RESUMO
Sexual maturation of Anastrepha fraterculus is a long process. Methoprene (a mimic of juvenile hormone) considerably reduces the time for sexual maturation in males. However, in other Anastrepha species, this effect depends on protein intake at the adult stage. Here, we evaluated the mating competitiveness of sterile laboratory males and females that were treated with methoprene (either the pupal or adult stage) and were kept under different regimes of adult food, which varied in the protein source and the sugar:protein ratio. Experiments were carried out under semi-natural conditions, where laboratory flies competed over copulations with sexually mature wild flies. Sterile, methoprene-treated males that reached sexual maturity earlier (six days old), displayed the same lekking behaviour, attractiveness to females and mating competitiveness as mature wild males. This effect depended on protein intake. Diets containing sugar and hydrolyzed yeast allowed sterile males to compete with wild males (even at a low concentration of protein), while brewer´s yeast failed to do so even at a higher concentration. Sugar only fed males were unable to achieve significant numbers of copulations. Methoprene did not increase the readiness to mate of six-day-old sterile females. Long pre-copulatory periods create an additional cost to the management of fruit fly pests through the sterile insect technique (SIT). Our findings suggest that methoprene treatment will increase SIT effectiveness against A. fraterculus when coupled with a diet fortified with protein. Additionally, methoprene acts as a physiological sexing method, allowing the release of mature males and immature females and hence increasing SIT efficiency.
Assuntos
Proteínas Alimentares , Hormônios Juvenis , Metoprene , Controle Biológico de Vetores/métodos , Comportamento Sexual Animal/efeitos dos fármacos , Tephritidae , Ração Animal , Animais , Suplementos Nutricionais , Feminino , Masculino , Controle Biológico de Vetores/economia , Pupa/crescimento & desenvolvimento , Maturidade Sexual/efeitos dos fármacos , Tephritidae/crescimento & desenvolvimentoRESUMO
Over the past years, progress has been made in understanding B cells and antibody recognition functions, particularly in the context of autoimmune diseases. In addition to the existence of "natural antibodies", recent studies suggest the existence of immunoglobulins with no apparent specificity that may acquire polyreactivity following a mild denaturation in inflammatory sites. They are called "silent antibodies". Together with related observations on B cell development, selection and signaling, the recent insights are providing clues into our understanding of autoimmunity.
Assuntos
Autoanticorpos/imunologia , Autoimunidade/imunologia , Linfócitos B/imunologia , Imunoglobulinas/imunologia , Especificidade de Anticorpos/imunologia , Reações Antígeno-Anticorpo/imunologia , Doenças Autoimunes/imunologia , Anergia Clonal/imunologia , Humanos , Imunidade Inata/imunologia , Inflamação/imunologia , Tolerância a Antígenos Próprios/imunologia , Linfócitos T/imunologiaRESUMO
In a previous study, it was shown that an intramuscular administration of amino acid PADRE-ELDKWA sequence induced a mucosal immune response to a conserved epitope of human immunodeficiency virus in mice. In the same model, here it is shown that this method can be used with a selected peptide from the M protein of group A streptococci. The PADRE-ASREAK sequence was injected in mice by the intramuscular route. Antibodies against M protein were detected in extracts of mucosal tissues and in serum. The repertoire isotypes of serum immunoglobulin G (IgG) and mucosal IgA and IgG antibodies varied, according to the dose of injected peptide. The highest mucosal IgA antibody response was obtained with 0.01 micro g of antigen per injection, whereas the systemic IgG antibody response increased with 10 micro g of antigen. Mucosal antibody production against streptococci was confirmed by immunofluorescence analysis. These results provide evidence that this novel approach of mucosal vaccination may be of advantage for bacterial systems and suggest a new field of investigation based on synthetic peptide analogues.
Assuntos
Antígenos de Bactérias , Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas , Proteínas de Transporte/imunologia , Vacinas Antimaláricas/imunologia , Mucosa/imunologia , Streptococcus pyogenes/imunologia , Sequência de Aminoácidos , Animais , Anticorpos/análise , Anticorpos Antibacterianos/imunologia , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito T/imunologia , Feminino , Imunofluorescência , Injeções Intramusculares , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Confocal , Dados de Sequência MolecularRESUMO
Optically stimulated luminescence dosimetry (OSL-D) used in conjunction with fibre optics enables a remote measurement of dose, for the purpose of radioprotection in the nuclear industry and in medicine (radiology, radiotherapy). Alumina OSL crystals are used because of their low Z, low fading and optical transparency, which improves the sensitivity. An optoelectronic portable dosemeter has been designed and tested that shows a dose detection of 50 microGy with a 20 metre-long fibre. Following irradiation, all trapped electrons are released under light stimulation while the OSL is integrated to provide dose-equivalent measurements. A compensation technique is designed with the help of the MCNP4b code, so that both angular and photon energy characteristics comply with international standards (CEI 61066) for photon dose equivalent Hp(10). Two sensors are described that allow measurements over a wide solid angle (95% of 4piSr), for photon energies ranging from 15 keV to 3 MeV.
Assuntos
Óxido de Alumínio/efeitos da radiação , Dosimetria Termoluminescente/métodos , Óxido de Alumínio/química , Carbono/química , Cristalização , Desenho de Equipamento , Tecnologia de Fibra Óptica , Raios gama , Fótons , Proteção Radiológica/instrumentação , Proteção Radiológica/métodos , Proteção Radiológica/estatística & dados numéricos , Radioquímica , Dosimetria Termoluminescente/instrumentação , Dosimetria Termoluminescente/estatística & dados numéricosRESUMO
To improve the mucosal antibody response against a short amino acid (aa) sequence (ELDKWA) of HIV gp41, we have investigated a construction including this peptide in-line with the Pan DR epitope (PADRE). ELDKWA is a conserved peptide playing a key role in the pathogenicity of HIV transmission. PADRE is a non-natural peptide with multipotential immunogenic properties. The results show striking differences between mucosal and systemic immune systems, with a preferential response of the mucosal organs. In contrast with most mucosal immunizations, the intracellular response persists for over two months after the last injection. This strongly suggests that further investigations of conserved key epitopes from various pathogens may lead to safe and chemically defined mucosal vaccines with synthetic peptides. These candidate vaccines with free peptides may be suitable for mass campaigns even in developing countries.
Assuntos
Epitopos de Linfócito B/imunologia , Anticorpos Anti-HIV/imunologia , Proteína gp41 do Envelope de HIV/imunologia , Peptídeos/imunologia , Adjuvantes Imunológicos , Compostos de Alúmen , Animais , Feminino , Anticorpos Anti-HIV/sangue , Humanos , Imunidade nas Mucosas , Imunoglobulina A/sangue , Imunoglobulina A/imunologia , Injeções Intramusculares , Camundongos , Camundongos Endogâmicos BALB C , Vagina/imunologiaAssuntos
Anticorpos Monoclonais/imunologia , Imunoglobulina M/imunologia , Animais , Anticorpos Monoclonais/química , Especificidade de Anticorpos , Antígenos , Autoanticorpos/química , Autoanticorpos/imunologia , Sítios de Ligação de Anticorpos , Reações Cruzadas , Humanos , Imunoglobulina M/química , CamundongosRESUMO
Mucosal antibodies consist of a variety of molecules, including secretory IgA and local IgG, involved in the first immune barrier of defence against pathogens. They account for the majority of daily synthesized immunoglobulins in the body and mostly depend on the secretory immune system which is independent from its systemic counterpart. Acting by immune exclusion and immune elimination, these immunoglobulins correspond to preimmune poly-reactive natural antibodies and to antigen-induced antibodies. Recent progress in this field have suggested new approaches of mucosal vaccines preventing the entry of pathogens in the body.
Assuntos
Imunoglobulina A Secretora/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Mucosa/imunologia , Animais , Formação de Anticorpos , Linfócitos B/imunologia , Previsões , Humanos , Imunidade Inata , Imunoglobulina A Secretora/biossíntese , Imunoglobulina A Secretora/química , Imunoglobulina D/imunologia , Imunoglobulina G/biossíntese , Imunoglobulina G/química , Cadeias J de Imunoglobulina/química , Cadeias J de Imunoglobulina/imunologia , Imunoglobulina M/biossíntese , Imunoglobulina M/química , Modelos Imunológicos , Nódulos Linfáticos Agregados/imunologia , Vacinação/métodosRESUMO
To investigate a possible dysregulation of the autoantibody network in AIDS patients, the relative activity of representative natural antibodies was measured in serum IgG and IgM. These immunoglobulins were purified from two cohorts of 20 HIV-infected patients undergoing, or not, a triple combination therapy. A cohort of 20 normal patients was used as a control. Marked alterations of the natural antibody repertoire were observed, varying according to the isotype and specificity of the antibody studied. For the classical self-protein antigens, human actin and myosin, the changes observed in the untreated cohort were absent in the treated cohort. In contrast, no changes, or even increased changes of the activity of antibodies to special antigens, DNA and TNP, occurred in the treated cohort. The differences were highly significant, indicating that this repertoire is regulated and not randomly modified by the disease. These results suggest the presence of different factors of dysregulation of the B cell repertoire of natural antibodies associated with the disease as well as with the treatment. These major dysregulations may favor the autoimmune phenomena observed during HIV infection.
Assuntos
Infecções por HIV/imunologia , Infecções por HIV/terapia , Imunoglobulina G/metabolismo , Imunoglobulina M/metabolismo , Actinas/imunologia , Adulto , Idoso , Antígenos/imunologia , Estudos de Coortes , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Miosinas/imunologiaRESUMO
The induction of a mucosal immunity provides an additional principle of vaccination by preventing the entry of pathogens in the body. Albeit the fact that intensive research has been conducted on local vaccines, the major mucosal vaccine commercially available for human use remains the oral polio vaccine. We have previously demonstrated that parenteral vaccination in humans with tetanus toxoid (TT) results in a genital immunoglobulin (Ig)G antibody (Ab) response. Here, we show that injections of TT with no adjuvant induces an anti-TT response in the mucosal tissues of normal BALB/c mice. The response is multiregional, involves both IgG and IgA isotypes, and is long-lasting. Similarly, injections of haptens coupled to TT or to other diffusible proteins may induce mucosal Abs. These results led us to immunize normal BALB/c mice with a viral peptide coupled to TT by disulfide bridging. The hapten was a 17 amino acid peptide containing the ELDKWA sequence of human immunodeficiency virus (HIV)-1 gp41. A significant IgG and IgA Ab response to the immunizing peptide was induced in various mucosal tissues despite the presence of a suboptimal Ab response in the spleen. The results indicate that mucosal immunity to peptides that are candidates for human vaccinations may be achieved by parenteral adjuvant-free immunization with peptide coupled to TT.
Assuntos
Antígenos/administração & dosagem , Imunidade nas Mucosas , Imunoglobulina A/biossíntese , Imunoglobulina G/biossíntese , Sequência de Aminoácidos , Animais , Epitopos/administração & dosagem , Epitopos/química , Feminino , Proteína gp41 do Envelope de HIV/administração & dosagem , Proteína gp41 do Envelope de HIV/química , Haptenos/administração & dosagem , Humanos , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Injeções Intramusculares , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Solubilidade , Toxoide Tetânico/administração & dosagem , Distribuição Tecidual , Vacinação , Vagina/imunologiaRESUMO
Treatment of normal polyclonal human IgG and of F(ab')2 fragments of IgG with 6.0 M urea, 1.3 M sodium thiocyanate or with acidic buffers (pH 2.0), resulted in a dramatic and selective enhancement of the preexisting antibody reactivity with self antigens. Enhanced antibody activity revealed by the dissociating agents was inhibited by the addition of an excess of the relevant soluble antigen. Human monoclonal IgG, including four different IgG1m(1) V(H)3+ and V(K)3+ paraproteins differing only in their CDRs, exhibited different changes in reactivity following urea treatment indicating major involvement of CDR sequences. The calculated dissociation constant of the binding reaction of normal IgG to the self antigen actin was 10(-6) M, whether IgG had been treated or not, indicating that the treatment increased the proportion of available self-reactive molecules instead of increasing the affinity of the preexisting natural autoantibodies. Enhanced autoreactivity was not due to aggregation of Ig, unmasking of the antibody site by removal of low MW antigens, nor to the denaturation of natural Id-anti-Id complexes. Taken together, these results suggest that treatment of Ig with dissociating agent results in the exposure of basic polyreactive antibody structures. The enhancement of reactivity may be of relevance in physiology of mucosal immunity and in therapeutic immunomodulation.
Assuntos
Autoanticorpos/imunologia , Fragmentos Fab das Imunoglobulinas/imunologia , Imunoglobulina G/imunologia , Imunoglobulinas Intravenosas/imunologia , Tiocianatos , Ureia , Actinas/imunologia , Complexo Antígeno-Anticorpo/imunologia , Autoanticorpos/sangue , Autoantígenos/imunologia , Soluções Tampão , Humanos , Imunoglobulinas Intravenosas/uso terapêuticoRESUMO
Four different monoclonal Ig (MIg) (IgA1kappa, IgG1kappa, IgG2kappa and IgG4kappa) displaying anti-tubulin activity were detected in the serum from a lymphoma patient. The complete sequence of three of these MIg showed identical V(H) and V(L) domains and the presence of mutations compatible with an antigen-driven process. Surprisingly, despite complete homology in their variable domains, IgA1kappa, IgG1kappa, or their Fab fragments bound to a common motif recognized in beta tubulin, with significant differences in affinity (IgA1kappa 1.52x10(-8) M, and IgG1kappa 2.09x10(-7) M). To substantiate these results, the V(H) and V(L) domains from IgA1kappa were cloned and introduced into expression vectors containing the constant kappa exon and either the mu or the gamma1 constant exon, and complete recombinant IgMkappa and IgG1kappa were obtained. Like the IgA1kappa, the IgMkappa construction bound to the tubulin epitope with consistent affinity (7.7x10(-9) M), whereas the IgG1kappa construction displayed a significantly lower affinity (3.28x10(-7) M). These results provide definitive evidence that isotype can influence binding affinity to antigen and suggest that malignant transformation occurred at the germinal center once the mutational process was achieved and the switch process was still active.
Assuntos
Reações Antígeno-Anticorpo , Switching de Imunoglobulina , Sequência de Aminoácidos , Sequência de Bases , Epitopos , Humanos , Imunoglobulina A/imunologia , Imunoglobulina G/classificação , Imunoglobulina G/imunologia , Cadeias Pesadas de Imunoglobulinas/química , Cadeias Pesadas de Imunoglobulinas/genética , Cadeias Leves de Imunoglobulina/química , Cadeias Leves de Imunoglobulina/genética , Região Variável de Imunoglobulina/química , Região Variável de Imunoglobulina/genética , Dados de Sequência Molecular , Ressonância de Plasmônio de Superfície , Tubulina (Proteína)/imunologiaRESUMO
Human heart mast cells (HHMC) have been identified in heart tissue, perivascularly, and in the intima of coronary arteries. In vitro activation of isolated HHMC induces the release of vasoactive and proinflammatory mediators (histamine, tryptase, and cysteinyl leukotriene C(4) [LTC(4)]). We investigated the effects of several bacterial proteins on HHMC activation in vitro. HHMC released histamine, tryptase, and LTC(4) in response to Staphylococcus aureus Cowan 1 and the immunoglobulin (Ig)-binding protein A, but not to S. aureus Wood 46, which does not synthesize protein A. The effect of protein A was inhibited by preincubation with monoclonal IgM V(H)3(+). Some strains of Peptostreptococcus magnus express an Ig light chain-binding surface protein called protein L. Such bacteria and soluble protein L stimulated the release of preformed and newly synthesized mediators from HHMC. Preincubation of HHMC with either protein A or protein L resulted in complete cross-desensitization to a subsequent challenge with the heterologous stimulus or anti-IgE. Monoclonal IgE (kappa chains) blocked protein L-induced release, whereas IgE (lambda chains) had no effect. Streptococcal protein G, formyl-containing tripeptide, and pepstatin A did not activate HHMC. Bacterial products protein A and protein L and intact bacteria (S. aureus and P. magnus) activate HHMC by acting as Ig superantigens.
Assuntos
Proteínas de Bactérias/imunologia , Imunoglobulina E/imunologia , Mastócitos/imunologia , Miocárdio/imunologia , Proteína Estafilocócica A/imunologia , Superantígenos/imunologia , Adulto , Idoso , Liberação de Histamina , Humanos , Imunoglobulina E/metabolismo , Região Variável de Imunoglobulina/metabolismo , Leucotrieno C4/biossíntese , Mastócitos/metabolismo , Pessoa de Meia-Idade , Peptostreptococcus/imunologia , Peptostreptococcus/metabolismo , Serina Endopeptidases/metabolismo , Staphylococcus aureus/imunologia , Staphylococcus aureus/metabolismo , TriptasesRESUMO
We have taken advantage of the property of IgA1-proteases to selectively cleave the human IgA1 subclass into Fabalpha and Fcalpha-J chain-secretory component (Fcalpha-J-SC) fragments in order to design a novel ELISA method for measuring the two secretory IgA (S-IgA) subclasses in secretions. The assay is based on the loss of detection of S-IgA1 by a combination of peroxidase-labelled antibodies to secretory component and Fab following IgA1-protease treatment. The specificity is that of the protease and the sensitivity of the detection is 5 ng/ml. Moreover, the use of purified S-IgA1 and S-IgA2 controls is not necessary. The assay has been successfully applied to the analysis of colostral S-IgA antibodies (Abs) to HIV-1-gp160 from HIV-1 positive women. The major subclass of colostral S-IgA antibodies to gp160 was found to be of the alpha1 isotype but the specific activity of anti-HIV-gp160 S-IgA2 was, however, higher than that of S-IgA1.
Assuntos
Especificidade de Anticorpos , Ensaio de Imunoadsorção Enzimática/métodos , Imunoglobulina A Secretora/análise , Serina Endopeptidases/metabolismo , Adulto , Colostro/imunologia , Feminino , Anticorpos Anti-HIV/isolamento & purificação , Proteína gp160 do Envelope de HIV/imunologia , Soropositividade para HIV/imunologia , HIV-1/imunologia , Humanos , Saliva/imunologia , Especificidade por SubstratoRESUMO
PROBLEM: Except for the description of a secretory immunoglobulin (S-Ig) of a low size, no recent study has investigated the molecular status of antibodies in the human amniotic fluid. METHOD: After separation with a high performance chromatography, we analyzed the different isotypes of amniotic Igs by immunoblotting and ELISA. RESULTS: IgG is found to be the major isotype and to contain mother-derived tetanus antitoxins. IgA is much less abundant, whereas no IgM can be detected. IgA is monomeric, with a low level of secretory IgA and with various amounts of free secretory component (SC). The presence of a low level of SC-containing immunoglobulin of a low size is confirmed during the last trimester of pregnancy. This molecule contains no alpha chain but includes a Fabgamma fragment noncovalently associated with SC. IgG, IgA, and SC are detected in the fetal urine and, therefore, can reach the amniotic fluid by this route. CONCLUSION: In addition to the predominant maternal IgG, the amniotic fluid contains different molecular forms of fetal immunoglobulins. Their function as an immune barrier against infection and against mother-derived autoantibodies is discussed.
Assuntos
Líquido Amniótico/química , Líquido Amniótico/imunologia , Isotipos de Imunoglobulinas/análise , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Feminino , Feto/imunologia , Humanos , Immunoblotting , Imunoglobulina A/análise , Imunoglobulina A/urina , Imunoglobulina A Secretora/análise , Imunoglobulina A Secretora/urina , Fragmentos de Imunoglobulinas/análise , Fragmentos de Imunoglobulinas/urina , Imunoglobulina G/análise , Imunoglobulina G/urina , Isotipos de Imunoglobulinas/urina , Peso Molecular , GravidezRESUMO
BACKGROUND: Protein Fv is an endogenous protein, synthesized in liver and largely released in the digestive tract during acute and chronic viral hepatitis, that binds to immunoglobulin (Ig) from various mammalian and nonmammalian species. METHODS: Basophils obtained from normal subjects were purified by a double Percoll gradient and elutriation. The secretion of histamine induced by protein Fv was assayed by a fluorometric technique, the extracellular protein levels of IL-4 and IL-13 were measured by ELISA, and IL-4 mRNA levels were evaluated by RT-PCR. RESULTS: Protein Fv concentration-dependently induced histamine and IL-4 and IL-13 release from purified basophils. IL-4 mRNA, constitutively present in basophils, was increased after stimulation by protein Fv. Histamine and IL-4 secretion from basophils, but not histamine and IL-13 release, activated by protein Fv was significantly correlated (rs = 0.70, p<0.001). Basophils from which IgE had been dissociated by brief exposure to lactic acid no longer released IL-4 in response to protein Fv and anti-IgE. Two preparations of human VH3(+) monoclonal IgM inhibited protein Fv-induced secretion of IL-4 and histamine from basophils. In contrast, VH6(+) monoclonal IgM did not inhibit the release of mediators caused by protein Fv. CONCLUSIONS: These results indicate that protein Fv, which acts as an endogenous superallergen, interacts with the VH3 domain of IgE to induce the synthesis and release of IL-4, IL-13 and the secretion of histamine from basophils.
Assuntos
Basófilos/imunologia , Citocinas/imunologia , Imunoglobulina E/imunologia , Linfocinas/imunologia , Sialoglicoproteínas/imunologia , Citocinas/metabolismo , Humanos , Fragmentos de Imunoglobulinas/imunologia , Cadeias Pesadas de Imunoglobulinas/imunologia , Região Variável de Imunoglobulina/imunologiaAssuntos
Fármacos Anti-HIV/uso terapêutico , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Cadeias Pesadas de Imunoglobulinas/fisiologia , Região Variável de Imunoglobulina/fisiologia , Adulto , Idoso , Criança , Pré-Escolar , Regulação para Baixo , Quimioterapia Combinada , Feminino , Inibidores da Protease de HIV/uso terapêutico , Humanos , Fragmentos de Imunoglobulinas , Imunoglobulina G/sangue , Imunoglobulina G/metabolismo , Cadeias Pesadas de Imunoglobulinas/sangue , Região Variável de Imunoglobulina/sangue , Linfocinas/metabolismo , Masculino , Pessoa de Meia-Idade , Inibidores da Transcriptase Reversa/uso terapêutico , Sialoglicoproteínas/metabolismo , Superantígenos/imunologiaRESUMO
We show that the natural autoantibody activity of amniotic IgG dramatically increases after purification, and that the IgG-depleted fraction can suppress the activity of IgG natural antibodies from amniotic fluid or from the maternal serum. This suppression is also observed towards serum IgG from unrelated adults but does not impair the tetanus antitoxin activity of serum-derived IgG. Absorption experiments and immunoglobulin separation by gel permeation demonstrate that this suppression is due to monomeric immunoglobulins of the IgA isotype. The inhibition is associated with an anti-F(ab')2 activity of the amniotic IgA, involving hypervariable regions of the IgG as demonstrated by different reactivities towards monoclonal IgG sharing the same family of VH and Vkappa domains. These results indicate that the inhibition of natural autoantibodies not only occurs with fetal and adult serum IgM, as reported by other groups, but also with amniotic IgA, suggesting a general and important phenomenon. In the case of the amniotic fluid, IgA could protect the fetus against maternal IgG autoantibodies without interfering with simultaneously translocated antigen-induced IgG antibodies to pathogens.