RESUMO
Human epidemiological studies have suggested that social variables can modulate the effects of stress on the immune system, and this concept has been gaining increasing attention with positive results emerging from empirical studies using nonhuman primates over the last two decades. Results from a previous study in rhesus monkeys suggested that receiving grooming positively affected recovery of T-helper and T-suppressor cells following the initial stress associated with group formation, and this co-varied with high dominance rank. Thus, the present study was undertaken in order to determine: (1) if the stress effect of formation could be replicated in another species and (2) if social behaviors or dominance rank, given that formation is a stressor, might independently correlate with physiological recovery from the stressor. Eight adult female pigtail macaques were moved from individual cages and simultaneously introduced into an outdoor enclosure along with an adult male, while eight weight-matched controls remained in individual caging. Behavioral data were collected during the introduction and over 4 weeks thereafter. Blood samples were collected prior to and at intervals for 4 weeks following formation. Compared to control subjects, the test subjects showed an increase in basal cortisol secretion (+28.9%) and a significant decrease in T-helper cells (-33.6%), T-suppressor cells (-30.8%), and B cells (-22.5%), while there was a significant increase in white blood cells (+29.5%) 24 hr following formation. When dominance rank and seven behavioral categories were analyzed, only the frequency of receiving grooming significantly predicted change, with animals who received a greater frequency of grooms showing a lesser negative percent change from baseline in the absolute number of T-helper cells 1 week following formation. The establishment of a dominance hierarchy, apparent within 1 week, was accomplished with no serious fighting and a complete absence of wounding or trauma, suggesting that psychosocial stress was responsible for the physiological changes observed. © 1996 Wiley-Liss, Inc.
RESUMO
The absence of detectable levels of CSF-1 in op/op mice results in a marked deficiency of macrophage colony-stimulating activity (M-CSA) in both the unstimulated and postendotoxin sera of these animals. These deficiences are not secondary to the presence of an inhibitor of macrophage formation. In contrast, various organs, particularly the organs of endotoxin-treated op/op mice, released amounts of M-CSA comparable to those of normal mice. Similarly, mitogen-stimulated lymphoid cells from op/op mice released either similar or increased amounts of M-CSA compared to mitogen stimulated +/+ lymphoid cells. On the other hand, conditioned media from cultures of fibroblastoid cells obtained from primary and secondary cultures of op/op organs were nearly totally devoid of M-CSA. However, incubation of these op/op fibroblasts with endotoxin in vitro induced the easily and readily detectable levels of M-CSA. These data suggest that CSF-1 is most likely a major source of M-CSA in serum and postendotoxin serum, while its contribution to soluble M-CSA in other organs may be only partial. In addition, in specific circumstances, the induced release of other macrophage growth factors may partially compensate for CSF-1 deficiency. Furthermore, it appears that the generalized macrophage deficiency in op/op mice cannot be fully explained by the deficiency of soluble M-CSA (soluble CSF-1) and argues for an in vivo role of membrane-bound CSF-1. These data may be interpreted as supporting a model in which the regulation of CSF-1-dependent and CSF-1-independent macrophage production is carried out by partly unrelated mechanisms.
Assuntos
Fator Estimulador de Colônias de Granulócitos e Macrófagos/sangue , Fator Estimulador de Colônias de Macrófagos/deficiência , Macrófagos/metabolismo , Animais , Medula Óssea/metabolismo , Meios de Cultivo Condicionados , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Rim/metabolismo , Pulmão/metabolismo , Macrófagos/química , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Músculos/metabolismo , Técnicas de Cultura de Órgãos , Pele/metabolismo , Baço/metabolismo , Timo/metabolismoRESUMO
Osteopetrosis and the absence of colony-stimulating factor 1 (CSF-1) in op/op mice are associated with decreased cellularity of the bone marrow (to one tenth of the normal), a very significant reduction in the number of cells recovered from peritoneal, pleural, and alveolar lavages, moderate leukopenia, and a slight decrease in the number of cells per spleen and thymus. Furthermore, op/op mice possess deficiencies in the number of macrophages in various organs. These cells are apparently absent in the bone marrow, severely reduced (5%-15% of the normal number) in peritoneal and pleural cavities and in the lungs. In addition, a marked decrease in the frequency and total number of circulating monocytes is present (5% of the normal). The deficiency of macrophages is less severe in the liver, spleen, and thymus of op/op mice (approximately 30% of those seen in normal). There is a concomitant redistribution of macrophage progenitor cells (granulocyte-macrophage colony-forming units, CFU-GM) in op/op mice from the marrow to the spleen and liver, associated with an increased sensitivity to interleukin 3 (IL-3). Their total number is decreased at least threefold compared to control mice. Moreover, op/op mice have at least a fivefold reduction in the total number of day-11 spleen colony-forming units (CFU-S) associated with their redistribution to the spleen and liver. These data suggest that the macrophage system in op/op mice is reduced at all levels tested, that is, at the level of mature macrophages, the level of progenitors, and the level of stem cells, whereas the redistribution of progenitor and stem cells could be viewed as a secondary consequence of osteopetrosis. Furthermore, these data suggest that macrophage dependency in vivo on CSF-1 is limited and different in various organs. Particularly in the liver, spleen, and thymus, other growth factors may significantly compensate for CSF-1 deficiency. Based on the relative decrease in the number of CFU-GM in the op/op mice, it appears that the population size of these progenitors is less dependent on CSF-1 than the hematopoietic stem cell population size as evidenced by the day-11 CFU-S assay. The day-11 CFU-S population is severely reduced in op/op mice, suggesting a physiological involvement of CSF-1 in expanding its size. These data provide evidence that CSF-1, besides acting on the final and intermediate stages of macrophage maturation, may also play a role in early stages of hematopoiesis.
Assuntos
Fator Estimulador de Colônias de Macrófagos/deficiência , Macrófagos/citologia , Camundongos Mutantes/fisiologia , Animais , Contagem de Células , Diferenciação Celular/fisiologia , Divisão Celular/fisiologia , Feminino , Hematopoese/fisiologia , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/fisiologia , Fator Estimulador de Colônias de Macrófagos/fisiologia , Macrófagos/fisiologia , Masculino , Camundongos , Osteopetrose/genética , Osteopetrose/patologia , Células-Tronco/citologia , Células-Tronco/fisiologiaRESUMO
Psychosocial stress associated with the removal of six naive juvenile rhesus monkeys from their natal social group to peer housing resulted in increased basal cortisol secretion and significant decrements in the absolute numbers of the T lymphocyte subsets in the peripheral blood. Six subjects matched for age and social rank remained in the group of 80 animals serving as controls. Baseline immune and cortisol measurements were obtained before the six test subjects were removed from the group and housed together in an outdoor circular enclosure. Blood samples were taken 24 h following removal of the test subjects from the group and at intervals thereafter through 11 weeks. Compared to controls, test subjects showed a significant decrease in the absolute numbers of CD4+ (-56.9%) and CD8+ T cells (-57.6%) and a significant increase in basal cortisol levels (+43.9%) 24 h following removal to peer housing. Group difference in the absolute numbers of most immune cells persisted through 11 weeks, whereas cortisol differences lasted only through 2 weeks. These data, when compared to an earlier study employing an identical protocol, with the exception that subjects were housed in indoor individual cages following separation, fail to demonstrate a modulating effect of randomly chosen peer-mates on the stress effects produced by social separation.
Assuntos
Ansiedade de Separação/imunologia , Hidrocortisona/sangue , Meio Social , Estresse Psicológico/imunologia , Subpopulações de Linfócitos T , Animais , Ansiedade de Separação/sangue , Ansiedade de Separação/fisiopatologia , Feminino , Abrigo para Animais , Contagem de Leucócitos , Macaca mulatta/imunologia , Macaca mulatta/fisiologia , Masculino , Grupo Associado , Apoio Social , Estresse Psicológico/sangue , Estresse Psicológico/fisiopatologiaRESUMO
Removal of juvenile rhesus monkeys from their natal social group to indoor individual caging resulted in increased basal cortisol secretion and significant decrements in the frequency of lymphoid subpopulations. Fourteen juvenile rhesus monkeys, which had never been removed from the group, were studied. Baseline immune and cortisol measurements were obtained before seven of the subjects were removed from social housing to standard individual cages. The remaining seven subjects, matched for age, sex, weight, and rank, remained in the social group throughout the study serving as controls. Blood samples were taken 24 hours after removal of the test subjects from the group and at specific intervals thereafter through 11 weeks. At 24 hours after the separation test subjects showed a significant increase in basal cortisol levels (40%) and a significant decrease in several immune parameters, with absolute numbers of total T cells declining 72 +/- 12%. Significant group differences in immune parameters persisted through 11 weeks. Eighteen weeks following removal, the test subjects were returned to the group which produced a cortisol rise in both test and controls at the 24-hour postreturn sample. Although there were no group differences in the frequency of lymphoid subsets 24 hours after return, some test subjects showed marked decrements which were inversely related to cortisol and were predicted by behavioral events. These data demonstrate that the removal of naive juvenile rhesus monkeys from their natal social group to individual indoor caging is a potent psychosocial stressor and that the behavioral interactions which characterize the return of the individual subjects to the natal group may predict physiological response.
Assuntos
Contagem de Leucócitos , Meio Social , Isolamento Social , Estresse Psicológico/imunologia , Animais , Nível de Alerta/fisiologia , Linfócitos B/imunologia , Relação CD4-CD8 , Hidrocortisona/sangue , Imunofenotipagem , Macaca mulatta , Monócitos/imunologiaRESUMO
Social stress associated with the formation of a new group of rhesus monkeys resulted in increased basal cortisol secretion and significant decreases in immunological parameters. Eight adult female rhesus monkeys, all of which had been raised in social groups, but with no common social history, were simultaneously introduced into an outdoor enclosure along with an adult male. Behavioral data were collected during the introduction and over 9 weeks thereafter. Blood samples were collected prior to and at intervals for 9 weeks following formation. The establishment of a dominance hierarchy, apparent within 48 h, was accomplished with no serious fighting and a complete absence of wounding or trauma. Overall, the group showed a significant increase in cortisol and a significant decrease in the absolute number of total lymphocytes and CD4+ and CD8+ T cells at 24 h postformation, but not thereafter. However, when partitioned into high and low dominance rank, differences in CD4+ and CD8+ T cells were evident for up to 9 weeks with low ranking subjects showing significantly lower values. The housing condition of the subjects immediately prior to introduction, either indoors in individual caging or outdoors in social groups, may have influenced behavior, rank acquisition, and possibly differences in immune parameters. These data demonstrate that social group formation is a potent psychosocial stressor in primates, since stress-sensitive changes were observed in the absence of serious aggression and wounding.
Assuntos
Macaca mulatta/psicologia , Sistema Hipófise-Suprarrenal/fisiopatologia , Comportamento Social , Estresse Psicológico/fisiopatologia , Agressão , Animais , Relação CD4-CD8 , Feminino , Asseio Animal , Abrigo para Animais , Hidrocortisona/sangue , Contagem de Leucócitos , Macaca mulatta/imunologia , Macaca mulatta/fisiologia , Predomínio Social , Estresse Psicológico/imunologia , Subpopulações de Linfócitos TRESUMO
Human cardiac myocytes do not express detectable levels of major histocompatibility complex (MHC) class II antigens and express low levels, if any, of MHC class I antigens. During rejection episodes, cardiac biopsies show massive increases of MHC antigens, which are thought to be induced by cytokines released by donor-sensitized recipient mononuclear cells. In efforts to determine the nature of the cytokines that induce MHC expression on cardiac myocytes, human fetal cardiac myocyte cultures were established. Interferon-alpha (IFN-alpha), interferon-gamma (IFN-gamma), interleukin (IL)-1, IL-2, IL-3, IL-4, and tumor necrosis factor (TNF)-alpha were added to these cultures and dose/kinetics of MHC class I/II induction quantitated. Data show that IFN-gamma induces both MHC class I and II expression, and all the other cytokines (except IL-2) induce only MHC class I but not class II. Cytokines used in combination showed that IFN-alpha with TNF-alpha was the only combination that induced MHC class II expression. Addition of immunosuppressive drugs such as cytoxan, azathioprine, cyclosporine-A, and FK-506, even when added at the initiation of the cultures, did not appreciably affect the ability of the appropriate cytokines to induce MHC expression by the myocytes in vitro.
Assuntos
Citocinas/farmacologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Antígenos de Histocompatibilidade Classe I/metabolismo , Imunossupressores/farmacologia , Miocárdio/imunologia , Anticorpos Monoclonais , Células Cultivadas , Feto , Humanos , Técnicas Imunoenzimáticas , Miocárdio/citologia , Miocárdio/metabolismoRESUMO
Techniques such as polyclonal B cell activation with pokeweed mitogen (PWM) and polymerase chain reaction (PCR) analysis have documented the existence of simian immunodeficiency virus (SIV)-and human immunodeficiency virus type 1-seronegative but infected humans and nonhuman primates. To establish whether blood from such seronegative but PWM- and PCR-positive monkeys can transmit infection, naive macaques were transfused with whole blood (n = 2) or cultured cells and supernatant fluid (n = 2) from two seronegative but PWM- and PCR-positive sooty mangabeys. After transfusion, three of the four recipients seroconverted, and peripheral blood mononuclear cells from all four recipients secreted SIV-reactive antibodies upon polyclonal activation in vitro and were SIV-positive by PCR that used highly specific gag primer pairs and probe. In addition, CD8+ cells from all four recipients markedly inhibited replication of SIV in autologous cells in vitro. These data suggest caution in the sole use of serologic tests for the detection of retroviral infection and document the ability of such blood samples to transmit infection.
Assuntos
Transfusão de Sangue , Síndrome de Imunodeficiência Adquirida dos Símios/transmissão , Animais , Anticorpos Antivirais/sangue , Western Blotting , Cercopithecidae , DNA Viral/análise , Ensaio de Imunoadsorção Enzimática , Ativação Linfocitária , Macaca mulatta , Macaca nemestrina , Reação em Cadeia da Polimerase , Vírus da Imunodeficiência Símia/genética , Vírus da Imunodeficiência Símia/imunologia , Vírus da Imunodeficiência Símia/fisiologia , Replicação ViralRESUMO
Human cardiac myocytes undergo degeneration, cytolysis, and necrosis in a number of clinical disease conditions such as myocarditis, dilated cardiomyopathy, and during episodes of cardiac allograft rejection. The precise cellular, biochemical, and molecular mechanisms that lead to such abnormalities in myocytes have been difficult to investigate because at present it is not possible to obtain and maintain viable cell cultures of human adult cardiac myocytes in vitro. However, human fetal cardiac myocytes are relatively easy to maintain and culture in vitro, but their limited availability and growth, variability from one preparation to another, and varying degrees of contamination with endothelial and epithelial cell types have made it difficult to obtain reliable data on the effect of cardiotropic viruses and cardiotoxic drugs on such myocytes. These thoughts prompted us to attempt to derive a cell line of human cardiac origin. Highly enriched human fetal cardiac myocytes were transfected with the plasmids pSV2Neo and pRSVTAg and gave rise to a cell line (W1) which has been maintained in culture for 1 yr. Morphologic and phenotypic analyses of W1 cells by flow microfluorometry and immunoperoxidase techniques indicate that the W1 cell line shares many properties of human fetal cardiac myocytes, but appears not to react with specific antibodies known to react with markers unique to human endothelial, epithelial, skeletal muscle, and dendritic cells. These preliminary data suggest that the W1 cells may provide a unique source of an established cell line that shares many properties ascribed to human cardiac myocytes.
Assuntos
Miocárdio/citologia , Northern Blotting , Divisão Celular , Linhagem Celular , Membrana Celular/imunologia , Creatina Quinase/metabolismo , Técnicas de Cultura/métodos , Feto , Antígenos HLA/análise , Coração/fisiologia , Humanos , L-Lactato Desidrogenase/metabolismo , Complexo Principal de Histocompatibilidade , Microscopia Eletrônica , Miocárdio/metabolismo , Miocárdio/ultraestrutura , Miosinas/biossíntese , RNA/genética , RNA/isolamento & purificaçãoRESUMO
While normal cardiac tissue expresses low levels of MHC-class I, undetectable levels of MHC-class II antigens, and no mononuclear cell infiltrates, posttransplantation allogeneic donor cardiac tissue demonstrates dramatic increases of MHC-class I/class II expression coincident with the infiltration of the tissue with mononuclear cells. Results of this study demonstrate that the kinetics of MHC-class I/II antigen expression and the phenotype of mononuclear cell infiltrate are influenced, to a great degree, by the genetic H-2, intra-H-2 and non-H-2 incompatibility between donor and recipient strains of mice. Increases of MHC-class I precede class II expression in cells from donor cardiac tissue from completely allogeneic BALB/c, H-2-disparate B10.D2, B10.BR, and K, I-A and I-E-disparate B10.T (6R) strains of mice implanted in B10 recipients. In contrast, increase in the level of MHC-class II precedes MHC-class I increases in donor cardiac tissue from H-2-identical but non-H-2-incompatible A. By and the I-E + H-2D end-different B10.A(5R) donor tissue. The completely allogeneic, H-2-disparate or K, I-A, I-E-disparate donor cardiac tissue induced the infiltration of predominantly CD8+ T cells, whereas the non H-2 and I-E + H-2D end-different donor cardiac tissue induced the infiltration of predominantly CD4+ T cells. Finally, whereas bm1 donor cardiac tissue is rejected by B6 recipients by day 32, the (bm1 x bm12)F1 allografts are rejected by day 20, and both express MHC-class I antigens followed by MHC-class II antigens, and contain predominantly CD8+ T cells. In contrast, bm12 allografts are not rejected by B6 recipients, express chronic low levels of both MHC-class I and II antigens, and contain predominantly CD4+ T cells. Of interest is our preliminary finding that bm12 allografts placed in one ear of B6 recipients appear to modify the kinetics of MHC antigen expression and the predominant phenotype of mononuclear cell infiltrates in bm1 allografts placed in the opposite ear. Cumulatively, these data suggest that the type of genetic disparity between cardiac donor and recipient greatly influences the quantitative and qualitative host responses.
Assuntos
Transplante de Coração/imunologia , Antígenos de Histocompatibilidade Classe II/imunologia , Antígenos de Histocompatibilidade Classe I/imunologia , Complexo Principal de Histocompatibilidade , Animais , Antígenos de Diferenciação de Linfócitos T/análise , Antígenos Ly/análise , Sobrevivência de Enxerto , Antígenos H-2/genética , Antígenos H-2/imunologia , Histocompatibilidade , Antígenos de Histocompatibilidade Classe I/análise , Antígenos de Histocompatibilidade Classe I/genética , Antígenos de Histocompatibilidade Classe II/análise , Antígenos de Histocompatibilidade Classe II/genética , Técnicas Imunoenzimáticas , Memória Imunológica , Leucócitos Mononucleares/imunologia , Camundongos , Camundongos Endogâmicos/genética , Miocárdio/imunologia , Radioimunoensaio/métodos , Fatores de TempoRESUMO
Osteopetrotic (op/op) mutant mice suffer from congenital osteopetrosis due to a severe deficiency of osteoclasts. Furthermore, the total number of mononuclear phagocytes is extremely low in affected mice. Serum, 11 tissues, and different cell and organ conditioned media from op/op mice were shown to be devoid of biologically active colony-stimulating factor 1 (CSF-1), whereas all of these preparations from littermate control +/+ and +/op mice contained the growth factor. The deficiency was specific for CSF-1 in that serum or conditioned media from op/op mice possessed elevated levels of at least three other macrophage growth factors. Partial correction of the op/op defect was observed following intraperitoneal implantation of diffusion chambers containing L929 cells, which in culture produce CSF-1 as their sole macrophage growth factor. No rearrangement of the CSF-1 gene in op/op mice was detected by Southern analysis. However, in contrast to control lung fibroblasts, which contained 4.6- and 2.3-kilobase CSF-1 mRNAs, only the 4.6-kilobase species was detected in op/op cells. An alteration in the CSF-1 gene is strongly implicated as the primary defect in op/op mice because they do not contain detectable CSF-1, their defect is correctable by administration of CSF-1, the op locus and the CSF-1 gene map within the same region of mouse chromosome 3, their CSF-1 mRNA biosynthesis is altered, and the op/op phenotype is consistent with the phenotype expected in a CSF-1 deficient mouse.
Assuntos
Medula Óssea/patologia , Fatores Estimuladores de Colônias/metabolismo , Macrófagos/citologia , Osteopetrose/genética , Animais , Células da Medula Óssea , Células Cultivadas , Fatores Estimuladores de Colônias/genética , Cruzamentos Genéticos , Meios de Cultura , DNA/análise , Feminino , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/patologia , Fator Estimulador de Colônias de Macrófagos , Masculino , Camundongos , Camundongos Mutantes , Osteopetrose/patologia , RNA Mensageiro/análise , RNA Mensageiro/genética , Valores de ReferênciaRESUMO
The measurement of cell-mediated immunity against the etiologic agent of human AIDS (HIV) in the non-human primate model of AIDS (simian immunodeficiency virus, SIV) has been difficult. In general, culture of peripheral blood mononuclear cells from HIV-1- and SIV-infected humans and monkeys, respectively, with purified inactivated HIV and SIV virus preparations has given inconsistent or negative proliferative responses. However, we describe herein an assay which consists of coculturing monocytes that have been pulsed with inactivated SIVsmm with nylon-wool-purified autologous T cells, leading to antigen-specific T-cell proliferation. The proliferative response, which predominantly occurs in CD4+ T cells, is major histocompatibility complex (MHC) class II-restricted and requires antigen processing. This assay will greatly facilitate the identification of the immunodominant epitopes recognized by T cells in sooty mangabeys, which are naturally infected but remain clinically asymptomatic, and in rhesus macaques, in which experimental infection leads to clinical symptomatology similar to human AIDS, eventually resulting in death.
Assuntos
Antígenos Virais/imunologia , Ativação Linfocitária , Infecções por Retroviridae/imunologia , Vírus da Imunodeficiência Símia/imunologia , Linfócitos T/imunologia , Animais , Anticorpos Monoclonais/imunologia , Células Apresentadoras de Antígenos/imunologia , Antígenos de Diferenciação de Linfócitos T/análise , Antígenos CD4/análise , Antígenos CD8 , Cercopithecidae , Citomegalovirus/imunologia , Macaca mulattaRESUMO
Identification of human immunodeficiency virus type 1 (HIV-1)-infected individuals is of paramount importance for the control of the spread of AIDS worldwide. Currently, the vast majority of screening centers throughout the world rely on serological techniques. As such, clinically asymptomatic but HIV-infected, seronegative individuals are rarely identified. In this report we show that 18% (30/165) of seronegative individuals who were considered to be a unique cohort of patients at high risk for HIV infection had circulating B cells that, upon in vitro polyclonal activation with pokeweed mitogen, produced antibodies reactive with HIV. Furthermore, polymerase chain reaction analysis of DNA obtained from aliquots of the peripheral blood mononuclear cells from these seronegative but pokeweed mitogen assay-positive individuals tested revealed the presence of HIV-specific sequences in a significant number of samples. In addition, depletion of CD8+ T cells from peripheral blood mononuclear cells of HIV-1-seronegative individuals prior to in vitro culture with pokeweed mitogen resulted in increased sensitivity for detecting HIV-reactive antibodies. This assay has obvious epidemiological implications, especially in the case of high-risk groups, and also provides a simple technique to enhance detection of HIV-infected individuals. Of further interest is the determination of the mechanisms related to the lack of HIV-specific antibodies in the serum of these infected individuals.
Assuntos
Linfócitos B/imunologia , Anticorpos Anti-HIV/análise , Soropositividade para HIV , HIV-1/imunologia , Ativação Linfocitária , Antígenos de Diferenciação de Linfócitos T/análise , Western Blotting , Antígenos CD8 , Células Cultivadas , DNA/genética , DNA/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , HIV-1/genética , Humanos , Mitógenos de Phytolacca americana , Reação em Cadeia da PolimeraseRESUMO
Immunization with recombinant human pyruvate dehydrogenase (PDH)-E2, the major autoantigen of primary biliary cirrhosis, readily induces a vigorous murine antibody response but does not generate hepatic disease. To determine the fine specificity of this response, 18 mAb were generated from three strains of mice and the reactive epitopes mapped. An initial examination of mAb suggested that they behaved similarly to the antimitochondrial autoantibodies in primary biliary cirrhosis (PBC) because i) all polyclonal antisera and 2 of 18 mAb reacted with all species of mammalian PDH-E2 examined including mouse PDH-E2, ii) 15 of 18 mAb inhibited PDH enzyme function, and iii) the reactivity of mAb toward rPDH-E2 were blocked by PBC sera. However, fine examination of the reactive sequences of the PDH-E2 complex revealed that antibodies identical to those in PBC patients were not produced by experimental immunization. In contrast to PBC, none of the mAb or murine polyclonal sera were able to react with protein X, a lipoic acid-containing component of the PDH complex previously shown to cross-react with PDH-E2 when probed with PBC sera. Although the epitopes for 12 mAb were localized within the inner lipoyl domain, none reacted with mouse PDH-E2 or cross-reacted with the outer lipoyl domain as observed in PBC. In addition, the epitopes of the two mAb which did react with all mammalian species of mitochondria were not localized within the PBC epitope. These findings indicate the highly immunogenic nature of the inner lipoyl domain of PDH-E2. The inability to elicit antibodies of the same specificity in mice, considered together with the highly localized autoantibody response in humans, suggests that antimitochondrial autoantibodies are most likely the result of specific breakdown of tolerance to a unique autoepitope.
Assuntos
Anticorpos Monoclonais/imunologia , Autoanticorpos/imunologia , Autoantígenos/imunologia , Cirrose Hepática Biliar/imunologia , Mitocôndrias/imunologia , Complexo Piruvato Desidrogenase/imunologia , Acetiltransferases/imunologia , Sequência de Aminoácidos , Animais , Epitopos , Humanos , Camundongos , Dados de Sequência Molecular , Proteínas Recombinantes/imunologia , Mapeamento por Restrição , Especificidade da EspécieRESUMO
While cell-mediated immunity is known to play an important role in controlling viral infections, its role in human and experimental animal models of human AIDS has not been established. To address this issue, four juvenile rhesus macaques were infected with simian immunodeficiency virus SIVMAC. Freshly isolated peripheral blood mononuclear cells from these SIVMAC-infected macaques and four uninfected control macaques were assessed for T-cell proliferative activity to SIV, monthly, for 10 consecutive months. T cells from SIV-infected monkeys failed to proliferate in response to SIV added directly to the culture. However, when SIV was processed by autologous antigen-presenting cells prior to culture with purified T cells, proliferative responses were uniformly demonstrated in SIV-infected monkeys, but not in uninfected controls. Proliferation in response to heat-inactivated SIV was mediated by CD4+ T cells and was shown to be MHC class II-restricted. However, the proliferative response to infectious SIV was mediated by both CD4+ and CD8+ T cells and was MHC class-restricted. As disease progressed, a decline in the T-cell proliferative response was observed.
Assuntos
Ativação Linfocitária , Infecções por Retroviridae/imunologia , Vírus da Imunodeficiência Símia/imunologia , Linfócitos T/imunologia , Animais , Antígenos de Diferenciação de Linfócitos T , Antígenos CD4 , Antígenos CD8 , Feminino , Antígenos de Histocompatibilidade Classe II , Técnicas In Vitro , Macaca mulatta , Masculino , Fatores de TempoRESUMO
The histologic diagnosis of active myocarditis is frequently difficult to establish. A nonhistologic marker of immune activation would be clinically useful in identifying cases of immune-mediated myocarditis. A viral etiology with subsequent autoimmunity to cardiac antigens has been implicated in human myocarditis. Because autoimmunity and viral disease are commonly associated with increased expression of major histocompatibility complex (MHC) antigens on targeted tissue, we examined endomyocardial biopsy samples from patients with active myocarditis for abnormal levels of MHC antigen expression. Thirteen patients with active myocarditis and eight control patients with other well-defined cardiac diagnoses (coronary disease, amyloidosis or neoplasm) were studied. A sensitive radioimmunoassay was developed that utilized monoclonal antibodies to human MHC class I and class II antigens in order to quantitate the expression of both of these antigens within each biopsy. Abnormal MHC class I and class II antigen expression was present in 11 of 13 myocarditis specimens and 1 of 8 control samples (specificity 88%, sensitivity 84.6%). Active myocarditis samples had approximately a 10-fold increase in MHC class I and class II expression. Immunoperoxidase staining localized abnormal MHC expression primarily within microvascular endothelium and along myocyte surfaces (11 of 13). This study is the first to demonstrate a marked increase in major histocompatibility complex antigen expression within the myocardium of patients with active myocarditis. The identification of abnormal histocompatibility antigen expression within an endomyocardial biopsy may prove a useful adjunct to the histologic diagnosis of myocarditis.
Assuntos
Antígenos HLA/análise , Antígenos HLA-D/análise , Miocardite/diagnóstico , Adulto , Idoso , Anticorpos Monoclonais , Autoanticorpos/análise , Biomarcadores/análise , Endotélio Vascular/imunologia , Feminino , Humanos , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-Idade , Miocardite/imunologia , Valor Preditivo dos Testes , RadioimunoensaioRESUMO
Initial studies have revealed subtle differences in the T cell proliferative response to whole SIV antigen in the peripheral blood mononuclear cells (PBMC) from sooty mangabeys and rhesus macaques. Preliminary findings utilizing the cellular Western blot assay are described.
Assuntos
Proteínas dos Retroviridae/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/imunologia , Linfócitos T/imunologia , Animais , Anticorpos Antivirais/biossíntese , Antígenos Virais/imunologia , Western Blotting , Cercopithecidae , Ativação Linfocitária , Macaca mulattaRESUMO
Several investigators have demonstrated the ability of CD8+ T cells from HIV-1 infected humans and SIV infected rhesus macaques to inhibit viral replication in vitro. In this report we show that CD8+ cells from naturally SIV infected sooty mangabeys also have the ability to inhibit viral replication in vitro. In addition, initial experiments which seek to elucidate the mechanism and antigen specificity of CD8-mediated suppression are described.
Assuntos
Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/imunologia , Linfócitos T Reguladores/imunologia , Animais , Linfócitos B/imunologia , Linfócitos B/microbiologia , Antígenos CD4/genética , Linhagem Celular Transformada , Cercopithecidae , HIV-2/imunologia , HIV-2/fisiologia , Herpesvirus Humano 4 , Humanos , DNA Polimerase Dirigida por RNA/metabolismo , Vírus da Imunodeficiência Símia/enzimologia , Vírus da Imunodeficiência Símia/fisiologia , Replicação ViralRESUMO
Monocytes from SIV/SMM infected sooty mangabeys and rhesus macaques were incubated in vitro with live SIV/SMM. The reverse transcriptase (RT) activity in the supernatant fluids of the monocyte cultures of the former species was higher than the RT activity in the latter species. No differences were found in the supernatant fluid of similar cultures of CD4+ T cells from both these species. Autologous (but not allogeneic) CD8+ T cells from SIV infected mangabeys and rhesus macaques inhibited SIV replication in vitro. The suppression appeared more marked in monocytes from the mangabey species. These in vitro differences may relate to the clinically asymptomatic state of the sooty mangabeys and the disease-susceptible state of the rhesus macaques.
Assuntos
Linfócitos T CD4-Positivos/microbiologia , Macrófagos/microbiologia , Monócitos/microbiologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Vírus da Imunodeficiência Símia/fisiologia , Animais , Células Cultivadas , Cercopithecidae , Cinética , Macaca mulatta , DNA Polimerase Dirigida por RNA/metabolismo , Vírus da Imunodeficiência Símia/enzimologia , Vírus da Imunodeficiência Símia/imunologia , Linfócitos T Reguladores/imunologia , Replicação ViralRESUMO
Four juvenile rhesus macaques were infected with simian immunodeficiency virus (SIV)MAC-Freshly isolated peripheral blood mononuclear cells (PBMC) from these SIVMAC-infected and from uninfected control macaques were assessed for cytotoxic T-lymphocyte (CTL) activity monthly for 7 consecutive months, beginning 2 months after infection. Target cells consisted of major histocompatibility complex (MHC) haploidentical parental PBMC which were stimulated with mitogen and then pulsed with heat-killed SIVMAC. CTL activity was demonstrated on all four infected animals. The effector cells are T cells which mediate cytotoxicity against SIVMAC-pulsed target cells in an MHC-restricted manner. Furthermore, the cytotoxicity is virus specific and predominantly, if not exclusively, mediated by CD8+ T cells; it is also MHC class-I restricted. Incubation of target cells with leupeptin prior to the cytotoxic assay inhibited target cell generation, suggesting that viral antigens are processed via an endocytic pathway.