RESUMEN
Avermectin-binding proteins from the free-living nematode worm Caenorhabditis elegans and from the fruitfly Drosophila melanogaster were purified to homogeneity via a three-step procedure. The binding proteins were covalently labelled using a radioactive photoaffinity probe and then partially purified on a Sephacryl S-300 gel-filtration column. The radiolabelled binding proteins were then purified by immunoaffinity chromatography using a monoclonal antibody to avermectin covalently attached to Protein A-Sepharose beads. Three affinity-labelled Drosophila proteins with molecular masses between 45 and 50 kDa were isolated in this way and then separated from each other by electroelution. This three-step protocol provides a rapid technique for receptor purification which may be of use in the purification of other binding proteins.
Asunto(s)
Caenorhabditis elegans/química , Proteínas Portadoras/aislamiento & purificación , Drosophila melanogaster/química , Ivermectina/análogos & derivados , Animales , Anticuerpos Monoclonales/inmunología , Proteínas Portadoras/inmunología , Proteínas Portadoras/metabolismo , Cromatografía de Afinidad , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Ivermectina/metabolismo , Pruebas de PrecipitinaRESUMEN
Polyclonal anti-idiotype 1073 (anti-Id 1073), raised against a monoclonal antibody specific for the protective epitope(s) of Eimeria tenella sporozoites, induced cell-mediated immune (CMI) responses in bursectomized chickens. Whereas alhydrogel-adsorbed anti-Id 1073 was sufficient to engender the CMI response at 4 h after injection, induction of the CMI response at 24 h required both alhydrogel and muramyl dipeptide sterol. Exposure of immunized chickens to live parasites prompted a dichotomous effect on the CMI response engendered by anti-Id in that the 4-h CMI response was preferentially stimulated and the 24-h CMI response was down regulated. Both types of CMI response were transferable to naive chickens by T cells from anti-Id 1073 immune donors or by parasite-specific T cells from clones 21 and 27. These T-cell clones were generated from chickens immunized by repeated infections with E. tenella and showed in vitro proliferative responses to anti-Id 1073. The abilities of T cells from clone 21 to selectively transfer the 4-h CMI response and to generate gamma interferon to activate macrophages for their cytotoxic effects on Eimeria sporozoites correlate with the preferential stimulation by parasites of the 4-h CMI response in chickens immunized with anti-Id 1073. These data show that anti-Id 1073 mimics the protective epitope(s) of the parasite and primes chickens for protective CMI responses. Cytotoxic T cells, equivalent to the mammalian T-cell subset of the Lyt2+ phenotype, appear to be the primary effector T cells in the CMI response engendered by anti-Id 1073 against Eimeria parasites.
Asunto(s)
Anticuerpos Antiidiotipos/fisiología , Coccidiosis/inmunología , Hipersensibilidad Tardía/inmunología , Idiotipos de Inmunoglobulinas/inmunología , Linfocitos T/parasitología , Animales , Anticuerpos Antiidiotipos/administración & dosificación , Antígenos de Protozoos/inmunología , Pollos , Coccidiosis/prevención & control , Epítopos/inmunología , Inmunidad Celular , Inmunización Pasiva , Cinética , Linfocitos T/inmunologíaRESUMEN
Anti-idiotypic antibodies were raised in rabbits against four monoclonal antibodies with specificity for the surface antigenic determinants of Eimeria tenella sporozoites, the infective stage of the coccidial parasite. Two of the monoclonal antibodies (1073 and 15-1) transferred passive protection in chickens against E. tenella infection. The polyclonal anti-idiotype antibody preparations against protective monoclonal antibodies contained specificities for the paratope-associated idiotypes of these monoclonal antibodies, as assessed by the competitive inhibition of binding of the homologous idiotype-anti-idiotype by the sporozoite antigen. Competitive inhibition of binding of homologous idiotype-anti-idiotype by the parasite antigen was not observed when the anti-idiotype antibody preparations against monoclonal antibodies 1546 and 1096 were tested. The anti-idiotype 1073 and 15-1 antibodies functioned as surrogate antigens in vivo when used for vaccination of young chickens, as evidenced by the induction of partial protective immunity against subsequent challenge infection with virulent parasites and induction of antisporozoite antibodies. These data clearly support the view that anti-idiotypic antibodies raised against the paratope-associated idiotypes can mimic pathogen antigens and therefore can provide a possible alternative approach for the vaccination of chickens against coccidiosis.
Asunto(s)
Anticuerpos Antiprotozoarios/inmunología , Antígenos de Protozoos/inmunología , Coccidiosis/prevención & control , Eimeria/inmunología , Idiotipos de Inmunoglobulinas/inmunología , Enfermedades de las Aves de Corral/prevención & control , Vacunas/inmunología , Animales , Anticuerpos Antiidiotipos/inmunología , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Pollos , Inmunización Pasiva , RatonesAsunto(s)
Anticuerpos Antiidiotipos/inmunología , Anticuerpos Monoclonales/inmunología , Pollos/inmunología , Eimeria/inmunología , Idiotipos de Inmunoglobulinas/inmunología , Animales , Formación de Anticuerpos , Especificidad de Anticuerpos , Hipersensibilidad Tardía/inmunología , Inmunidad Celular , Inmunización PasivaAsunto(s)
Eimeria/inmunología , Linfocitos T/citología , Animales , Pollos , Células Clonales , Interleucina-2/farmacología , Activación de Linfocitos , Cooperación Linfocítica , Especificidad de la Especie , Linfocitos T/inmunología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunologíaRESUMEN
T lymphocyte clones reacting specifically with the antigenic components of Eimeria tenella were generated from splenic lymphocytes of immunized chickens and were maintained for 12 to 14 wk in vitro. These T cell growth factor-dependent T lymphocyte clones from bursectomized and normal chickens proliferated in vitro when stimulated with antigens from different developmental stages of homologous but not heterologous species of the parasite. Specific proliferative responses of the cloned T cells showed an absolute requirement for antigen presentation by histocompatible antigen-presenting cells. Some of the T cell clones exhibited functionally discrete interactions with syngeneic primed B cells; 25% of the T cell clones from immunized normal chickens and 7% of those obtained from immunized bursectomized chickens showed antigen-dependent helper activity and induced specific antibody production by syngeneic primed B cells. Of the T cell clones from immunized normal chickens, 19% showed suppression of in vitro antibody production in comparison to 7% of those isolated from immunized bursectomized chickens. The frequency of cloned T cells with ability to induce cytotoxic activity in macrophages against the sporozoites of E. tenella was much higher in those isolated from bursectomized chickens (80%) than in those isolated from normal chickens. Because both bursectomized and normal chickens can be immunized by repeated infections, differences in the distribution among cloned T cells suggest different effector mechanisms of immunity against coccidiosis in these chickens. Lack of B cells seem to affect the development of T cell immunity as reflected by slower development of immunity and enhanced activation of cytotoxic T cell function.
Asunto(s)
Pollos/inmunología , Eimeria/inmunología , Linfocitos T/inmunología , Animales , Formación de Anticuerpos , Antígenos de Protozoos/inmunología , Bolsa de Fabricio/inmunología , Células Clonales/inmunología , Eimeria/crecimiento & desarrollo , Inmunidad Celular , Interleucina-2/farmacología , Activación de Linfocitos , Activación de Macrófagos , Mitógenos , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Reguladores/inmunologíaRESUMEN
Although responses of BALB/c mice to TNP-Ficoll or TNP-Brucella abortus are usually decreased by injection of allo anti-IgD (anti-Igh-5a) given 1 day before antigen, increased responses are obtained if a lymphokine mixture (SN) containing IL 2 is also injected. Simultaneous injection of anti-IgD and SN 4 days after priming with TNP-KLH induces an increase in antibody production similar to that induced by a second antigen injection. Injected together with a second injection of TNP-KLH at that time, anti-IgD and SN cause a synergistic enhancement of the secondary response. In allotype heterozygous (BALB/c X SJL)F1 mice injected with anti-IgD directed against one allotype, this enhancement of the secondary response is seen predominantly in the alternate allotype, because the IgG response of linked allotype specificity is slightly suppressed by the anti-IgD alone and is less enhanced than the alternate allotype by anti-IgD plus SN. Cells from unprimed heterozygous mice, incubated with anti-Igh-5a in vitro and transferred, together with antigen, to TNP-KLH-primed recipients, cause a much greater enhancement of the IgG responses of the Igb than of the Iga allotype in recipients. If, however, SN is also injected into the recipients, the anti-TNP response of both IgG allotypes is greatly enhanced.
Asunto(s)
Anticuerpos Antiidiotipos/administración & dosificación , Formación de Anticuerpos , Linfocitos B/inmunología , Inmunoglobulina D/inmunología , Inmunoglobulina D/fisiología , Isoanticuerpos/administración & dosificación , Animales , Anticuerpos Antiidiotipos/fisiología , Antígenos T-Independientes/administración & dosificación , Hemocianinas/administración & dosificación , Técnica de Placa Hemolítica , Tolerancia Inmunológica , Inmunización Secundaria , Inmunoglobulina D/administración & dosificación , Isoanticuerpos/fisiología , Linfocinas/administración & dosificación , Linfocinas/farmacología , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Bazo/citologíaRESUMEN
The responses to 2,4,6-trinitrophenyl conjugates of different Ficoll preparations differ with respect to the magnitude of the accompanying auto-anti-idiotype (Id) response in both mice and chickens. Evidence is presented that reduced auto-anti-Id production in the chicken is due to the activation of suppressor activity by some preparations of Ficoll.
Asunto(s)
Autoanticuerpos/biosíntesis , Ficoll/inmunología , Idiotipos de Inmunoglobulinas/inmunología , Nitrobencenos/inmunología , Polisacáridos/inmunología , Trinitrobencenos/inmunología , Animales , Células Productoras de Anticuerpos/metabolismo , Pollos , Ficoll/administración & dosificación , Ficoll/análogos & derivados , Ficoll/farmacología , Haptenos/inmunología , Técnica de Placa Hemolítica , Inmunosupresores/farmacología , Cinética , Activación de Linfocitos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos BALB C , Bazo/citología , Linfocitos T Reguladores/inmunología , Trinitrobencenos/administración & dosificación , Trinitrobencenos/farmacologíaRESUMEN
The spontaneous production of auto-anti-idiotype (Id) was demonstrated after injection of chickens with trinitrophenylated Ficoll (TNP-F) by: (a) the presence of hapten-augmentable plaque-forming cells (PFC), (b) the ability of serum and of hapten eluates from immune spleen cells to cause hapten-reversible inhibition of anti-TNP plaque formation, and (c) an enzyme-linked immunosorbent assay (ELISA). Tests for anti-Id using the ELISA and hapten-reversible inhibition of PFC correlated very well. As in the mouse, the incidence of hapten-augmentable PFC was reduced by thymectomy and increased by the transfer of TNP-F-immune spleen cells. Hapten-augmentable PFC were also observed during the immune response of chickens to p-azobenzene arsonate-conjugated Brucella abortus.
Asunto(s)
Autoanticuerpos/biosíntesis , Ficoll/inmunología , Idiotipos de Inmunoglobulinas/inmunología , Nitrobencenos/inmunología , Polisacáridos/inmunología , Trinitrobencenos/inmunología , Animales , Células Productoras de Anticuerpos/metabolismo , Autoanticuerpos/fisiología , Unión Competitiva , Pollos , Ensayo de Inmunoadsorción Enzimática , Ficoll/administración & dosificación , Ficoll/análogos & derivados , Haptenos/inmunología , Técnica de Placa Hemolítica , Inmunización Pasiva , Bazo/citología , Bazo/inmunología , Trinitrobencenos/administración & dosificaciónRESUMEN
The maturation of the C57BL/6 B cell population to be able to re-express surface immunoglobulin (sIg) after its removal by treatment with rabbit antimouse Ig (RAMIg) was studied in a cell transfer system. It was found that thymus cells were required for the maturation of a subset of the B cell population to be able to re-express sIg. The B cell population of irradiated, thymectomized mice reconstituted with spleen cells from donors under 2 wk of age remained deficient in their ability to re-express sIg even after 4 wk residence in the cell transfer recipient. In contrast, if adult thymus cells were transferred together with the immature B cells, the B cell population matured to be able to re-express sIg after treatment with RAMIg. Approximately one-third of the B cell population appears to require thymus cells for this maturation. The maturation of the thymus cell population to be capable of mediating this maturation of the B cell population occurs in two steps: between 2 and 3 and between 3 and 4 wk of age. This timing corresponds to the age at which the B cell population of C57BL/6 mice normally acquires the capacity to re-express sIg, which we have previously shown to also occur in two steps. Thymus cells from 3-wk-old donors can mediate the first step in B cell maturation to be able to re-express sIg, but cannot mediate the second step in this maturation of the B cell population. Thymus cells from 4-wk-old donors can mediate both steps in the maturation of the B cell population. The results suggest that thymus cells are involved in regulating some aspects of B cell differentiation.
Asunto(s)
Anticuerpos Antiidiotipos/fisiología , Linfocitos B/citología , Receptores de Antígenos de Linfocitos B/biosíntesis , Timo/citología , Envejecimiento , Animales , Linfocitos B/inmunología , Linfocitos B/metabolismo , Diferenciación Celular , Femenino , Inmunización Pasiva , Hígado/citología , Hígado/embriología , Masculino , Ratones , Ratones Endogámicos C57BL , Embarazo , Quimera por Radiación , Trasplante de Células Madre , Timo/fisiologíaRESUMEN
Aging NZB X SJL (NS) female mice provide a unique model of thymus pathology characterized by the intrathymic accumulation of large numbers of mature T and B cells. The purpose of the present work was to examine the possibility that this phenomenon results from the invasion of the thymus by cells from the periphery. Lymphoid cells labeled with chromium-51 or indium-111 were injected into syngeneic recipients to study their patterns of in vivo migration. Lymph node (LN) or spleen cells were found to localize significantly (1-2% of injected radioactivity) into the thymus of 12-month-old NS females but not into that of young recipients or of old NS males. However, intrathymic localization of injected LN cells was observed in castrated NS males which exhibit the same thymus pathology as NS females. Both radiolabeled T and B cells were found to enter the thymus of aged NS females but the latter cells about three times less efficiently than the former. Moreover, while thymocytes from young NS females were unable to recirculate to LN, those of old NS females showed increased LN-seeking capacity and part (1%) of them did migrate back into the thymus of old but not young NS females. In additional cell transfer experiments, the intrathymic migration of B cells into old NS females was further documented by using the antibody response to sheep erythrocytes as a tracer. Taken together, these observations indicate that the thymus of aging NS female mice is permeable to recirculating lymphocytes, suggesting that at least part of the mature T and B cells detected in this thymus are migrants from the periphery.
Asunto(s)
Linfocitos B/fisiología , Linfocitos T/fisiología , Timo/citología , Envejecimiento , Animales , Movimiento Celular , Femenino , Ganglios Linfáticos/citología , Masculino , Ratones , Ratones Endogámicos CBA , Ratones Endogámicos NZB , Ratones Endogámicos , Bazo/citología , Distribución TisularAsunto(s)
Anticuerpos Antiidiotipos/biosíntesis , Autoanticuerpos/biosíntesis , Idiotipos de Inmunoglobulinas/inmunología , Animales , Células Productoras de Anticuerpos/inmunología , Pollos , Ficoll/análogos & derivados , Ficoll/inmunología , Haptenos , Conejos , Linfocitos T/inmunología , Trinitrobencenos/inmunologíaRESUMEN
Previous studies have shown that anti-IgD-suppressed mice give normal primary and secondary splenic plaque-forming cell responses following i.v. challenge, although mice suppressed by the injection of anti-IgD from birth lack IgD-bearing cells in all lymphoid tissue examined. The present studies show that, in contrast, secondary immune responses in regional lymph nodes of such mice, even after i.v. priming with trinitrophenylated B. abortus, respond to a challenge injection in the footpad up to only less than 10% of control levels. When compared with respect to B cell numbers transferred, primed spleen cells from control and anti-IgD-suppressed mice are about equally effective in producing adoptive secondary plaque-forming cell responses in the spleens of recipient mice. Lymph nodes in recipients of anti-IgD-suppressed primed spleen cells show much lower responses than do lymph nodes in recipients of control primed cells, both upon immediate and delayed challenge with antigen in the footpads. It is concluded that the immunodeficiency caused by suppression with anti-IgD is much more marked in peripheral lymph nodes than in the spleen. The possible relationship of these results to the migratory properties of IgD+ as compared to IgD-B cells is discussed.
Asunto(s)
Envejecimiento , Anticuerpos Antiidiotipos/administración & dosificación , Inmunoglobulina D/fisiología , Ganglios Linfáticos/inmunología , Animales , Animales Recién Nacidos , Formación de Anticuerpos , Linfocitos B/inmunología , Femenino , Inmunoglobulina D/biosíntesis , Inmunoglobulina D/inmunología , Memoria Inmunológica , Terapia de Inmunosupresión , Ganglios Linfáticos/fisiología , Masculino , Ratones , Ratones Endogámicos BALB CAsunto(s)
Formación de Anticuerpos , Linfocitos B/inmunología , Inmunoglobulina D/inmunología , Animales , Anticuerpos Antiidiotipos , Antígenos T-Independientes/inmunología , Homocigoto , Alotipos de Inmunoglobulinas/inmunología , Memoria Inmunológica , Linfocinas/inmunología , Ratones , Receptores de Antígenos de Linfocitos B/inmunologíaAsunto(s)
Formación de Anticuerpos , Linfocitos B/inmunología , Inmunoglobulina D/inmunología , Animales , Anticuerpos Antiidiotipos , Heterocigoto , Alotipos de Inmunoglobulinas/inmunología , Memoria Inmunológica , Linfocinas/inmunología , Ratones , Receptores de Antígenos de Linfocitos B/inmunologíaAsunto(s)
Refuerzo Inmunológico de Injertos , Linfoma no Hodgkin/inmunología , Linfocitos T/inmunología , Animales , Trasplante de Médula Ósea , Células Cultivadas , Técnicas In Vitro , Activación de Linfocitos , Ratones , Ratones Endogámicos , Sarcoma Experimental/inmunología , Timectomía , Factores de Tiempo , Irradiación Corporal TotalRESUMEN
The role of delta-positive cells in the immune response was studied by comparing the effects of treatment with allotype-specific IgD hybridoma antibody on homozygous BALB/c or SJL/J and heterozygous (BALB x SJL)F1 mice. Homozygous mice, injected from birth with the relevant anti-delta antibody, made primary or secondary immune responses to intravenously injected trinitrophenyl (TNP)-Brucella abortus, TNP-Ficoll, and TNP-keyhole limpet hemocyanin, which did not differ significantly from those of control mice, despite the fact that IgD+ cells were depleted and Ig+ cells were markedly reduced in the spleens of treated mice. Responses in nodes draining a local injection of TNP-Brucella abortus were, however, significantly suppressed. Heterozygous mice, injected from birth with either anti-Ig-5a or anti-Ig-5b, showed a marked reduction in the number cells producing IgG antibody of linked allotype specificity in the secondary response to intravenously injected sheep erythrocytes. A corresponding decrease in the amount of serum IgG2a of that allotype specificity was also noted. However, in agreement with the results obtained in homozygotes, heterozygotes injected simultaneously with anti-IgD directed against each of the allotypes made normal, if not enhanced, plaque-forming cell responses of both allotype specificities. Similarly, serum IgG2a levels were normal in all but one mouse treated in this fashion. These results indicate that IgD+ cells are not essential for an immune response in vivo. Although the delta-positive cell is used preferentially under normal conditions, it appears that an alternative mechanism exists by which, in the absence of these cells, the animal is able to make a normal immune response.
Asunto(s)
Anticuerpos Antiidiotipos/fisiología , Linfocitos B/inmunología , Inmunoglobulina D/inmunología , Inmunoglobulina D/fisiología , Activación de Linfocitos , Animales , Formación de Anticuerpos , Heterocigoto , Tolerancia Inmunológica , Ratones/genéticaRESUMEN
(SJL X BALB)F1 suppressed mice have, in their lymphoid tissues, a population of suppressor T cells directed specifically against a paternal gamma G2a allotype (Ig-1b). Spleen or lymph node cells from these mice were injected into syngeneic nude mice and the effect on thymus-independent synthesis of Ig-1b in the athymic recipients was determined. After the injection of suppressor cells, Ig-1b disappeared from the serum of the recipients in a time course similar to that seen in normal mice. These results indicate that suppression occurs in the absence of thymus-derived helper cells, and they suggest that Ig-1b-producing B cells are the target of allotype-suppressor cells.