RESUMEN
Ginseng, the dry extract prepared from the Panax ginseng C.A. Mayer-root contain immunomodulators named ginsenosides, which in the pig enhance the antibody response to viral and bacterial antigens. The enhancing effect of ginseng was demonstrated vaccinating pigs against porcine parvovirus (PPV) and Erysipelothrix rhusiopathiae infections, using commercially available vaccines. The potency of the licensed, aluminium hydroxide adjuvanted; vaccines were compared with those supplemented with ginseng. The antibody response to PPV was measured by the haemagglutination inhibition (HI) test whereas the mouse potency test and ELISA evaluated the immune response to E. rhusiopathiae. Antibodies to the 64-66 kDa glycoprotein of the E. rhusiopathiae were demonstrated by immunoblotting. The qualitative antibody responses were evaluated by means of ELISA(s) using monoclonal antibodies to swine IgG1 and IgG2. The addition of 2mg ginseng per vaccine dose, potentiate the antibody response of the commercial vaccines without altering their safety. Significantly higher (P<0.001) antibody titres were achieved to both PPV and to E. rhusiopathiae by the supplementation with ginseng. Aluminium hydroxide adjuvanted vaccines favoured the production of IgG1 antibodies. Interestingly, the vaccines supplemented with ginseng favoured IgG2. The vaccines used in the evaluations varied in their immunogenic potency. However, after the addition of ginseng the less immunogenic vaccine proved to be as potent as the better one without ginseng. Thus, the use of ginseng as a co-adjuvant provides a simple, safe and cheap alternative for improving the potency of aluminium hydroxide adjuvanted vaccines.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Hidróxido de Aluminio/farmacología , Vacunas Bacterianas/inmunología , Erysipelothrix/inmunología , Panax , Parvovirus Porcino/inmunología , Extractos Vegetales/farmacología , Vacunas Virales/inmunología , Animales , Anticuerpos Antibacterianos/sangre , Anticuerpos Antivirales/sangre , Ensayo de Inmunoadsorción Enzimática , Femenino , Pruebas de Inhibición de Hemaglutinación , Ratones , PorcinosRESUMEN
Kappa (kappa)-light (L) chain-deficient (Ckappa-/-) mice readily mount a T cell-dependent antibody response after immunization with kappa-containing proteins. Such antibody responses areabsent in normal (Ckappa+/+) animals because of tolerance due to the abundance of kappa-L chains expressed on more than 95% of all B cells and serum immunoglobulins. When heterozygous kappa-sufficient (Ckappa+/-) females are bred with homozygous kappa-deficient (Ckappa-/-) males, half of their offspring will become kappa-deficient but have received kappa-L chain containing maternal Ig, mainly IgG and IgA, through placental and intestinal transmission. The kappa-containing maternal Ig persists for more than 2 months in the circulation of the offspring. Starting from week 15-20 of age, a spontaneous antibody response towards the maternal kappa-L chains can be recorded. The time of onset, as well as the magnitude of the responses differ among individuals of the same litter. Invariably, once a response has been initiated, it transits into an IgG-type of response, which upon injection with kappa-containing protein shows the features of a secondary type of immune response.
Asunto(s)
Inmunidad Materno-Adquirida , Inmunoglobulina G/inmunología , Cadenas kappa de Inmunoglobulina/genética , Cadenas kappa de Inmunoglobulina/inmunología , Animales , Animales Recién Nacidos , Femenino , Inmunización , Inmunoglobulina G/biosíntesis , Cinética , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , EmbarazoRESUMEN
The organization of immunoglobulin heavy (H) chain genes in teleosts resembles that of mammals and amphibians, whereas light (L) chain genes are arranged in multiple clusters of variable (VL), joining (JL), and constant (CL) region segments. Sequence analysis of two Atlantic cod genomic clones (14,966 and 13,116 bp in length) revealed a very compact IgL chain locus with the VL genes in opposite transcriptional orientation to the JL and the CL genes. This suggests the possibility of rearrangements between clusters by inversion. Each cluster spans approximately 2.1 kb and distances between clusters vary between 2.1 and 4.8 kb. To gain insight into the transcriptional regulation of this complex, multiclustered locus, chloramphenicol acetyl transferase reporter constructs containing 14 different DNA segments from the two genomic clones were transfected into channel catfish B and non-B-cell lines, as well as into mouse B-cell lines. These studies showed strong enhancer activity downstream of the CL region in three out of six L chain gene clusters when assayed in fish, but not in mouse B cells. Interestingly, both mouse and human lambda enhancers exhibited strong activity in the fish B cells, while the mouse 3' kappa enhancer did not. This suggests that transcription factors similar to those involved in mammalian lambda expression are present in B cells from teleosts.
Asunto(s)
Elementos de Facilitación Genéticos , Peces/genética , Cadenas Ligeras de Inmunoglobulina/genética , Transcripción Genética , Animales , Secuencia de Bases , Línea Celular , Clonación Molecular , ADN Complementario , Peces/inmunología , Humanos , Ictaluridae , Regiones Constantes de Inmunoglobulina/genética , Región de Unión de la Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Masculino , Ratones , Datos de Secuencia Molecular , Familia de Multigenes , Análisis de Secuencia de ADN , Distribución TisularRESUMEN
This study was undertaken to determine if a lack of V(L) domain variability could explain, in part, the failure of Atlantic cod to respond to immunization with the production of specific antibodies. The variability of cod V(L) regions was studied in 33 cDNA and two genomic clones. The variability of the CDRs was estimated by the Shannon entropy method and compared with that in other species. It was found to be lowest in the little skate (Raja erinacea), higher in cod, and highest in Xenopus and mouse. While the variability of the CDRs is slightly lower in cod than in Xenopus and mouse, it is spread over broader areas of the amino acid sequence. The length of CDR1 and CDR3 in cod is equal to or exceeds that found in skate, Xenopus, chicken and mammals. Isoelectric points and hydrophobicity vary substantially among the studied Ig light chain domains. Thus, neither the length, nor the variability, nor the physicochemical properties (pI and hydrophobicity) of the L chain CDRs can explain the absence of antibody response to immunization in cod.
Asunto(s)
Peces/inmunología , Variación Genética , Cadenas Ligeras de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , ADN Complementario , Peces/genética , Isotipos de Inmunoglobulinas , Ratones , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Xenopus laevisRESUMEN
Two different rainbow trout cDNA sequences encoding a heavy chain secreted Ig (Hs) and a part of a membrane-bound heavy chain Ig (Hm) are reported. The sequences were most similar to those encoding the Ig heavy chains (IgH) of other teleost fish. As in the Hm of the other teleost fish the rainbow trout Hm results from the splicing of the 3' end of the third constant exon (CH3) to the sequence encoding the membrane-bound domain. Analysis of a rainbow trout IgH genomic clone revealed that a joining heavy chain (JH) segment, different to the one observed in the cDNA, is located 825 bp 5' of the CH1 exon. The sequence also contains possible enhancer-like and octamer-like motifs.
Asunto(s)
Genes de Inmunoglobulinas , Trucha/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Southern Blotting , ADN/genética , Biblioteca de Genes , Intrones , Proteínas de la Membrana/inmunología , Datos de Secuencia Molecular , Oligodesoxirribonucleótidos/química , Alineación de Secuencia , Trucha/inmunologíaRESUMEN
Antibody screening and colony hybridization of cDNA libraries have been used to isolate clones of the immunoglobulin light (IgL) chain from Atlantic cod (Gadus morhua L.) and rainbow trout (Oncorhynchus mykiss Walbaum). Sequence analysis shows dissimilarities in the constant part of the molecule (CL) within each species. Comparisons of the amino acid sequences of the constant parts of the IgL chains show a 55% identity between the two teleost species. When compared with other species the highest similarities are found to the constant domain of the IgL chain from mammals (30%-37%), but the teleost IgL chain can be classified neither as kappa nor lambda. The VL domain in Atlantic cod and rainbow trout is also more similar to those of mammals than to those of other animal species, but no difference between kappa and lambda was noticed. Genomic Southern blots hybridized with fragments coding for the constant part of IgL gave several bands larger than 2 kilobases and a similar pattern was obtained with fragments coding for the variable part. These results show that the locus of the IgL chain has a multiple organization in teleost fish and that the locus has an organization similar to that of sharks. Several of the cDNA clones isolated from both the head kidney and the spleen represent nonrearranged or nonspliced mRNA, and northern blot analysis shows that such transcripts are present in both the head kidney and the spleen.