RESUMEN

This paper analyzes the technology-related outputs from The National Institute of Disability, Independent Living, and Rehabilitation Research (NIDILRR). We seek to answer the questions: What are the types and frequency of assistive technology (AT) technology transfer (ATTT) outputs from NIDILRR grants? How does NIDILRR's ATTT generation compare to other granting organizations? What types of ATTT outputs occur, how, and what is the relative productivity of the most frequently funded universities and small businesses performing with funding by NIDILRR grants? An online search was conducted for indications of ATTT from grants funded from 1983-2021 through publicly available databases, the National Rehabilitation Information Center (NARIC), and the internet. This data was then categorized across relevant output types and analyzed. NIDILRR funded 662 organizations and 951 different investigators from 1983 to 2021. The NIDILRR-funded portfolio includes 6,996 papers, 438 informational websites, 163 patents, 120 software products, and 29 hardware products. Compared to the National Institutes of Health (NIH), NIDILRR produced slightly more products per dollar. Our results highlight the substantial portfolio of technology-related outputs generated with NIDILRR funding and demonstrate how productivity measures can be calculated to guide future funding strategies.

RESUMEN

To explore global trends in manual wheelchair service provision knowledge across geographic, professional, and socioeconomic domains. A secondary analysis of a dataset from the International Society of Wheelchair Professionals' Wheelchair Service Provision Basic Knowledge Test was conducted. The dataset included test takers from around the world and was extracted from Test.com and International Society of Wheelchair Professionals' Wheelchair International Network. Participants 2,467 unique test takers from 86 countries. Interventions Not applicable. International Society of Wheelchair Professionals' Wheelchair Service Provision Basic Knowledge Test. We identified significant inverse associations between pass rate and the following variables: education (high school and some college), test taker motivation (required by academic program or employer), and country income setting (low and middle). There were significant positive associations between pass rate and the following variables: training received (offered by Mobility India or 'other NGO'), and age group served (early childhood). Global wheelchair knowledge trends related to key variables such as training, occupation, and income setting have been preliminarily explored. Future work includes further validation of the primary outcome measure and recruitment of a larger sample size to further explore significant associations between additional test taker variables.

Asunto(s)

Silla de Ruedas , Preescolar , Humanos , Universidades , Demografía , India

RESUMEN

PURPOSE: The objectives of this mixed-methods study were to gather survey and interview data about the barriers and facilitators from grantees funded by the National Institute on Disability, Independent Living, and Rehabilitation Research (NIDILRR) and to extract themes that could inform program changes that would increase technology translation (TT) success in assistive technology (AT). MATERIALS AND METHODS: We developed a TT Barriers and Facilitators survey consisting of Likert scale and multiple-choice questions about barriers and facilitators to TT. With survey respondents who were willing, we conducting a semi-structured interview and asked pointed questions to expand upon survey response rankings and perceived barriers and facilitators. The questions were framed to explore the grantee's personal experience with ATTT and what helped and hindered their individualised processes. RESULTS: Across survey and interview respondents, the three most common themes when exploring the barriers and facilitators of TT were funding, incentives, and collaboration. CONCLUSIONS: Results indicate that there is a need for increased collaboration and access to additional resources such as funding for pilot grants, support to assess technology marketability, help to navigate regulatory and legal aspects, and assistance in establishing goals to help grantees successfully transfer assistive technologies to consumers. IMPLICATIONS FOR REHABILITATIONA large amount of research and development into assistive technology does not lead to tech transfer which means that these technologies are not getting to the people that need them.Educating tech transfer offices at universities about how to transfer AT would improve outcomes greatly.Creating a community of practice where grantees can find academic or industry partners would also increase the likelihood of tech transfer.Some tools to catalyse these improvements are: mentoring, access to consultants, podcasts, and online training.

RESUMEN

Assistive products outcomes are needed globally to inform policy, practice, and drive investment. The International Society of Wheelchair Professionals developed a Minimum Uniform Dataset (MUD) for wheelchair services worldwide with the intent to gather data that is comparable globally. The MUD was developed with the participation of members from around the globe and its feasibility piloted at 3 sites. Three versions of the MUD are now available-a short form with 29 data points (available in English, Spanish, and French) and a standard version with 38 data points in English. Future work is to validate and complete the translation cycles followed by promoting the use of the MUD globally so that the data can be leveraged to inform policy, practice and direct investments.

Asunto(s)

Recolección de Datos/normas , Personas con Discapacidad/rehabilitación , Encuestas y Cuestionarios/normas , Análisis y Desempeño de Tareas , Silla de Ruedas/normas , Humanos , Agencias Internacionales , Traducción

RESUMEN

PURPOSE: Combined immunodeficiency (CID) presents a unique challenge to clinicians. Two patients presented with the prior clinical diagnosis of common variable immunodeficiency (CVID) disorder marked by an early age of presentation, opportunistic infections, and persistent lymphopenia. Due to the presence of atypical clinical features, next generation sequencing was applied documenting RAG deficiency in both patients. METHODS: Two different genetic analysis techniques were applied in these patients including whole exome sequencing in one patient and the use of a gene panel designed to target genes known to cause primary immunodeficiency disorders (PIDD) in a second patient. Sanger dideoxy sequencing was used to confirm RAG1 mutations in both patients. RESULTS: Two young adults with a history of recurrent bacterial sinopulmonary infections, viral infections, and autoimmune disease as well as progressive hypogammaglobulinemia, abnormal antibody responses, lymphopenia and a prior diagnosis of CVID disorder were evaluated. Compound heterozygous mutations in RAG1 (1) c256_257delAA, p86VfsX32 and (2) c1835A>G, pH612R were documented in one patient. Compound heterozygous mutations in RAG1 (1) c.1566G>T, p.W522C and (2) c.2689C>T, p. R897X) were documented in a second patient post-mortem following a fatal opportunistic infection. CONCLUSION: Astute clinical judgment in the evaluation of patients with PIDD is necessary. Atypical clinical findings such as early onset, granulomatous disease, or opportunistic infections should support the consideration of atypical forms of late onset CID secondary to RAG deficiency. Next generation sequencing approaches provide powerful tools in the investigation of these patients and may expedite definitive treatments.

Asunto(s)

Inmunodeficiencia Variable Común/genética , Proteínas de Homeodominio/genética , Mutación , Agammaglobulinemia/diagnóstico , Agammaglobulinemia/etiología , Biopsia , Preescolar , Inmunodeficiencia Variable Común/complicaciones , Inmunodeficiencia Variable Común/diagnóstico , Análisis Mutacional de ADN , Resultado Fatal , Femenino , Humanos , Inmunohistoquímica , Enfermedades Pulmonares Intersticiales/diagnóstico , Enfermedades Pulmonares Intersticiales/etiología , Linfopenia/diagnóstico , Linfopenia/etiología , Tomografía Computarizada por Rayos X , Adulto Joven

RESUMEN

BACKGROUND: The role of T-helper 17 cells (Th17) in neonatal host defense remains to be fully elucidated. Interleukin (IL)-17 plays an important role in the immune response to bacterial and fungal pathogens by promoting inflammation. METHODS: We examined neonatal production of IL-17 in mixed mononuclear cells (MMCs) isolated from umbilical cord blood for comparison with adult peripheral blood mononuclear cell controls. RESULTS: IL-17 production was profoundly diminished in MMCs isolated from cord blood when compared with MMCs from adult blood. This was associated with a marked reduction in the population of CCR6+ IL-17(+) T-cells in the neonatal cord blood. We also found diminished intracellular formation of IL-17, and diminished IL-17 responses to both group B streptococci (GBS) and Escherichia coli. Neonatal mononuclear cells were found to adequately phosphorylate signal transducer and activator of transcription 3, pY705, and pS727. We and others have reported markedly reduced interferon-γ production by neonate mononuclear cells exposed to GBS. Here, we correct that profound abnormality with added IL-17. CONCLUSION: Our results suggest that profound deficiency of IL-17 production associated with a marked decrease in Th17 cells likely contributes significantly to the increased susceptibility of human neonates to invasive bacterial and fungal infections.

Asunto(s)

Sangre Fetal/metabolismo , Interleucina-17/metabolismo , Células Th17/metabolismo , Adulto , Recuento de Linfocito CD4 , Células Cultivadas , Regulación hacia Abajo , Escherichia coli/inmunología , Escherichia coli/patogenicidad , Sangre Fetal/citología , Sangre Fetal/inmunología , Interacciones Huésped-Patógeno , Humanos , Recién Nacido , Interferón gamma/metabolismo , Fosforilación , Receptores CCR6/metabolismo , Factor de Transcripción STAT3/metabolismo , Streptococcus/inmunología , Streptococcus/patogenicidad , Células Th17/inmunología , Células Th17/microbiología

RESUMEN

Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by antibody deficiency, poor humoral response to antigens, and recurrent infections. To investigate the molecular cause of CVID, we carried out exome sequence analysis of a family diagnosed with CVID and identified a heterozygous frameshift mutation, c.2564delA (p.Lys855Serfs(∗)7), in NFKB2 affecting the C terminus of NF-κB2 (also known as p100/p52 or p100/p49). Subsequent screening of NFKB2 in 33 unrelated CVID-affected individuals uncovered a second heterozygous nonsense mutation, c.2557C>T (p.Arg853(∗)), in one simplex case. Affected individuals in both families presented with an unusual combination of childhood-onset hypogammaglobulinemia with recurrent infections, autoimmune features, and adrenal insufficiency. NF-κB2 is the principal protein involved in the noncanonical NF-κB pathway, is evolutionarily conserved, and functions in peripheral lymphoid organ development, B cell development, and antibody production. In addition, Nfkb2 mouse models demonstrate a CVID-like phenotype with hypogammaglobulinemia and poor humoral response to antigens. Immunoblot analysis and immunofluorescence microscopy of transformed B cells from affected individuals show that the NFKB2 mutations affect phosphorylation and proteasomal processing of p100 and, ultimately, p52 nuclear translocation. These findings describe germline mutations in NFKB2 and establish the noncanonical NF-κB signaling pathway as a genetic etiology for this primary immunodeficiency syndrome.

Asunto(s)

Inmunodeficiencia Variable Común/genética , Mutación de Línea Germinal , Subunidad p52 de NF-kappa B/genética , Transducción de Señal , Adolescente , Adulto , Secuencia de Aminoácidos , Animales , Linfocitos B/citología , Linfocitos B/metabolismo , Línea Celular , Niño , Inmunodeficiencia Variable Común/patología , Modelos Animales de Enfermedad , Femenino , Pruebas Genéticas , Heterocigoto , Humanos , Inmunoglobulina A/sangre , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Microscopía Confocal , Datos de Secuencia Molecular , Subunidad p52 de NF-kappa B/metabolismo , Linaje , Fenotipo , Adulto Joven

Asunto(s)

Síndromes de Inmunodeficiencia/genética , Timoma/genética , Proteína Activadora Transmembrana y Interactiva del CAML/genética , Femenino , Humanos , Masculino

RESUMEN

Chronic granulomatous disease (CGD) is a rare congenital disorder in which phagocytes cannot generate superoxide (O(2)(-)) and other microbicidal oxidants due to mutations in one of the five components of the O(2)(-)-generating NADPH oxidase complex. The most common form is caused by mutations in CYBB on the X chromosome, encoding gp91phox, the enzymatic subunit of the phagocyte NADPH oxidase. Here, we report two rare cases of male X-linked CGD patients, one caused by a 5.7-kb duplication of a region containing CYBB exons 6 to 8 and the other caused by a deletion of this same region. We found both the duplication in patient 1 and the deletion in patient 2 to be bordered by a GT repeat. Indeed, in control DNA, the 3' part of CYBB intron 5 contains a GT repeat and the 5' part of intron 8 also contains such a repeat. Duplication of exons 6, 7 and 8 in patient 1 was probably caused by a non-homologous crossing over between the two GT repeats. The deletion found in patient 2 probably arose from a similar misalignment. The results found in these patients were confirmed by multiplex ligation-dependent probe amplification. The clinical profile of XCGD is severe in both patients.

Asunto(s)

Enfermedades Genéticas Ligadas al Cromosoma X/genética , Enfermedad Granulomatosa Crónica/genética , Glicoproteínas de Membrana/genética , NADPH Oxidasas/genética , Adolescente , Niño , Exones , Duplicación de Gen , Predisposición Genética a la Enfermedad , Humanos , Masculino , Mutación , NADPH Oxidasa 2 , Eliminación de Secuencia

Asunto(s)

Recolección de Datos , Grupos Diagnósticos Relacionados/clasificación , Mecanismo de Reembolso , Regulación Gubernamental , Casas de Salud/legislación & jurisprudencia , Diseño de Software

RESUMEN

The Job or hyper-immunoglobulinemia E syndrome is a primary immunodeficiency that is usually inherited in an autosomal dominant fashion. With the discovery of mutations in the STAT3 gene in the majority of autosomal dominant cases, it is now possible to make a molecular diagnosis of hyper-IgE syndrome. Both primary and secondary immunodeficiencies, including hyper-IgE syndrome, may predispose for malignancies, especially lymphomas, mainly mature B cell lymphomas, and classical Hodgkin lymphoma. Here, we report of a 48-year-old male with hyper-IgE syndrome who developed a primary parotid gland diffuse large B cell lymphoma. Analysis for STAT3 mutations demonstrated that the causal mutation of hyper-IgE syndrome, R382Q, arose de novo in the patient and it was transmitted to three of his five children, all three of whom are clinically affected. We review the literature regarding lymphoma in hyper-IgE syndrome and the possible etiologic relationship with STAT3 mutations.

Asunto(s)

Síndrome de Job/genética , Linfoma de Células B/genética , Factor de Transcripción STAT3/genética , Adolescente , Niño , Trastornos de los Cromosomas , Humanos , Síndrome de Job/complicaciones , Síndrome de Job/inmunología , Síndrome de Job/fisiopatología , Linfoma de Células B/etiología , Linfoma de Células B/inmunología , Linfoma de Células B/fisiopatología , Masculino , Persona de Mediana Edad , Mutación/genética

RESUMEN

High-resolution melting analysis was applied to X-linked chronic granulomatous disease, a rare disorder resulting from mutations in CYBB. Melting curves of the 13 PCR products bracketing CYBB exons were predicted by Poland's algorithm and compared with observed curves from 96 normal individuals. Primer plates were prepared robotically in batches and dried, greatly simplifying the 3- to 6-hour workflow that included DNA isolation, PCR, melting, and cycle sequencing of any positive products. Small point mutations or insertions/deletions were detected by mixing the hemizygous male DNA with normal male DNA to produce artificial heterozygotes, whereas detection of gross deletions was performed on unmixed samples. Eighteen validation samples and 22 clinical kindreds were analyzed for CYBB mutations. All blinded validation samples were correctly identified. The clinical probands were identified after screening for neutrophil oxidase activity. Nineteen different mutations were found, including seven near intron-exon boundaries predicting splicing defects, five substitutions within exons, three small deletions predicting premature termination, and four gross deletions of multiple exons. Ten novel mutations were found, including (c.) two missense (730T>A, 134T>G), one nonsense (90C>A), four splice site defects (45 + 1G>T, 674 + 4A>G, 1461 + 2delT, and 1462-2A>C), two small deletions (636delT, 1661_1662delCT), and one gross deletion of exons 6 to 8. High-resolution melting can provide timely diagnosis at low cost for effective clinical management of rare, genetic primary immunodeficiency disorders.

Asunto(s)

Genes Ligados a X/genética , Enfermedad Granulomatosa Crónica/genética , Reacción en Cadena de la Polimerasa/métodos , Femenino , Humanos , Masculino , Glicoproteínas de Membrana/genética , NADPH Oxidasa 2 , NADPH Oxidasas/genética , Análisis de Secuencia de ADN

RESUMEN

With the recent discovery of mutations in the STAT3 gene in the majority of patients with classic Hyper-IgE syndrome, it is now possible to make a molecular diagnosis in most of these cases. We have developed a PCR-based high-resolution DNA-melting assay to scan selected exons of the STAT3 gene for mutations responsible for Hyper-IgE syndrome, which is then followed by targeted sequencing. We scanned for mutations in 10 unrelated pedigrees, which include 16 patients with classic Hyper-IgE syndrome. These pedigrees include both sporadic and familial cases and their relatives, and we have found STAT3 mutations in all affected individuals. High-resolution melting analysis allows a single day turn-around time for mutation scanning and targeted sequencing of the STAT3 gene, which will greatly facilitate the rapid diagnosis of the Hyper-IgE syndrome, allowing prompt and appropriate therapy, prophylaxis, improved clinical outcome, and accurate genetic counseling.

Asunto(s)

Análisis Mutacional de ADN/métodos , Síndrome de Job , Factor de Transcripción STAT3/genética , Exones , Femenino , Humanos , Síndrome de Job/genética , Síndrome de Job/inmunología , Síndrome de Job/fisiopatología , Masculino , Datos de Secuencia Molecular , Mutación , Linaje , Análisis de Secuencia de ADN/métodos

RESUMEN

BACKGROUND: The human neonate's increased susceptibility to bacterial infections is not completely understood. Toll-like receptors (TLRs) have been recognized as pattern-recognition receptors critical to the innate immune response. TLR function in neonates, however, remains incompletely defined. OBJECTIVE: To examine regulatory and proinflammatory cytokine responses to TLR-1-6 stimulation of cord blood compared to adult blood. METHODS: We stimulated cord blood with ligands for each of TLRs 1-6 and compared these responses to adult controls. The following TLR ligands were utilized: Pam3CSK4 (TLR-1 and 2), zymosan (TLR-2 and 6), poly I:C (TLR-3), LPS (TLR-4), and flagellin (TLR-5). Cytokine production was measured with an assay developed in-house utilizing multi-analyte technology. RESULTS: TLR-1-6 stimulation produced higher concentrations of proinflammatory cytokines (IL-1beta, IL-6, and IL-8) in cord blood compared to adult blood, with the exception of TLR-4-stimulated TNF-alpha production, which was significantly lower in cord blood (319 pg/ml) compared to adult blood (645 pg/ml; p = 0.027). In contrast, TLR-1-6 stimulation resulted in decreased concentrations of Th1 and Th2 cytokines in cord blood compared to adult blood, with significantly diminished production of IL-12 (TLRs 1/2, 2/6, 3 and 4), IL-13 (TLR-1-6), and IL-10 (TLR-4). CONCLUSION: Cord blood production of regulatory Th1 and Th2 cytokines following TLR stimulation is decreased compared to that of adult blood. In contrast, TLR-stimulated proinflammatory cytokine production was markedly higher in neonates than in adults, with the exception of TLR-4-induced TNF-alpha production. The human neonate's increased susceptibility to bacterial infections may be related to abnormal TLR responsiveness, with enhanced proinflammatory and decreased regulatory cytokine production.

Asunto(s)

Análisis Químico de la Sangre/métodos , Citocinas/análisis , Citocinas/metabolismo , Receptores Toll-Like/fisiología , Adulto , Citocinas/sangre , Sangre Fetal/inmunología , Sangre Fetal/metabolismo , Flagelina/farmacología , Humanos , Inmunidad Innata/fisiología , Inmunoensayo/métodos , Recién Nacido , Ligandos , Lipopéptidos/farmacología , Lipopolisacáridos/farmacología , Poli I-C/farmacología , Pruebas Serológicas/métodos , Células TH1/inmunología , Células TH1/metabolismo , Células Th2/inmunología , Células Th2/metabolismo , Receptores Toll-Like/agonistas , Zimosan/farmacología

RESUMEN

Chronic granulomatous disease (CGD) is a rare inherited immunodeficiency disorder. The clinical presentation is varied depending on the degree of involvement of the NADPH oxidase system responsible for the oxidative burst of neutrophils. We present 3 cases of variant X-linked CGD in an effort to introduce the disease and highlight the importance and limitations of CGD screening. The variant X-linked form of CGD results in a less severe phenotype and frequently presents later in life. Variant X-linked CGD is difficult to diagnose, but is becoming more readily recognized based on improved testing methods. A high index of suspicion in the setting of unusual infections such as Burkholderia cepacia pneumonia is essential to make the diagnosis. Family screening can lead to early intervention, prophylaxis, and appropriate genetic counseling.

Asunto(s)

Enfermedad Granulomatosa Crónica , Adolescente , Adulto , Líquido del Lavado Bronquioalveolar/microbiología , Infecciones por Burkholderia/microbiología , Burkholderia cepacia/aislamiento & purificación , Niño , Preescolar , Familia , Femenino , Tamización de Portadores Genéticos , Variación Genética , Enfermedad Granulomatosa Crónica/diagnóstico , Enfermedad Granulomatosa Crónica/genética , Humanos , Masculino , Glicoproteínas de Membrana/genética , NADPH Oxidasa 2 , NADPH Oxidasas/genética , Neutrófilos/metabolismo , Linaje , Rodaminas/metabolismo

RESUMEN

Hyperimmunoglobulin E syndrome (HIES) is a rare primary immunodeficiency characterized by recurrent skin and lung infections. We report the first case of Coccidioides immitis meningitis in a patient with HIES. Coccidioides should be included in the differential diagnosis for central nervous system infections in HIES patients.

Asunto(s)

Coccidioides/aislamiento & purificación , Coccidioidomicosis/diagnóstico , Síndrome de Job/complicaciones , Síndrome de Job/diagnóstico , Meningitis Fúngica/diagnóstico , Mutación Missense , Factor de Transcripción STAT3/genética , Adolescente , Sustitución de Aminoácidos/genética , Anfotericina B/uso terapéutico , Anticuerpos Antifúngicos/líquido cefalorraquídeo , Antifúngicos/uso terapéutico , Líquido Cefalorraquídeo/microbiología , Coccidioidomicosis/tratamiento farmacológico , Coccidioidomicosis/microbiología , Coccidioidomicosis/patología , Exones/genética , Femenino , Humanos , Inmunoglobulina G/líquido cefalorraquídeo , Síndrome de Job/tratamiento farmacológico , Síndrome de Job/patología , Meningitis Fúngica/tratamiento farmacológico , Meningitis Fúngica/microbiología , Meningitis Fúngica/patología , Análisis de Secuencia de ADN

RESUMEN

Acute rheumatic fever (ARF) is an autoimmune disease occurring in individuals following untreated group A streptococcal infection believed to be triggered by antibodies to bacterial components that cross-react with human tissues. We developed a multiplexed immunoassay for the simultaneous quantitation of antibodies to nine streptococcal-related antigens including streptolysin O (SLO), DNase B, collagen I and IV, fibronectin, myosin, group A carbohydrate, M6 protein and streptococcal C5a peptidase. Utilizing this method, we examined serum from 49 ARF, 58 pharyngitis patients and age- and sex-matched controls in samples collected at initial disease onset, and at 4 weeks, 6 months and 1 year after diagnosis. Antibody responses were significantly higher for SLO, DNase B, M6 protein, group A carbohydrate and the cross-reactive antigens collagen I and myosin in ARF compared with pharyngitis patients (P

Asunto(s)

Anticuerpos Antibacterianos/sangre , Formación de Anticuerpos/inmunología , Fiebre Reumática/inmunología , Infecciones Estreptocócicas/inmunología , Enfermedad Aguda , Adolescente , Adulto , Anticuerpos Antibacterianos/inmunología , Antígenos Bacterianos/inmunología , Niño , Preescolar , Colágeno/inmunología , Reacciones Cruzadas/inmunología , Femenino , Humanos , Masculino , Miocarditis/inmunología , Cardiopatía Reumática/inmunología , Streptococcus/inmunología , Adulto Joven

RESUMEN

Lymphocyte proliferation in response to mitogens, phytohemagglutinin (PHA), concanavalin A, pokeweed, and/or specific antigens has been the method of choice for in vitro diagnosis of cell-mediated immune dysfunction. Recently, an assay to measure intracellular adenosine triphosphate (ATP) production in response to PHA has been developed that requires a shorter, overnight incubation. We compared a standard 5- to 7-day lymphocyte mitogen stimulation assay utilizing tritiated thymidine (3H-thy) incorporation to one in which ATP production in response to PHA by CD4-positive cells is measured in a luminometer that requires only 18-24 hr. A total of 20 patient samples suspected of having decreased cell-mediated immunity submitted for mitogen induced lymphocyte proliferation and 21 normal controls were tested in both assays. A comparison of these two methods has demonstrated that the screening ATP assay has a sensitivity at 24 hr of 100% in detecting decreased PHA induced lymphocyte proliferation at 5 days and a specificity of 85% in the samples obtained from normal controls. The data indicate that the ATP assay may be a useful screening tool for more rapid detection of blood samples with decreased cell-mediated immune responses. However, a positive screen should always be confirmed by 3H-thy uptake using mitogens and recall antigens like candida and tetanus.

Asunto(s)

Adenosina Trifosfato/sangre , Síndromes de Inmunodeficiencia/diagnóstico , Activación de Linfocitos , Mitógenos/farmacología , Fitohemaglutininas/farmacología , Adolescente , Adulto , Proliferación Celular/efectos de los fármacos , Niño , Preescolar , Femenino , Humanos , Inmunidad Celular/fisiología , Síndromes de Inmunodeficiencia/inmunología , Pruebas Inmunológicas/métodos , Lactante , Activación de Linfocitos/efectos de los fármacos , Linfocitos/citología , Linfocitos/efectos de los fármacos , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad

RESUMEN

The host immunologic response to group A streptococcal infections gives rise to numerous antibodies directed against cellular and extracellular bacterial antigens. For determining individual immune status, or studying the pathogenesis of group A streptococcal associated diseases, such as acute rheumatic fever (ARF), an assay capable of determining antibodies responses to multiple antigens would be of great advantage. We have developed a microsphere based, multiplexed immunoassay for the simultaneous quantitation of antibodies to nine different extracellular, ARF related tissue and group A streptococci specific antigens using only 5 microl of sample. Through the selection of microspheres and serum diluent, non-specific antibody binding was reduced by 17%. Different formulations of the coupling buffer were found to greatly influence the efficiency of coupling antigens to the carboxylated microspheres. Monoclonal antibodies against the different antigens demonstrated assay specificity as well as sensitivities of less than 1 ng/ml of antibody. This multiplexed assay should be a powerful research and clinical tool in determining antibody responses to group A streptococcal infections and in potentially determining the role of a variety of cross-reactive antigens in rheumatic fever and rheumatic heart disease.

Asunto(s)

Anticuerpos Antibacterianos/sangre , Citometría de Flujo/métodos , Fiebre Reumática/inmunología , Infecciones Estreptocócicas/inmunología , Streptococcus pyogenes/inmunología , Anticuerpos Antibacterianos/inmunología , Especificidad de Anticuerpos , Antígenos Bacterianos/inmunología , Humanos , Microesferas , Fiebre Reumática/microbiología , Infecciones Estreptocócicas/sangre , Infecciones Estreptocócicas/microbiología

RESUMEN

D8/17, an alloantigen found on B lymphocytes, has been reported to be elevated in patients susceptible to rheumatic fever and may be associated with autoimmune types of neuropsychiatric disorders. The pediatric-autoimmune-neuropsychiatric-disorders-associated-with-streptococci model is a putative model of pathogenesis for a group of children whose symptoms of obsessive-compulsive disorder and Tourette's disorder (TD) are abrupt and may be triggered by an infection with group A streptococci. As a test of this model, we have examined D8/17 levels on the B cells of patients with TD and acute rheumatic fever (ARF) along with those on the B cells of normal controls by flow cytometry. We have utilized several different preparations of D8/17 antibody along with a variety of secondary antibodies but have been unable to show an association with an elevated percentage of D8/17-positive, CD19-positive B cells in either ARF or TD. We did find, however, that the percentages of CD19-positive B cells in ARF and TD patients were significantly elevated compared to those in normal controls. Group A streptococcal pharyngitis patients also had an elevated percentage of CD19 B cells, however. These studies failed to confirm the utility of determining the percentage of B cells expressing the D8/17 alloantigen in ARF patients or our sample of TD patients. In contrast, the percentage of CD19-positive B cells was significantly elevated in ARF and TD patients, as well as group A streptococcal pharyngitis patients, suggesting a role for inflammation and/or autoimmunity in the pathogenesis of these disorders.

Asunto(s)

Antígenos CD19/metabolismo , Linfocitos B/metabolismo , Isoantígenos/metabolismo , Fiebre Reumática/inmunología , Síndrome de Tourette/inmunología , Enfermedad Aguda , Antígenos CD19/inmunología , Autoinmunidad/inmunología , Linfocitos B/inmunología , Citometría de Flujo , Humanos , Isoantígenos/inmunología , Faringitis/inmunología , Infecciones Estreptocócicas/inmunología