antibodies for these patients were 80.5% (range 61%-95%) and 83.0% (range 42%-94%), respectively. Patients (81.8%) had a positive T-cell and/or B-cell CDCXM and 4 (18.2%) had a positive T-cell and/or B-cell FCXM. Patients underwent transplantation after a median of 6 (range 3-16). The median donor-specific antibody mean fluorescence intensity sum was 5057 (range 2246-11,691) before and 1389 (range 934-2492) after desensitization (p = 0.0001). Mean patient follow-up time after transplantation was 60.5 (SD, 36.8) months. Nine patients (45.0%). Death-censored graft survival at 1, 3, and 5 years after transplant was 86.4, 86.4, and 79.2%, respectively and patient survival was 95.5, 95.5, and 83.7%, respectively. Conclusions: Desensitization using IVIG alone is an effective strategy, allowing successful transplantation in 87.9% of these highly sensitized patients.

Resumo Introdução: Sensibilização HLA é uma barreira ao transplante em pacientes sensibilizados. Há poucos dados publicados sobre dessensibilização utilizando somente imunoglobulina intravenosa humana polivalente (IgIV). Métodos: Revisamos retrospectivamente prontuários de 45 pacientes com prova cruzada positiva por citotoxicidade dependente do complemento (CDCXM) ou citometria de fluxo (FCXM) contra doadores vivos, de Janeiro/2003-Dezembro/2014. Destes, excluímos 12. 33 pacientes receberam infusões mensais de IgIV (2 g/kg) apenas até apresentarem FCXM células T e B negativa. Resultados: Durante dessensibilização, 22 pacientes (66,7%) realizaram transplante renal com doador vivo, 7 (21,2%) receberam enxerto de doador falecido, 4 (12,1%) não realizaram transplante. A mediana do painel de reatividade de anticorpos classes I e II para estes pacientes foi 80,5% (intervalo 61%-95%) e 83,0% (intervalo 42%-94%), respectivamente. 18 pacientes (81,8%) apresentaram CDCXM célula T e/ou B positiva; 4 (18,2%) apresentaram FCXM célula T e/ou B positiva. Pacientes realizaram transplante após mediana de 6 (intervalo 3-16) infusões. A mediana da somatória da intensidade média de fluorescência do anticorpo específico contra o doador foi 5057 (intervalo 2246-11.691) antes e 1389 (intervalo 934-2492) após dessensibilização (p = 0,0001). O tempo médio de acompanhamento do paciente pós transplante foi 60,5 (DP, 36,8) meses. Nove pacientes (45,0%) não apresentaram rejeição e 6 (27,3%) apresentaram rejeição mediada por anticorpos. Sobrevida do enxerto censurada para óbito em 1, 3, 5 anos após transplante foi 86,4; 86,4; 79,2%, respectivamente, e sobrevida do paciente foi 95,5; 95,5; 83,7%, respectivamente. Conclusões: Dessensibilização utilizando apenas IgIV é uma estratégia eficaz, permitindo transplante bem-sucedido em 87,9% destes pacientes altamente sensibilizados.

The relevance of a bank with genotyped platelets donors

ABSTRACT Objective: Describe the clinical and laboratory characteristics and the transfusion strategy of patients at Hospital Israelita Albert Einstein with platelet refractoriness and identify their etiological characteristics. Standardize the platelet immunofluorescence technique by flow cytometry as a test for platelet compatibility in immune platelet refractoriness in transfusion support. Methods: Review of medical records of refractory platelet patients followed at HIAE from January 2011 to May 2017. Clinical-demographic data, laboratory data and identification of the use of compatible genotyped platelets for patients in need of transfusion therapy were collected. The analyzed patients were classified according to the etiology of their platelet refractoriness. To standardize the FC-PIFT technique, blood group O platelets were incubated with serum from blood group AB donors and anti-IgG monoclonal antibody to determine the negative control. In order to verify the influence of the ABO system, monoclonal anti-IgG antibodies were incubated with blood group A or B platelets and with blood group O donor serum with isohemagglutinins below and above 1/64. Results: A total of 47 patients were evaluated, a 51% (24/47) preponderance of associated immune and non-immune factors (NIPR + IPR). The most common causes of NIPR + IPR were splenomegaly (54%) and the development of HLA antibodies (88%), consistent with the literature. For patients who required therapeutic transfusion, only a small portion received compatible genotyped platelets. Conclusion: Although 60% of patients could benefit from the therapeutic transfusion of genotyped platelets, only 10% were actually transfused with this type of blood component. This reaffirms the need for investments in a bank of genotyped platelet donors.

Bullous pemphigoid and milia: prevalence and clinical laboratory findings in a Brazilian sample

Abstract Background: Bullous pemphigoid (BP) associated with milia lesions has been increasingly reported, but its prevalence has not been reported in a Brazilian BP population yet. Objectives: To describe the occurrence and clinical-laboratorial findings of BP-milia association in a southeastern Brazilian sample. Methods: A descriptive study based on the medical charts of 102 BP patients was accomplished. Clinical and laboratory data of BP-milia patients were compiled. Total serum IgE measurements, immunoblot assays based on basement membrane zone antigens, and HLA-DQ alleles typing were performed. Results: Milia was evident in 8 (7.8%) BP patients, five males, aged between 46 and 88 years. Increased total IgE levels were determined in 7 (87.5%) of the eight patients. In five of eight patients, immunoblotting showed IgG reactivity against the BP180-NC16a domain but not against collagen VII or laminin-332; it also revealed reactivity against the BP180 C-terminal domain or LAD-1, or both in four of them. The HLA-DQB1*03:01 and HLA-DQA1*05:05 alleles were identified in three of five BP-milia patients. Moreover, three of five cases presented the HLA-DQB1*06 allelic group. Study limitations: HLA determination was performed in five patients. Conclusions: Milia formation in BP patients seems to be less uncommon than previously admitted. Laboratory data revealed increased IgE; autoantibodies against the BP180 C-terminal domain or LAD-1, or both; and the HLA-DQB1*06 allelic group, described for the BP-milia association. Careful determination of antibodies against basement membrane zone molecules and HLA characterization in different populations may provide further insights into this association. © 2022 Sociedade Brasileira de Dermatologia. Published by Elsevier España, S.L.U. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/).

Aspectos técnicos y clínicos de la prueba cruzada de histocompatibilidad en el trasplante de órganos solidos / Technical and clinical aspects of the histocompatibility crossmatch assay in solid organ transplantation

La presencia de anticuerpos dirigidos contra los antígenos leucocitarios humanos (Human Leukocyte Antigens, HLA) que se expresan en las células del donante, es uno de los factores de riesgo más importantes asociados con las complicaciones clínicas después del trasplante. La prueba cruzada es una de las pruebas de histocompatibilidad más eficaces para la detección de anticuerpos específicos contra el donante en los receptores de injertos. En los primeros métodos de la prueba cruzada, se utilizaba la citotoxicidad dependiente del complemento, que es útil para detectar dichos anticuerpos responsables del rechazo hiperagudo del injerto, pero carece de la sensibilidad adecuada. Por ello, se desarrollaron métodos de pruebas cruzadas más sensibles, entre ellas, la prueba cruzada por citometría de flujo que hoy se considera el método preferido. En este artículo se revisa la evolución de la prueba cruzada y los factores más importantes que deben tenerse en cuenta al realizarla y al interpretar los resultados de esta prueba fundamental para la supervivencia a largo plazo del injerto.

The presence of antibodies directed against human leukocyte antigens (HLA) expressed on donor cells is a significant risk factor for serious clinical complications after transplantation. The crossmatch assay is one of the most important tests available for the detection of donor-specific antibodies in potential allograft recipients. Early crossmatch methods utilized complement-dependent cytotoxicity, which is useful for detecting the donor-specific anti- HLA antibodies responsible for hyperacute allograft rejection but lacks adequate sensitivity. Consequently, more sensitive crossmatch methods have been developed, ultimately leading to the flow cytometry crossmatch as the currently preferred methodology. Herein, we review the evolution of the crossmatch assay and the most important factors to consider when performing and interpreting the results of this fundamental assay for ensuring the long-term survival of the transplanted organ.

Síndrome DRESS inducido por ácido valproico en una niña que respondió a la ciclosporina con análisis de HLA / Valproic acid-induced DRESS in a child responding to cyclosporine with HLA analysis

El síndrome de erupción medicamentosa con eosinofilia y síntomas sistémicos (drug reaction with eosinophilia and systemic symptoms, DRESS), también conocido como síndrome de hipersensibilidad inducida por medicamentos, es una reacción rara potencialmente mortal que causa una erupción grave y que puede provocar insuficiencia multiorgánica. Como con otras erupciones medicamentosas graves, los linfocitos T específicos para un medicamento tienen una función crucial en el síndrome DRESS. El modelo de hapteno/pro-hapteno, el modelo de interacción farmacológica y el modelo alterado de repertorio de péptidos son tres modelos diferentes desarrollados para describir la relación/interacción entre un medicamento o sus metabolitos y el sistema inmunitario. Analizamos nuestra experiencia con el tratamiento con ciclosporina en un caso de síndrome DRESS resistente a esteroides causado por ácido valproico en una niña y sus resultados clínicos, de laboratorio y de antígeno leucocitario humano (HLA).

Drug reaction with eosinophilia and systemic symptoms (DRESS), also known as drug-induced hypersensitivity syndrome, is a potentially life-threatening rare reaction that causes a severe rash and can lead to multiorgan failure. As in other severe drug eruptions, drug-specific T lymphocytes play a crucial role in DRESS. The hapten/pro-hapten model, pharmacological interaction model, and altered peptide repertoire model are three different models developed to describe the relationship/interaction between a medication or its metabolites and the immune system. We discuss our experience with cyclosporine treatment in a steroid-resistant DRESS syndrome caused by valproic acid in a girl, as well as her clinical, laboratory, and human leukocyte antigens (HLA) study results

Significance of expression of serum sHLA-G, ɑ-HBDH and STAT1 in acute myeloid leukemia and their relationship with prognosis / 中国基层医药

Objective:To investigate the significance of expression of serum soluble HLA-G (sHLA-G), alpha-hydroxybutyrate dehydrogenase (α-HBDH) and signal transducer and activator of transcription 1 (STAT1) in acute myeloid leukemia (AML) and their relationship with prognosis.Methods:A total of 107 patients with AML who received treatment in Xiaoshan Hospital, Wenzhou Medical University, from June 2016 to June 2019 were included in the AML group. An additional 100 healthy controls who concurrently received physical health examination in the same hospital were included in the control group. Serum samples were collected from both patients with AML and healthy controls. Serum concentration of sHLA-G concentration was determined using the double-antibody sandwich enzyme-linked immunosorbent assay. Serum α-HBDH concentration was determined using the rate method. Peripheral blood was collected from patients with AML and healthy controls. The relative expression of STAT1 was determined by quantitative real-time polymerase chain reaction.Results:sHLA-G and α-HBDH concentrations in the AML group were (13.26 ± 2.19) μg/L and (362.17 ± 38.47) U/L, respectively, which were significantly higher than those in the control group [(5.08 ± 0.43) μg/L, (108.94 ± 12.19) U/L, t = 36.69, 62.93, both P < 0.05]. The relative expression of STAT1 in the AML group was significantly higher than that in the control group [(3.17 ± 0.25) vs. (1.01 ± 0.01), t = 86.32, P < 0.05] Among patients with Hb ≤ 80 g/L, the number of patients with high sHLA-G expression was higher than that of patients with low sHLA-G expression ( χ2 = 7.15, P < 0.05). The number of patients with white blood cells > 10 × 10 9/L was significantly higher than that of patients with low sHLA-G expression ( χ2 = 17.31, P < 0.05). Among patients with Hb ≤ 80 g/L, the number of patients with high α-HBDH expression was significantly higher than that of patients with low α-HBDH expression ( χ2 = 10.76, P < 0.05). The number of patients with white blood cells > 10 × 10 9/L was significantly higher than that with low α-HBDH expression ( χ2 = 13.17, P < 0.05). Among patients with Hb ≤ 80 g/L, the number of patients with high STAT1 expression was significantly higher than that of patients with low STAT1 expression ( χ2 = 18.14, P < 0.05). The number of patients with white blood cells > 10 × 10 9/L was significantly higher than that of patients with low STAT1 expression ( χ2 = 18.51, P < 0.05). The overall survival time in patients with high sHLA-G, α-HBDH expression and STAT1 expression was (17.92 ± 1.72) months, (19.34 ± 1.57) months, and (16.36 ± 2.08) months, respectively, which were significantly lower than those in patients with low sHLA-G, α-HBDH and STAT1 expression [(28.93 ± 1.46) months, (27.53 ± 1.89) months, (30.48 ± 1.12) months, t = 35.08, 24.07, 38.32, all P < 0.05]. Conclusion:sHLA-G, α-HBDH and STAT1 are abnormally expressed in patients with AML. Higher sHLA-G, α-HBDH and STAT1 expression indicates poorer prognosis of AML. Therefore, sHLA-G, α-HBDH and STAT1 can be used as the potential targets for AML treatment and prognosis prediction. Findings from this study are highly innovative and scientific.

Negative effects of donor specific anti-HLA antibody on poor hematopoietic recovery in patients with hematological diseases receiving haploidentical stem cell transplantation and rituximab for desensitization / 中华内科杂志

Objective:To investigate the incidences and risk factors of poor hematopoietic reconstitution (PHR) in patients with hematological diseases who underwent haploidentical allograft and were treated with rituximab for desensitization.Methods:Eight-three donor specific anti-HLA antibody (DSA, 2000 ≤MFI<10 000) positive patients who underwent haploidentical allograft were prospectively enrolled. Rituximab (375 mg/m 2) was used for desensitization day-3 of conditioning regimen. Incidence and factors associated with PHR, including primary poor graft function and prolonged thrombocytopenia, were investigated. Results:There were 22 males and 61 females with a median age of 39(range: 1-65) years. Kaplan-Meier analysis showed that the 100 day cumulative incidences of neutrophil and platelet engraftment were 93.0% and 90.7%, respectively. The incidences of PHR were 14.7%. The 3-year relapse rate, non-relapse mortality (NRM) rate, event-free survival (EFS), leukemia-free survival (DFS) and overall survival (OS) were 6.5%, 15.1%, 70.8%, 79.4% and 79.4%, respectively. Patients with DSA MFI<5 000 (group A, n=46) experienced lower PHR (4.4% vs. 27.5%, P=0.003), and higher 3-year EFS (79.5% vs. 59.8%, P=0.020) compared to those with DSA MFI≥5 000 (group B, n=37). Multivariate analysis showed that DSA MFI≥5 000 was correlated with PHR ( HR=6.101, P=0.021). PHR was associated with higher NRM ( HR=4.110, P=0.026), lower DFS ( HR=3.656, P=0.019) and OS ( HR=3.656, P=0.019). Conclusion:Our data suggest that high pre-transplant DSA level is a risk factor for PHR in patients with hematological diseases receiving haploidentical allograft and rituximab for desensitization.

Lamotrigine-induced drug eruptions / 中华皮肤科杂志

Lamotrigine is a new type of drug for the treatment of epilepsy, bipolar affective disorder and neuralgia. Drug eruptions are the common adverse reaction to lamotrigine, such as fixed drug eruption, acute generalized exanthematous pustulosis, toxic epidermal necrolysis, drug-induced hypersensitivity syndrome, etc. The pathogenesis of lamotrigine-induced drug eruptions includes immune and non-immune mechanisms. HLA alleles are related to drug eruptions caused by lamotrigine, and their relationship varies with the race of populations and type of drug eruptions. This review summarizes the etiology, pathogenesis and clinical characteristics of drug eruptions caused by lamotrigine, aiming to guide clinical treatment.

Frequency of alleles associated with celiac disease in patients with autoimmune thyroid disease / Frequência dos alelos associados com a doença celíaca em portadores de doenças autoimunes da tireoide

ABSTRACT Objective To determine the frequency of Human leukocyte antigen alleles and to verify the association of the presence of these alleles with symptoms and other diseases related to celiac disease in patients with autoimmune thyroid diseases. Methods A questionnaire on the symptoms and diseases associated with celiac disease was applied. Genomic deoxyribonucleic acid was extracted by collecting cells from the oral mucosa. The alleles (DQA1*0501; DQB1*0201; DRB1*04) were identified by means of the polymerase chain reaction. Results A total of 110 patients with autoimmune thyroid diseases participated in this study. It was observed that 66.4% of the individuals carried at least one of the alleles assessed and that 58.2% of the individuals were positive for at least one of the DQ2 alleles (DQA1*0501; DQB1*0201) and out of these 18.2% were positive for both DQ2 alleles (DQA1*0501; DQB1*0201). With regard to DQ8 (DRB1*04), 21.8% of the studied population was positive for this allele and 3.6% was positive for both DQ2 (DQA1*0501; DQB1*0201) and DQ8 (DRB1*04). A significant association was found between the presence of the DRB1*04 allele and gastrointestinal symptoms (p=0.02). A significant association of the DRB1*04 allele with type 1 diabetes mellitus (p=0.02) was observed. Conclusion The genetic profiles most commonly associated with celiac disease, such as DQ2 (DQA1*0501; DQB1*0201) and DQ8 (DRB1*04) were around 20.0% prevalent in the studied population. These are risk haplotypes for celiac disease especially when symptoms and diseases related to celiac disease are present. Therefore, it is important to screen patients to investigate a potential diagnosis for celiac disease.

RESUMO Objetivo Determinar a frequência dos alelos do Human leukocyte antigen e verificar a associação da presença desses alelos com sintomas e outras doenças relacionados à doença celíaca em portadores de doenças autoimunes da tireoide. Métodos Aplicou-se um questionário relacionado aos sintomas e doenças associados à doença celíaca. O ácido desoxirribonucleico genômico foi extraído por meio da coleta das células da mucosa bucal. Os alelos (DQA1*0501; DQB1*0201; DRB1*04) foram identificados por meio da reação em cadeia da polimerase. Resultados Participaram deste estudo 110 portadores de doenças autoimunes da tireoide. Observou-se que 66,4% dos indivíduos carregavam pelo menos um dos alelos estudados e que 58,2% dos indivíduos eram positivos para pelo menos um dos alelos DQ2 (DQA1*0501; DQB1*0201) e destes 18,2% foram positivos para ambos alelos do DQ2(DQA1*0501; DQB1*0201). Com relação ao DQ8 (DRB1*04), 21,8% da população estudada eram positivos para esse alelo e 3,6% eram positivos tanto para o DQ2 (DQA1*0501; DQB1*0201) quanto para o DQ8 (DRB1*04). Foi encontrada associação significativa da presença do alelo DRB1*04 com os sintomas gastrointestinais (p=0,02). Houve associação significativa do alelo DRB1*04 com diabetes mellitus tipo 1 (p=0,02). Conclusão O perfil genético mais fortemente associado à doença celíaca, tais como DQ2 (DQA1*0501; DQB1*0201) e DQ8 (DRB1*04) estavam presentes em torno de 20,0% da população estudada, estes são haplótipos de risco para doença celíaca e principalmente na presença de sintomas e doenças relacionadas à doença celíaca. Sendo assim, é importante realizar o rastreamento para investigar um possível diagnóstico para doença celíaca.

Neutropenia aloimune neonatal em gêmeas idênticas / Neonatal alloimmune neutropenia in identical twins

A neutropenia aloimune neonatal (NAN) é uma patologia causada pelo antagonismo imunológico, como a doença hemolítica do recém-nascido ou a trombocitopenia aloimune neonatal, mas relacionada aos neutrófilos, em vez de glóbulos vermelhos ou plaquetas. Descreveremos um caso clínico de duas gêmeas idênticas nascidas a termo, com Apgar de 8 e 9, sendo que após algumas horas do nascimento apresentaram febre. Um exame de sangue revelou neutropenia grave que resultou em sepse. O diagnóstico da NAN foi realizado clinicamente e por testes de histocompatibilidade. A prova cruzada por citometria de fluxo foi positiva, usando soro da mãe e suspensões celulares (granulócitos e linfócitos) das gêmeas e do pai. Este teste não fornece informações sobre para qual sistema genético os anticorpos foram positivos, se contra os antígenos específicos de neutrófilos humanos (HNA) ou contra os antígenos leucocitários humanos (HLA). Para o esclarecimento, realizamos o teste de aglutinação de granulócitos (GAT) com um painel de doadores fidelizados e com antígenos HNA1-5 conhecidos, utilizando o soro materno como reagente. Foi também realizada a pesquisa de anticorpos anti-HLA e anti-HNA no soro materno. Os genótipos HLA e HNA foram identificados, permitindo conhecer as especificidades dos anticorpos maternos contra os antígenos dos neutrófilos do marido e das filhas. O diagnóstico de NAN não é realizado na maioria dos hospitais de nosso país e do exterior, devido à dificuldade de execução dos testes de histocompatibilidade, no entanto a prova cruzada por citometria de fluxo pode facilmente ser implantada nos laboratórios clínicos, sendo que está descrita detalhadamente nesse caso clínico.

Neonatal alloimmune neutropenia (NAN) is a disease caused by immunological antagonism, such as hemolytic disease of the newborn or neonatal alloimmune thrombocytopenia, but related to neutrophils rather than to red blood cells or platelets. We will describe a clinical case of two identical twins born with Apgar 8 and 9 that started with fever few hours after delivery. A blood test revealed severe neutropenia, which was followed by sepsis. The diagnosis of NAN was done clinically and by histocompatibility testing. Flow cytometry crossmatch was positive, using mother serum and cell suspensions (granulocytes and lymphocytes) from the twin girls and from the father. This test did not provide information about the genetic system for which the antibodies are positive, if against human neutrophil antigens (HNA) or human leucocyte antigens (HLA). To clear this, the granulocyte agglutination test (GAT) was performed with a panel of control donors with known HNA1-5 antigens, using the maternal serum as a reagent. We did also a Luminex screening assay for detection of anti-HLA and anti-HNA antibodies in the mother serum. The HLA and HNA genotypes were identified, which allowed to define specificities in mother's antibodies against the neutrophil surface antigens from her husband and from the twins. The diagnosis of NAN diagnose is not done in most hospitals worldwide, mainly by the difficulty in executing the histocompatibility test. However, the crossmatch by flow cytometry could be easily done in clinical laboratories following the method described in this article.

Renal transplant patients with preformed anti-HLA antibodies: early biopsy findings and clinical outcomes / Pacientes transplantados renais com anticorpos anti-HLA pré-formados: achados precoces de biópsia e desfechos clínicos

Abstract Introduction: Renal fibrosis is the end point of a process that begins at transplant, with ischemia reperfusion and early inflammation, and progresses over time with immunological and non-immunological phenomena. Early identification of morphological markers and intervention could improve graft function and survival. Objective: to evaluate the correlation between intensity and specificity of pre-transplant anti-HLA antibodies and kidney allograft pathology in order to identify early risk factors or markers of allograft dysfunction. Methods: A retrospective cohort of kidney transplant recipients with pre-transplant anti-HLA antibodies who underwent graft biopsy within the first two years post-transplant was divided into two groups according to the specificity of anti-HLA antibodies: nonspecific (non-DSA, n = 29) and specific (DSA+, n = 16). Kidney graft pathology, renal function, and proteinuria were analyzed. Results: general characteristics were similar in both groups, except for the higher dose of thymoglobulin in DSA+ group (p < 0.05). The non-DSA group had higher scores for glomerulosclerosis, interstitial inflammation (i) and interstitial fibrosis (ci) (p < 0.05) and higher incidence of cell-mediated acute rejection. No statistical difference in incidence of antibody-mediated rejection, renal function, and proteinuria was observed during follow up. Discussion and conclusions: the difference in inflammation scores and interstitial fibrosis may be associated to the higher incidence of acute cell-mediated rejection and polyomavirus nephropathy in the Non-DSA group. We also should take into account the protective effect of higher doses of thymoglobulin, reducing ischemia reperfusion injury in the DSA+ group. The short follow-up might have been insufficient to detect long-term changes in allograft tissue, renal function, and proteinuria.

Resumo Introdução: A fibrose renal é o desfecho de um processo iniciado no transplante, com reperfusão, isquemia e inflamação precoce, que progride ao longo do tempo com fenômenos imunológicos e não imunológicos. A identificação de marcadores morfológicos e a intervenção precoce poderiam melhorar a função e a sobrevida do enxerto. Objetivo: Avaliar a correlação entre intensidade e especificidade de anticorpos anti-HLA pré-transplante alterações histológicas do enxerto renal, de forma a identificar fatores de risco ou marcadores de disfunção precoces do aloenxerto. Métodos: O presente estudo incluiu uma coorte retrospectiva de receptores de transplante renal sensibilizados com anticorpos anti-HLA no pré-transplante submetidos a biópsia de enxerto nos primeiros dois anos após o transplante. Os grupos foram divididos em função da especificidade dos anticorpos anti-HLA: sem anticorpos doador-específicos (não-DSA, n = 29) e com anticorpos doador-específicos (DSA+, n = 16). Alterações histológicas do enxerto renal, função renal e proteinúria foram analisados. Resultados: Os dois grupos tinham características gerais semelhantes, exceto pela dose mais elevada de timoglobulina administrada nos indivíduos do grupo DSA+ (p < 0,05). O grupo não-DSA teve escores mais elevados de glomeruloesclerose, inflamação intersticial (i) e fibrose intersticial (ci) (p < 0,05), além de maior incidência de rejeição celular aguda (RCA). Não foi observada diferença estatística na incidência de rejeição mediada por anticorpos, função renal ou proteinúria durante o seguimento. Discussão e Conclusões: A diferença nos escores de inflamação e fibrose intersticial pode estar associada à maior incidência de RCA e nefropatia por poliomavírus no grupo não-DSA. Devemos considerar ainda o efeito protetor das doses mais elevadas de timoglobulina na redução da lesão por isquemia-reperfusão no grupo DSA+. O curto período de seguimento pode ter sido insuficiente para detectar alterações de longo prazo no tecido do aloenxerto, função renal e proteinúria.

Current status of immunogenetic studies on primary biliary cholangitis / 临床肝胆病杂志

Primary biliary cholangitis (PBC) has the pathological feature of progressive intrahepatic cholestasis caused by immune-mediated apoptotic necrosis of small biliary epithelial cells, with a risk of progression to bile duct fibrosis, liver cirrhosis, and hepatocellular carcinoma. PBC has immunogenetic characteristics, and the abnormal genetic regulation of immune response in patients with PBC includes abnormal immune response of T and B lymphocytes involving human leukocyte antigen (HLA) and non-HLA genes and taking pyruvate dehydrogenase complex-E2 in mitochondria of intrahepatic small biliary epithelial cells as the antigen. More than 30% of PBC patients have poor response to ursodeoxycholic acid treatment, and therefore, clarifying the mechanism of abnormal immune regulation in PBC has great clinical significance in guiding the immunotherapy for PBC.

Soluble Human Leukocyte Antigen Molecules Detected in Orofacial Cleft Patients: A Case-Control Study

Abstract Objective: To compare soluble HLA-C and HLA-DR molecules present in the plasma of orofacial cleft and non-orofacial cleft populations. Material and Methods: Orofacial cleft patients were recruited using an accidental sampling approach (n=15). Peripheral blood was collected from the participants and processed for Enzyme Linked Immunosorbent Assay (ELISA) against HLA-C and HLA-DR with specific antibodies. The absorbance was calculated utilizing ELISA reader. Data were statistically analyzed using an independent t-test to compare the disease and control groups. Results: The levels of soluble HLA-C and HLA-DR were significantly higher in the diseased group compared to the control group (p<0.05). Conclusion: The role of HLA molecules in non-communicable disease and congenital anomalies, particularly orofacial cleft, remains speculative despite the positive results of this study and those of previous investigations. It suggests that the variables examined may affect specific pathways involved in the pathogenesis of orofacial cleft, and predispose the individuals concerned to the oral cleft.

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)

HLA-alleles class I and II associated with genetic susceptibility to neuromyelitis optica in Brazilian patients / Alelos HLA classes I e II associados à suscetibilidade genética a neuromielite óptica em pacientes brasileiros

ABSTRACT Objective: To study the genetic susceptibility to neuromyelitis optica (NMO) as well as the relationship between HLA genotypes and susceptibility to the disease in the southern Brazilian population. Methods: We analyzed patients with NMO, who met criteria for Wingerchuk's diagnosis of NMO, with detected serum anti-AQP4-IgG antibody. The HLA genotyping was performed by high-resolution techniques (Sanger sequencing) in patients and controls. The HLA genotypes were statistically compared with a paired control population. Results: The HLA genotyping revealed the diversity of the southern Brazilian population whose HLA profile resembled European and Asian populations. Some alleles had statistical correlations with a positive association (increased susceptibility) with NMO, particularly the HLA-DRB1*04:05 and *16:02. Conclusions: In our study, the HLA genotype was different to that previously reported for other Brazilian populations. Although our study had a small cohort, HLA genotypes were associated with increased susceptibility to NMO for HLA-DRB1*04:05 and *16:02. The alleles of HLA class I HLA-A*02:08 and *30:09, HLA-B*08:04 and *35:04 showed an association before the Bonferroni correction.

RESUMO Objetivo: Estudar a suscetibilidade genética a neuromielite óptica (NMO) assim como sua relação com o genótipo HLA na população do sul do Brasil. Métodos: Nós analisamos pacientes com NMO que preenchiam os critérios diagnósticos de Wingerchuk para NMO, com presença do anticorpo anti-AQP4-IgG no soro. O genótipo HLA foi realizado usando técnicas de alta resolução (sequenciamento de Sanger) em pacientes e controles. Genótipos HLA foram estatisticamente comparados com uma população controle pareada. Resultados: Genotipagem HLA revelou a diversidade da população sul brasileira cujo perfil HLA lembra as populações europeia e asiática. Alguns alelos tiveram correlação estatística com associação positiva (suscetibilidade aumentada) com NMO, particularmente o HLA-DRB1*04:05 e *16:02. Conclusões: Em nosso estudo, o genótipo HLA foi diferente do previamente relatado em outras populações brasileiras. Embora o número de pacientes tenha sido pequeno, HLA específicos foram associados com suscetibilidade aumentada a NMO para HLA-DRB1*04:05, *16:02. Os alelos HLA classe I HLA*02:08 e *30:09, HLA-B*08:04 e *35:04 tiveram associação antes da correção de Bonferroni.

Loss of Human Leukocyte Antigen Class I Expression Is Associated with Poor Prognosis in Patients with Advanced Breast Cancer

BACKGROUND: Human leukocyte antigen class I (HLA-I) molecules play important roles in regulating immune responses. Loss or reduction of HLA-I expression has been shown to be associated with prognosis in several cancers. Regulatory T-cells (Tregs) also play critical functions in immune response regulation. Evaluation of HLA-I expression status by the EMR8-5 antibody and its clinical impact in breast cancer have not been well studied, and its relationship with Tregs remains unclear. METHODS: We evaluated HLA-I expression and Treg infiltration by immunohistochemistry in 465 surgically resected breast cancer samples. We examined the correlation between HLA-I expression and Treg infiltration and clinicopathologic characteristics and survival analyses were performed. RESULTS: Total loss of HLA-I expression was found in 84 breast cancer samples (18.1%). Univariate survival analysis revealed that loss of HLA-I expression was significantly associated with worse disease-specific survival (DSS) (p = .029). HLA-I was not an independent prognostic factor in the entire patient group, but it was an adverse independent prognostic factor for DSS in patients with advanced disease (stage II–IV) (p = .031). Treg numbers were significantly higher in the intratumoral stroma of HLA-I–positive tumors than in HLA-I–negative tumors (median 6.3 cells/high power field vs 2.1 cells/high power field, p < .001). However, Tregs were not an independent prognostic factor in our cohort. CONCLUSIONS: Our findings suggest that the loss of HLA-I expression is associated with poor prognosis in breast cancer patients, highlighting the role of HLA-I alterations in immune evasion mechanisms of breast cancer. HLA-I could be a promising marker that enables the application of more effective and precise immunotherapies for patients with advanced breast cancer.

Attach importance to the immunological mechanism of preeclampsia / 中国医师杂志

Preeclampsia is one of the main causes of high morbidity and mortality of pregnant women and perinatal infants worldwide. Affected by many factors, preeclampsia has a complex pathogen-esis and can cause involvement of multiple organs and systems. Its pathogenesis is still unclear. Current research suggests that maternal immune system indirectly involved in the pathophysiology of preeclampsia change, that is, abnormal activation of innate immune cells and unbalanced differentiation of T helper cell subsets interfere with normal immune regulation, and interact with inflammatory response of the body, which produces cytotoxic environment at the maternal-fetal interface and affects trophoblast inva-sion. Therefore, clarifying the role of the immune system can not only clarify the pathogenesis of pre-eclampsia, but also contribute to the development of diagnosis and treatment of preeclampsia. This paper reviews the research status of immune system in preeclampsia, including innate immunity and adaptive immunity . The immune mechanism of preeclampsia is elaborated mainly from immune regulation mediated by T lymphocyte, natural killer ( NK) cell, macrophage and human leukocyte antigen.

Expression of Immunoproteasome Subunit LMP7 in Breast Cancer and Its Association with Immune-Related Markers / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: In the presence of interferon, proteasome subunits are replaced by their inducible counterparts to form an immunoproteasome (IP) plays a key role in generation of antigenic peptides presented by MHC class I molecules, leading to elicitation of a T cell‒mediated immune response. Although the roles of IP in other cancers, and inflammatory diseases have been extensively studied, its significance in breast cancer is unclear. MATERIALS AND METHODS: We investigated the expression of LMP7, an IP subunit, and its relationship with immune system components in two breast cancer cohorts. RESULTS: In 668 consecutive breast cancer cohort, 40% of tumors showed high level of LMP7 expression, and tumors with high expression of LMP7 had more tumor-infiltrating lymphocytes (TILs) in each subtype of breast cancer. In another cohort of 681 triple-negative breast cancer patients cohort, the expression of LMP7 in tumor cells was significantly correlated with the amount of TILs and the expression of interferon-associated molecules (MxA [p < 0.001] and PKR [p < 0.001]), endoplasmic reticulum stress-associated molecules (PERK [p=0.012], p-eIF2a [p=0.001], and XBP1 [p < 0.001]), and damage-associated molecular patterns (HMGN1 [p < 0.001] and HMGB1 [p < 0.001]). Patients with higher LMP7 expression had better disease-free survival outcomes than those with no or low expression in the positive lymph node metastasis group (p=0.041). CONCLUSION: Close association between the TIL levels and LMP7 expression in breast cancer indicates that better antigen presentation through greater LMP7 expression might be associated with more TILs.