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1.
J Clin Apher ; 36(3): 291-298, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33253430

RESUMO

BACKGROUND: In organ transplantation, apheresis is frequently used for removal of anti-HLA antibodies. However, it is unclear whether plasmapheresis (PP) or semi-selective immunoadsorption (IA) should be employed, and the optimal number of apheresis sessions required to reach post-treatment objectives is also unknown. METHODS: We enrolled 43 patients from Bordeaux University Hospital who were treated with PP (n = 29) or IA (n = 14) for antibody-mediated rejection or pre-transplant desensitization. Using Luminex single-antigen flow beads, we assessed the initial mean fluorescence intensity (MFI) of 1416 positive beads with MFIs obtained after 7 to 8 apheresis sessions (extended protocol) and, if a serum was available, after the first four sessions (short protocol). RESULTS: MFI reduction after extended apheresis protocol was stronger with IA [87% (61%-100%)] than with PP [73% (22%-100%)] (P < .001). Indeed, 59% of the beads had a final MFI < 2000 with IA, whereas only 38% with PP (P < .001). The efficacy of removal depended on initial MFI but not on HLA specificity. A short protocol of apheresis showed excellent results without superiority of IA over PP for antibodies with an initial MFI < 3000. For antibodies showing MFI ≥2000 after four sessions, the residual MFI predicted the effectiveness of four additional sessions. CONCLUSION: Monitoring the MFI of anti-HLA antibodies before and during apheresis protocol can guide physicians in the selection of apheresis technique and the number of sessions to be performed.


Assuntos
Antígenos HLA/imunologia , Técnicas de Imunoadsorção , Isoanticorpos/isolamento & purificação , Plasmaferese/métodos , Adulto , Feminino , Fluorescência , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos
2.
Arq. ciências saúde UNIPAR ; 24(3): 133-138, set-dez. 2020.
Artigo em Português | LILACS | ID: biblio-1129455

RESUMO

Quando um indivíduo é exposto a antígenos eritrocitários não próprios, ocorre uma resposta imunológica, que leva à produção de anticorpos irregulares voltados contra esses antígenos. Esse processo é conhecido como aloimunização eritrocitária e acontece em decorrência de transfusões de sangue ou gestações incompatíveis. Na medicina transfusional a pesquisa de anticorpos irregulares é fundamental, pois a falha na detecção de um aloanticorpo pode provocar reações transfusionais, aloimunizações, anemias hemolíticas autoimunes e doença hemolítica perinatal. Este estudo tem por objetivo analisar a frequência de anticorpos irregulares de pacientes atendidos no Hemocentro Regional de Francisco Beltrão, Paraná, no ano de 2017. Os dados foram coletados a partir da revisão de registros em arquivos do Laboratório de Imunohematologia do Hemonúcleo. Foram avaliados dados de 49 protocolos de pacientes que apresentaram dificuldades transfusionais no ano de 2017. Dentre os pesquisados, 37 pacientes (75,5%) apresentaram anticorpos irregulares. Dentre os anticorpos anti-eritrocitários observados neste estudo, evidenciou-se a presença de doze pacientes com anti-D (27,2%), seis pacientes com anti-K (13,6%), quatro pacientes com anti-C (9,0%) e em seis pacientes (13,6%) foi observada a presença de autoanticorpos. Este estudo indica que, nos pacientes transfundidos, os anticorpos mais frequentes foram os aloanticorpos Anti-D do Sistema Rh, provavelmente devido ao seu alto grau de imunogenicidade. A prevalência desses anticorpos é semelhante a vários estudos encontrados na literatura.


When an individual is exposed to not-self red blood cell antigens, an immune response occurs, which leads to the production of irregular antibodies directed against these antigens. This process is known as erythrocyte alloimmunization and occurs as a result of blood transfusions or incompatible pregnancies. In transfusion medicine, the search for irregular antibodies is essential, since failure to detect an alloantibody can cause transfusion reactions, alloimmunizations, autoimmune hemolytic anemias, and perinatal hemolytic disease. This study aims at analyzing the frequency of irregular antibodies of patients seen at the Regional Blood Center of Francisco Beltrão, Paraná, in 2017. The data were collected from the review of records in files of the Immunohematology Laboratory of Hemonúcleo. Data from 49 protocols of patients who had transfusion difficulties in 2017 were evaluated. Among those surveyed, 37 patients (75.5%) had irregular antibodies. Among the anti-erythrocyte antibodies observed in this study, the presence of twelve patients with anti-D (27.2%), six patients with anti-K (13.6%), four patients with anti-C (9.0 %), and in six patients (13.6%) with the presence of autoantibodies were observed. This study indicates that, in transfused patients, the most frequent antibodies were the Rh System Anti-D alloantibodies, probably due to their high degree of immunogenicity. The prevalence of these antibodies is similar to several studies found in the literature.


Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Autoanticorpos/imunologia , Isoanticorpos/imunologia , Autoanticorpos/isolamento & purificação , Transfusão de Sangue , Estudos Retrospectivos , Distribuição por Sexo , Distribuição por Idade , Eritrócitos/imunologia , Reação Transfusional/imunologia , Isoanticorpos/isolamento & purificação , Anticorpos/isolamento & purificação , Anticorpos/imunologia
3.
Clin Lab ; 66(6)2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32538044

RESUMO

BACKGROUND: Red blood cell (RBC) alloantibody titration is a quasi-quantitative method to assess antibody concentration and is considered a useful means of estimating maternal alloimmunization during pregnancy. Traditionally, titration is performed using conventional tube test (CTT). The gel microcolumn agglutination-based method (GMA) has been proven reliable for many immunohematology tests. Our study compared CTT with GMA of two different, commercially available GMA systems for RBC alloantibody titration. METHODS: Serum samples with significant RBC-alloantibodies were evaluated in our study. Each sample was titrated concurrently with CTT, with ID-DiaMed-GmbH, Cressier, Switzerland (GMA1), and with DG Gel Coombs Diagnostic Grifols, Passeig Fluvial, Spain (GMA2). RESULTS: One hundred thirty-seven titration tests including 50 anti-D, 25 anti-Kell, 10 anti-E, 9 anti-Jka, 8 anti-c, 5 anti-Cw, 5 anti-Fya, 7 anti-M, 6 anti-Kpa, 3 anti-Lua, 1 anti-e, 3 anti-G, and 2 anti-Cha were performed and evaluated. Samples tested by CTT versus GMA1 and GMA2 generated mostly equal or higher titers by GMAs. The results of both comparisons were in good agreement (W = 0.91, p < 0.0001, and W = 0.92, p < 0.0001, respectively). For all antibody specificities, the mean absolute difference in titers ranged from 1 - 3 for both GMA1 and GMA2 versus CTT. Samples tested by GMA1 vs. GMA2 were in almost perfect agreement (W = 0.95, p < 0.0001). CONCLUSIONS: Although both GMAs were found slightly more sensitive than CTT for alloantibody titration, the differences were not significant and the agreement between all methods was very good, possibly indicating GMA as a suitable alternative to CTT in RBC antibody titration.


Assuntos
Antígenos de Grupos Sanguíneos/imunologia , Eritrócitos/imunologia , Isoanticorpos , Feminino , Testes de Hemaglutinação/métodos , Humanos , Testes Imunológicos/métodos , Isoanticorpos/análise , Isoanticorpos/isolamento & purificação , Gravidez , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
4.
J Immunol Res ; 2019: 2754920, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31223627

RESUMO

Transferon® is an immunomodulator made of a complex mixture of peptides from human dialyzable leucocyte extracts (hDLEs). Development of surrogate antibodies directed to hDLE is an indispensable tool for studies during process control and preclinical trials. These antibodies are fundamental for different analytical approaches, such as identity test and drug quantitation, as well as to characterize its pharmacokinetic and mechanisms of action. A previous murine study showed the inability of the peptides of Transferon® to induce antibody production by themselves; therefore, in this work, two approaches were tested to increase its immunogenicity: chemical conjugation of the peptides of Transferon® to carrier proteins and the use of a rabbit model. Bioconjugates were generated with Keyhole Limpet Hemocyanin (KLH) or Bovine Serum Albumin (BSA) through maleimide-activated carrier proteins. BALB/c mice and New Zealand rabbits were immunized with Transferon® conjugated to KLH or nonconjugated Transferon®. Animals that were immunized with conjugated Transferon® showed significant production of antibodies as evinced by the recognition of Transferon®-BSA conjugate in ELISA assays. Moreover, rabbits showed higher antibody titers when compared with mice. Neither mouse nor rabbits developed antibodies when immunized with nonconjugated Transferon®. Interestingly, rabbit antibodies were able to partially block IL-2 production in Jurkat cells after costimulation with Transferon®. In conclusion, it is feasible to elicit specific and functional antibodies anti-hDLE with different potential uses during the life cycle of the product.


Assuntos
Isoanticorpos/imunologia , Fator de Transferência/efeitos adversos , Adjuvantes Imunológicos , Animais , Formação de Anticorpos , Especificidade de Anticorpos/imunologia , Antígenos/administração & dosagem , Antígenos/imunologia , Ensaio de Imunoadsorção Enzimática , Humanos , Imunização , Imunoglobulina G/imunologia , Imunoglobulina G/isolamento & purificação , Isoanticorpos/isolamento & purificação , Masculino , Camundongos , Peptídeos/administração & dosagem , Peptídeos/imunologia , Coelhos , Fator de Transferência/imunologia , Fator de Transferência/uso terapêutico
5.
Rev. esp. anestesiol. reanim ; 65(7): 403-406, ago.-sept. 2018.
Artigo em Espanhol | IBECS | ID: ibc-177137

RESUMO

Describimos el manejo de una paciente programada para esofagectomía por neoplasia a la que durante el proceso de reserva de hemoderivados le fueron detectados aloanticuerpos, que prácticamente imposibilitaban la obtención de sangre compatible. El manejo de la anemia perioperatoria («patient blood management») se debe realizar rutinariamente en los pacientes quirúrgicos con riesgo de transfusión. Esta estrategia se ha considerado como una de las medidas a tener en cuenta en la rehabilitación multimodal quirúrgica o programa de recuperación intensificada


A description is presented on the management of a patient with an oesophageal neoplasm scheduled for oesophagectomy. Alloantibodies were detected during a blood components reservation procedure, which made it almost impossible to obtain compatible blood. Peri-operative anaemia management or "Patient Blood Management" should be routinely performed in all patients at transfusion risk. This strategy has been considered as one of the actions to bear in mind in fast-track surgery or enhanced recovery after surgery


Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Anemia/diagnóstico , Transfusão de Sangue Autóloga/normas , Recuperação de Sangue Operatório/normas , Esofagectomia/métodos , Neoplasias Esofágicas/cirurgia , Período Perioperatório , Anemia/complicações , Segurança do Sangue/tendências , Isoanticorpos/isolamento & purificação
6.
Liver Transpl ; 24(8): 1101-1108, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30142248

RESUMO

Formation of de novo donor-specific antibodies (dn-DSAs) has been associated with longterm immunologic complications after liver transplantation (LT). We hypothesized that human leukocyte antigen (HLA) epitope/eplet mismatch (MM) is a marker of immunogenicity and a risk factor for dn-DSA formation. Sera from 80 LT recipients were prospectively screened for dn-DSA by a Luminex single-antigen test (One Lambda, Inc., Canoga Park, CA) at 1, 2, 3, 6, 12, 18, 24, and 36 months after LT. HLA typing of the recipients and donors was performed using polymerase chain reaction (PCR)-SSP and PCR-SSOP Luminex low-resolution methods (One Lambda, Inc.). The HLAMatchmaker computer algorithm was used for identification of MM eplets at HLA-DRB1 and -DQA1/B1 loci. Luminex single-antigen bead solid phase assay was used for antibody analysis. Standard immunosuppression included thymoglobulin-rituximab induction and tacrolimus maintenance. There were 27 (34%) patients who developed dn-DSA. There were no episodes of antibody-mediated rejection, and 9 (11%) developed acute cellular rejection (ACR). A positive crossmatch status and a higher number of HLA-A, -B, -DR, and -ABDR MMs were not associated with dn-DSA formation. Patients developing dn-DSA had a significantly higher number of total (38 ± 2.7 versus 28 ± 2.3; P = 0.01) and antibody-verified (AbVer; 14 ± 1.1 versus 10 ± 1; P = 0.015) class II MM eplets. By a multivariate regression analysis, the number of class II MM eplets was strongly associated with risk of class II dn-DSA formation (odds ratio [OR], 1.2; P < 0.01). Patients with ACR had a significantly higher number of total (20.2 ± 1.3 versus 13.9 ± 0.9; P < 0.01) as well as AbVer (10.7 ± 1.1 versus 7.5 ± 0.6; P = 0.03) class I MM eplets. In conclusion, donor-recipient HLA epitope MM is associated with a risk of dn-DSA formation and rejection after LT. However, further studies are required to evaluate the clinical utility of epitope matching in LT.


Assuntos
Rejeição de Enxerto/diagnóstico , Antígenos de Histocompatibilidade Classe II/imunologia , Teste de Histocompatibilidade , Isoanticorpos/sangue , Transplante de Fígado/efeitos adversos , Adulto , Idoso , Doença Hepática Terminal/cirurgia , Epitopos/imunologia , Feminino , Rejeição de Enxerto/sangue , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Sobrevivência de Enxerto , Humanos , Imunossupressores/uso terapêutico , Isoanticorpos/imunologia , Isoanticorpos/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Prognóstico , Estudos Prospectivos , Tacrolimo/uso terapêutico , Doadores de Tecidos
7.
Liver Transpl ; 24(8): 1091-1100, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29665189

RESUMO

The incidence and impact of anti-human leukocyte antigen donor-specific alloantibodies (DSAs) developing after liver transplantation (LT) remains controversial and not extensively studied. The aim of the present study was to assess the incidence of DSAs, to identify risk factors for the development of DSAs, and to understand the impact of DSAs in a large population of adult LT recipients. This single-center retrospective study included all adult patients who underwent a first LT between 2000 and 2010 in our center. The study population mainly consisted of male patients, the mean age was 52.4 years, and the main indication was alcoholic cirrhosis (54.1%). From the 297 patients included in the cross-sectional study, 14 (4.7%) had preformed DSAs, and 59 (19.9%) presented de novo DSAs (12.2% at 1 year, 13.4% at 5 years, and 19.5% at 10 years). Multivariate analysis found that female donor sex (hazard ratio [HR], 1.50; 95% confidence interval [CI], 1.12-2.01; P = 0.01) and delay between LT and DSA screening (HR, 1.10; 95% CI, 1.01-1.20; P = 0.03) were associated with occurrence of de novo DSAs. From the 190 patients included in the subgroup longitudinal analysis, exposure to tacrolimus (mean trough level during the periods 0-2 years and 0-3 years) was significantly lower for patients having DSAs at 5 years. Concerning histology, only acute rejection (P = 0.04) and portal fibrosis ≥2 (P = 0.02) were more frequent at 1 year for patients with DSAs. Patient survival and graft survival were not significantly different according to the presence or not of DSAs at 1 year. Among the 44 patients who had de novo or persistent preformed DSAs, the diagnosis of antibody-mediated rejection was made in 4 (9.1%) patients after 1, 47, 61, and 74 months following LT. In conclusion, the results of the present study suggest that DSAs are observed in a minority of LT adult patients, with limited overall impact on graft and patient outcome.


Assuntos
Rejeição de Enxerto/epidemiologia , Isoanticorpos/sangue , Cirrose Hepática Alcoólica/cirurgia , Transplante de Fígado/efeitos adversos , Complicações Pós-Operatórias/epidemiologia , Adulto , Estudos Transversais , Feminino , Seguimentos , Rejeição de Enxerto/sangue , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Sobrevivência de Enxerto , Antígenos HLA/imunologia , Humanos , Imunossupressores/uso terapêutico , Incidência , Isoanticorpos/imunologia , Isoanticorpos/isolamento & purificação , Cirrose Hepática Alcoólica/mortalidade , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/sangue , Complicações Pós-Operatórias/imunologia , Complicações Pós-Operatórias/prevenção & controle , Prevalência , Estudos Retrospectivos , Fatores de Risco , Análise de Sobrevida , Tacrolimo/uso terapêutico , Transplantados , Resultado do Tratamento
8.
Lab Med ; 48(1): 24-28, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28007780

RESUMO

Non-specific antibodies or antibodies of undetermined significance (AUS) often pose problems for a blood bank technologist and physician. It is well known that antibodies can weaken and evanesce over time, thus eluding detection by routine blood bank techniques. Special enhancement techniques exist (eg, ficin treatment); however, they are often underutilized due to concerns over expense. Ficin is known to enhance reactivity caused by antibodies in the ABO, Rh, Kidd, Lewis, I, and P blood group systems, while destroying reactivity of antibodies in the Duffy, and MNS blood group systems. Herein, we discuss our protocol for using ficin treatment to determine the specificity of antibodies that would otherwise be classified as AUS. Of the 97 AUS specimens that were treated with ficin, we were able to identify 25 new alloantibodies that would have otherwise been missed without ficin treatment. Thus, we believe our protocol enhances transfusion safety, while minimizing additional workload and cost.


Assuntos
Eritrócitos , Ficina/metabolismo , Isoanticorpos/isolamento & purificação , Testes Sorológicos/métodos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Segurança do Sangue , Criança , Pré-Escolar , Eritrócitos/química , Eritrócitos/citologia , Eritrócitos/imunologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Adulto Jovem
9.
Transfusion ; 55(3): 553-62, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25234110

RESUMO

BACKGROUND: RhIG is obtained from hyperimmunized healthy anti-D donors (HIDs) boosted with D+ red blood cells (RBCs). One hypothesis for its mechanism of action is fast clearance of opsonized D+ RBCs through Fcγ receptor (FcγR)III. Levels of immunoglobulin (Ig)G Fc-fucosylation influence interactions with FcγRIII, with less Fc-fucosylation strengthening the interaction. STUDY DESIGN AND METHODS: Anti-D IgG1 Fc-glycosylation patterns in 93 plasma samples from 28 male and 28 female Dutch HIDs and RhIG were analyzed with mass spectrometry. The Fc-glycosylation profiles of HIDs were evaluated with regard to their immunization history. RESULTS: HID sera demonstrated clearly lowered anti-D Fc-fucosylation compared to normal IgG fucosylation (93%); this was more pronounced for female than for male HIDs (47% vs. 65%, p = 0.001). RhIG preparations from seven manufacturers varied greatly in the level of Fc-fucosylation (56%-91%). The level of fucosylation slightly increased upon repeated immunization, although it remained fairly constant over time. The RhIG from the different manufacturers all demonstrated increased Fc-galactosylation (64%-82%) compared to total IgG (38%-51%). CONCLUSION: RhIG preparations vary in Fc-fucosylation and all demonstrate increased galactosylation. Despite not knowing the exact working mechanism, immunoprophylaxis could perhaps be optimized by selection of donors whose anti-D have low amounts of Fc-fucose, to increase the clearance activity of anti-D preparations, as well as high amounts of galactosylation, for anti-inflammatory effects. Implementing a biologic assay in the standardization of RhIG preparations might be considered.


Assuntos
Fucose/sangue , Imunoglobulina G/química , Isoanticorpos/química , Adulto , Doadores de Sangue , Feminino , Galactose/sangue , Glicosilação , Humanos , Imunização , Fragmentos Fc das Imunoglobulinas/química , Imunoglobulina G/isolamento & purificação , Isoanticorpos/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Gravidez , Processamento de Proteína Pós-Traducional , Receptores de IgG/metabolismo , Isoimunização Rh/terapia , Imunoglobulina rho(D) , Caracteres Sexuais , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
10.
Indian J Med Res ; 140(3): 401-5, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25366208

RESUMO

BACKGROUND & OBJECTIVES: The development of alloantibodies can significantly complicate transfusion therapy and results in difficulties in cross-matching of blood. Most literature on alloimmunization is limited to multitransfused individuals, with very few studies on the general hospital patients. This study was aimed at assessing the frequency and type of unexpected red cell antibodies in the general patient population at a multispecialty tertiary care centre in New Delhi, India. METHODS: The results of 49,077 antibody screening tests carried out on patients, from January 2009 to December 2012 were analyzed. The clinical and transfusion records were reviewed. The data were compiled and statistically analysed. RESULTS: A total of 49,077 (29,917; 60.96% males and 19,160; 39.04% females) patient samples were screened for the presence of unexpected antibodies. Antibody screening was positive in 403 patients (0.82%). In the serum samples of 164 patients only autoantibodies were identified, 27 revealed autoantibodies with one or more underlying alloantibodies, while 212 patients had only alloantibody/ies in their serum. The overall alloimmunization rate was 0.49 per cent. Antibodies against the Rh system were the most frequent (64.1%), the most common alloantibody identified being anti E (37.2%), followed by anti D (19.2%). INTERPRETATION & CONCLUSIONS: Since clinically significant antibodies are frequently detected in our patient population, antibody screening and if required, identification is the need of the hour. Since antibodies against the common Rh and Kell blood group antigens are the most frequent, provision of Rh and Kell matched red cells may be of protective value.


Assuntos
Isoanticorpos/imunologia , Sistema do Grupo Sanguíneo de Kell/imunologia , Sistema do Grupo Sanguíneo Rh-Hr/imunologia , Antígenos de Grupos Sanguíneos/imunologia , Transfusão de Sangue , Feminino , Humanos , Índia , Isoanticorpos/sangue , Isoanticorpos/isolamento & purificação , Masculino , Imunoglobulina rho(D) , Centros de Atenção Terciária
11.
Immunohematology ; 30(3): 113-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25695435

RESUMO

Elution is a procedure for recovery of antibody attached to intact,immunoglobulin-coated red blood cells (RBCs) by disrupting the antigen-antibody bonds. The recovered antibody is collected in an inert diluent and is referred to as an eluate. Testing of an eluate may be desired to identify antibody(ies) coating the RBCs of patients with a positive direct antiglobulin test. Many types of elution procedures have been developed and described; however,·an acid elution is suitable for antibody recovery in most cases, such as recovery of alloantibodies and warm-reactive autoantibodies.Studies have compared methods such as xylene, chloroform, digitnin acid, dichloromethane, citric acid, and Immucor Elu-KitII (cold acid elution). The ELU-Kit II has been shown to be quick and effective at eluting a wide range of alloantibodies as well as autoantibodies without the use of hazardous chemicals or costly reagent preparation time that some methods use. It is for these reasons that the ELU-Kit II is a very popular method for the elution of immunoglobulin G (IgG) antibodies.


Assuntos
Autoanticorpos/isolamento & purificação , Ácido Cítrico/química , Eritrócitos/química , Imunoglobulina G/isolamento & purificação , Isoanticorpos/isolamento & purificação , Kit de Reagentes para Diagnóstico , Autoanticorpos/química , Humanos , Imunoglobulina G/química , Isoanticorpos/química
12.
Immunohematology ; 29(3): 101-4, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24325170

RESUMO

Our facility changed antibody screening methods from a gel microcolumn-based test (ID-Micro Typing System Gel TEst; Ortho Clinical Diagnostics, Inc., Raritan, NJ) to an automated solid-phase test (Galileo/Capture-R Ready Screen [I and II], Immucor, Inc., Norcross, GA). To determine whether detection rates for commonly encountered clinically significant red blood cell antibodies differed as a consequence of this change, preimplementation and postimplementation antibody identification records were retrospectively reviewed. A statistically significant difference in the percentage of positive screening tests during the gel microcolumn testing period (73,903 total screens, 1.56% confirmed positive) versus the solid0-phase screening period (80,242 total screens, 1.81% confirmed positive; p< 0.0002) was observed . The number of antibodies to K identified was significantly lower with solid phase that with gel (27% decrease; p=0.004). It is unknown whether there is a statistical difference in delayed or hemolytic transfusion reaction rates as this was not evaluated.


Assuntos
Incompatibilidade de Grupos Sanguíneos/prevenção & controle , Tipagem e Reações Cruzadas Sanguíneas/estatística & dados numéricos , Eritrócitos/imunologia , Isoanticorpos/isolamento & purificação , Especificidade de Anticorpos , Transfusão de Sangue , Eritrócitos/citologia , Ensaios de Triagem em Larga Escala/instrumentação , Humanos , Estudos Retrospectivos
13.
Curr Opin Nephrol Hypertens ; 22(6): 681-8, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24076558

RESUMO

PURPOSE OF REVIEW: Humoral sensitization to antigens of the human leukocyte antigen and ABO systems remains one of the largest barriers to further expansion in renal transplantation. This barrier translates into prolonged waiting time and a greater likelihood of death. The number of highly sensitized patients on the renal transplant waiting list continues to increase. This review focuses on the options available to these patients and speculates on future directions for incompatible transplantation. RECENT FINDINGS: Desensitization protocols (to remove antibodies), kidney-paired donation (to circumvent antibodies) or a hybrid technique involving a combination of both have broadened the access to transplantation for patients disadvantaged by immunologic barriers. However, the risk of antibody-mediated rejection may be increased and warrants caution. Technical advances in antibody characterization using sensitive bead immunoassays and the C1q assay and therapeutic modalities such as complement inhibitors and proteasome inhibitors have been used to avoid or confront these antibody incompatibilities. SUMMARY: A growing body of knowledge and literature indicates that these diagnostic and therapeutic modalities can facilitate a safer and more successful treatment course for these difficult-to-treat patients. Rigorous investigations into newer interventions will help in broadening the options for these patients and also expand the living donor pool.


Assuntos
Transplante de Rim/métodos , Imunologia de Transplantes , Sistema ABO de Grupos Sanguíneos , Dessensibilização Imunológica/métodos , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/prevenção & controle , Rejeição de Enxerto/terapia , Antígenos HLA , Teste de Histocompatibilidade , Humanos , Imunidade Humoral , Isoanticorpos/isolamento & purificação , Transplante de Rim/efeitos adversos , Doadores de Tecidos , Obtenção de Tecidos e Órgãos
14.
Immunohematology ; 29(1): 5-10, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24046916

RESUMO

Routine adsorption procedures to remove autoantibodies from patients' serum often require many hours to perform. This time-consuming process can create significant delays that affect patient care. This study modified the current adsorption method to reduce total adsorption time to 1 hour. A ratio of one part serum to three parts red blood cells (RBCs; 1:3 method) was maintained for all samples. The one part serum was split into three tubes. Each of these three aliquots of serum was mixed with one full part RBCs, creating three adsorbing tubes. All tubes were incubated for 1 hour with periodic mixing. Adsorbed serum from the three tubes was harvested, combined, and tested for reactivity. Fifty-eight samples were evaluated using both the current method and the 1:3 method. Forty-eight (83%) samples successfully adsorbed using both methods. Twenty (34.5%) samples contained underlying alloantibodies. The 1:3 method demonstrated the same antibody specificities and strengths in all 20 samples. Eight samples failed to adsorb by either method. The 1:3 method found previously undetected alloantibodies in three samples. Two samples successfully autoadsorbed but failed to alloadsorb by either method. The 1:3 method proved to be efficient and effective for quick removal of autoantibodies while allowing for the detection of underlying alloantibodies.


Assuntos
Autoanticorpos/sangue , Autoanticorpos/isolamento & purificação , Testes Hematológicos/métodos , Técnicas de Imunoadsorção , Isoanticorpos/sangue , Isoanticorpos/isolamento & purificação , Adsorção , Autoanticorpos/imunologia , Eritrócitos , Humanos , Isoanticorpos/imunologia , Temperatura
15.
Transpl Int ; 26(2): 121-30, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23145861

RESUMO

Protocols for recipient desensitization may allow for successful kidney transplantation across major immunological barriers. Desensitized recipients, however, still face a considerable risk of antibody-mediated rejection (AMR), which underscores the need for risk stratification tools to individually tailor treatment. Here, we investigated whether solid phase detection of complement-fixing donor-specific antibodies (DSA) has the potential to improve AMR prediction in high-risk transplants. The study included 68 sensitized recipients of deceased donor kidney allografts who underwent peritransplant immunoadsorption for alloantibody depletion (median cytotoxic panel reactivity: 73%; crossmatch conversion: n = 21). Pre and post-transplant sera were subjected to detection of DSA-triggered C4d deposition ([C4d]DSA) applying single-antigen bead (SAB) technology. While standard crossmatch and [IgG]SAB testing failed to predict outcomes in our desensitized patients, detection of preformed [C4d]DSA (n = 44) was tightly associated with C4d-positive AMR [36% vs. 8%, P = 0.01; binary logistic regression: odds ratio: 10.1 (95% confidence interval: 1.6-64.2), P = 0.01]. Moreover, long-term death-censored graft survival tended to be worse among [C4d]DSA-positive recipients (P = 0.07). There were no associations with C4d-negative AMR or cellular rejection. [C4d]DSA detected 6 months post-transplantation were not related to clinical outcomes. Our data suggest that pretransplant SAB-based detection of complement-fixing DSA may be a valuable tool for risk stratification.


Assuntos
Anticorpos/isolamento & purificação , Complemento C4b/metabolismo , Antígenos HLA/metabolismo , Transplante de Rim/imunologia , Fragmentos de Peptídeos/metabolismo , Adsorção , Adulto , Estudos de Coortes , Feminino , Rejeição de Enxerto , Teste de Histocompatibilidade , Humanos , Imunoglobulina G/isolamento & purificação , Isoanticorpos/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Análise de Regressão , Fatores de Risco , Fatores de Tempo , Resultado do Tratamento
16.
J Clin Apher ; 27(4): 173-7, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22411078

RESUMO

Approximately 10% of renal transplant recipients experience acute antibody-mediated rejection (AMR) due to alloimmunization against human leukocyte antigen (HLA) molecules and other antigens. While therapeutic apheresis is included in most treatment protocols for acute kidney allograft rejection, these protocols have been derived mainly from single center experience rather than controlled trials. This concise review focuses on the role of therapeutic apheresis in AMR treatment. Two groups have recently reported treating acute AMR using drug-only strategies without therapeutic apheresis in particular situations, namely in clinically less severe cases or in resource-limited situations without testing for donor specific antibodies. A randomized controlled trial, designed to test the efficacy of immunoadsorption apheresis in AMR treatment, was terminated early and suggested a benefit of apheresis. An observational study suggested efficacy of plasmapheresis in acute AMR treatment, but all patients who received plasmapheresis also received rituximab. As new therapeutic modalities are becoming available, therapeutic apheresis continues to play a role in the treatment of acute kidney allograft rejection.


Assuntos
Remoção de Componentes Sanguíneos/métodos , Rejeição de Enxerto/etiologia , Rejeição de Enxerto/terapia , Transplante de Rim/efeitos adversos , Doença Aguda , Anticorpos Monoclonais Murinos/uso terapêutico , Rejeição de Enxerto/imunologia , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Técnicas de Imunoadsorção , Imunossupressores/uso terapêutico , Isoanticorpos/isolamento & purificação , Transplante de Rim/imunologia , Plasmaferese , Ensaios Clínicos Controlados Aleatórios como Assunto , Rituximab , Doadores de Tecidos
17.
Transfusion ; 52(4): 860-9, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21981750

RESUMO

BACKGROUND: According to Landsteiner's law, alloantibodies are prevalent and autoantibodies are absent in the ABO blood group system. However, one study (Spalter et al., Blood 1999;93:4418-24) has suggested that low-affinity ABO autoantibodies, mitigated by anti-idiotypic immunoglobulins are also prevalent, while another publication (Rieben et al., Eur J Immunol 1992;22:2713-7) shows that humans do not have B-lymphocytes capable of producing immunoglobulin G ABO autoantibodies. STUDY DESIGN AND METHODS: We used hapten-specific chromatography to isolate allo- and autoantibodies from pools of A or B serum and then characterized the resultant antibodies against a wide range of ABO and related glycoconjugates. RESULTS: We found that the apparent autoantibodies are directed against blood group A or B disaccharides, without consideration for the presence of fucose, but requiring the absence of elongating sugar X in composition of Gal(NAc)α1-3(Fucα1-2)Galß1-X-terminated carbohydrate chain. In contrast, ABO alloantibodies required a minimum trisaccharide Gal(NAc)α1-3(Fucα1-2)Gal epitope and recognize the elongated type-specific tetrasaccharides. Furthermore, alloantibodies appear to be a small set of specific yet crossreactive antibodies that detect all backbone types of A or B antigens, rather than being a collection of specific antibodies, each of which detects a different type of A or B antigen. CONCLUSION: Apparent ABO autoantibodies appear to have no natural human target.


Assuntos
Sistema ABO de Grupos Sanguíneos/imunologia , Autoanticorpos/imunologia , Epitopos , Isoanticorpos/imunologia , Adulto , Autoanticorpos/isolamento & purificação , Humanos , Isoanticorpos/isolamento & purificação , Polissacarídeos/metabolismo
18.
Transfusion ; 52(3): 613-21, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21883263

RESUMO

BACKGROUND: Alloantibodies against human neutrophil antigen-3 (HNA-3) are responsible for the fatalities reported in transfusion-related acute lung injury. Consequently, reliable detection of these alloantibodies is mandatory to improve blood transfusion safety. In this study, we developed stable cell lines for the detection of HNA-3 antibodies. STUDY DESIGN AND METHODS: HEK293T were transfected with HNA-3a or HNA-3b constructs and sorted by flow cytometry according to high surface expression. Transfected cells were tested with sera containing HNA-3 antibodies in flow cytometry and antibody capture assay (ACA). The results were compared with granulocyte agglutination test and granulocyte immunofluorescence test. RESULTS: In flow cytometry, 12 of 14 HNA-3a sera reacted specifically with HNA-3aa cells. One serum sample showed positive reaction with HNA-3bb cells. All HNA-3b sera recognized HNA-3bb cells. No reaction was observed with broad reactive antibodies against HLA Class I. In ACA, all HNA-3a sera (12/12) showed positive reactivity with HNA-3aa cells with no cross-reactivity with HNA-3bb cells. Again, all HNA-3b sera reacted with HNA-3bb cells only. Furthermore, genotyping of 249 individuals detected a new HNA-3 allele caused by a nucleotide substitution C>T at Position 457 leading to L(153)F mutation in choline transporter-like protein-2. This mutation impairs polymerase chain reaction with sequence-specific primers based HNA-3a typing. However, analysis with cells expressing F(153) isoform showed that this mutation did not alter the binding of HNA-3 antibodies. CONCLUSIONS: This study demonstrated that HEK293T cells expressing stable recombinant HNA-3 are suitable for the detection of HNA-3 alloantibodies allowing reliable screening of blood products.


Assuntos
Transfusão de Componentes Sanguíneos/normas , Citometria de Fluxo/métodos , Isoanticorpos/imunologia , Isoanticorpos/isolamento & purificação , Isoantígenos/genética , Isoantígenos/imunologia , Lesão Pulmonar Aguda/etiologia , Lesão Pulmonar Aguda/imunologia , Lesão Pulmonar Aguda/prevenção & controle , Testes de Aglutinação/métodos , Transfusão de Componentes Sanguíneos/efeitos adversos , Citometria de Fluxo/normas , Imunofluorescência/métodos , Genótipo , Células HEK293 , Humanos , Programas de Rastreamento/métodos , Programas de Rastreamento/normas , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/imunologia , Mutação Puntual/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Transfecção
19.
Transpl Int ; 24(12): 1142-55, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21831227

RESUMO

Antibody-mediated rejection (AMR), which is commonly caused by preformed and/or de novo HLA alloantibodies, has evolved as a leading cause of early and late kidney allograft injury. In recent years, effective treatment strategies have been established to counteract the deleterious effects of humoral alloreactivity. One major therapeutic challenge is the barrier of a positive pretransplant lymphocytotoxic crossmatch. Several apheresis- and/or IVIG-based protocols have been shown to enable successful crossmatch conversion, including a strategy of peritransplant immunoadsorption for rapid crossmatch conversion immediately before deceased donor transplantation. While such protocols may increase transplant rates and allow for acceptable graft survival, at least in the short-term, it has become evident that, despite intense treatment, many patients still experience clinical or subclinical AMR. This reinforces the need for innovative strategies, such as complementary allocation programs to improve transplant outcomes. For acute AMR, various studies have suggested efficiency of plasmapheresis- or immunoadsorption-based protocols. There is, however, no established treatment for chronic AMR and the development of strategies to reverse or at least halt chronic active rejection remains a big challenge. Major improvements can be expected from studies evaluating innovative therapeutic concepts, such as proteasome inhibition or complement blocking agents.


Assuntos
Rejeição de Enxerto/prevenção & controle , Rejeição de Enxerto/terapia , Isoanticorpos/sangue , Transplante de Rim/efeitos adversos , Transplante de Rim/imunologia , Soro Antilinfocitário/efeitos adversos , Soro Antilinfocitário/uso terapêutico , Inativadores do Complemento/uso terapêutico , Dessensibilização Imunológica , Rejeição de Enxerto/imunologia , Antígenos HLA/sangue , Teste de Histocompatibilidade , Humanos , Imunoglobulinas Intravenosas/administração & dosagem , Técnicas de Imunoadsorção , Isoanticorpos/isolamento & purificação , Plasmaferese , Inibidores de Proteassoma , Doadores de Tecidos
20.
Transplantation ; 92(1): 12-7, 2011 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-21512428

RESUMO

BACKGROUND: We have demonstrated that immunodominant donor-specific antibody (DSA) more than 100 mean fluorescence intensity (MFI) at the time of transplant is associated with a significantly higher risk of rejection. We now present short-term outcomes of DSA-based desensitization (DSZ) strategies in patients with a negative complement-dependent cytotoxicity crossmatch. METHODS: Between January 1, 2009, and January 1, 2010, live-donor kidney transplant recipients were divided into three protocols based on their immunodominant DSA MFI pretransplant (D1: 100-500, D2: 501-1000, and D3: 1001-3000). Deceased donor kidney transplant recipients were stratified into two protocols (D4: 501-1000 and D5: 1001-3000). The intensity of the conditioning treatment increased with DSA levels and included thymoglobulin induction, plasmapheresis, and intravenous immunoglobulin in the highest risk groups. We compared outcomes between desensitized patients (DSZ) and those undergoing no DSZ (or D0) during the same interval. RESULTS: Forty-eight of 249 (23%) kidney transplants underwent DSZ (n=20, 4, 3, 4, and 17 in D1-D5 protocols, respectively). There was more retransplantation (50% vs. 18%, P<0.001) and live donor transplantation (56% vs. 30%, P<0.001) in the DSZ group. In this group, mean peak panel reactive antibody and MFI at transplant were 51% ± 7% and 960 ± 136, respectively. The incidence of antibody-mediated rejection (25% vs. 12.5%, P=0.008) and acute cellular rejection (23% vs. 14%, P=0.02) was greater in the DSZ group. However, mixed rejection (8%), graft loss (0 vs. 6), patient death (0 vs. 3), cytomegalovirus infection (15% vs. 12%), and 1-year serum creatinine (1.4 ± 0.5 and 1.4 ± 0.4 mg/dL) were similar between DSZ and no-DSZ groups. CONCLUSION.: Long-term follow-up is needed to determine the role of Luminex-based strategies in current preconditioning regimens.


Assuntos
Transplante de Rim/imunologia , Transplante de Rim/métodos , Condicionamento Pré-Transplante/métodos , Adulto , Soro Antilinfocitário/uso terapêutico , Protocolos Clínicos , Feminino , Teste de Histocompatibilidade , Hospitais Universitários , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Isoanticorpos/sangue , Isoanticorpos/isolamento & purificação , Doadores Vivos , Masculino , Pessoa de Meia-Idade , Plasmaferese , Estudos Retrospectivos , Doadores de Tecidos , Resultado do Tratamento , Wisconsin
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