RESUMEN
The preferential recognition of certain amino acid sequences from foreign protein antigens by T cells is referred to as T cell epitope immunodominance. To determine the mechanisms underlying this phenomenon, we have studied the correlation between the interaction of a series of synthetic peptides encompassing the entire hen egg-white lysozyme (HEL) sequence with class II molecules of the H-2k haplotype, and T cell responsiveness to these peptides. After HEL priming, three immunodominant T cell epitopes were found: two, included in the HEL sequences 51-61 and 112-129, were recognized in association with I-Ak molecules, and one, included in sequence 1-18, in association with I-Ek molecules. Accordingly, these peptides bound to the appropriate class II molecule, as demonstrated by competition for antigen presentation. Several other HEL peptides, although capable of associating with class II molecules, were not immunodominant. The absence of immunodominance has been shown to arise by three different mechanisms: (a) competition by an immunodominant peptide for presentation in vivo, (b) failure to generate the peptide during antigen processing, and (c) an inherently poor capacity of the T cell repertoire to respond to a particular peptide-MHC complex.
Asunto(s)
Epítopos , Antígenos de Histocompatibilidad Clase II/inmunología , Muramidasa/inmunología , Linfocitos T/inmunología , Animales , Activación de Linfocitos , Ratones , Ratones Endogámicos C3H , Péptidos/inmunologíaRESUMEN
The synthetic nonapeptide VQGEESNDK, corresponding to the fragment 163-171 of human IL-1 beta, showed in vivo immunomodulatory capacities qualitatively and quantitatively comparable to those of the mature human IL-1 beta protein. In fact, both IL-1 beta and the 163-171 fragment stimulated the immune response of normal mice and restored immune reactivities of immunocompromised animals. In addition, the synthetic IL-1 peptide was as efficient as the entire protein in inducing tumor rejection and radioprotection. On the other hand, the 163-171 fragment did not cause any of several inflammation-associated metabolic changes inducible by the whole IL-1 beta molecule in vivo: hypoferremia, hypoglycemia, hyperinsulinemia, increase in circulating corticosterone, SAA and fibrinogen, decrease in hepatic drug-metabolizing enzymes. Furthermore, at variance with IL-1 beta, the 163-171 peptide did not show the toxic effects causing shock and death in adrenalectomized mice. Thus, these results confirm our previous in vitro observations that functional domains are identifiable within the multipotent cytokine IL-1 beta, and demonstrate the biological relevance of this finding in a variety of in vivo systems. The identification of a selectively active fragment of a cytokine may thus represent a significant step towards a better directed and more rational immunotherapeutic approach.
Asunto(s)
Citotoxicidad Inmunológica , Inmunización , Interleucina-1/farmacología , Neoplasias Experimentales/inmunología , Choque/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , 7-Alcoxicumarina O-Dealquilasa , Adrenalectomía , Animales , Femenino , Hormonas/sangre , Interleucina-1/síntesis química , Hígado/enzimología , Masculino , Ratones , Ratones Endogámicos , Oxigenasas/metabolismoRESUMEN
A specific colorimetric DNA detection method based on oligonucleotide functionalized gold-silver-alloy nanoparticles (AuAg-alloy-nanoprobes) is presented. The AuAg-alloy-nanoprobes were then used for the specific detection of a DNA sequence from TP53-a gene involved in cancer development. The AuAg-alloy-nanoprobes were then used in combination with Au-nanoprobes for a one-pot dual-colour detection strategy that allowed for the simultaneous differential detection of two distinct target sequences. This system poses an unprecedented opportunity to explore the combined use of metal nanoparticles with different composition towards the development of a multiplex one-pot colorimetric assay for DNA detection.
Asunto(s)
Colorimetría/métodos , Aleaciones de Oro/química , Nanopartículas del Metal/química , Hibridación de Ácido Nucleico/métodos , Plata/química , Sondas de ADN/química , Sondas de ADN/metabolismo , ADN de Cadena Simple/análisis , ADN de Cadena Simple/metabolismo , Proteína p53 Supresora de Tumor/genéticaRESUMEN
The essential oils from leaves of Hyptis fruticosa (Lamiaceae) Salzm., H. pectinata (Lamiaceae) Poit., and Lippia gracilis (Verbenaceae) HBK were investigated for their larvicidal activity against Aedes aegypti and analyzed by GC/MS. Fifty-nine compounds, representing 91.28-98.39% of the essential oils, have been identified. A standard solution was used to make 20 mL solutions ranging from 30 to 2000 ppm. Twenty larvae between third and fourth stages were added to the essential oil solution. A mortality count was conducted 24 h after treatment. Essential oils LC50 and their confidence limits at 95% probability were calculated by the methods of Reed-Muench and Pizzi, respectively. The essential oil of Lippia gracilis showed potent insecticidal effect against Aedes aegypti larvae, the vector of dengue fever. Carvacrol and caryophyllene oxide were the main responsible for the activity of L. gracilis and H. pectinata. Minor compounds are probably acting synergistically to achieve H. fruticosa activity.
Asunto(s)
Aedes/crecimiento & desarrollo , Insecticidas/farmacología , Larva/crecimiento & desarrollo , Aceites Volátiles/farmacología , Aedes/efectos de los fármacos , Animales , Brasil , Hyptis , Larva/efectos de los fármacos , Aceites Volátiles/aislamiento & purificación , Hojas de la Planta , Preparaciones de Plantas/aislamiento & purificación , Preparaciones de Plantas/farmacología , Polvos , SeguridadRESUMEN
Biozzi mice selected for high (H) or low (L) antibody responsiveness to natural antigens have been followed for their entire life-span to examine their pathology at death. As previously found in selection I, shorter life-span and higher lymphoma incidence were observed in L responder mice than in H responder mice selected for antibody responsiveness to sheep red blood cells (selection II). In mice selected for antibody responsiveness to Salmonella flagellar antigens (selection III), similar life-span and similar lymphoma incidence were found in H and L responder mice. Natural killer (NK) cell activity, as assessed in spleen cells from young mice, was lower in L than in H responder mice of selection I but higher in L than in H responder mice of both selections II and III. All these results indicate that longevity and lymphoma incidence at death are independent of NK cell activity in mice selected for H or L antibody responsiveness to natural antigens. Furthermore, genetic selection for antibody responsiveness does not always appear to influence life-span and lymphoma incidence.
Asunto(s)
Formación de Anticuerpos , Células Asesinas Naturales/inmunología , Longevidad , Neoplasias Experimentales/epidemiología , Animales , Antígenos Bacterianos/inmunología , Eritrocitos/inmunología , Femenino , Linfoma/epidemiología , Masculino , Ratones , Factores Sexuales , Ovinos , Bazo/inmunologíaRESUMEN
Biozzi mice selected for high (Hi) or low (Lo) responsiveness to phytohemagglutinin (PHA) have been followed for their entire life-span to examine their pathology at death. Spontaneous lymphomas were found to exhibit higher incidence and faster development in Lo/PHA than in Hi/PHA females, whereas a similar difference between the two lines did not attain the level of statistical significance in male mice. The incidence of solid tumors was higher in Lo/PHA than in Hi/PHA males but the same in females of the two lines, yet the probability of dying from solid tumors was slightly increased in Lo/PHA mice of both sexes. All these results indicate that T-cell-mediated immunity influences mainly the spontaneous incidence of lymphomas and, to a lesser degree, the appearance of other solid tumors.
Asunto(s)
Linfoma/veterinaria , Ratones/inmunología , Fitohemaglutininas/farmacología , Enfermedades de los Roedores/inmunología , Linfocitos T/inmunología , Animales , Bronconeumonía/epidemiología , Bronconeumonía/veterinaria , Femenino , Longevidad , Linfoma/epidemiología , Linfoma/genética , Linfoma/inmunología , Masculino , Ratones/genética , Neoplasias/epidemiología , Neoplasias/veterinaria , Enfermedades de los Roedores/epidemiología , Enfermedades de los Roedores/genética , Factores SexualesRESUMEN
Cutaneous mast cell tumours (cMCTs) are one of the most common cutaneous tumours in ferrets (Mustela putorius furo). However, limited information is available regarding cytological and histological features of these tumours and studies evaluating KIT expression are lacking in this species. The aims of this prospective study were to describe the most common clinical, cytological and histological features of cMCTs in ferrets and to compare the usefulness of different staining techniques in the diagnosis of these tumours in ferrets as well as evaluating KIT expression in neoplastic mast cells (MCs) by immunohistochemistry. Macroscopically, the tumours were small, round to plaque-like and frequently associated with surface crusting. The most common locations were the extremities and the trunk. MC granules were stained in all cases using toluidine blue (TB) and Wright-Giemsa stains in cytological specimens, but none stained with modified Wright's stain. Haematoxylin and eosin and TB on histological sections failed to stain MC granules in all the cases. Cytological and histological examination revealed low to moderate anisocytosis and anisokaryosis. An infiltrative rather than a delineated or encapsulated growth pattern was noted histologically in all cases. Eosinophilic infiltration was not uncommon and 'collagenolysis' was detected on cytological and histological examination. KIT expression was detected in all cases evaluated. In approximately one third of the cases the MCs exhibited KIT labelling pattern I and in the remaining ferrets, KIT pattern III. No correlation was found between KIT expression pattern and biological behaviour.
Asunto(s)
Hurones , Mastocitoma Cutáneo/veterinaria , Animales , InmunohistoquímicaRESUMEN
The treatment with infliximab is employed successfully in Crohn's disease (CD) but predictors of efficacy are lacking. Activation of the transcription factor NF-kB has been demonstrated in CD and its inhibition is one of the mechanisms by which anti-inflammatory agents exert their effects. We evaluated the production of TNFalpha by peripheral blood mononuclear cells (PBMC) and the levels of NF-kappaB family molecules in the intestinal mucosa during infliximab therapy in 12 patients. TNFalpha was assayed on supernatants of PBMC culture stimulated with PHA or LPS. Immunohistochemistry was also done on intestinal biopsies. In six patients, Western blot analysis of the NF-kappaB subunit Rel-A, and its inhibitors IkappaBalpha and IkappaBgamma was performed on intestinal biopsies and PBMC. The TNFalpha production by LPS stimulated PBMC showed mild changes, while it was increased by PHA-stimulated PBMC after treatment. The number of inflammatory cells in the intestinal mucosa was reduced (p<0.002) by the treatment. In five out of six cases we detected an increase of the IkappaBalpha and IkappaBgamma)inhibitor levels in intestinal biopsies after treatment. An increase of IkappaB inhibitors levels could be one of the mechanisms by which infliximab decreases NF-kappaB activity and exerts its anti-inflammatory effects.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Enfermedad de Crohn/tratamiento farmacológico , Enfermedad de Crohn/metabolismo , Fármacos Gastrointestinales/uso terapéutico , Proteínas I-kappa B/metabolismo , Mucosa Intestinal/metabolismo , FN-kappa B/antagonistas & inhibidores , Fragmentos de Péptidos/metabolismo , Factores de Transcripción/metabolismo , Adulto , Anciano , Western Blotting , Femenino , Humanos , Inmunohistoquímica , Infliximab , Masculino , Persona de Mediana Edad , Monocitos/inmunología , Inhibidor NF-kappaB alfa , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
A simple and ready-to-use program to analyze hydrophilicity and amphipathicity characteristics of protein sequences is described. Three of the most commonly used hydrophilicity scales can be selected and the block length can also be varied. The program is written in BASIC language, can be run on any microcomputer and does not require computer experience. The program output is exemplified by a sample of lysozyme hydrophilicity and amphipathicity analysis.
Asunto(s)
Computadores , Epítopos , Microcomputadores , Proteínas/inmunología , Programas Informáticos , Muramidasa/inmunología , Periodicidad , AguaRESUMEN
In our previous studies aimed at designing appropriate strategies to accelerate recovery of the immune system after irradiation, we found that the hematopoietic cytokine recombinant murine (rmu) interleukin (IL)-3 was able to induce differentiation and growth of thymocytes and splenic T and B lymphocytes in mice exposed to x-rays (200-500 cGy). The recovery, however, was complete at 7 days only after a dose of 200 cGy, whereas 2, 3, and 4 weeks were necessary to achieve full recovery after 300, 400, and 500 cGy, respectively. These studies were extended to investigate the effects of another hematopoietic cytokine, recombinant human (rhu) IL-11, a bone marrow stromal-derived cytokine, administered together with IL-3 to irradiated mice. The synergistic effect of the two cytokines was evident when relatively small doses of rhu IL-11 were injected with an optimal dose of rmu IL-3.
Asunto(s)
Linfocitos B/efectos de la radiación , Interleucina-11/farmacología , Interleucina-3/farmacología , Linfocitos T/efectos de la radiación , Irradiación Corporal Total , Animales , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/efectos de la radiación , Linfocitos T CD4-Positivos/efectos de la radiación , Linfocitos T CD8-positivos/efectos de la radiación , Ciclo Celular/efectos de los fármacos , Ciclo Celular/efectos de la radiación , Citocinas/farmacología , Relación Dosis-Respuesta en la Radiación , Hematopoyesis/efectos de la radiación , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Índice Mitótico/efectos de los fármacos , Índice Mitótico/efectos de la radiación , Proteínas Recombinantes/farmacología , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/efectos de la radiación , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Timo/efectos de los fármacos , Timo/inmunología , Timo/efectos de la radiación , Rayos XRESUMEN
Age-related alterations of the immune system affect both antibody and cell-mediated immune responses, T-cell responses being more severely affected than B-cell responses. Within the T-cell population, aging leads to replacement of virgin by memory cells and to accumulation of cells with signal transduction defects. Changes in T-cell subsets and in cytokine production profiles may produce suitable conditions for T-cell-mediated disregulation of antibody responses characterized by the production of low affinity and self-reactive antibodies. Also B-cells exhibit intrinsic defects and natural killer (NK) cell activity a profound loss in old mice. Whether age-related immune disfunctions influence life span and tumor incidence has been examined in mice genetically selected for high or low antibody responsiveness. It has been found that genetic selection of vigorous antibody responses in most cases produces mice with longer life span and lower lymphoma incidence. Moreover, the results of genetic segregation experiments indicate that antibody responsiveness and life span are polygenic traits regulated by a small number of the same or closely linked loci. Mice genetically selected for high or low mitotic responsiveness to PHA exhibit low or high tumor incidence, respectively, but no difference in life span, suggesting that T-cell activity is restricted to immune surveillance of neoplastic transformation. Studies on mice genetically selected for resistance or sensitivity to chemical carcinogenesis have uncovered loci that control both resistance to tumor induction and longevity while have no effects on immunity and disease incidence. Thus, the relative role of the immune system in conditioning the duration and the biological quality of life remains to be determined.
Asunto(s)
Envejecimiento/genética , Sistema Inmunológico/fisiología , Neoplasias/genética , Envejecimiento/inmunología , Animales , Formación de Anticuerpos , Inmunidad Celular/genética , Incidencia , Longevidad/inmunología , Ratones , Neoplasias/epidemiología , Neoplasias/inmunologíaRESUMEN
Young and old mice have been lethally irradiated and injected with syngeneic bone marrow cells from young or old donors to investigate whether self reactivity in old mice results from age-related damage of the radioresistant stromal cells and/or of the bone marrow hematopoietic cells. Thymus and spleen cell repopulations and mitotic responses at 3 months after irradiation are lower in old than in young recipients, suggesting age-related accumulation of stromal cell damage in the thymus as well as in other central and peripheral lymphoid tissues. The same efficiency of bone marrow cells from young and old donors to repopulate the thymus and spleen in recipients of equal age rules out the detrimental effects of aging on stem cells as well as T and B cell precursors. The serum concentration of auto-antibody and glomerular lesions at 3 and 9 months after irradiation were more pronounced in old than in young recipients and displayed no difference in recipients of equal age, regardless of the age of the bone marrow cell donors. These findings support the possibility that age-related damage of stromal cells induces disregulation of the immune system leading to autoimmune phenomena.
Asunto(s)
Envejecimiento/inmunología , Autoinmunidad , Trasplante de Médula Ósea/inmunología , Quimera por Radiación/inmunología , Animales , Autoinmunidad/efectos de la radiación , Femenino , Sistema Hematopoyético/citología , Sistema Hematopoyético/inmunología , Sistema Hematopoyético/efectos de la radiación , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Linfocitos T/inmunología , Linfocitos T/efectos de la radiación , Trasplante IsogénicoRESUMEN
We investigated the production of IL-2 and IFN-gamma (Th1 type) and IL-4 (Th2 type) cytokines by mitogen-activated spleen cells from young, adult and old mice. Cytokine production was evaluated in culture supernatants by CTLL proliferation (IL-2), ELISA (IFN-gamma), CT4.S proliferation (IL-4) and in mRNA extracted from activated CD4+ cells by RT-PCR (IL-2, IFN-gamma and IL-4). Results show that the production of IL-2, as protein and mRNA, is profoundly depressed by aging, whereas that of IFN-gamma, as protein and mRNA, firstly declines and then increases with age. The production of IL-4, as protein, monotonically declines with aging whereas, as mRNA, firstly decreases and then increases above the level in young mice. Spleen cells in culture were also incubated with mitogens and with a recombinant cytokine (IL-1 beta, IL-2, IL-3, IL-4, IL-12 or IFN-gamma) at various concentrations. It was found that recombinant cytokines by and large enhance cytokine production when the level induced by mitogens only is low. This conclusion applies to IL-2 and IFN-gamma production as protein and mRNA. The addition of recombinant cytokines also increases the production of IL-4 at the protein level in spleen cells from old mice but, at the mRNA level, only in spleen cells from young mice. This finding suggests age-related changes in IL-4-specific mRNA transcription rate and post-transcriptional half-life as well as translation kinetics.
Asunto(s)
Envejecimiento/inmunología , Citocinas/biosíntesis , Citocinas/farmacología , Regulación del Desarrollo de la Expresión Génica , Linfocitos T/inmunología , Animales , Linfocitos T CD4-Positivos/inmunología , Células Cultivadas , Femenino , Interferón gamma/biosíntesis , Interleucina-2/biosíntesis , Interleucina-4/biosíntesis , Activación de Linfocitos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/biosíntesis , Proteínas Recombinantes/farmacología , Bazo/inmunología , Linfocitos T/efectos de los fármacos , Células TH1/inmunología , Células Th2/inmunología , Transcripción GenéticaRESUMEN
Twenty-six week-old BDF1 mice were gonadectomized and grafted with thymus from irradiated (8.5 Gy) newborn, 6-week-old, or 26-week-old mice. One month later, grafted thymuses were recovered and examined in terms of thymocyte numbers, subpopulations and proliferative responses to Concananavlin A (Con A). The growth of the irradiated thymus was significantly higher in gonadectomized (Gx) than in sham-operated (Sham) mice and the magnitude of thymic growth was apparently age-dependent, as it was greater for newborns than for older mice. Con A response of thymocytes was also significantly higher in Gx mice than in Sham mice, and the magnitude of the response declined with advancing age of the thymus donors. Flow cytometric analysis revealed that a significant increase in the percentage of CD4+CD8- was observed in thymus grafts showing high Con A responses. However, this effect of Gx on the thymus graft was dependent on age of the thymus donor. Namely, newborn thymus grafts could grow equally well in both Gx and Sham recipients, whereas thymus grafts from 6- and 26-week-old mice could grow well only in Gx, but not in Sham recipients. The number of thymocytes was comparable in thymus grafts from 6- and 26-week-old mice, but the proliferative response to Con A was higher in the former than in the latter graft. Collectively, Gx appeared to promote immigration of thymocyte precursors into the thymus and to enhance proliferation and differentiation of thymocytes towards CD4+CD8- T cells, in an age-related manner.
Asunto(s)
Envejecimiento/patología , Linfocitos T/citología , Testículo/fisiología , Hiperplasia del Timo/patología , Animales , Animales Recién Nacidos , Relación CD4-CD8 , Diferenciación Celular/fisiología , Estudios de Evaluación como Asunto , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Orquiectomía , Células del Estroma/fisiología , Timo/trasplanteRESUMEN
The effects of aging on the activation of the cytoplasmic tyrosine protein kinase p56(lck) have been investigated in PBL from adult and elderly subjects upon activation with mitogens or different co-stimuli. Results show that the amount and phosphorylation of p56(lck) are reduced in PBL from elderly as compared to adult subjects. This finding suggests that alterations in p56(lck) may contribute to the age-associated loss of some T cell functions, such as proliferation and IL-2 production, which are found decreased in PBL from old individuals. However, p56(lck) seems irrelevant to the production of IFN-gamma and IL-4 which were both found increased in the PBL from old subjects, as expected from the relative expansion of memory versus naive T cell subpopulations in aging.
Asunto(s)
Envejecimiento/metabolismo , Leucocitos Mononucleares/metabolismo , Proteína Tirosina Quinasa p56(lck) Específica de Linfocito/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , División Celular , Células Cultivadas , Humanos , Interferón gamma/biosíntesis , Interleucina-2/biosíntesis , Interleucina-4/biosíntesis , Leucocitos Mononucleares/citología , Mitosis , FosforilaciónRESUMEN
Previous studies on DNA repair in ageing have demonstrated increased frequencies of single and double strand breaks in lymphocytes from elderly subjects and, as a consequence, decreased efficiency in DNA replication. We have investigated the relationship between cell proliferation and the nuclear expression of ku protein in a human population of 43 subjects of different ages. Ku is an heterodimeric protein composed of two subunits of 70 and 80 kDa, which is involved in the early steps of DNA damage recognition. In the present study, PBL from subjects of different ages were PHA-activated to evaluate the stimulation index and the production of Th1- and Th2-type cytokines. Moreover, nuclear extracts were obtained from activated lymphocytes to evaluate by a gel retardation assay the presence and the functional activity of the heterodimer ku 70/80. Our results indicate that ageing affects the mitotic responsiveness and cytokine production to a significant extent, but only marginally the expression of ku 70/80. These findings suggest that the age-related impairment in DNA repair mechanisms are only in part related to the reduced expression of ku protein able to recognize DNA damage.
Asunto(s)
Envejecimiento/metabolismo , Antígenos Nucleares , ADN Helicasas , Proteínas de Unión al ADN/metabolismo , Proteínas Nucleares/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Envejecimiento/fisiología , División Celular , Extractos Celulares , Núcleo Celular/metabolismo , Citocinas/biosíntesis , ADN/metabolismo , Dimerización , Humanos , Autoantígeno Ku , Leucocitos Mononucleares , MitosisRESUMEN
We have investigated the effects of an interleukin (IL)-6-type cytokine on the DNA-binding activity of ku and on unscheduled DNA repair in X-ray-treated peripheral blood mononuclear cells (PBMC) from human subjects of different ages. The cytokine used, called K-7/D-6, is an IL-6 variant with increased in vivo and in vitro biological activity compared to the wild type molecule. Ku is the DNA-binding component of the DNA-dependent protein kinase (DNA-PK). It binds the ends of various types of DNA discontinuity and is involved in the repair of DNA breaks caused by V(D)J recombination, isotype switching, physiological oxidation reactions, ionizing radiation and some chemotherapeutic drugs. The ku-dependent repair process, called non-homologous end joining, is the main DNA double strand break repair mechanism in irradiated mammalian cells. Results show that K-7/D-6 significantly increases DNA-binding activity of ku in irradiated PBMC from young but not from elderly subjects. However, K-7/D-6 is able to induce unscheduled DNA repair in irradiated PBMC from both young and elderly subjects. These effects of K-7/D-6 are relevant to the mechanisms of the cellular response to DNA damage.
Asunto(s)
Envejecimiento/sangre , Antígenos Nucleares , Daño del ADN/efectos de los fármacos , ADN Helicasas , Reparación del ADN/efectos de los fármacos , Interleucina-6/metabolismo , Monocitos/fisiología , Monocitos/efectos de la radiación , Adulto , Anciano , Anciano de 80 o más Años , Células Cultivadas , ADN/metabolismo , Proteínas de Unión al ADN/metabolismo , Relación Dosis-Respuesta en la Radiación , Humanos , Autoantígeno Ku , Proteínas Nucleares/metabolismo , Rayos XRESUMEN
IL-2 activity is commonly estimated in cell culture supernatants by an IL-2-dependent cell proliferation assay. This method is both reproducible and sensitive. However, it often appears from the literature that statistical analysis of the titration data either is disregarded or, when performed, is based on statistically incorrect assumptions. The proposed method is based on the principle of biological assay by parallel lines as applied to probit analysis of quantitative responses. The procedure has been embodied in a simple and interactive computer program which automatically estimates the IL-2 concentration in the biological sample, in terms of U/ml, and provides its standard error and confidence limits. This program is also suitable for quantitative determination of other biologically active substances that show a sigmoid dose/response relationship.
Asunto(s)
Interleucina-2/análisis , Animales , Bioensayo/normas , Relación Dosis-Respuesta Inmunológica , Femenino , Activación de Linfocitos , Ratones , Programas InformáticosRESUMEN
A sensitive and specific immunoenzyme assay (ELISA) for quantitation of total and cross-reactive idiotype-bearing (CRI) anti-ABA antibodies is described. Total anti-ABA antibodies are directly assessed in ABA-BGG coated polyvinyl wells with enzyme-labelled rabbit anti-mouse immunoglobulins. By interpolation on a standard curve absorbance values give the concentration of anti-ABA antibodies with a sensitivity of 30 ng/ml. CRI+ antibodies are quantitated by inhibition of enzyme-labelled monoclonal CRI+ antibody binding to solid-phase coated rabbit anti-CRI immunoglobulins. The concentration of CRI+ antibodies, evaluated by interpolation on a standard inhibition curve, can be measured at the level of 10 ng/ml. This highly sensitive, rapid, specific and reproducible assay is easily used, with minor modifications, to detect specific antibodies in any idiotype system.
Asunto(s)
Compuestos Azo/inmunología , Ensayo de Inmunoadsorción Enzimática , Técnicas para Inmunoenzimas , Idiotipos de Inmunoglobulinas/análisis , p-Azobencenoarsonato/inmunología , Animales , Especificidad de Anticuerpos , Sitios de Unión de Anticuerpos , Unión Competitiva , Reacciones Cruzadas , Idiotipos de Inmunoglobulinas/metabolismo , Ratones , Ratones Endogámicos A , ConejosRESUMEN
Data obtained from assays for the evaluation of helper or suppressor activity are analyzed by statistical techniques that increase the accuracy and amount of information. The analysis is performed by a set of simple and ready to use computer programs which do not require statistical expertise and are available on request.