RESUMO
BACKGROUND: The fucose-mannose ligand (FML) of Leishmania infantum is a complex glycoprotein which does not elicit adequate immunogenicity in humans. In recent years, adjuvant compounds derived from plants have been used for improving the immunogenicity of vaccines. Glycyrrhizin (GL) is a natural triterpenoid saponin that has known immunomodulatory activities. In the present study, we investigated the effects of co-treatment with FML and GL on the production of cytokines and nitric oxide (NO) by macrophages, in vitro. METHODS: Lipopolysaccharide (LPS) stimulated murine peritoneal macrophages were treated with FML (5 µg/ml) of L. infantum and various concentrations of GL (1 µg/ml, 10 µg/ml and 20 µg/ml). After 48 h of treatment, cell culture supernatants were recovered and the levels of TNF-α, IL-10, IL-12p70 and IP-10 were measured by sandwich ELISA and NO concentration by Griess reaction. RESULTS: Our results indicate that the treatment of activated macrophages with FML plus GL leads to enhanced production of NO, TNF-α and IL-12p70, and reduction of IL-10 levels in comparison with FML treatment alone. CONCLUSIONS: Therefore, we concluded that GL can improve the immunostimulatory effect of FML on macrophages and leads to their polarization towards an M1-like phenotype.
Assuntos
Citocinas/metabolismo , Ácido Glicirrízico/farmacologia , Lectinas/farmacologia , Leishmania infantum/metabolismo , Óxido Nítrico/metabolismo , Adjuvantes Imunológicos/farmacologia , Animais , Antígenos de Protozoários/imunologia , Combinação de Medicamentos , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Lectinas/metabolismo , Vacinas contra Leishmaniose/química , Leishmaniose Visceral/imunologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/metabolismo , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was developed for residue analysis of azoxystrobin in garden crops, for which the maximum residue limits (MRLs) are 0.5-50 mg/kg in Japan. For hapten synthesis, an ethyl carboxyl group was introduced to the 4-position of the 2-cyanophenoxy group in azoxystrobin, and its cyano group was changed to a methyl group. An anti-azoxystrobin monoclonal antibody was prepared from mice immunized with hapten-keyhole limpet hemocyanin conjugate. The dc-ELISA using prepared antibody showed 50-250-fold higher sensitivity compared to the MRLs. The working range of the dc-ELISA was 10-200 ng/mL. The dc-ELISA showed high specificity to azoxystrobin. When methanol extracts from nine kinds of garden crops spiked with azoxystrobin ranging near the MRLs were analyzed, the determined results by the dc-ELISA agreed well with the results of their controls. In addition, azoxystrobin spiked in garden crops homogenates was satisfactorily extracted by methanol solution and easily analyzed. The recovery rate of dc-ELISA was 96-109% and correlated well with the results obtained by HPLC analysis.
Assuntos
Produtos Agrícolas/química , Ensaio de Imunoadsorção Enzimática/métodos , Fungicidas Industriais/análise , Metacrilatos/análise , Resíduos de Praguicidas/análise , Pirimidinas/análise , Animais , Anticorpos Monoclonais/biossíntese , Japão , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Extratos Vegetais/química , EstrobilurinasRESUMO
Oil-in-water emulsions have shown promise as safe and effective adjuvant formulations for vaccines. In particular, formulations consisting of metabolizable oils such as shark-derived squalene and detergents such as egg phosphatidylcholine have been used to produce stable vaccine emulsion formulations. However, there is an emphasis in pharmaceutical regulatory bodies on using synthetic or plant-derived components from sustainable sources instead of animal-derived components. This study compares the physicochemical properties and biological efficacy of emulsions consisting of oil and detergent components from animal, plant, and synthetic sources. In particular, effects of component structure and source on emulsion stability and biological activity are examined. It is shown that oil-in-water emulsions using animal-derived components can be substituted with synthetic or plant-derived materials while still exhibiting satisfactory physicochemical and biological properties.
Assuntos
Adjuvantes Imunológicos/química , Emulsões/química , Óleos/química , Água/química , Animais , Estabilidade de Medicamentos , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Fosfatidilcolinas/química , Poloxâmero/química , Polissorbatos/química , Esqualeno/química , Tensoativos/química , Triglicerídeos/químicaRESUMO
The in vivo immunogenicity of a new interferon (IFN) beta-1a product (Rebif New Formulation; RNF) was compared with that of two approved recombinant human IFN beta-1a products (Rebif and Avonex). Immunogenic potential was assessed based on time to development of neutralizing antibodies (NAbs) and NAb titer. Female BALB/c mice (six in each group) received RNF, Rebif or Avonex (1.0 microg/mL subcutaneously three times weekly), and serum samples collected on Days 7, 21, and 35 (Study 1), or 28, 42, 49, and 60 (Study 2) were assayed for NAbs. In Study 1, no mice had NAbs at Day 7, but by Day 21 one mouse in the RNF group had NAbs, compared with three and four mice in the Rebif and Avonex groups, respectively. Results were similar in Study 2. All control mice were NAb negative; all actively treated mice had NAbs by day 35 or 42. Throughout Study 1, NAb titers were lowest in the RNF group and highest in the Avonex group, and at day 35, NAb titers were significantly lower in the RNF group than the Rebif group (p = 0.037). Results indicate that, on a gram-for-gram basis, RNF appears less immunogenic than Rebif or Avonex.
Assuntos
Anticorpos/sangue , Imunização , Interferon beta/imunologia , Animais , Esquema de Medicação , Avaliação Pré-Clínica de Medicamentos , Feminino , Injeções Subcutâneas , Interferon beta-1a , Interferon beta/administração & dosagem , Camundongos , Camundongos Endogâmicos BALB C/sangue , Camundongos Endogâmicos BALB C/imunologia , Esclerose Múltipla/terapia , Testes de Neutralização , Fatores de TempoRESUMO
BACKGROUND: Nasal instillation is an effective method for inducing antigen-specific immune tolerance. However, it is not clear how a tolerization scheme established in one mouse strain will perform when used in a mouse of a different haplotype. OBJECTIVES: To compare the antigen-specific recall responses in four mouse strains--BALB/c, C57BL/6, NOD, and B10.PL--that were pretreated nasally with 50 micrograms of hen egg-white lysozyme prior to parenteral immunization with homologous antigen. METHODS: Mice were nasally treated with a prototype antigen, HEL, and then immunized with the same antigen emulsified in complete Freund's adjuvant. Spleens and lymph nodes were assayed for T cell proliferation measured by tritiated thymidine incorporation. Cytokine production was measured using ELISPOT assay. Serum antibody response to HEL was measured using an enzyme-linked immunosorbent assay. RESULTS: Proliferative recall responses to HEL in B10.PL, C57BL/6, and BALB/c were greatly reduced compared to control mice, but non-obese diabetic mice were resistant to the tolerization regime. Despite their susceptibility to nasally induced suppression, the mechanisms responsible for tolerance induction differed in BALB/c and C57BL/6 mice. CONCLUSIONS: Our findings demonstrate that while mucosal contacts with specific antigen consistently affect the outcome of subsequent exposure to the same antigen, the observed response will vary non-predictably, depending on the genetic background of the animal.
Assuntos
Doenças Autoimunes/imunologia , Doenças Autoimunes/terapia , Modelos Animais de Doenças , Tolerância Imunológica/imunologia , Imunização/métodos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos C57BL/imunologia , Camundongos Endogâmicos NOD/imunologia , Muramidase/imunologia , Muramidase/uso terapêutico , Animais , Doenças Autoimunes/induzido quimicamente , Avaliação Pré-Clínica de Medicamentos , Epitopos/genética , Epitopos/imunologia , Haplótipos/genética , Haplótipos/imunologia , Tolerância Imunológica/genética , Imunidade nas Mucosas/imunologia , Instilação de Medicamentos , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos BALB C/genética , Camundongos Endogâmicos C57BL/genética , Camundongos Endogâmicos NOD/genética , Mucosa Nasal , Linfócitos T/imunologiaRESUMO
Los antígenos del sistema ABO de grupos sanguíneos son oligosacáridos presentes en la membrana celular que forman parte de las glicoproteínas y glicolípidos. La introducción de la tecnología de hibridomas murinos para la obtención de anticuerpos monoclonales (AcM) con especificidad para los antígenos ABO ha permitido iniciar en Cuba la producción de reactivos hemoclasificadores basados en AcM. El perfeccionamiento de los reactivos requiere la búsqueda de nuevos clones de hibridomas, para lo que se necesita la sensibilización de ratones Balb/c con los antígenos. Se presentan los resultados de la evaluación de dos vías de inoculación (intraperitoneal e intraesplénica) y tres formas de presentación del antígeno (hematíes completos del grupo B, sustancia específica B y sustancia B tratada con alúmina) en ratones Balb/c a los que se les determinó el título de anticuerpos hemaglutinantes por la técnica de microplacas frente a un panel de hematíes de los grupos A, B y O antes y durante el período de inmunización. La vía intraperitoneal con hematíes completos del grupo B resultó adecuada para alcanzar altos títulos de anticuerpos aglutinantes contra hematíes B, aunque también reaccionaron con los hematíes de otros grupos.
Assuntos
Animais , Camundongos , Humanos , Sistema ABO de Grupos Sanguíneos , Anticorpos Monoclonais/imunologia , Antígenos de Grupos Sanguíneos , Hemaglutinação , Hibridomas , Indicadores e Reagentes/síntese química , Imunização/métodos , Camundongos Endogâmicos BALB C/imunologia , Etanol/administração & dosagem , Adjuvante de Freund/administração & dosagem , Óxido de Alumínio/administração & dosagem , Interpretação Estatística de DadosRESUMO
Los antígenos del sistema ABO de grupos sanguíneos son oligosacáridos presentes en la membrana celular que forman parte de las glicoproteínas y glicolípidos. La introducción de la tecnología de hibridomas murinos para la obtención de anticuerpos monoclonales (AcM) con especificidad para los antígenos ABO ha permitido iniciar en Cuba la producción de reactivos hemoclasificadores basados en AcM. El perfeccionamiento de los reactivos requiere la búsqueda de nuevos clones de hibridomas, para lo que se necesita la sensibilización de ratones Balb/c con los antígenos. Se presentan los resultados de la evaluación de dos vías de inoculación (intraperitoneal e intraesplénica) y tres formas de presentación del antígeno (hematíes completos del grupo B, sustancia específica B y sustancia B tratada con alúmina) en ratones Balb/c a los que se les determinó el título de anticuerpos hemaglutinantes por la técnica de microplacas frente a un panel de hematíes de los grupos A, B y O antes y durante el período de inmunización. La vía intraperitoneal con hematíes completos del grupo B resultó adecuada para alcanzar altos títulos de anticuerpos aglutinantes contra hematíes B, aunque también reaccionaron con los hematíes de otros grupos. (AU)
Assuntos
Animais , Camundongos , Humanos , Antígenos de Grupos Sanguíneos , Hibridomas , Anticorpos Monoclonais/imunologia , Camundongos Endogâmicos BALB C/imunologia , Sistema ABO de Grupos Sanguíneos , Imunização/métodos , Hemaglutinação , Indicadores e Reagentes/síntese química , Óxido de Alumínio/administração & dosagem , Adjuvante de Freund/administração & dosagem , Etanol/administração & dosagem , Interpretação Estatística de DadosRESUMO
PGE2 has been shown to play a prominent role in regulating Th1 and Th2 type responses. We studied the role of PGE2 in IFN-gamma production by Staphylococcus aureus Cowan I-stimulated spleen cells from several mouse strains such as BALB/c, C3H/HeN, and C57BL/6. When spleen cells were pretreated with indomethacin (cyclooxygenase (COX)-1 and COX-2 inhibitor) or NS-398 (COX-2-specific inhibitor), S. aureus Cowan I -induced IFN-gamma production was increased more markedly in spleen cells from BALB/c mice than from C3H/HeN and C57BL/6 mouse. However, PGE2 production was not significantly different among spleen cells from three mouse strains. When various concentrations of PGE2 were exogeneously added to spleen cells, PGE2 showed a stronger suppressive effect on IFN-gamma production in spleen cells from BALB/c mice than from other strains of mice. This suppressive effect of PGE2 in BALB/c mice mainly depended on IL-12p70 production by APCs. More PGE2 binding sites were found in BALB/c spleen cells than in C3H/HeN spleen cells, indicating that the sensitivity difference to the suppressive effect of PGE2 was due to the difference of the number of PGE2 receptors. The administration of NS-398 into BALB/c mice enhanced Ag-specific IFN-gamma production, but not IL-4 production. This effect is the same as IL-12 administration in vivo. From these results, we propose that the modulation of PGE2 is important for Th1 activation via IFN-gamma and IL-12p70 production in vitro and in vivo and that PGE2 is one of the pivotal factors in the Th2-dominant immune response in BALB/c mice.
Assuntos
Dinoprostona/farmacologia , Imunossupressores/farmacologia , Camundongos Endogâmicos BALB C/imunologia , Células Th2/imunologia , Adjuvantes Imunológicos/administração & dosagem , Animais , Células Apresentadoras de Antígenos/imunologia , Sítios de Ligação/imunologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Células Cultivadas , Ciclo-Oxigenase 2 , Citocinas/fisiologia , Dinoprostona/administração & dosagem , Dinoprostona/metabolismo , Imunidade Celular/efeitos dos fármacos , Imunossupressores/administração & dosagem , Injeções Intramusculares , Injeções Intraperitoneais , Interferon gama/antagonistas & inibidores , Interferon gama/biossíntese , Interleucina-10/farmacologia , Isoenzimas/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Óxido Nítrico/fisiologia , Nitrobenzenos/administração & dosagem , Prostaglandina-Endoperóxido Sintases/farmacologia , Baço/citologia , Baço/efeitos dos fármacos , Baço/imunologia , Baço/metabolismo , Sulfonamidas/administração & dosagem , Células Th1/citologia , Células Th1/efeitos dos fármacos , Células Th1/enzimologia , Células Th2/efeitos dos fármacos , Células Th2/metabolismoRESUMO
A novel form of adjuvant-neuroantigen formulation was established which was highly encephalitogenic in previously resistant BALB/c mice. The antigen formulation contained mouse whole spinal cord homogenate (MSCH), mycobacteria, and mineral oil, identically to the conventional preparation, but emulsification was completed by sonication instead of extrusion. Sonication of MSCH alone did not render a conventionally prepared emulsion encephalitogenic. The novel adjuvant formulation showed reduced water-oil droplet size, and the neuroantigen was located on the surface of the droplets as well as in the intermicellar space, while in the extruded formulation the material was buried in the droplet interior. Mice inoculated with the sonicated emulsion showed strong brain and spinal cord infiltration of lymphoid cells. The sonicated emulsion was highly encephalitogenic in all six BALB/c substrains tested. The results suggest that availability of the neuroantigen is of critical importance for the development of clinical EAE in the BALB/c mouse.
Assuntos
Encefalomielite Autoimune Experimental/imunologia , Camundongos Endogâmicos BALB C/imunologia , Proteínas do Tecido Nervoso/imunologia , Adjuvantes Imunológicos/química , Animais , Emulsões , Feminino , Camundongos , Microscopia Eletrônica , Proteínas do Tecido Nervoso/química , Sonicação , Medula Espinal/imunologiaRESUMO
Three monoclonal antibodies with either specificity to taxol or baccatin III, or cross-reactivity with several common taxanes have been prepared and used to develop sensitive competitive-inhibition enzyme immunoassays. The hybridomas producing these monoclonal antibodies were obtained by fusing P3X63Ag8.653 plasmacytoma cells and splenocytes from mice hyperimmunized with keyhole limpet hemocyanin-7-succinyltaxol or -7-succinylbaccatin III conjugates. Direct and indirect competitive inhibition enzyme immunoassays were developed with these monoclonal antibodies and microtiter plates coated with bovine serum albumin conjugates of the complementary hapten. Detection limits for the direct competitive inhibition enzyme immunoassays, conducted in buffer containing 10% MeOH, were 0.6 nM taxol for 3C6 (anti-taxol); 1.1 nM baccatin III for 3H5 (anti-baccatin III); and 0.6 nM taxol or baccatin III for 8A10 (anti-taxane). The immunoassays accurately detected taxol, baccatin III, and "total taxanes" in crude MeOH extracts of Taxus brevifolia bark and in hplc fractions of these extracts.
Assuntos
Alcaloides/análise , Anticorpos Monoclonais , Hidrocarbonetos Aromáticos com Pontes/análise , Medicamentos de Ervas Chinesas/análise , Paclitaxel/análise , Plantas Medicinais/química , Taxoides , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/isolamento & purificação , Especificidade de Anticorpos , Reações Cruzadas , Feminino , Técnicas Imunoenzimáticas , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Extratos Vegetais/química , Folhas de Planta/químicaRESUMO
A new strategy for generating abzyme was developed. Glutathione peroxidase (GPX, EC 1.11.1.9) is one of the important members of antioxidation enzyme system; it catalyzes the reductions of a variety of hydroperoxides in presence of glutathione(GSH). We have first prepared the monoclonal antibody (McAb) with GSH binding sites, then incorporated GPX catalytic group selenocystein (SeCys) into the antibody combining sites by using chemical mutation. Thus the mutated antibody displays high GPX activity, which approaches the magnitude level of native GPX, exhibits the kinetic behavior similar to native GPX, and has some advantages over native GPX.
Assuntos
Glutationa Peroxidase/metabolismo , Mutagênese , Selênio , Animais , Anticorpos Monoclonais/metabolismo , Sítios de Ligação de Anticorpos , Ensaio de Imunoadsorção Enzimática , Glutationa/análogos & derivados , Glutationa/síntese química , Glutationa/metabolismo , Glutationa Peroxidase/análise , Glutationa Peroxidase/biossíntese , Haptenos , Imunoglobulina G/metabolismo , Cinética , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Coelhos/imunologia , Selenocisteína/metabolismoRESUMO
PMEA was found to influence the immune system of the mouse and rat in vivo in different ways. PMEA administered after injection of parental splenocytes to F1 recipients reduced the development of local graft-versus-host reaction (GVHR). However, PMEA treatment of donors of splenocytes had no influence on GVHR. Modifications in the immune system triggered by PMEA were confirmed in the rat model by evaluation of subsets of white blood cells isolated from peripheral blood by a set of monoclonal antibodies. Enhanced formation of nitric oxide (NO) was found in both unconditioned and LPS-stimulated macrophage cultures on day 14 of the drug treatment of rats.
Assuntos
Adenina/análogos & derivados , Adjuvantes Imunológicos/farmacologia , Organofosfonatos , Adenina/imunologia , Animais , Anticorpos Monoclonais , Células Cultivadas , Reação Enxerto-Hospedeiro , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos C3H/imunologiaRESUMO
Naturally occurring factors that regulate the infectivity of P. berghei infected rodent hosts to the mosquito vector in vivo have been compared in T.O., Balb/C and immunodeficient SCID mice. No detectable differences in infectivity were observed suggesting B and T cell mediated factors are not involved. Further studies investigated roles for macrophage colony stimulating factors, the cytokines IFN gamma and TNF alpha, of neutrophils, and of nitric oxide in the SCID mouse, but have failed to demonstrate an important role in vivo for any factor examined. Differences between these results and those obtained in vitro on the human and primate parasites must therefore be explained by biological differences between the parasite/host combinations, or by technical differences in experimental designs. Induced immunity to the ookinete surface antigen Pbs 21 of P. berghei can totally block the transmission of the parasite from the gametocyte infected host to the vector. We have cloned the gene encoding Pbs 21 and shown it bears striking structural similarities to Pfs 25, Pgs 25 and more particularly Pgs 28 in that it has a high cysteine content (9.5%), 4 EGF-like domains and hydrophobic amino-'signal'--and carboxyl-'anchor' sequences. The encoding gene is on chromosome 5 and is found also in P. chabaudi, P. yoelii and P. vinckei.
Assuntos
Anopheles/parasitologia , Antígenos de Protozoários/imunologia , Insetos Vetores/parasitologia , Malária/transmissão , Plasmodium berghei/fisiologia , Proteínas de Protozoários/imunologia , Aminoácido Oxirredutases/antagonistas & inibidores , Sequência de Aminoácidos , Animais , Antígenos de Protozoários/genética , Arginina/análogos & derivados , Arginina/farmacologia , Sequência de Bases , Clonagem Molecular , Sequência Consenso , DNA Complementar/genética , DNA de Protozoário/genética , Feminino , Interações Hospedeiro-Parasita , Lipopolissacarídeos/farmacologia , Malária/complicações , Malária/imunologia , Malária/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos BALB C/parasitologia , Camundongos SCID/imunologia , Camundongos SCID/parasitologia , Dados de Sequência Molecular , Óxido Nítrico Sintase , Fases de Leitura Aberta , Plasmodium/genética , Plasmodium berghei/imunologia , Proteínas de Protozoários/genética , Reprodução , Imunodeficiência Combinada Severa/complicações , Especificidade da Espécie , ômega-N-MetilargininaRESUMO
Five Shigella vaccine candidates (EcSf2a-1, EcSf2a-2, Sfl124, T32-Istrati and SMD) were tested for safety and efficacy in Balb/cJ mice using an intranasal challenge model. Experiments in this model suggest that (i) the relative attenuation of vaccines can be determined in mice by intranasal inoculation, (ii) all vaccines tested elicited antibacterial mucosal immunity protecting against pulmonary infection with Shigella flexneri 2a, (iii) protection was associated with serum IgA and/or IgG antibody recognizing the 2a somatic antigen.
Assuntos
Anticorpos Antibacterianos/biossíntese , Vacinas Bacterianas , Shigella flexneri/imunologia , Administração Intranasal , Animais , Anticorpos Antibacterianos/imunologia , Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/imunologia , Vacinas Bacterianas/toxicidade , Avaliação Pré-Clínica de Medicamentos , Disenteria Bacilar/prevenção & controle , Ensaio de Imunoadsorção Enzimática , Feminino , Isotipos de Imunoglobulinas/biossíntese , Isotipos de Imunoglobulinas/imunologia , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Pneumonia/microbiologia , Pneumonia/prevenção & controle , SegurançaRESUMO
Glutathione peroxidase (GSH-Px) is an important selenium-containing enzyme which protects cells from oxidative damage. Two hybridoma clones (GPX-121 and GPX-347), producing mouse IgG1 monoclonal antibodies specific for GSH-Px, were established. Immunoblot analysis revealed that GPX-347 was specific for human GSH-Px, while GPX-121 cross-reacted with human, rat, mouse and rabbit GSH-Px. Correlation between GSH-Px content and its enzymatic activity was investigated in erythrocytes of 76 humans and in human lung adenocarcinoma PC-9 cells by using a sandwich type ELISA. The results indicated that GSH-Px activity was expressed higher than expected from GSH-Px content especially in the range of low GSH-Px concentration. PC-9 cells selenium depleted medium did not stain but the cytoplasm of PC-9 cells grown in medium supplemented with selenium stained strongly.
Assuntos
Anticorpos Monoclonais/imunologia , Glutationa Peroxidase/imunologia , Imunoglobulina G/imunologia , Adenocarcinoma/enzimologia , Adulto , Idoso , Animais , Anticorpos Monoclonais/isolamento & purificação , Reações Cruzadas , Meios de Cultura , Ensaio de Imunoadsorção Enzimática , Eritrócitos/enzimologia , Feminino , Glutationa Peroxidase/sangue , Humanos , Hibridomas/imunologia , Neoplasias Pulmonares/enzimologia , Masculino , Mamíferos/imunologia , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Pessoa de Meia-Idade , Proteínas de Neoplasias/imunologia , Selênio/farmacologia , Especificidade da Espécie , Células Tumorais Cultivadas/efeitos dos fármacosRESUMO
We describe here the utilization of a modified enzyme-linked immunospot assay (ELISPOT) in order to detect an autologous antiidiotypic response in mice at the level of single antibody-forming cells (AFC). Severals assays have been routinely used to detect anti-Id producing cells; however, these approaches often produce contrasting data. We present results obtained with the modified ELISPOT, using as a model system the antiidiotypic response in mice after immunization with a vaccine from Streptococcus pneumoniae R36a, expressing the immunodominant epitope phosphorylcholine (PC). The response to PC is mediated by a large fraction of antibodies bearing the public idiotype T15. Mice of different genetics make up were immunized with a single injection of the vaccine. We observed that one mouse strain (D1.LP) out of three was able to mount a significant anti-T15 response during the primary anti-phosphorylcholine response. BALB/c and C57BL/6 mice did not produce significant levels of anti-T15 antibody following a single injection of the antigen. In contrast, BALB/c mice which were repeatedly stimulated showed a specific anti-Id response. Experimental controls were performed using either specific anti-T15 monoclonal antibodies (mAb) or splenocytes from mice immunized with TEPC15 myeloma protein in complete Freund's adjuvant.
Assuntos
Anticorpos Anti-Idiotípicos/imunologia , Anticorpos Antibacterianos/biossíntese , Células Produtoras de Anticorpos/imunologia , Antígenos de Bactérias/imunologia , Técnicas Imunoenzimáticas , Animais , Vacinas Bacterianas , Epitopos Imunodominantes/imunologia , Idiótipos de Imunoglobulinas/imunologia , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos C57BL/imunologia , Fosforilcolina/imunologia , Streptococcus pneumoniae/imunologia , VacinaçãoRESUMO
The effect of glycyrrhizin (GR), a Chinese herbal drug extracted from licorice roots, on the host resistance to tumors was investigated in a murine system. Administration of GR to BALB/c mice, which were inoculated s.c. with 1 x 10(6) cells/mouse of Meth A tumors, resulted in either no antitumor effect (8/20, 40%), a delay in tumor growth (9/20, 45%), or elimination of tumor growth (3/20, 15%) in these mice. In addition, the incidence of Meth A solid tumors was inhibited by GR when mice were inoculated with 1.5 x 10(4) cells/mouse or less of Meth A tumor cells, but not 7.5 x 10(4) cells/mouse or more. These results indicate that in this murine tumor system GR has a very weak antitumor effect. When GR at a dose of 20 mg/kg was administered to mice immunized with allogeneic lymphocytes 1 to 9 days after the immunization, the generation of suppressor macrophages (S-M phi) was clearly reduced as compared with that of S-M phi generated in immunized controls. In addition, when allospecific CTL (allo-CTL), which were generated in alloimmunized mice treated with GR followed by in vitro stimulation with allogeneic lymphocytes in a mixed lymphocyte reaction, were adoptively transferred to tumor-bearing mice treated with GR, the antitumor activity of allo-CTL derived from immunized mice treated with GR was markedly enhanced as compared with that of allo-CTL from immunized mice. Furthermore, established solid tumors were completely eliminated when interleukin-2 immunotherapy was performed in these mice in combination with GR treatments, but not interleukin-2 or GR alone.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Ácido Glicirretínico/análogos & derivados , Imunoterapia , Macrófagos/efeitos dos fármacos , Sarcoma Experimental/terapia , Animais , Feminino , Ácido Glicirretínico/farmacologia , Ácido Glicirretínico/uso terapêutico , Ácido Glicirrízico , Tolerância Imunológica/efeitos dos fármacos , Imunização , Imunoterapia Adotiva , Interleucina-2/uso terapêutico , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Camundongos Endogâmicos C57BL/imunologia , Transplante de Neoplasias , Sarcoma Experimental/imunologia , Baço/imunologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/transplanteRESUMO
Balb/c mice were immunized against papain-treated fetal erythrocytes and splenocytes were fused with Sp2/0-Ag-14 myeloma cells. Several hybrids secreting antibodies directed against antigenic determinants predominantly exposed on fetal and cord cells were selected and cloned twice. Antibodies NaM61-1A2 and NaM61-768 (IgM class) were shown to be specific for an endo-beta-galactosidase-sensitive oligosaccharide chain. The antigen, strongly expressed on fetal and cord cells, was identified as the i blood group antigen. The antibodies represent powerful blood group reagents to be use in conventional agglutination techniques as well as in the gel typing system and in indirect flow cytometry. The antibody NaM46-4A8 (IgG class) is specific for an antigenic structure expressed on fetal cells and accessible only after papain, ficin, bromelin and endo-beta galactosidase treatment. The antigen was not identified.
Assuntos
Anticorpos Monoclonais/imunologia , Membrana Eritrocítica/imunologia , Sangue Fetal/imunologia , Glicosídeo Hidrolases , Hemaglutininas/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Adulto , Animais , Especificidade de Anticorpos , Bromelaínas/farmacologia , Sequência de Carboidratos , Membrana Eritrocítica/efeitos dos fármacos , Feminino , Ficina/farmacologia , Citometria de Fluxo , Testes de Hemaglutinação , Humanos , Recém-Nascido/sangue , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Dados de Sequência Molecular , Papaína/farmacologia , Gravidez , beta-Galactosidase/farmacologiaRESUMO
A peptide isolated from Viscum album extract (Iscador) has been earlier reported to have cytotoxic and tumour reducing activity. Administration of the peptide (2 micrograms/ml) was found to produce increased natural killer cell activity (NK-activity) in the normal animals and tumour bearing animals. The peak activity was observed on the 3rd day after the administration of the peptide. Administration of the peptide also stimulated antibody dependent cellular cytotoxicity (ADCC) which was expressed maximally on the fourth day. There was also an increase in antibody forming cells in the spleen, and antibody titers were increased in the animals treated with the peptide. Activity of the crude plant extract coincided with the activity of the peptide, indicating that the isolated peptide is mainly responsible for the immunostimulatory activity present in Viscum album extract.
Assuntos
Adjuvantes Imunológicos/farmacologia , Formação de Anticorpos/efeitos dos fármacos , Citotoxicidade Celular Dependente de Anticorpos/efeitos dos fármacos , Células Matadoras Naturais/efeitos dos fármacos , Extratos Vegetais/química , Proteínas de Plantas/farmacologia , Adjuvantes Imunológicos/isolamento & purificação , Animais , Células Produtoras de Anticorpos/efeitos dos fármacos , Carcinoma de Ehrlich/imunologia , Citotoxicidade Imunológica/efeitos dos fármacos , Feminino , Células Matadoras Naturais/imunologia , Leucemia Eritroblástica Aguda/patologia , Camundongos , Camundongos Endogâmicos BALB C/imunologia , Proteínas de Plantas/isolamento & purificação , Células Tumorais CultivadasRESUMO
Fractions of bovine colostrum were prepared and their ability to support the growth of mouse-mouse hybridomas in culture was tested. Whey was prepared from defatted colostrum by removal of casein using acid precipitation. An ultrafiltrate was obtained from cleared whey by filtration through membranes with a nominal molecular mass cut-off of 100,000 Da. Colostrum ultrafiltrate contained 1.16 milligrams protein, 0.24 milligrams immunoglobulin G (IgG) and less than 0.24 EU (endotoxin unit)/ml endotoxins. The effect of defatted colostrum, whey and ultrafiltrate as serum substitutes was examined by cultivation of hybridoma cells in minimal essential medium containing different concentrations of the supplements. Under optimal conditions in ultrafiltrate-supplemented medium, the maximal cell concentration was 35-40% of that obtained using 10% foetal bovine serum, and IgG production per cell was equal to that achieved using serum. In 1% defatted colostrum the maximum hybridoma concentration was about 30% of that in 10% serum, but at higher concentrations hybridoma growth was significantly reduced. The growth-promoting activity of whey was low. The results show that bovine colostrum ultrafiltrate provides a very attractive alternative to serum for production of monoclonal antibodies.