RESUMO
10 different compounds, including natural and synthetic polypeptides, proteins, polysaccharides, amino acids, and steroid hormones, were assayed for their capacity to form complexes with peritoneal exudate cell RNA. Only molecules carrying negatively charged groups were able to do so. The formation of RNA-antigen complexes was unrelated to the immuno-potency of the "antigen," was not an enzyme-dependent reaction, did not require the synthesis of RNA following introduction of the antigen, did not seem to involve antigen-specific RNAs, was not specific for macrophages, since HeLa cells could be used as effectively, and occurred when purified RNA was mixed with antigen only in the presence of divalent cations. The complexes were very stable, once formed, but could be dissociated by exhaustive dialysis against buffers containing a chelating agent. The macrophage RNA-antigen complex therefore appears to be a chelate between anionic groups on the two components. Based on the total absence of a relationship between immunogenicity and the capacity to form such complexes, as well as the nonspecific nature of complex formation at every level examined, it appears unlikely that RNA-antigen complexes play a physiologically significant role in immune induction.
Assuntos
Reações Antígeno-Anticorpo , Macrófagos/imunologia , RNA/metabolismo , Aminoácidos/metabolismo , Animais , Soluções Tampão , Isótopos de Carbono , Fenômenos Químicos , Química , Dactinomicina/farmacologia , Dextranos , Diálise , Exsudatos e Transudatos/imunologia , Glutamatos/metabolismo , Cobaias , Células HeLa/imunologia , Magnésio/farmacologia , Mieloma Múltiplo , Peptídeos/metabolismo , Peritônio/imunologia , Ligação Proteica , RNA/biossíntese , RNA/isolamento & purificação , Coelhos , Esteroides/metabolismo , Temperatura , Ultracentrifugação , Uridina/metabolismoRESUMO
In central equatorial Africa the frequency of uninterpretable or atypical Western blots (WB)--ie. antibodies to gag proteins only--can represent up to 50% of enzyme-linked immunosorbent assay (ELISA)-positive samples. To date the significance of such serology remains unknown. Nevertheless, an unusual HIV-1 strain has been isolated from the blood of a healthy Gabonese individual who presented an atypical WB. This virus, identified as isolated HIV-1OYl, grew to low titres of reverse transcriptase activity (less than 50,000 cpm/ml) and was not obviously cytopathic. Radioimmunoprecipitation and peptide ELISA studies indicated that the lack of env-specific reactivity was probably due to the absence of antibodies to the viral glycoproteins, rather than the virus encoding a highly divergent envelope protein. Molecular cloning and sequencing of the provirus proved it to be a string of HIV-1 which was genetically closer to European and North American than to African strains. Furthermore the envelope protein sequence contained all the features of a typical HIV-1 env gene. However, the tat gene derived from the proviral clone was functionally defective. Site-directed mutagenesis of this gene showed that this was due to the substitution of an essential cysteine residue for a serine. Polymerase chain reaction amplification of the tat gene, as well as parts of the gag and env gene sequences of HIV-1OYl, showed that essentially all of the proviruses were defective. These data emphasize the need to view HIV isolates as populations of distinct genomes capable of complementing each other.
Assuntos
Síndrome da Imunodeficiência Adquirida/microbiologia , Genes Virais , Genes env , Genes gag , Genes tat , HIV-1/genética , Síndrome da Imunodeficiência Adquirida/diagnóstico , Sequência de Aminoácidos , Sequência de Bases , Western Blotting , Gabão , Produtos do Gene env/genética , Produtos do Gene gag/genética , Produtos do Gene tat/genética , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Provírus/genética , Homologia de Sequência do Ácido Nucleico , Ativação Transcricional , Produtos do Gene tat do Vírus da Imunodeficiência HumanaRESUMO
We tested ability of 11 different lectins to stimulate DNA synthesis in bovine peripheral blood lymphocytes, using a wide range of experimental conditions. Five of the 6 lectins which induced DNA synthesis (concanavalin A, succinyl-concanavalin A, phytohemagglutinin M, pokeweed mitogens and lipopolysaccharide) did so under conditions similar to those optimal for stimulation of murine lymphocytes. The other lectin (peanut agglutinin) stimulated normal bovine lymphocytes whereas it does not stimulate normal mouse, rat, guinea pig or human lymphocytes. The binding of 5 different lectins to bovine peripheral blood lymphocytes was measured by fluorescence microscopy. Four of the lectins bound to various proportions of cells. Double labeling experiments using a rhodamine-labeled goat anti-bovine immunoglobulin reagent and fluorescein-labeled lectins showed that both peanut agglutinin and soybean agglutinin bound to lymphocyte populations which were negative for surface immunoglobulin. The majority of lymphocytes negative for surface immunoglobulin bound peanut agglutinin, indicating that this lectin may bind specifically to bovine 'T' lymphocytes.
Assuntos
Bovinos/sangue , DNA/biossíntese , Lectinas/farmacologia , Linfócitos/imunologia , Animais , Arachis , Imunofluorescência , Ativação Linfocitária , Linfócitos/metabolismo , Microscopia de Fluorescência , Lectinas de PlantasRESUMO
We describe detailed methods for derivation and cloning of myeloma hybrids which secrete antibodies specific for antigens of protozoan parasites. The methods were designed to enable the derivation of large numbers of specific monoclonal antibodies and to give high cloning efficiencies of desired hybrids. Although special attention is paid to derivation and detection of anti-parasite antibodies, the methods can be applied to many different antibody-antigen systems. Using the described methods we have isolated more than 90 myeloma hybrids which secrete antibodies specific for antigens of African trypanosomes and Theileria parasites, thus illustrating their effectiveness.
Assuntos
Anticorpos/análise , Apicomplexa/imunologia , Theileriose/parasitologia , Trypanosoma/imunologia , Animais , Especificidade de Anticorpos , Bovinos , Fusão Celular , Eucariotos/imunologia , Métodos , Camundongos , Theileriose/imunologiaRESUMO
A rabbit antimouse brain theta reagent was made specific for cells of the T lineage by absorption in vivo in nude mice. When used in double fluorescence together with an antimouse immunoglobulin reagent, four types of cells were found in spleen and lymph nodes of both normal and nude mice: Ig+thetaBr-, Ig-thetaBr+, Ig-thetaBr-, and Ig+thetaBr+. The data show that about 20% of nude mouse spleen lymphocytes are definitely of T lineage (Ig-thetaBr+). On these cells, the detection of the "thetaBr" determinant, which is identical or very close to the "theta" determinant, depends on the large amplification produced by indirect immunofluorescence, which suggests a low density of theta antigen. Similar experiments suggest the presence of cells that express some TL antigen in the spleen of nudes made congenic to a TL+ strain (BALB/c). It is proposed that the T-cell precursor that will further differentiate in the thymus already expresses a low density of theta and, in TL+ strains, TL antigen.
Assuntos
Diferenciação Celular , Imunofluorescência , Linfócitos T/imunologia , Animais , Especificidade de Anticorpos , Encéfalo/imunologia , Membrana Celular/imunologia , Cromatografia DEAE-Celulose , Feminino , Soros Imunes , Imunoglobulina G , Leucemia/imunologia , Linfonodos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Coelhos , Ovinos/imunologia , Baço/imunologia , Timo/imunologia , Timo/transplante , Transplante IsogênicoRESUMO
Zinc, copper and magnesium levels were determined by atomic absorption spectrophotometry in the serum of 32 cattle (Zebus and Baoulés) which were proven to be sensitive to African trypanosomiasis under field challenge and 45 cattle (Baoulés and Ndamas/Baoulés) which were proven to be resistant. Copper and magnesium levels were similar in all animals but zinc levels were higher in sensitive animals (1.50 ppm) than in resistant ones (1.10 ppm) (p less than 10(-5)); the reported normal levels of serum zinc is 1.00 ppm. These differences persisted on repeated measurements and whether individuals were infected with trypanosomes or not. Elevated levels of zinc depressed the stimulation of bovine T cells by trypanosomes in vitro and is reported to inhibit antigen presentation by macrophages. Zinc levels may be an influencial factor determining susceptibility or resistance of West African cattle to trypanosomiasis.
Assuntos
Tripanossomíase Bovina/imunologia , Zinco/sangue , Animais , Bovinos , Cobre/sangue , Feminino , Magnésio/sangue , Masculino , Espectrofotometria Atômica , Linfócitos T/imunologia , Tripanossomíase Africana/sangue , Tripanossomíase Africana/imunologia , Tripanossomíase Africana/veterinária , Tripanossomíase Bovina/sangueRESUMO
Cattle were exposed to natural trypanosome challenge in an area of high Glossina density (Samandéni, Burkina Faso) for various periods of time during 1982, 1983, 1984 and 1985. All of 30 Zebu proved to be sensitive to trypanosomiasis i.e. they died or were treated in extremis in 10 +/- 4 weeks. Twenty-one (31%) Baoulé were as sensitive as the Zebu while 47 (69%) were resistant i.e. they survived in good condition. Twenty Ndama/Baoulé crosses, indigenous to Samandéni were all resistant. Weekly blood samples were taken (2,317 in total) for the determination of parasitaemia and packed cell volume (PCV) as a measure of anaemia, the most important pathological feature of cattle trypanosomiasis. In both Zebu and sensitive Baoulé 59% of the blood samples showed positive parasitaemia, of which 38% and 52% respectively were T. congolense the major cattle pathogen in the area considered. In resistant Baoulé and Ndama/Baoulé 11% and 10% of the samples were positive for trypanosomes of which only 4% and 2% were T. congolense respectively. PCV decreased from 35 to 20 in Zebu, 39 to 20 in sensitive Baoulé and 40 to 34 in resistant Baoulé, there was no change in the indigenous Ndama/Baoulé. Six Ndama/Baoulé indigenous to Samandéni remained resistant to trypanosomiasis when moved to another area of high Glossina challenge. Seven Ndama/Baoulé calves, conceived in Samandéni but born and kept for 2 1/2 years in a Glossina-free area, also proved to be resistant to challenge. Twelve Baoulé calves, born from cattle selected under natural field challenge, and which had not come in contact with trypanosomes for the first 10 months of their life, proved to be resistant when exposed in the field. These observations show that some, but not all, cattle from the Baoulé breed are naturally resistant to African trypanosomiasis, that this resistance does not need repeated exposure to trypanosomes early in life but appears to be inherited and functional against many types of antigenically different trypanosomes. Thus, selective breeding of trypanoresistant animals and their successful introduction, without trypanocidal drug protection, into areas of high Glossina density appears feasible.
Assuntos
Seleção Genética , Tripanossomíase Bovina/imunologia , Animais , Burkina Faso , Bovinos , Imunidade Inata , Trypanosoma congolense , Tripanossomíase Africana/imunologiaRESUMO
Possible endocrinological repercussions of infection with Loa loa and Mansonella perstans filariae were studied in Gabonese subjects. Microfilaremic males were compared with amicrofilaremic controls. In the infected group 13/105 subjects (12%) presented only abnormally low serum levels of testosterone (less than 4 ng/ml), 25/105 (24%) only abnormally high serum levels of gonadotrophins, FSH (greater than 15 mIU/ml) and LH (greater than 20 mIU/ml), and 22/105 (21%) presented anomalies in both testosterone and gonadotrophin levels. One out of 68 control subjects had 3.6 ng/ml seric testosterone and all had normal levels of gonadotrophins. Ecdysteroids were detected (greater than 0.025 ng/ml) in the serum of 87/97 (90%) microfilaremic subjects (GM 0.123 ng/ml) compared to 12/64 (19%) controls (GM 0.030 ng/ml). Ecdysteroids were detected in the urine of all subjects, infected (GM 8.468 ng/ml) as well as control (GM 1.245 ng/ml). The hormonal perturbations were correlated with the levels of Loa loa microfilaremia but not with those of serum and urinary ecdysteroids. These results demonstrate that microfilaremic subjects often show endocrinal signs of hypogonadism and present appreciable levels of ecdysteroids in serum and urine. A direct role for parasitic ecdysteroids in hypogonadism remains to be demonstrated.
Assuntos
Filariose/metabolismo , Hipogonadismo/etiologia , Hormônios de Invertebrado/biossíntese , Loíase/metabolismo , Mansonelose/metabolismo , Adulto , Animais , Ecdisteroides , Hormônio Foliculoestimulante/sangue , Humanos , Hipogonadismo/metabolismo , Hormônios de Invertebrado/sangue , Hormônios de Invertebrado/urina , Loa/isolamento & purificação , Loíase/complicações , Hormônio Luteinizante/sangue , Masculino , Mansonella/isolamento & purificação , Mansonelose/complicações , Microfilárias/isolamento & purificação , Pessoa de Meia-Idade , Testosterona/sangueRESUMO
Ten animals from a reputedly trypanosensitive bovine breed, the Zebu, and ten from a reputedly trypanotolerant breed, the Baoulé, were exposed to natural trypanosome challenge in an area of high Glossina density. The ten Zebus and five of the Baoulés died within 6 to 11 weeks with heavy parasitaemia and severe anaemia. The five other Baoulés showed little or no parasitaemia, no anaemia and were all in good condition after weeks of exposure: Seven animals of a herd of Baoulé/Ndama crosses indigenous to the same area were followed concurrently and showed very rare or no patent parasitaemia. Clearly, only some individuals within a "trypanotolerant breed" are able to withstand natural fly challenge. Selection of trypanotolerant animals has to be done on an individual basis and not on the biometrical characteristics of the breed.
Assuntos
Tripanossomíase Bovina/imunologia , Animais , Cruzamento , Burkina Faso , Bovinos , Imunidade Inata , Masculino , Tripanossomíase Africana/imunologia , Tripanossomíase Africana/veterinária , Moscas Tsé-TséRESUMO
We have raised monoclonal antibodies to produce reagents specific for bovine lymphocyte subpopulations. Spleen cells from mice immunized with bovine peripheral blood lymphocytes were fused with X63-Ag8 myeloma cells and eleven myeloma-hybrids which secreted antibody specific for bovine lymphocytes were doubly cloned. Five of the hybrids secreted antibodies which bound to the majority of bovine lymphocytes. Two of these antibodies were specific for polymorphic antigens. One antibody bound to B lymphocytes and serum IgM molecules. The remaining five bound to subpopulations of lymphocytes. Four monoclonal antibodies bound only to bovine cells while six also bound to lymphocytes from other bovidae. None bound to human lymphocytes. We discuss the difficulty of correlating the specificities of monoclonal antibodies to functional lymphocyte subpopulations in outbred animals where few other defined markers are available.
Assuntos
Anticorpos Monoclonais/imunologia , Bovinos/imunologia , Linfócitos/imunologia , Tecido Linfoide/imunologia , Animais , Especificidade de Anticorpos , Bovinos/sangue , Células Clonais , Feminino , Humanos , Hibridomas/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Distribuição AleatóriaRESUMO
The nature and usefulness of trypanoresistance in West African Baoule cattle was studied by exposing animals in areas of high Glossina density. Under such conditions, Zebu and some Baoule died soon with high parasitaemia while resistant Baoule showed little patent parasitemia, almost no anaemia and thrived. Subsequently the role that specific antibody may play in such trypanoresistance was analyzed. In the first instance we examined protective antibody titres during T. congolense infection of inbred mice strains, comparing results obtained in trypanosome-mouse combinations where the host controls parasitemia and survives with those obtained when the host fails to control parasitemia and dies. We then attempted to extend these observations to cattle by following the disease course and appearance of neutralizing antibodies in animals of known sensitivity to natural Glossina challenges, following artificial challenge with T. congolense infected Glossina.
Assuntos
Anticorpos Antiprotozoários/imunologia , Doenças dos Bovinos/imunologia , Trypanosoma congolense/imunologia , Tripanossomíase Bovina/imunologia , Animais , Bovinos/classificação , Bovinos/genética , Bovinos/imunologia , Doenças dos Bovinos/genética , Interações Hospedeiro-Parasita , Imunidade Inata , Insetos Vetores/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos/genética , Camundongos Endogâmicos/imunologia , Tripanossomíase Africana/genética , Tripanossomíase Africana/imunologia , Tripanossomíase Africana/veterinária , Tripanossomíase Bovina/genética , Moscas Tsé-Tsé/parasitologiaRESUMO
Following tsetse-transmitted infection with Trypanosoma congolense, major differences in development of localised skin reactions, the ability to control parasitaemia, the degree of anaemia and in antibody response to trypanosomes were found between the reputedly trypanotolerant breeds of cattle (N'Dama, N'Dama/Baoule crosses, Baoule) and the trypanosusceptible West African Zebu. The local skin reactions that developed in the Zebu were large and severe while those that occurred in the other breeds were smaller and less severe or mild. The timing of appearance of parasitaemia and the height of the first peaks were similar in all the animals, but the Zebu were less able to control subsequent waves of parasitaemia. Possibly reflecting these events, it was only in the Zebu that significant anaemia developed. Neutralizing antibody against homologous metacyclic trypanosomes developed between 14 to 18 days after infection in all breeds of cattle; however, marked differences were found when antibody to trypanosomes derived from first peak parasitaemias were tested in the Zebu and Baoule. Neutralizing antibody against these parasites appeared in the Baoule on day 24 but were not detected in Zebu until day 51. Furthermore, the antibody titres were 3 log2 higher in the Baoule. It was concluded that the trypanotolerance exhibited by the West African taurine cattle might be related to a) their ability to control trypanosome numbers in the skin and in the bloodstream, an outcome that was possibly brought about by the earlier and superior immune response and b) failure to develop anaemia which might be associated with their capacity to control parasitaemia.
Assuntos
Bovinos/imunologia , Tripanossomíase Africana/imunologia , Moscas Tsé-Tsé/parasitologia , África Ocidental , Anemia/complicações , Animais , Formação de Anticorpos , Masculino , Pele/parasitologia , Pele/patologia , Trypanosoma congolense/imunologia , Tripanossomíase Africana/sangueRESUMO
Spleen cells from mice infected with T. congolense strongly suppressed lymphocyte stimulation induced in normal spleen cells by incubation with mitogens or allogeneic cells. Cell dilution studies showed that suppressor activity was extremely strong. Suppressor cell activity was markedly reduced by treatment of spleen cell populations with mitomycin-C and was unaffected by treatment with anti-Thy.1 sera and complement. Removal of cells which bound carbonyl iron or which bound to nylon columns, decreased but did not abolish suppressor activity.
Assuntos
Linfócitos T/imunologia , Tripanossomíase Africana/imunologia , Animais , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Mitomicinas/farmacologia , Baço/imunologiaRESUMO
Seven out of 11 bovines infected with different clones of Trypanosoma brucei showed 2 peaks of antibody activity against the infecting clone within 7 weeks, as measured by immunofluorescence, radioimmunoassay, and neutralization of infectivity tests. Using other clones from an unrelated Stock, antibodies to these clones were not detectable, indicating that the antibodies produced were specific to the infecting organisms. These results suggest that there was a reappearance or increase in numbers of the infecting organisms or of organisms with variable surface antigens similar to those of the infecting clones. The reappearance of variable antigen types in the presence of specific antibodies would imply that antibody plays a selective rather than an inductive role in the process of antigenic variation in African trypanosomes.
Assuntos
Antígenos , Trypanosoma brucei brucei/imunologia , Tripanossomíase Bovina/imunologia , Animais , Anticorpos/análise , Bovinos , Epitopos , Tripanossomíase Bovina/sangueAssuntos
Antígenos , Epitopos , Peritônio/imunologia , Ligação Proteica , RNA , Adjuvantes Imunológicos , Animais , Formação de Anticorpos , Linfócitos B/imunologia , Sítios de Ligação de Anticorpos , Sedimentação Sanguínea , Células Cultivadas , AMP Cíclico , Escherichia coli/imunologia , Exsudatos e Transudatos/imunologia , Cobaias , Células HeLa/imunologia , Tecido Linfoide/imunologia , Modelos Biológicos , Linfócitos T/imunologiaAssuntos
Tripanossomíase Africana/genética , Animais , Cruzamentos Genéticos , Feminino , Hibridização Genética , Terapia de Imunossupressão , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos A , Camundongos Endogâmicos AKR , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Tripanossomíase Africana/mortalidadeRESUMO
Glossina infected with African trypanosomes infest 10(7) Km2 of intertropical Africa. Ten thousand cases of human sleeping sickness are officially recorded each year and 35 X 10(6) human beings are at risk. Animal trypanosomiasis impedes the use of 7 X 10(6) Km2 of land adequate for cattle raising and constitutes a major constraint to increasing protein production in Africa. Several approaches are used to combat trypanosomiasis (a) vector eradication has been successful in certain defined situations but cannot be realistically extended to the whole area at risk, (b) prophylactic drugs, which are too toxic for humans, are widely used to protect cattle. This practice has led to field resistance to many drugs used in the past (reviewed in Holmes & Scott 1982) and the appearance of resistance to the last available prophylactic drug, isometamidium, is being reported (Bourn & Scott 1978, Küpper & Wolters 1983, Pinder & Authié 1984), (c) the hope for a vaccine has been largely abandoned, in the present state of knowledge, due to the considerable extent of antigenic diversity in trypanosomes (reviewed in Doyle 1977, Turner 1982, Roelants & Pinder 1984), (d) consequently, the possible use of certain West African breeds of cattle, which appear resistant to trypanosomiasis, has been emphasized as a solution to this problem in domestic animals. The analysis of this natural resistance is the subject of the present essay.