RESUMEN
OBJECTIVE: This was an open-label, phase 1/2 dose-escalation, safety trial of intrathecal recombinant human heparan-N-sulfatase (rhHNS) administered via intrathecal drug delivery device (IDDD) for treating mucopolysaccharidosis IIIA (NCT01155778). STUDY DESIGN: Twelve patients received 10, 45, or 90mg of rhHNS via IDDD once monthly for a total of 6 doses. Primary endpoints included adverse events (AEs) and anti-rhHNS antibodies. Secondary endpoints included standardized neurocognitive assessments, cortical gray matter volume, and pharmacokinetic/pharmacodynamic analyses. RESULTS: All patients experienced treatment-emergent AEs; most of mild-to-moderate severity. Seven patients reported a total of 10 serious AEs (SAEs), all but one due to hospitalization to revise a nonfunctioning IDDD. No SAEs were considered related to rhHNS. Anti-rhHNS antibodies were detected in the serum of 6 patients and in the cerebrospinal fluid (CSF) of 2 of these. CSF heparan sulfate levels were elevated at baseline and there were sustained declines in all tested patients following the first rhHNS dose. No impact of anti-rhHNS antibodies on any pharmacodynamic or safety parameters was evident. 4 of 12 patients showed a decline in developmental quotient, 6 were stable, and 2 patients had only a single data point. No dose group showed a clearly different response pattern. CONCLUSIONS: rhHNS administration via IDDD appeared generally safe and well tolerated. Treatment resulted in consistent declines in CSF heparan sulfate, suggesting in vivo activity in the relevant anatomical compartment. Results of this small study should be interpreted with caution. Future studies are required to assess the potential clinical benefits of rhHNS and to test improved IDDD models.
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Heparitina Sulfato/líquido cefalorraquídeo , Mucopolisacaridosis III/tratamiento farmacológico , Sulfatasas/administración & dosificación , Adolescente , Anticuerpos/sangre , Anticuerpos/líquido cefalorraquídeo , Niño , Preescolar , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Inyecciones Espinales/instrumentación , Masculino , Mucopolisacaridosis III/líquido cefalorraquídeo , Sulfatasas/efectos adversos , Sulfatasas/inmunología , Resultado del Tratamiento , Adulto JovenRESUMEN
The Sanfilippo Behavior Rating Scale (SBRS), a 68 item questionnaire, has been developed to assess the behavioral phenotype of children with Sanfilippo syndrome and its progression over time. Fifteen scales rate orality, movement/activity, attention/self-control, emotional function including anger and fear, and social interaction. Items within scales intercorrelate; measures of internal consistency are adequate. Twelve scales are grouped into 4 abnormality clusters: Movement, Lack of fear, Social/emotional and Executive Dysfunction. A Loess age-trajectory analysis showed that Lack of Fear, Social/Emotional and Executive Dysfunction increased steadily with age; Orality and Mood/Anger/Aggression leveled off. Movement peaked around 6years, then declined as children's excessive/purposeless actions stopped. Compared with standard scales, SBRS Movement was appropriately associated with the Vineland Motor scale; SBRS Lack of Fear had significant associations with the Autism Diagnostic Observation Schedule (ADOS), indicating a symptom overlap between Sanfilippo syndrome and autism. This suggests that reduced fearfulness may be the most salient/sensitive SBRS marker of disease progression. Volumetric MRI showed that increased Lack of Fear was significantly associated with reduced amygdala volume, consistent with our hypothesis that the behavior seen in Sanfilippo syndrome is a variant of Klüver-Bucy syndrome. Hippocampal volume loss had twice the effect on Social-Emotional Dysfunction as amygdala loss, consistent with a hippocampal role in attachment and social emotions. In conclusion, the SBRS assesses the Sanfilippo behavioral phenotype; it can measure behavior change that accompanies disease progression and/or results from treatment.
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Escala de Evaluación de la Conducta , Conducta , Mucopolisacaridosis III/psicología , Adolescente , Amígdala del Cerebelo/patología , Trastorno Autístico/diagnóstico , Trastorno Autístico/psicología , Niño , Preescolar , Miedo , Femenino , Humanos , Lactante , Masculino , Mucopolisacaridosis III/diagnóstico , Mucopolisacaridosis III/patología , Neuroimagen , Fenotipo , Conducta Social , Encuestas y CuestionariosRESUMEN
Understanding mechanisms of hepatocellular damage may lead to new treatments for liver disease, and genome-wide association studies (GWAS) of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) serum activities have proven useful for investigating liver biology. Here we report 100 loci associating with both enzymes, using GWAS across 411,048 subjects in the UK Biobank. The rare missense variant SLC30A10 Thr95Ile (rs188273166) associates with the largest elevation of both enzymes, and this association replicates in the DiscovEHR study. SLC30A10 excretes manganese from the liver to the bile duct, and rare homozygous loss of function causes the syndrome hypermanganesemia with dystonia-1 (HMNDYT1) which involves cirrhosis. Consistent with hematological symptoms of hypermanganesemia, SLC30A10 Thr95Ile carriers have increased hematocrit and risk of iron deficiency anemia. Carriers also have increased risk of extrahepatic bile duct cancer. These results suggest that genetic variation in SLC30A10 adversely affects more individuals than patients with diagnosed HMNDYT1.
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Alanina Transaminasa/sangre , Aspartato Aminotransferasas/sangre , Proteínas de Transporte de Catión/genética , Estudio de Asociación del Genoma Completo , Manganeso/sangre , Mutación/genética , Proteínas de Transporte de Catión/metabolismo , Regulación de la Expresión Génica , Ligamiento Genético , Sitios Genéticos , Genoma Humano , Células HeLa , Hematócrito , Heterocigoto , Homeostasis , Humanos , Hígado/patología , Manganeso/metabolismo , Anotación de Secuencia Molecular , Fenotipo , Reproducibilidad de los ResultadosRESUMEN
Lepromatous leprosy is a model of immune evasion wherein pathogen-specific IL-10-secreting T cells and concomitant failure of Th-1 immunity permit uncontrolled proliferation of the intracellular pathogen, Mycobacterium leprae (M. leprae). The mechanism of this immune escape is unknown. Here, the authors report that phenolic glycolipid-1 (PGL-1), a major and distinguishing feature of the M. leprae cell wall, is expressed in the cell membrane of M. leprae-infected human dendritic cells, where it can activate complement in human serum. The authors demonstrate that PGL-1 and the C3 component of complement colocalize in lipid rafts in the dendritic cell membrane, and enter the immune synapse upon co-culture of M. leprae-infected DCs and T cells. Hence, activated C3 is strategically located to costimulate naïve T cells via the complement regulatory protein, CD46, a process known to stimulate the differentiation of IL-10-secreting regulatory T cells. These observations suggest a potential novel mechanism of immune evasion, wherein M. leprae may subvert host natural immunity to provoke an adaptive response that favors bacillary survival.
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Lepra/inmunología , Mycobacterium leprae/inmunología , Inmunidad Adaptativa , Antígenos Bacterianos/metabolismo , Activación de Complemento , Complemento C3/inmunología , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Células Dendríticas/microbiología , Glucolípidos/metabolismo , Humanos , Interleucina-10/metabolismo , Mycobacterium leprae/metabolismo , Linfocitos T/inmunologíaRESUMEN
Leprosy and human immunodeficiency virus-1 (HIV-1) are examples of human infections where interactions between the pathogen and the host cellular immunity determine the clinical manifestations of disease. Hence, a significant immunopathological interaction between HIV-1 and leprosy might be expected. In the present study we explored several aspects of cellular immunity in patients co-infected with HIV-1 and Mycobacterium leprae. Twenty-eight individuals were studied, comprising four groups: healthy controls, HIV-1 and M. leprae co-infection, HIV-1 mono-infection, and M. leprae mono-infection. Subjects in the mono-infection and co-infection groups were matched as far as possible for bacillary load and HIV disease status, as appropriate. Peripheral blood mononuclear cells (PBMC) were analysed using six- and seven-colour flow cytometry to evaluate T-cell subpopulations and their activation status, dendritic cell (DC) distribution phenotypes and expression of IL-4 by T cells. The co-infected group exhibited lower CD4 : CD8 ratios, higher levels of CD8(+) T-cell activation, increased V delta : V delta 2 T cell ratios and decreased percentages of plasmacytoid DC, compared with HIV-1 mono-infected subjects. Across infected groups, IL-4 production by CD4(+) T lymphocytes was positively correlated with the percentage of effector memory CD4(+) T cells, suggesting antigenically driven differentiation of this population of T cells in both HIV-1 and M. leprae infections. Co-infection with M. leprae may exacerbate the immunopathology of HIV-1 disease. A T helper 2 (Th2) bias in the CD4(+) T-cell response was evident in both HIV-1 infection and leprosy, but no additive effect was apparent in co-infected patients.
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Infecciones por VIH/complicaciones , VIH-1 , Lepra/complicaciones , Adulto , Relación CD4-CD8 , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Diferenciación Celular/inmunología , Células Dendríticas/inmunología , Femenino , Infecciones por VIH/inmunología , VIH-1/aislamiento & purificación , Humanos , Inmunidad Celular , Interleucina-4/biosíntesis , Lepra/inmunología , Activación de Linfocitos/inmunología , Masculino , Persona de Mediana Edad , ARN Viral/sangre , Subgrupos de Linfocitos T/inmunología , Carga ViralRESUMEN
We report a case of acquired immunodeficiency syndrome (AIDS)-associated, acyclovir-refractory genital herpes infection treated with topical imidazoquinoline therapy. The patient's plasmacytoid dendritic cells made a robust interferon- alpha response following in vitro stimulation with imidazoquinoline but not with herpes simplex virus. We hypothesize that disease resulting from defective herpes simplex virus-stimulated interferon- alpha may be overcome by stimulating intact alternative pathways.
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Aminoquinolinas/uso terapéutico , Células Dendríticas/patología , Infecciones por VIH/inmunología , Herpes Genital/tratamiento farmacológico , Inductores de Interferón/uso terapéutico , Infecciones Oportunistas Relacionadas con el SIDA/tratamiento farmacológico , Infecciones Oportunistas Relacionadas con el SIDA/virología , Aciclovir/farmacología , Adulto , Antivirales/farmacología , Infecciones por VIH/complicaciones , VIH-1/inmunología , Herpesvirus Humano 2/efectos de los fármacos , Humanos , Imiquimod , MasculinoRESUMEN
We report a practical technique to assess peripheral blood dendritic cell (DC) maturation and function in whole blood (WB), which requires minimal blood volumes and minimizes ex vivo manipulations of clinical specimens. We determined optimal conditions for flow cytometric analysis of markers of DC maturation, including CCR7, CD25, CD80 and CD83, and of the intracellular cytokines, tumor necrosis factor-alpha (TNF-alpha) and interferon-alpha (IFN-alpha), in both myeloid (mDC) and plasmacytoid (pDC) lineages. We demonstrate concentration-dependent production of these cytokines by DC following short-term stimulation with ligands to Toll-like receptors (TLRs) 2/1, 3, 4, 7, 8 and 9. Kinetic studies revealed maximal TNF-alpha and IFN-alpha protein expression at 2 to 3 h after stimulation with certain TLR ligands. Finally, utilizing cells from a cohort of eight healthy donors, we compared DC responses to TLR activation in WB, freshly isolated peripheral blood mononuclear cells (PBMC) and cryopreserved PBMC. We found that TNF-alpha responses were essentially preserved, but IFN-alpha responses were profoundly diminished or entirely abrogated following cryopreservation. In conclusion, we propose that WB analysis of peripheral DC function is a rapid, reliable and simple method to evaluate TLR function in clinical specimens, which obviates artifact-prone cell purification. The major impact of cryopreservation on some DC responses further strengthens the case for a rapid method that uses fresh blood.
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Células Dendríticas/inmunología , Leucocitos Mononucleares/inmunología , Receptores Toll-Like/inmunología , Adulto , Antígeno B7-1/inmunología , Citometría de Flujo/métodos , Humanos , Imidazoles/inmunología , Interferón gamma/inmunología , Ligandos , Lipopolisacáridos/inmunología , Poli I-C/inmunología , Receptores CCR7 , Receptores de Quimiocina/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Quantification of gene expression is important to confirm changes in levels of gene expression in disease. Prior quantification methods include standard curves, absolute quantification, and relative quantification. This paper describes an analytic method for the relative quantification of Presenilin-1 (PS-1) in neurons and Forkhead-box (FOX) p3 in PBMNCs using real-time PCR analytic techniques. A comparative Ct method (deltadelatCt) is described in which the quantity of target normalized to a normalizer gene reference is given by 2-deltadelatCt where deltadelatCt = [Ct of the gene of interest in the unknown specimen - Ct normalizer gene in the unknown specimen] - [Ct of the gene of interest in the calibrator specimen - Ct normalizer gene in the calibrator specimen]. The calibrator specimen is ideally from a non-treated control specimen and is analyzed on every assay plate with the unknown specimens of interest. The use of the deltadelatCt methodology allows for a higher throughput and a more economical approach to investigate gene expression. We applied this methodology to the quantification of PS-1 and FOXp3 genes and compare the levels of expression by normalizing to different normalizer genes using the deltadelatCt methodology. We find that use of GAPDH is the optimum normalizer gene for the genes analyzed in neurons from human brain and in PBMNCs.
Asunto(s)
Factores de Transcripción Forkhead/sangre , Proteínas de la Membrana/análisis , Neuronas/metabolismo , Animales , Encéfalo/citología , Expresión Génica , Humanos , Presenilina-1 , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
In humans, innate immune recognition of mycobacteria, including Mycobacterium tuberculosis and Mycobacterium leprae, involves toll-like receptor-2 (TLR-2), expressed on immature dendritic cells (DCs), and the T-cell gammadelta receptor expressed by a subpopulation of T cells that utilize Vdelta2 (Vdelta2 T cells). To investigate modulatory relationships between these host-cell populations in a microbial context, in vitro experiments were performed with human DCs and Vdelta2 T cells stimulated with model TLR-2 ligands and phosphoantigens, respectively. We observed that TLR-2-stimulated DCs enhanced interferon-gamma (IFN-gamma) production by Vdelta2 T cells; conversely, activated Vdelta2 T cells enhanced TLR-2-induced DC maturation via soluble factors including IFN-gamma, which costimulated interleukin-12 (IL-12) p70 secretion by DCs. Exposure of DCs to activated Vdelta2 T cells was critical for Th1 T-cell priming when TLR-2 stimulation was limiting. These results suggest that Vdelta2 T cells may play an adjuvant role in priming protective antimycobacterial immunity when TLR-2 stimulation is lacking, as may occur if the infectious inoculum is small, or if the pathogen is an intrinsically weak activator of DCs.
Asunto(s)
Comunicación Celular/inmunología , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Inmunidad Activa , Receptores de Antígenos de Linfocitos T gamma-delta/metabolismo , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismo , Adulto , Diferenciación Celular/inmunología , Células Cultivadas , Técnicas de Cocultivo , Células Dendríticas/citología , Humanos , Activación de Linfocitos/inmunología , Mycobacterium leprae/inmunología , Mycobacterium tuberculosis/inmunología , Receptores de Antígenos de Linfocitos T gamma-delta/inmunología , Receptor Toll-Like 2/inmunología , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 2/fisiologíaRESUMEN
OBJECTIVE: To utilize the potent antigen-presenting capacity of mature dendritic cells (MDC) in order to develop a rapid, sensitive method for quantifying antigen-specific CD8 T cells present at low frequency in peripheral blood. DESIGN: Peripheral blood mononuclear cells (PBMC) were obtained from seven HIV-1-positive individuals with low to moderate CD8 T cell responses, including five on highly active antiretroviral therapy (HAART). IFN-gamma ELISPOT assays were performed using either monocytes or MDC to present antigens expressed by recombinant vaccinia viruses (r-VV). METHODS: Peripheral blood-derived monocytes were cultured for 5-6 days in the presence of IL-4 and granulocyte macrophage colony-stimulating factor, then matured in monocyte-conditioned medium. MDC were infected with r-VV and co-cultured in an ELISPOT assay with autologous monocyte-depleted PBMC. RESULTS: Relative to autologous monocytes, MDC amplified detection of antigen-specific CD8 T cells by 2-30-fold in response to antigens from HIV-1, Epstein-Barr virus and cytomegalovirus. Furthermore, antigenic specificities were revealed that had not been detected using standard ELISPOT of PBMC. CONCLUSION: This assay will prove useful for the detection of memory T cells present at low frequency, and may be of interest for identifying subdominant cytotoxic T lymphocyte epitopes. This method may have broad applications for the detection of antiviral CD8 T cell responses in patient populations in whom such responses have been difficult to detect, including HIV-1-seropositive individuals with advanced disease or undergoing HAART.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Células Dendríticas/inmunología , Infecciones por VIH/inmunología , VIH-1/inmunología , Adulto , Presentación de Antígeno/inmunología , Reacciones Falso Positivas , Femenino , Citometría de Flujo , Congelación , Vectores Genéticos , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/terapia , Herpesvirus Humano 4/inmunología , Humanos , Proteínas Inmediatas-Precoces/genética , Proteínas Inmediatas-Precoces/inmunología , Inmunoterapia/métodos , Leucocitos Mononucleares/inmunología , Masculino , Persona de Mediana Edad , Monocitos/inmunología , Fosfoproteínas/genética , Fosfoproteínas/inmunología , Recombinación Genética , Transactivadores/genética , Transactivadores/inmunología , Virus Vaccinia , Proteínas de la Matriz Viral/genética , Proteínas de la Matriz Viral/inmunología , Proteínas ViralesRESUMEN
We optimized a whole blood intracellular cytokine assay to quantitate the frequency of specific CD4+ and CD8+ T cells in small volumes of whole blood from infants from developing countries. The assay is performed in two steps. First, whole blood is stimulated in the presence of specific antigens for 6-18 h, ending with cryopreservation of fixed white cells. These stimulation steps were specifically adapted to be practical and reliable in a rural, developing country field setting. Later, in a more resourceful setting, interferon-gamma producing CD4+ or CD8+ T cells are detected by flow cytometry. The assay proved sensitive and specific for detecting mycobacteria-specific immunity 10 weeks after Bacillus Calmette-Guerin (BCG) vaccination of newborns from a rural field site.
Asunto(s)
Citocinas/sangre , Espacio Intracelular/química , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/metabolismo , Vacuna BCG/inmunología , Recolección de Muestras de Sangre , Criopreservación , Países en Desarrollo , Citometría de Flujo , Infecciones por VIH/sangre , Infecciones por VIH/inmunología , Humanos , Memoria Inmunológica/inmunología , Lactante , Interferón gamma/análisis , Interferón gamma/metabolismo , Mycobacterium tuberculosis/inmunología , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Subgrupos de Linfocitos T/inmunología , Factores de TiempoRESUMEN
Childhood tuberculous meningitis is associated with serious long-term sequelae, including mental retardation, behavior disturbances, and motor handicap. Brain damage in tuberculous meningitis results from a cytokine-mediated inflammatory response, which causes vasculitis and obstructive hydrocephalus. Thalidomide, a potent tumor necrosis factor alpha inhibitor, was well tolerated and possibly showed some clinical benefit in children with tuberculous meningitis during a pilot study. The purpose of the present study was to assess the effect of adjunctive thalidomide in addition to standard antituberculosis and corticosteroid therapy on the outcome of tuberculous meningitis. Thalidomide (24 mg/kg/day orally) or placebo was administered in a double-blind randomized fashion for 1 month to patients with stage 2 or 3 tuberculous meningitis. The study was terminated early because all adverse events and deaths occurred in one arm of the study (thalidomide group). Thirty of the 47 children enrolled received adjunctive thalidomide, of whom 6 (20%) developed a skin rash, 8 (26%) hepatitis, and 2 (6%) neutropenia or thrombocytopenia. Four deaths (13%) occurred in patients with very severe neurologic compromise at baseline; two deaths were associated with a rash. Motor outcome after 6 months of antituberculosis therapy was similar in the two groups, even though the thalidomide group showed greater neurologic compromise on admission. In addition, the mean IQ of the two treatment groups did not differ significantly (mean IQ thalidomide group 57.8 versus mean IQ control group 67.5; P = .16). These results do not support the use of adjunctive high-dose thalidomide therapy in the treatment of tuberculous meningitis.
Asunto(s)
Inmunosupresores/uso terapéutico , Talidomida/uso terapéutico , Tuberculosis Meníngea/tratamiento farmacológico , Corticoesteroides/uso terapéutico , Antiinflamatorios/efectos adversos , Antiinflamatorios/uso terapéutico , Antituberculosos/uso terapéutico , Quimioterapia Adyuvante , Niño , Preescolar , Estudios de Cohortes , Coma/etiología , Citocinas/sangre , Citocinas/líquido cefalorraquídeo , Método Doble Ciego , Exantema/inducido químicamente , Hepatitis/etiología , Humanos , Inmunosupresores/efectos adversos , Lactante , Inteligencia/efectos de los fármacos , Paresia/etiología , Sudáfrica , Estadísticas no Paramétricas , Talidomida/efectos adversos , Resultado del Tratamiento , Tuberculosis Meníngea/mortalidadRESUMEN
OBJECTIVES: (1) Develop a methodology for obtaining reliable cognitive and developmental data in children with neurodegenerative disease and cognitive impairment and in turn monitor disease state and treatment outcomes. (2) Demonstrate validity of age-equivalent scores. METHODS: We present guidelines for obtaining accurate test scores in low-functioning and behaviorally disruptive pediatric patients, followed by a method validation study: (1) using disease-specific protocols to assess salient aspects of the known phenotype, (2) selecting appropriate tests, (3) managing behavior, and (4) using age-equivalent scores on standardized tools. We used the Bayley Scales of Infant Development-III or Kaufman Assessment Battery for Children-II with a group of 25 children with mucopolysaccharidosis type IIIA (MPS IIIA or Sanfilippo syndrome type A) with dementia. To demonstrate concurrent validity, we used the Vineland Adaptive Behavior Scales-II, comparing parent-reported age-equivalent scores (AEs) with those of the cognitive measures. RESULTS: We were successful in obtaining cognitive age-equivalents for 25 patients with MPS IIIA including those with severe behavioral disruption and a correlation of 0.95 was obtained comparing scores on the parent measure with cognitive age-equivalents validating the age-equivalent approach. CONCLUSION: An approach to the assessment of severely impaired children including those with behavioral disruption was implemented and is applicable to children with other severe neurological diseases. This approach will enhance the assessment of disease progression and monitoring of treatment outcome in clinical trials.
RESUMEN
The immunomodulatory drug thalidomide is the treatment of choice for erythema nodosum leprosum (ENL), an inflammatory cutaneous and systemic complication of multibacillary leprosy. To elucidate the mechanism of action of thalidomide in this syndrome, we prospectively investigated 20 patients with ENL who were treated with thalidomide for 21 days. All patients responded to treatment, with the majority of them having complete resolution of cutaneous lesions within 7 days. This response was associated with a marked but transient increase in ex vivo mitogen-induced expression of interleukin (IL)-2 and interferon- gamma by CD4(+) and CD8(+) T cells that was observed on treatment day 7, but these returned to pretreatment levels by day 21. Plasma tumor necrosis factor- alpha levels were not high at baseline, and they increased modestly during treatment. Plasma levels of IL-12 increased steadily during thalidomide treatment. Hence, the therapeutic effect of thalidomide in ENL appears to be associated with transient immune stimulation, which suggests that the drug may promote an active immunoregulatory response.
Asunto(s)
Eritema Nudoso/tratamiento farmacológico , Eritema Nudoso/inmunología , Lepra Lepromatosa/tratamiento farmacológico , Lepra Lepromatosa/inmunología , Talidomida/uso terapéutico , Adolescente , Adulto , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Citocinas/sangre , Eritema Nudoso/patología , Citometría de Flujo , Expresión Génica , Humanos , Factores Inmunológicos/farmacología , Factores Inmunológicos/uso terapéutico , Interferón gamma/biosíntesis , Leprostáticos/farmacología , Leprostáticos/uso terapéutico , Lepra Lepromatosa/patología , Masculino , Persona de Mediana Edad , Estudios Prospectivos , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Piel/patología , Talidomida/farmacologíaRESUMEN
CD8(+) T cell immunity is critical for protection from viral disease, such as that caused by the human immunodeficiency virus (HIV) or cytomegalovirus (CMV). It is therefore important to identify therapies that can boost antiviral immunity. The recent finding that thalidomide acts as a T cell costimulator suggested that this drug may boost antiviral CD8(+) T cell responses. In this in vitro study, in a human autologous CD8(+) T cell/dendritic cell (DC) coculture system, thalidomide and a potent thalidomide analogue were shown to enhance virus-specific CD8(+) T cell cytokine production and cytotoxic activity. The drug-enhanced antiviral activity was noted in cells from both healthy donors and persons chronically coinfected with HIV and CMV. This stimulatory effect was directed at CD8(+) T cells, and not DCs. These results suggest an application for thalidomide and the thalidomide analogue as a novel immune-adjuvant therapy in chronic viral infections.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Linfocitos T CD8-positivos/efectos de los fármacos , Talidomida/farmacología , Linfocitos T CD8-positivos/inmunología , División Celular/efectos de los fármacos , Técnicas de Cocultivo , Citocinas/análisis , Citocinas/biosíntesis , Infecciones por Citomegalovirus/inmunología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Relación Dosis-Respuesta Inmunológica , Citometría de Flujo , Infecciones por VIH/inmunología , Humanos , Técnicas de Inmunoadsorción , Lenalidomida , Talidomida/análogos & derivadosRESUMEN
To induce effector immunity, dendritic cells (DCs) must differentiate into fully mature cells. We show that, after human monocyte-derived DCs were infected with virulent Mycobacterium tuberculosis, up-regulation of cellular-surface maturation markers was minimal and reversible. In the presence of a potent stimulus for maturation (tumor necrosis factor [TNF]-alpha, interleukin [IL]-1beta, and prostaglandin E2 [PGE2]), M. tuberculosis inhibited phenotypic DC maturation. M. tuberculosis-infected DCs had an impaired ability to induce allogeneic lymphoproliferation and activated autologous memory CD4+ and CD8+ T cells optimally only in the presence of TNF-alpha, IL-1beta, and PGE2. Thus, virulent M. tuberculosis inhibits phenotypic and functional maturation of human monocyte-derived DCs. This mechanism, which has been described elsewhere for various viruses and for the virulent mycobacterium M. leprae, may be a novel mechanism that this pathogen uses to evade the host's immune response.
Asunto(s)
Células Dendríticas/citología , Células Dendríticas/microbiología , Monocitos/citología , Monocitos/microbiología , Mycobacterium tuberculosis/fisiología , Biomarcadores , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular , Células Dendríticas/efectos de los fármacos , Dinoprostona/farmacología , Humanos , Memoria Inmunológica , Interleucina-1/farmacología , Activación de Linfocitos , Fenotipo , Linfocitos T/inmunología , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
This study investigated whether peripheral nerve damage in patients with leprosy impairs local cellular immune responses, thereby reducing wound healing and leading to chronic skin ulceration. Anesthetic and contralateral sensitive skin sites in 42 patients with leprosy were compared for delayed-type hypersensitivity responses to purified protein derivative (PPD) of tuberculin. Leukocyte recruitment, epidermal activation, keratinocyte proliferation, and rates of wound healing after skin biopsy were compared. No significant differences in PPD-induced induration, epidermal activation and thickening or numbers of total T cells, CD8+ T cells, CD1a+ Langerhans cells, and proliferating Ki67+ keratinocytes were observed between anesthetic and sensitive skin sites. Similarly, rates of wound healing over 5 days after skin biopsy did not differ significantly. Thus, local leprosy-associated anesthesia does not appear to contribute to local immune compromise or impaired wound healing. Rather, chronic cutaneous ulceration in leprosy most likely results from repeated trauma associated with loss of sensation.