RESUMEN
Live attenuated Listeria monocytogenes (LM) is a promising bacterial vector able to induce a T-cell response to tumor-associated antigens and demonstrates great potential for use in vaccine development. A novel recombinant LM-based vaccine (Lmdd (LM ΔdalΔdat)-MPFG (multiple peptide fusing genes)) was developed with the ability to express and secrete hepatocellular carcinoma (HCC)-related tumor-associated antigens fragments due to the insertion of hepatitis B virus (HBV)-X protein (HBx)-derived epitopes HBx(52-60) and HBx(140-148), the universal T-helper epitope, alpha-fetoprotein (AFP) epitope AFP(158-166), and melanoma antigen gene (MAGE)-3(271-279) into the HBV core protein. Following immunization with the Lmdd-MPFG vaccine, macrophages exhibited uptake of the bacteria; the vaccine was then nearly cleared 3 days after the first administration. It disappeared even more quickly following subsequent vaccinations. However, recombinant Lmdd-MPFG allowed for the full development of an antitumor response towards the human leukocyte antigen (HLA)-A0201 epitopes of MPFG. Each epitope stimulated an augmented T-cell proliferation and enhanced the supernatant level of interferon (IFN)-γ in vitro. In addition, IFN-γ-producing CD8(+) T cells as well as in vivo cytolytic activity were significantly increased in HLA-A2 transgenic mice. Additionally, the Lmdd-MPFG developed a strong antitumor response, as indicated by the significant resistance of immunized mice to MPFG-positive Hepa1-6 cell challenge in both a prophylactic and therapeutic setting. Tumor regression was accompanied by an enhanced cytotoxic T lymphocyte response and a decrease of regulatory T cells in the tumor. Collectively, these results suggest that utilizing attenuated LM as a vaccine vector, able to carry the MPFG gene, presents a potentially feasible strategy for prevention of HCC.
Asunto(s)
Vacunas contra el Cáncer/uso terapéutico , Carcinoma Hepatocelular/terapia , Listeria monocytogenes/inmunología , Neoplasias Hepáticas/terapia , Animales , Antígenos de Neoplasias/inmunología , Vacunas contra el Cáncer/inmunología , Carcinoma Hepatocelular/inmunología , Línea Celular Tumoral , ADN Recombinante , Epítopos , Estudios de Factibilidad , Humanos , Interferón gamma/metabolismo , Neoplasias Hepáticas/inmunología , Activación de Linfocitos/inmunología , Ratones , Linfocitos T/inmunología , Transactivadores/inmunología , Trasplante Heterólogo , Proteínas Reguladoras y Accesorias ViralesRESUMEN
Vaccines designed to control chronic infections by intracellular agents such as human immunodeficiency virus type 1 (HIV-1) require the induction of cell-mediated immune responses to rid the host of pathogen-infected cells. Listeria monocytogenes has characteristics that make it an attractive vaccine vector for use against such infections. Here we show that parenteral immunization with a new highly attenuated strain of this organism provided complete protection against systemic and mucosal challenges with a recombinant vaccinia virus expressing HIV-1 gag. Immunization also generated a strong, long-term memory cytotoxic-T-lymphocyte (CTL) response in spleen, mesenteric lymph nodes, and Peyer's patches directed against the gag protein. Oral immunization with this attenuated strain also produced complete, long-lasting protection against the recombinant virus but only against mucosal virus challenge. Curiously, oral immunization was associated with a transient CTL response in the three lymphoid tissues examined.
Asunto(s)
Productos del Gen gag/inmunología , Infecciones por VIH/inmunología , Inmunización , Listeria monocytogenes/genética , Listeria monocytogenes/inmunología , Vacunas contra el SIDA , Animales , Femenino , Productos del Gen gag/genética , Productos del Gen gag/metabolismo , VIH-1/genética , VIH-1/inmunología , VIH-1/metabolismo , Inmunidad Mucosa , Memoria Inmunológica , Listeria monocytogenes/metabolismo , Listeria monocytogenes/patogenicidad , Tejido Linfoide/inmunología , Ratones , Ratones Endogámicos BALB C , Recombinación Genética , Linfocitos T Citotóxicos/inmunología , Virus Vaccinia/genética , Virus Vaccinia/inmunología , Virus Vaccinia/metabolismoRESUMEN
Induction of cell-mediated immunity may be essential for an effective AIDS vaccine. Listeria monocytogenes is an attractive bacterial vector to elicit T-cell immunity to human immunodeficiency virus (HIV) because it specifically infects monocytes, key antigen-presenting cells, and because natural infection originates at the mucosa. Immunization with recombinant L. monocytogenes has been shown to protect mice from lymphocytic choriomeningitis virus, influenza virus, and tumor inoculation. L. monocytogenes expressing HIV gag elicits sustained high levels of Gag-specific cytotoxic T lymphocytes (CTLs) in mice. We have examined the ability of Listeria to infect human monocytes and present HIV antigens to CD8 T lymphocytes of HIV-infected donors to induce a secondary T-cell immune response. Using this in vitro vaccination protocol, we show that L. monocytogenes expressing the HIV-1 gag gene efficiently provides a strong stimulus for Gag-specific CTLs in HIV-infected donor peripheral blood mononuclear cells. Listeria expressing Nef also elicits a secondary in vitro anti-Nef CTL response. Since L. monocytogenes is a pathogen, before it can be seriously considered as a human vaccine vector, safety concerns must be addressed. We therefore have produced a highly attenuated strain of L. monocytogenes that requires D-alanine for viability. The recombinant bacteria are attenuated at least 10(5)-fold. We show that when these hyperattenuated bacteria are engineered to express HIV-1 Gag, they are at least as efficient at stimulating Gag-specific human CTLs in vitro as wild-type recombinants. These results suggest that attenuated Listeria is an attractive candidate vaccine vector to induce T-cell immunity to HIV in humans.
Asunto(s)
Linfocitos T CD8-positivos/inmunología , Antígenos VIH/metabolismo , VIH-1/inmunología , Listeria monocytogenes/genética , Listeria monocytogenes/inmunología , Vacunas contra el SIDA/genética , Vacunas contra el SIDA/inmunología , Alanina/genética , Alanina/metabolismo , Secuencia de Aminoácidos , Animales , Presentación de Antígeno , Citometría de Flujo , Productos del Gen gag/genética , Productos del Gen gag/inmunología , Productos del Gen gag/metabolismo , Productos del Gen nef/genética , Productos del Gen nef/inmunología , Productos del Gen nef/metabolismo , Vectores Genéticos , Antígenos VIH/genética , Antígenos VIH/inmunología , Infecciones por VIH/inmunología , VIH-1/genética , VIH-1/metabolismo , Humanos , Listeria/genética , Listeria/inmunología , Listeria/metabolismo , Listeria monocytogenes/metabolismo , Listeria monocytogenes/patogenicidad , Activación de Linfocitos , Ratones , Datos de Secuencia Molecular , Monocitos/inmunología , Monocitos/microbiología , Proteínas Recombinantes/inmunología , Recombinación Genética , Productos del Gen nef del Virus de la Inmunodeficiencia HumanaRESUMEN
The self-esteem of children with Attention Deficit Hyperactivity Disorder (ADHD) has been shown to be low. The effects of stimulant medication upon their self-esteem have not been systematically studied. The present study employed a reliable self-report instrument to measure the self-esteem of children with ADHD medicated with stimulants vs. those who were unmedicated. Results showed that stimulants were associated with significantly higher self-esteem. Children with ADHD prescribed stimulants reported feeling more intelligent and more popular than unmedicated children with ADHD. Children with ADHD and Oppositional Defiant Disorder (ODD) prescribed stimulants reported feeling better behaved. Significant correlations indicated that higher doses were associated with higher self-esteem. The present results suggest a need for a well-controlled study to determine if stimulants were responsible for the observed differences in self-esteem.
Asunto(s)
Trastorno por Déficit de Atención con Hiperactividad/tratamiento farmacológico , Trastorno por Déficit de Atención con Hiperactividad/psicología , Estimulantes del Sistema Nervioso Central/uso terapéutico , Metilfenidato/uso terapéutico , Grupo Paritario , Autoimagen , Análisis de Varianza , Déficit de la Atención y Trastornos de Conducta Disruptiva/tratamiento farmacológico , Déficit de la Atención y Trastornos de Conducta Disruptiva/psicología , Estimulantes del Sistema Nervioso Central/administración & dosificación , Niño , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Relaciones Interpersonales , Entrevistas como Asunto , Masculino , Metilfenidato/administración & dosificación , Encuestas y CuestionariosRESUMEN
Biological surface engineering using synthetic biological materials has a great potential for advances in our understanding of complex biological phenomena. We developed a simple system to engineer biologically relevant surfaces using a combination of self-assembling oligopeptide monolayers and microcontact printing (muCP). We designed and synthesized two oligopeptides containing a cell adhesion motif (RADS)n (n = 2 and 3) at the N-terminus, followed by an oligo(alanine) linker and a cysteine residue at the C-terminus. The thiol group of cysteine allows the oligopeptides to attach covalently onto a gold-coated surface to form monolayers. We then microfabricated a variety of surface patterns using the cell adhesion peptides in combination with hexa-ethylene glycol thiolate which resist non-specific adsorption of proteins and cells. The resulting patterns consist of areas either supporting or inhibiting cell adhesion, thus they are capable of aligning cells in a well-defined manner, leading to specific cell array and pattern formations.
Asunto(s)
Materiales Biocompatibles , Adhesión Celular/fisiología , Oligopéptidos/química , Células 3T3/citología , Secuencia de Aminoácidos , Animales , Materiales Biocompatibles/síntesis química , Materiales Biocompatibles/química , Carcinoma de Células Escamosas , Humanos , Ratones , Modelos Moleculares , Datos de Secuencia Molecular , Oligopéptidos/síntesis química , Conformación Proteica , Propiedades de Superficie , Células Tumorales CultivadasRESUMEN
Using a strain of Listeria monocytogenes that stably expresses and secretes HIV gag to deliver this Ag to the MHC class I pathway of Ag processing, we have identified the immunodominant CTL epitope to gag in the BALB/c mouse and shown that it is Kd restricted. The specific motif for the peptides that bind the MHC class I molecule H-2 Kd is believed to be a nonamer with residues tyrosine or phenylalanine in the second amino acid position and leucine or isoleucine in the carboxyl-terminal or ninth amino acid position as dominant anchoring positions. Surprisingly, the identified gag peptide, AMQMLKETI, does not contain an anchoring aromatic residue in position two although competition assays with other Kd-restricted epitopes indicated that it binds to Kd with comparable affinity. Using a theoretical molecular dynamics approach to probe the stability of peptide binding to MHC class I molecules, we show that the absence of an appropriate anchor residue at P2 in AMQMLKETI is compensated by favorable interactions of the glutamine at P3 with pocket D of Kd. These findings were verified experimentally, demonstrating the predictive power of this theoretical approach in analyzing MHC class I/peptide interactions. These studies also indicate that CTL epitope prediction that relies on dominant peptide motifs may not always identify the correct epitope.
Asunto(s)
Productos del Gen gag/inmunología , Glutamina/metabolismo , Antígenos H-2/inmunología , Anticuerpos Anti-VIH/biosíntesis , Epítopos Inmunodominantes/inmunología , Secuencia de Aminoácidos , Animales , Citotoxicidad Inmunológica , Femenino , Productos del Gen gag/administración & dosificación , Productos del Gen gag/metabolismo , Glutamina/química , Antígenos H-2/química , Antígenos H-2/metabolismo , Proteínas Hemolisinas/inmunología , Epítopos Inmunodominantes/metabolismo , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Listeria monocytogenes/inmunología , Ratones , Ratones Endogámicos BALB C , Modelos Moleculares , Datos de Secuencia Molecular , Fragmentos de Péptidos/inmunología , Fragmentos de Péptidos/metabolismo , Unión Proteica/inmunología , Conformación Proteica , Linfocitos T Citotóxicos/inmunologíaRESUMEN
Listeria monocytogenes is an intracellular bacterial pathogen that elicits a strong cellular immune response following infection and therefore has potential use as a vaccine vector. However, while infections by L. monocytogenes are fairly rare and can readily be controlled by a number of antibiotics, the organism can nevertheless cause meningitis and death, particularly in immunocompromised or pregnant patients. We therefore have endeavored to isolate a highly attenuated strain of this organism for use as a vaccine vector. D-Alanine is required for the synthesis of the mucopeptide component of the cell walls of virtually all bacteria and is found almost exclusively in the microbial world. We have found in L. monocytogenes two genes that control the synthesis of this compound, an alanine racemase gene (dal) and a D-amino acid aminotransferase gene (dat). By inactivating both genes, we produced an organism that could be grown in the laboratory when supplemented with D-alanine but was unable to grow outside the laboratory, particularly in the cytoplasm of eukaryotic host cells, the natural habitat of this organism during infection. In mice, the double-mutant strain was completely attenuated. Nevertheless, it showed the ability, particularly under conditions of transient suppression of the mutant phenotype, to induce cytotoxic T-lymphocyte responses and to generate protective immunity against lethal challenge by wild-type L. monocytogenes equivalent to that induced by the wild-type organism.
Asunto(s)
Alanina/metabolismo , Proteínas Bacterianas , Listeria monocytogenes/inmunología , Listeria monocytogenes/patogenicidad , Alanina Racemasa/genética , Alanina Racemasa/inmunología , Secuencia de Aminoácidos , Aminoaciltransferasas/genética , Aminoaciltransferasas/inmunología , Animales , Vacunas Bacterianas/inmunología , Secuencia de Bases , Pared Celular , Clonación Molecular , ADN Bacteriano , Femenino , Listeria monocytogenes/enzimología , Listeria monocytogenes/genética , Listeriosis/inmunología , Listeriosis/prevención & control , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Mutagénesis , Homología de Secuencia de AminoácidoRESUMEN
Assessment of parenting skills was based upon a standardized videotape of vignettes of parents reading a magazine while their children were playing, playing with their children, or trying to get them to clean up. Parents who viewed this videotape came from 40, 2-parent families referred to an outpatient clinic for behavior problems (Behavior Problem group) or 20 nonreferred 2-parent families (Nonclinic group). They were compared on the degree to which they selected: (1) positive responses to children's prosocial behavior (praise), (2) ignoring children's inappropriate behavior (ignore), and (3) punishing children's aversive behavior (punish). The Nonclinic group was more accurate than the Behavior Problem Group and mothers were more accurate than fathers in their selection of praise to children's prosocial behavior. Nonclinic mothers were more accurate than all other combinations of Group and Parent Sex in their selection of ignore for children's inappropriate behavior. Sensitivity calculations suggested that this instrument could potentially be useful as a screening tool in clinical pediatric practice and early intervention. Implications for nursing research and practice were discussed.
Asunto(s)
Crianza del Niño/psicología , Salud de la Familia , Evaluación en Enfermería/métodos , Enfermería Pediátrica , Grabación de Cinta de Video , Adulto , Niño , Preescolar , HumanosRESUMEN
OBJECTIVE: Previous research has demonstrated that peer rejection is a significant part of the clinical presentation of many children with attention-deficit hyperactivity disorder (ADHD). Outcome studies of treatment interventions have typically failed to show generalization of treatment gains to the home and classroom. This has been especially true for children who have comorbid oppositional defiant disorder (ODD). The present study was intended to demonstrate generalization of an outpatient social skills training program when parents were trained in skills relevant to their child's social adjustment. METHOD: Thirty-five children with ADHD and 14 children without ADHD were given 12 sessions of treatment (treatment group). Outcome was compared with 12 children with ADHD and 12 children without ADHD who were on a waitlist for treatment (waitlist group). Nineteen children with ODD were in the treatment group and five in the waitlist. Stimulant medication was prescribed for all children with ADHD. RESULTS: Subjects with ADHD showed improvement comparable with that of subjects without ADHD on all teacher- and parent-reported measures of peer adjustment and social skills, except teacher-reported withdrawal. Children with ODD had outcome comparable with that of children without ODD. Effect sized ranged from 0.93 to 1.34 indicating that the average treatment group subject was better off than 83.4% of waitlist subjects on outcome measures. CONCLUSIONS: The present results suggest that children with ADHD are best heiped by a combination of social skills training for themselves, collateral training for their parents and stimulant medication.
Asunto(s)
Trastorno por Déficit de Atención con Hiperactividad/terapia , Terapia Conductista/normas , Terapia Familiar/normas , Padres/educación , Psicoterapia de Grupo/normas , Trastorno de la Conducta Social/terapia , Socialización , Transferencia de Experiencia en Psicología , Análisis de Varianza , Trastorno por Déficit de Atención con Hiperactividad/complicaciones , Terapia Conductista/métodos , Estudios de Casos y Controles , Niño , Estudios Transversales , Terapia Familiar/métodos , Femenino , Humanos , Relaciones Interpersonales , Masculino , Grupo Paritario , Estudios Prospectivos , Psicoterapia de Grupo/métodos , Autocuidado/normas , Trastorno de la Conducta Social/complicaciones , Resultado del TratamientoRESUMEN
A standardized 12-week cognitive-behavioral social skills package in which parents assisted children with socialization homework assignments, was presented to 52 nonpsychotic outpatient boys, many of whom were diagnosed with DSM-III-R Attention Deficit-Hyperactivity Disorder and Oppositional Defiant Disorder. Results demonstrated that the Thought Problems factor of the mother-completed Child Behavior Checklist aided in predicting changes in teacher-rated aggression. DSM-III-R diagnosis of Oppositional Defiant Disorder predicted changes in teacher-rated withdrawal subsequent to treatment. The need for research to determine which children benefit from which treatment was discussed.
Asunto(s)
Terapia Cognitivo-Conductual/normas , Manuales como Asunto/normas , Evaluación de Resultado en la Atención de Salud/normas , Escalas de Valoración Psiquiátrica/normas , Psicometría/normas , Trastorno de la Conducta Social/terapia , Socialización , Agresión , Análisis de Varianza , Distribución de Chi-Cuadrado , Niño , Análisis Discriminante , Humanos , Funciones de Verosimilitud , Modelos Logísticos , Masculino , Trastornos Mentales/diagnóstico , Oportunidad Relativa , Pacientes Ambulatorios/estadística & datos numéricos , Selección de Paciente , Pronóstico , Deseabilidad Social , Resultado del TratamientoRESUMEN
An attempt was made by the authors of DSM-III to restrict its focus to the experimental, the observable, and the measurable. The intention was to free the nosology from the influence of unproven theories, and the philosophy was driven largely by the importance of research being able to identify diagnostic categories to facilitate the study of homogeneous groups. So it is of interest that the authors accepted dissociation-an ambiguous event linked to an explicit theoretical concept that had been introduced by Janet-as the basis for classification of clinical presentations that were formerly included under the rubric of hysteria, a similarly unclear category. Since DSM-III, there have been an increasing number of reports of dissociative experiences and dissociative identity disorder (formerly known as multiple personality disorder), but neither of these clinical presentations seems able to withstand the concern that it is dramatically influenced by environmental cues, e.g., the expectations of the therapist. Thus, a restricted phenomenological perspective does not fully appreciate the distorting potential of suggestibility and imagination on the nature of the emerging clinical picture. These factors might well have contributed to and laid the conceptual groundwork for the growth in the number of reports of dissociation.
Asunto(s)
Trastornos Disociativos/diagnóstico , Adulto , Niño , Trastornos Disociativos/clasificación , Trastornos Disociativos/terapia , Humanos , Hipnosis , Histeria/clasificación , Histeria/diagnóstico , Imaginación , Memoria , Modelos Psicológicos , Psicometría , Terminología como AsuntoRESUMEN
Cytolytic T cells, acting through cytokines or by direct lysis of infected target cells, have been shown to play a significant role in the control of viral infections and may be responsible for the prolonged asymptomatic phase following infection by HIV. Accordingly, methods that can generate strong cell-mediated immune responses may be useful in the development of prophylactic and therapeutic vaccines against HIV. Listeria monocytogenes is a Gram-positive intracellular microorganism that elicits strong cell-mediated immune responses against its own secreted proteins following infection. In this study we have modified the chromosome of L. monocytogenes so that it stably expresses and secretes the p55 HIV gag gene product and examined the cell-mediated immune response of BALB/c mice to infection with this recombinant organism. Infected animals were found to mount a specific, strong, long-lasting CD8+ cytolytic T cell response against a predominant epitope contained within the p24 fragment of the HIV Gag protein. This epitope previously has been shown to be recognized by CTLs obtained from some HIV-infected humans. Our results suggest that chromosomally modified strains of L. monocytogenes may provide valuable vaccine vectors for use against HIV.
Asunto(s)
Vacunas contra el SIDA , Productos del Gen gag/genética , Productos del Gen gag/inmunología , VIH-1 , Inmunidad Celular , Listeria monocytogenes/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Citotoxicidad Inmunológica , Técnicas de Transferencia de Gen , Humanos , Listeria monocytogenes/inmunología , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Linfocitos T/inmunologíaAsunto(s)
Hiperemesis Gravídica/terapia , Hipnosis , Terapia Combinada , Femenino , Humanos , Hiperemesis Gravídica/psicología , EmbarazoRESUMEN
A computer search of the literature for papers indexed under "flashbacks" produced a list of 70 references, many found in publications on the topics of substance abuse and trauma. Several of these were letters or papers written in languages other than English. In all, the author reviewed 55 papers. Although most of these papers contained comments that addressed the subject matter to some extent as recurrences or reminiscences of past happenings, the variability in the use of the term leaves many unresolved questions regarding the veridicality of the imagery. Nothing in the presentations reviewed by the author clearly demonstrates the unidimensional nature of flashbacks nor any recognizable neurophysiological correlate. The content of a flashback appears to be at least as likely to be the product of imagination as it is of memory.
Asunto(s)
Déjà Vu , Imaginación , Acontecimientos que Cambian la Vida , Recuerdo Mental , Represión Psicológica , Adulto , Niño , Trastornos de Combate/psicología , Femenino , Alucinógenos , Humanos , Masculino , Trastornos por Estrés Postraumático/psicología , Trastornos Relacionados con Sustancias/psicologíaRESUMEN
From the origins of mysticism and theatrics, trivialized by stage performers, psychics, and charlatans, modern medical hypnosis has struggled to achieve and maintain a sense of professional integrity. Many of the principles of dynamic psychiatry are deeply rooted in the work of the early healers. Yet hypnosis as a clinical entity continues to fall in and out of favor over the years; again it is now being pushed beyond the limits of that which is reasonable and valid.
Asunto(s)
Hipnosis/historia , Europa (Continente) , Historia del Siglo XVIII , Historia del Siglo XIX , Historia del Siglo XX , Humanos , Estados UnidosRESUMEN
This study examined the levels of hypnotizability and dissociation in an outpatient sample of 17 normal-weight bulimic women and 20 healthy controls. Bulimic patients were significantly more hypnotizable than controls (p < .003) and scored higher on a self-report scale of dissociative experiences (p < .02). The results of measures of hypnotizability and dissociation showed a significant positive correlation in the combined subject group. These results are consistent with previous reports on hospitalized patients and college students and suggest that psychological factors associated with hypnotizability might play a role in the etiology and treatment of bulimia nervosa.