RESUMO
Anti-RNA polymerase III (RNAP III) antibodies are highly specific for scleroderma (SSc) and associated with diffuse SSc and renal crisis. Coexistence of anti-RNAP III and other SSc autoantibodies is rarely documented. We report three cases with coexisting anti-RNAP III and anti-U1RNP. Autoantibodies in 3829 sera from rheumatology clinics were screened by immunoprecipitation. Anti-RNAP III-positive sera were also examined by immunofluorescence and anti-RNAP III ELISA. In total, 35 anti-RNAP III-positive sera were identified by immunoprecipitation, in which three had coexisting anti-U1RNP. All three were anti-RNAP III ELISA positive. Two had anti-RNAP I dominant (vs. RNAP III) reactivity and showed strong nucleolar staining. A case with anti-U1/U2RNP (U2RNP dominant) had systemic lupus erythematosus (SLE)-SSc overlap syndrome; however, the remaining two cases had SLE without signs of SSc. All three cases of anti-RNAP III + U1RNP fulfilled ACR SLE criteria but none in the group with anti-RNAP III alone (p = 0.0002). In contrast, only one case in the former group had sclerodermatous skin changes and Raynaud's phenomenon, vs. 92% with scleroderma in the latter (p < 0.05). Although anti-RNAP III is highly specific for SSc, cases with coexisting anti-U1RNP are not so uncommon among anti-RNAP III positives (8%, 3/35) and may be SLE without features of SSc.
Assuntos
Anticorpos Antinucleares/sangue , Autoanticorpos/sangue , Lúpus Eritematoso Sistêmico , RNA Polimerase III/imunologia , Ribonucleoproteína Nuclear Pequena U1/imunologia , Escleroderma Sistêmico/imunologia , Adulto , Feminino , Humanos , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/patologia , Escleroderma Sistêmico/sangueRESUMO
The NZM2410-derived Sle1a lupus susceptibility locus induces activated autoreactive CD4(+) T cells and reduces the number and function of Foxp3(+) regulatory T cells (Tregs). In this study, we first showed that Sle1a contributes to autoimmunity by increasing antinuclear antibody production when expressed on either NZB or NZW heterozygous genomes, and by enhancing the chronic graft versus host disease response indicating an expansion of the autoreactive B-cell pool. Screening two non-overlapping recombinants, the Sle1a.1 and Sle1a.2 intervals that cover the entire Sle1a locus, revealed that both Sle1a.1 and Sle1a.2 were necessary for the full Sle1a phenotype. Sle1a.1, and to a lesser extent Sle1a.2, significantly affected CD4(+) T-cell activation as well as Treg differentiation and function. Sle1a.2 also increased the production of autoreactive B cells. As the Sle1a.1 and Sle1a.2 intervals contain only 1 and 15 known genes, respectively, this study considerably reduces the number of candidate genes responsible for the production of autoreactive T cells. These results also show that the Sle1 locus is an excellent model for the genetic architecture of lupus, in which a major obligate phenotype results from the coexpression of multiple genetic variants with individual weak effects.
Assuntos
Predisposição Genética para Doença , Lúpus Eritematoso Sistêmico/genética , Linfócitos T/imunologia , Animais , Autoimunidade/genética , Autoimunidade/imunologia , Linfócitos B/imunologia , Linfócitos B/metabolismo , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Feminino , Doença Enxerto-Hospedeiro/genética , Doença Enxerto-Hospedeiro/imunologia , Lúpus Eritematoso Discoide/genética , Lúpus Eritematoso Discoide/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Lúpus Eritematoso Sistêmico/metabolismo , Lúpus Vulgar/genética , Lúpus Vulgar/imunologia , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Camundongos , Camundongos Congênicos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NZB , Camundongos Endogâmicos , Camundongos Knockout , Linfócitos T/metabolismo , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismoRESUMO
Mice homozygous for the lpr gene develop autoantibodies and polyclonal B cell activation similar to what is seen in human systemic lupus erythematosus patients. We have previously shown that an lpr-specific intrinsic B cell defect was necessary for autoantibody production in this model. In the current study, we have further defined these autoantibody-producing B cells. Two major subsets of B cells have been described. B-1 cells (CD5+ B cells) can be distinguished from conventional B cells on the basis of phenotype, cytokine secretion, gene expression, anatomical location, and function. In addition, B-1 cells have been implicated in autoimmunity in several murine and human studies. To address the question of which B cell subset produces autoantibodies in lpr mice, we used immunoglobulin heavy chain (Igh) allotype-marked peritoneal (B-1 cell source) and bone marrow (conventional B cell source) cells from lpr mice to establish B cell chimeras. We used two general approaches. In one, we reconstituted sublethally irradiated mice with B-1 cells of one allotype and bone marrow cells of another allotype. In the second method, we suppressed endogenous B cells in neonatal mice with allotype-specific anti-IgM antibody, and injected peritoneal cells of another allotype. After antibody treatment was stopped, the mouse's conventional B cells recovered, but the B-1 subset was only reconstituted by the donor. In both types of chimeras, antichromatin, rheumatoid factor, and anti-single stranded DNA (ssDNA) autoantibodies were produced by the conventional B cell bone marrow source. In addition, an age-related decrease in peritoneal B-1 cells was seen, even in unmanipulated lpr mice. These data show that lpr B-1 cells are not important producers of autoantibodies. Conventional B cells are the source of autoantibodies directed at chromatin, ssDNA, and IgG.
Assuntos
Autoanticorpos/biossíntese , Doenças Autoimunes/imunologia , Linfócitos B/imunologia , Transtornos Linfoproliferativos/imunologia , Envelhecimento , Animais , Camundongos , Camundongos Endogâmicos C57BL , Quimera por RadiaçãoRESUMO
Mice homozygous for the gene lpr develop marked lymphadenopathy and a spectrum of autoantibodies closely resembling that of human systemic lupus erythematosus. The unusual T cell phenotype of the expanded lymphocyte population and the T-dependence of several antibodies in this strain have suggested that primary T cell abnormalities underlie the autoimmune syndrome. Using double chimeras, we now show that expression of the lpr gene in B cells is absolutely necessary for autoantibody production. Combinations of anti-Thy 1.2 + C' treated bone marrow from congenic strains of C57BL/6 mice, differing only at the immunoglobulin heavy chain (Igh) and lpr loci, were transferred into lethally irradiated B6/lpr mice. Double chimerism was documented by allotype-specific surface IgD and IgM immunofluorescence assay of peripheral blood and by allotype-specific enzyme-linked immunosorbent assay for total IgM in serum. Despite the presence of both +/+ and lpr B cells, IgM and IgG2a anti-chromatin as well as IgM anti-IgG were entirely the products of lpr B cells. Total serum IgG2a and IgG1 were also dominated by the lpr phenotype but not to the same extent. A similar experiment using B6/lpr-Igha recipients confirmed these findings. Additional experiments in which B6/lpr recipients were infused with ratios of donor bone marrow favoring B6.C20 +/+ over B6/lpr showed that even though +/+ B cells were overrepresented, autoantibodies were only of the lpr allotype. In addition, in the presence of lpr B cells, normal B cells showed little response to an exogenous, T cell-dependent antigen. The data thus indicate that lpr B cells manifest an intrinsic abnormality which is essential for autoantibody production in the lpr model.
Assuntos
Autoanticorpos/imunologia , Linfócitos B/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Animais , Transplante de Medula Óssea , Camundongos , Camundongos Endogâmicos , Quimera por Radiação , Linfócitos T/imunologia , Trinitrobenzenos/imunologiaRESUMO
MHC-linked genes strongly influence susceptibility to autoimmune diseases and also regulate responses to exogenous antigens. To begin to understand the mechanism of this MHC effect on disease, we have investigated MHC-congenic mouse strains that develop spontaneous autoimmunity because of the lpr gene. C57BL6/lpr (B6/lpr) mice (H-2b) are known to have substantial levels of autoantibodies to chromatin, single stranded DNA (ssDNA3), and IgG of different murine subclasses (rheumatoid factor). We have crossed the H-2d and the H-2bm12 (la mutant) haplotypes onto the B6/lpr background. Surprisingly, levels of all the autoantibodies were markedly lower in B6/lpr.H-2d, but levels in B6/lpr.H-2bm12 were no different from those in B6/lpr mice. The downregulating influence of the H-2d allele was dominant, and there was no effect on autoantibody fine specificities. The genetics of the H-2d effect and its diffuse influence on multiple autoantibody specificities, in addition to the lack of effect of the bm12 mutation, which modifies the peptide-binding groove of I-A, together raise the question of whether MHC-linked genes other than classical (IR) genes may be responsible for MHC disease associations in this model.
Assuntos
Autoanticorpos/imunologia , Epitopos/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Complexo Principal de Histocompatibilidade/imunologia , Animais , Cromatina/imunologia , Cruzamentos Genéticos , DNA/imunologia , Antígenos H-2/imunologia , Haplótipos/imunologia , Imunização , Lúpus Eritematoso Sistêmico/genética , Linfonodos/patologia , Camundongos , Camundongos Endogâmicos C57BL/imunologia , Camundongos Endogâmicos/imunologia , Camundongos Mutantes/imunologia , Tamanho do Órgão , Baço/patologiaRESUMO
In viable motheaten mice, a mutation in the gene encoding the phosphatase, SHP1, causes severe immunodeficiency and autoimmunity. A defective phosphatase may result in modified phosphorylation of proteins involved in gene regulation. Since the NFkappaB/IkappaB proteins are regulated through phosphorylation, we wished to understand if the expression of these proteins was altered by the SHP1 defect. Splenic B cells from viable motheaten mice were isolated and assessed for purity by flow cytometry. Levels of each protein in isolated B cells were examined by Western blot analyses. Measurement of RNA levels for each protein was assessed by semi-quantitative RT-PCR. Western blots revealed that, in me(v) whole cell lysates, there were reduced levels of RelA and RelB proteins and increased levels of p50 and c-Rel. Furthermore, we analyzed the protein levels of IkappaBalpha and found that, in me(v), this inhibitor was significantly reduced, while the level of another member of the IkappaB family, IkappaBbeta, was not. To determine if these findings in me(v) were secondary to the autoimmune process, we evaluated NF-kappaB/IkappaB expression in the BXSB murine model of autoimmunity. Unlike me(v), B cells from BXSB/Yaa mice had NF-kappaB complexes composed of the RelA submit, and IkappaBalpha was readily detected. In addition, RNA for the RelA and IkappaBalpha proteins in me(v) and control littermates was detected by RT-PCR, indicating that the reduced amounts of these proteins was not exclusively due to transcriptional defects. We conclude that the differences in NF-kappaB/IkappaB proteins that we have described in me(v) are likely a consequences of the SHP1 defect and could contribute to the clinical disorder that characterizes me(v) mice.
Assuntos
Doenças Autoimunes/genética , Linfócitos B/metabolismo , Proteínas de Ligação a DNA/biossíntese , Regulação da Expressão Gênica/genética , Proteínas I-kappa B , Síndromes de Imunodeficiência/genética , NF-kappa B/biossíntese , Proteínas Tirosina Fosfatases/genética , Animais , Doenças Autoimunes/enzimologia , Doenças Autoimunes/imunologia , Proteínas de Ligação a DNA/genética , Células-Tronco Hematopoéticas/enzimologia , Células-Tronco Hematopoéticas/patologia , Síndromes de Imunodeficiência/enzimologia , Síndromes de Imunodeficiência/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Ligases/análise , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Modelos Biológicos , Inibidor de NF-kappaB alfa , NF-kappa B/análise , NF-kappa B/genética , Subunidade p50 de NF-kappa B , Fosforilação , Processamento de Proteína Pós-Traducional , Proteína Tirosina Fosfatase não Receptora Tipo 11 , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas Tirosina Fosfatases/deficiência , Proteínas Proto-Oncogênicas/análise , Proteínas Proto-Oncogênicas c-rel , RNA Mensageiro/análise , Baço/imunologia , Fator de Transcrição RelB , Fatores de Transcrição/análise , Transcrição GênicaAssuntos
Antígenos CD/análise , Subpopulações de Linfócitos B/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Animais , Formação de Anticorpos , Autoanticorpos/análise , Autoanticorpos/biossíntese , Antígenos CD5 , Quimera , Cromatina/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Imunoglobulina G/biossíntese , Imunoglobulina G/sangue , Imunoglobulina G/classificação , Imunoglobulina M/biossíntese , Imunoglobulina M/sangue , Camundongos , Camundongos Endogâmicos C57BL , Camundongos MutantesRESUMO
Measurement of high sensitivity C-reactive protein (hs-CRP), has been used in the assessment of disease activity in numerous rheumatic conditions including systemic lupus erythematosus (SLE). However, the utility of hs-CRP measurement in patients with lupus is uncertain. This study examined if hs-CRP can be used to assess disease activity, severity and cardiovascular risk in SLE. Serum samples from 601 visits of 213 SLE patients and 134 controls were analysed for hs-CRP by nephelometry. Detailed demographic data were obtained from all subjects and medication history and key laboratory parameters were collected. Disease activity was assessed using the SLEDAI. High sensitivity CRP was not associated with disease activity (SLEDAI), number of ACR SLE criteria or presence of any particular organ involvement. hs-CRP levels were significantly correlated with standard cardiovascular risk factors including body weight (P = 0.0002), hypertension (P = 0.001), and apolipoprotein A-I (P < 0.0001). Interestingly an inverse correlation was seen between hs-CRP levels and antimalarial use (P = 0.0018). Our results suggest that measurement of hs-CRP, though not valuable as marker of disease activity in SLE may be of some use in the assessment of cardiovascular risk. We speculate that antimalarials may help to reduce cardiovascular risk in patients with SLE.
Assuntos
Proteína C-Reativa/metabolismo , Doenças Cardiovasculares/etiologia , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/complicações , Apolipoproteínas/sangue , Biomarcadores/sangue , Estudos de Casos e Controles , Proteínas do Sistema Complemento/metabolismo , Feminino , Humanos , Lipoproteínas/sangue , Masculino , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
The lpr and gld murine models have been important contributors to our understanding of systemic autoimmune diseases. Mice homozygous for either of these autosomal recessive genes develop a phenotypically identical disease characterized by the accumulation of CD4-CD8- T-cells and the production of a wide spectrum of autoantibodies. The lpr (lymphoproliferation) mutation encodes a defective Fas apoptosis receptor gene. More recently, gld (generalized lymphadenopathy) has been shown to be a point mutation in the Fas ligand gene. Despite the molecular characterization of these mutations, the exact mechanism by which tolerance is lost is still unknown, although in vivo cell transfer studies have provided clues. Chimera studies, in which normal and lpr bone marrow were co-infused into lpr mice, demonstrated not only that the normal Fas receptor is functionally expressed in both T- and B-cells, but that the Fas mutation is required in both for full expression of the lpr phenotype. Conversely, in analogous experiments with gld mice, co-infusion of normal and gld bone marrow largely prevented the development of autoantibodies. Sporadic autoantibody titers were seen in some mice, but were derived from both donors. The effects on T-cells were subtly different: The CD4-CD8- T-cells were also greatly reduced in number, but all were of gld origin. These data indicate that the gld defect is extrinsic to B-cells but only partially extrinsic to T-cells, and suggest that Fas ligand in T-cells may have an autocrine and paracrine function.
Assuntos
Autoimunidade/genética , Linfócitos/imunologia , Transtornos Linfoproliferativos/genética , Glicoproteínas de Membrana/imunologia , Receptor fas/imunologia , Animais , Apoptose/imunologia , Autoanticorpos/biossíntese , Doenças Autoimunes/genética , Doenças Autoimunes/imunologia , Modelos Animais de Doenças , Proteína Ligante Fas , Expressão Gênica , Tolerância Imunológica , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos MRL lpr , Camundongos Mutantes , Linfócitos T/imunologia , Receptor fas/genéticaRESUMO
Total intensity, Rayleigh light scattering has been used to measure the rms radius, second virial coefficient, persistence length, and excluded volume of homogeneous T7 bacteriophage DNA as a function of Na+ concentration (0.005 to 3.0 M). All parameters decrease sharply as [Na+] increases, and tend to level off at high Na+. The variation of persistence length with [Na+] is consistent with predictions from counterion condensation theory.
Assuntos
DNA Viral/química , Sódio/farmacologia , Fagos T/metabolismo , DNA Viral/efeitos dos fármacos , Matemática , Modelos Teóricos , Conformação de Ácido Nucleico , Espalhamento de RadiaçãoRESUMO
Exposure to low doses of mercury chloride induces autoantibodies to the nucleolar protein fibrillarin in H-2s, but not in H-2b, mice. Surprisingly, F1 crosses between resistant and sensitive haplotypes are resistant. Previously, we have shown that the resistance in these F1 mice was due to coexpression of the resistant class II allele. Using adoptive transfer techniques we have examined several mechanisms by which the resistant haplotype could be down-regulating the antifibrillarin response in F1 (s/b) mice. Similar to other autoimmune models, mercury-induced autoimmunity requires cognate MHC-restricted T cell help. The absence of autoantibody production in F1 mice was not due to a difference in thymic education or to the absence of antifibrillarin-specific T cell help. These results suggest that the resistance is due to an intrinsic property of the haplotype-heterozygous B cells.
Assuntos
Doenças Autoimunes/genética , Subpopulações de Linfócitos B/imunologia , Heterozigoto , Cloreto de Mercúrio/imunologia , Animais , Anticorpos Antinucleares/biossíntese , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Subpopulações de Linfócitos B/metabolismo , Nucléolo Celular/imunologia , Proteínas Cromossômicas não Histona/imunologia , Cruzamentos Genéticos , Haplótipos , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Imunidade Inata/efeitos dos fármacos , Imunidade Inata/genética , Cooperação Linfocítica/genética , Cloreto de Mercúrio/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Quimera por Radiação/genética , Quimera por Radiação/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Linfócitos T Auxiliares-Indutores/metabolismo , Timo/imunologia , Timo/patologiaRESUMO
Mice homozygous for the gene Ipr develop a spectrum of autoantibodies closely resembling that of human SLE. Previous work has shown that the lpr defect must be expressed in the T cells that hyperproliferate and in the B cells that produce autoantibodies. Although autoantibody production in lpr mice requires T cells, it is not known whether these need to be lpr T cells. To ask whether normal (+/+) T cells can help lpr B cells produce autoantibodies, we have constructed chimeras containing mixtures of lpr-derived and normal-derived lymphoid cells, and have selectively eliminated the lpr-derived T cells by in vivo treatment with monoclonal anti-Thy-1 of the appropriate allotype. A mixture of T cell-depleted bone marrow from congenic strains of normal and lpr mice differentially marked by Ig H chain allotype and Thy-1 alleles was transferred into lethally irradiated lpr mice. The mice received weekly injections of either anti-Thy-1.2 to deplete specifically lpr T cells or an isotype-matched irrelevant control mAb. Absence of lpr-derived T cells in the experimental group was documented by immunofluorescence. In mice treated with control antibody, autoantibodies of Ipr origin were present in high titers, as determined by allotype-specific ELISA. In contrast, mice depleted of lpr-derived T cells had greatly reduced titers of antichromatin and rheumatoid factor. These mice also had increased levels of serum total IgM and IgG2a of +/+ origin. Parallel experiments were performed using a combination of two lpr marrow sources, also differentially marked by Ig H chain allotype and Thy-1 expression. Mice depleted of Thy-1.2-bearing T cells produced autoantibodies of both allotypes due to the presence of Thy-1.1-bearing T cells of Ipr origin. These data indicate that autoantibody production in lpr mice requires expression of the lpr gene in those T cells that provide help.
Assuntos
Autoanticorpos/biossíntese , Doenças Autoimunes/imunologia , Genes Recessivos , Transtornos Linfoproliferativos/imunologia , Linfócitos T/imunologia , Animais , Doenças Autoimunes/genética , Linfócitos B/fisiologia , Quimera , Alótipos de Imunoglobulina/biossíntese , Isoanticorpos/imunologia , Depleção Linfocítica , Transtornos Linfoproliferativos/genética , CamundongosRESUMO
One of the most striking features of exposure to low doses of mercury in mice is the high-titer haplotype-linked anti-nucleolar (ANoA) autoantibody response. Mice of H-2(s) haplotype have been high responders, while H-2(b) mice have been low. This pattern has been attributed to the class II molecule itself, but the poor response of F1 crosses between high and low responders raised the possibility that the anti-fibrillarin specificity was actually due to a closely linked dominant negative gene. To test the role of class II explicitly, F1 crosses between congenic B6.SJL (H-2(s)) and C57BL/6 (H-2(b)) mice with a targeted deletion of I-AbAbeta were generated, creating mice heterozygous for all MHC loci, but expressing only I-As. In comparison with B6.SJL, no diminution of ANoA titers was found, proving that I-As itself was responsible for susceptibility and I-Ab for downregulation. Unlike I-A, expression of the I-E class II molecule could not downregulate the response in an otherwise susceptible mouse. These results suggest a complicated role for class II in the regulation of a novel, environmentally induced autoimmune response.
Assuntos
Doenças Autoimunes/induzido quimicamente , Antígenos de Histocompatibilidade Classe II/genética , Cloreto de Mercúrio/toxicidade , Animais , Formação de Anticorpos , Autoanticorpos/imunologia , Autoimunidade/efeitos dos fármacos , Autoimunidade/genética , Feminino , Haplótipos , Imunidade Inata/genética , Masculino , Camundongos , Camundongos Endogâmicos A , Camundongos Endogâmicos C57BL , Escleroderma Sistêmico/genéticaRESUMO
lpr and gld mice develop phenotypically indistinguishable systemic autoimmune diseases and marked lymphadenopathy dominated by CD4-CD8- T cells. In vivo chimera experiments have demonstrated that both lpr T and lpr B cells are intrinsically defective. Analogous experiments were conducted using gld mice. Lethally irradiated gld mice were given mixtures of congenic gld and normal (+/+) bone marrow differentially marked by Ig heavy chain allotype. In sharp contrast to lpr-(+)/+ mixed chimeras, gld-(+)/+ chimeras had little autoantibody production at 5 months and minimal adenopathy at 6 months, indicating that the normal marrow-derived cells corrected the gld defect. Thus, aberrant autoantibody production is due to a defect extrinsic to the gld B cell and lymphoproliferation is due to a defect extrinsic to the gld T cell. These data support the hypothesis that gld mice lack an apoptosis-inducing ligand. The receptor for this ligand may be the Fas molecule, which is defective in lpr mice.
Assuntos
Antígenos de Superfície/genética , Apoptose , Doenças Autoimunes/genética , Transplante de Medula Óssea , Transtornos Linfoproliferativos/genética , Animais , Antígenos CD4/análise , Antígenos CD8/análise , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Camundongos , Camundongos Endogâmicos C57BL , Mutação , Receptor fasRESUMO
NF-kappaB is a potential target for immunosuppressive therapy. Two methods were evaluated to inhibit NF-kappaB: the antisense (AS) approach in which single-stranded oligodeoxynucleotides (ODNs) bind the mRNA for the RelA subunit of NF-kappaB and the transcription factor decoy (TFD) approach in which double-stranded ODNs bind the NF-kappaB protein. AS and TFD inhibited NF-kappaB binding and decreased total IgG and anti-dsDNA antibody production in splenocytes from the BXSB/Yaa autoimmune mouse strain. TNF-alpha expression was reduced by AS and TFD, as were the levels of IL-2. But AS effects did not last beyond 24 h, whereas TFD inhibited cytokine production after 72 h. AS had no effect upon IL-6, while the TFD reduced the secretion of IL-6. Therefore, the suppression of immune response mediators by AS or TFD, through inhibition of NF-kappaB, is substantial. These inhibitors can serve as novel choices for therapy in the treatment of autoimmune disorders.
Assuntos
Imunossupressores/farmacologia , NF-kappa B/antagonistas & inibidores , Oligonucleotídeos Antissenso/farmacologia , Tionucleotídeos/farmacologia , Animais , Células Cultivadas , Citocinas/biossíntese , Citocinas/genética , Citocinas/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Imunoglobulina G/metabolismo , Interleucina-2/metabolismo , Camundongos , NF-kappa B/biossíntese , NF-kappa B/genética , Baço/citologia , Baço/imunologia , Fator de Transcrição RelARESUMO
To define the functional consequences of the src-homology domain-1 protein (SHP-1) defect, we examined cytokine production and NF-kappa B activity in motheaten viable (Mev) mice. We found elevated levels of interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor (TNF), and interferon-gamma (IFN-gamma) in Mev mice sera and cultured B and T cells compared to littermate control adult mice. The levels of interleukin-2 (IL-2) detected in Mev sera and activated Mev T cells were decreased, but IL-2 receptor expression was increased. We then evaluated the activity of NF-kappa B and found that this protein is highly expressed in Mev B and T cells. To determine if NF-kappa B had a role in causing the elevated levels of cytokines in Mev mice, we treated activated Mev T cells with an NF-kappa B decoy and found that cell culture treatment with the decoy resulted in significant reduction of the secretion of IL-6, GM-CSF, and TNF, but not IFN-gamma. Therefore, our data show that Mev mice secrete elevated levels of inflammatory cytokines, which can be mediators in the development of the Mev clinical disorder, and that NF-kappa B has an important role in this process, impacting upon the regulation of the immune response.
Assuntos
NF-kappa B/fisiologia , Proteínas Tirosina Fosfatases/genética , Proteínas Tirosina Fosfatases/fisiologia , Animais , Doenças Autoimunes/genética , Doenças Autoimunes/imunologia , Subpopulações de Linfócitos B/efeitos dos fármacos , Subpopulações de Linfócitos B/metabolismo , Ligação Competitiva , Células Cultivadas , Citocinas/sangue , Citocinas/efeitos dos fármacos , Citocinas/metabolismo , Inflamação/sangue , Inflamação/imunologia , Peptídeos e Proteínas de Sinalização Intracelular , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Proteína Tirosina Fosfatase não Receptora Tipo 11 , Proteína Tirosina Fosfatase não Receptora Tipo 6 , Proteínas Tirosina Fosfatases/deficiência , Proteínas Tirosina Fosfatases Contendo o Domínio SH2 , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/metabolismo , Domínios de Homologia de src/genética , Domínios de Homologia de src/imunologiaRESUMO
Ipr and gld mice develop systemic autoimmune diseases with nearly indistinguishable manifestations, including the accumulation of massive numbers of CD4-CD8- T lymphocytes. In vivo chimera experiments have shown that the Ipr mutation is functionally expressed in both T and B cells. When lethally irradiated Ipr mice were given a combination of normal and Ipr bone marrow, only Ipr-derived B cells produced autoantibodies and only Ipr-derived T cells hyperproliferated. In contrast, analogous experiments with gld mice showed that the co-infusion of normal bone marrow greatly reduced autoantibody production. These results indicated that the gld B cell defect was extrinsic to those cells producing autoantibodies, in agreement with the recent molecular data showing that the normal gene products of the Ipr and gld loci form an interacting receptor-ligand pair. In the present study, we have extended our functional studies with gld mice using T cell-marked congenic donors. Lymphadenopathy was reduced three- to fourfold in gld mice given a combination of congenic normal and gld bone marrow compared with mice given gld bone marrow alone, and the absolute number of CD4-CD8- T cells was reduced by a factor of 7. Surprisingly, the residual CD4-CD8- T cells present in the mixed chimeras were derived entirely from the gld donor marrow. This suggests that the gld mutation results in both an extrinsic and intrinsic defect in T cells.
Assuntos
Doenças Autoimunes/terapia , Transplante de Medula Óssea , Transtornos Linfoproliferativos/terapia , Glicoproteínas de Membrana/fisiologia , Camundongos Mutantes/imunologia , Animais , Antígenos de Superfície/genética , Antígenos de Superfície/fisiologia , Autoanticorpos/biossíntese , Doenças Autoimunes/imunologia , Proteína Ligante Fas , Hipergamaglobulinemia/genética , Hipergamaglobulinemia/imunologia , Linfonodos/patologia , Cooperação Linfocítica , Contagem de Linfócitos , Linfócitos Nulos/imunologia , Transtornos Linfoproliferativos/imunologia , Glicoproteínas de Membrana/deficiência , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes/genética , Quimera por Radiação , Antígenos Thy-1 , Receptor fasRESUMO
Sle1 is a potent autoimmune susceptibility locus on chromosome 1 originally identified in a genome scan of testcross progeny between the systemic lupus erythematosus-prone NZM2410 strain and C57BL/6. We subsequently produced B6.NZMc1, a congenic strain carrying the NZM2410-derived Sle1 genomic interval on the B6 background and demonstrated that Sle1 mediated the loss of tolerance to chromatin in both the B and T cell compartments. In this communication, we show by adoptive transfer experiments that the autoimmune phenotypes of Sle1 are completely reconstituted in B6 radiation chimeras receiving B6.NZMc1 bone marrow but not by the reciprocal reconstitution, demonstrating that Sle1 is functionally expressed in B cells. In additional experiments, cotransfer of mixtures of bone marrow derived from B6.NZMc1 and nonautoimmune congenic B6 mice carrying allelic T and B cell markers showed that only B cells derived from B6.NZMc1 bone marrow produced anti-chromatin autoantibodies. In contrast, increased expression of CD69 was equivalent in CD4+ T cells derived from either B6.NZMc1 or congenic B6 bone marrow, suggesting that either T cell population could be activated subsequent to loss of tolerance in the B cell compartment. These findings indicate that the expression of Sle1 in B cells is essential for the development of autoimmunity.
Assuntos
Transferência Adotiva , Subpopulações de Linfócitos B/imunologia , Células da Medula Óssea/imunologia , Tolerância Imunológica/genética , Lúpus Eritematoso Sistêmico/etiologia , Lúpus Eritematoso Sistêmico/genética , Animais , Antígenos CD/biossíntese , Antígenos de Diferenciação de Linfócitos T/biossíntese , Autoanticorpos/biossíntese , Autoantígenos/biossíntese , Autoantígenos/imunologia , Subpopulações de Linfócitos B/metabolismo , Subpopulações de Linfócitos B/transplante , Antígeno B7-2 , Células da Medula Óssea/metabolismo , Transplante de Medula Óssea/imunologia , Linfócitos T CD4-Positivos/transplante , Regulação da Expressão Gênica/imunologia , Marcadores Genéticos/imunologia , Imunoglobulina G/biossíntese , Imunofenotipagem , Lectinas Tipo C , Lúpus Eritematoso Sistêmico/imunologia , Glicoproteínas de Membrana/biossíntese , Camundongos , Camundongos Congênicos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NZB , Nucleossomos/imunologia , Nucleossomos/metabolismo , Subpopulações de Linfócitos T/metabolismo , Quimeras de Transplante/genética , Quimeras de Transplante/imunologiaRESUMO
CD45 is involved in the regulation of lymphocyte activation, and it has been demonstrated that ligation of CD45 induces apoptosis of T and B lymphocytes. Recently anti-CD45RB antibody therapy was shown to block acute allograft rejection in a mouse model of transplantation. Therefore, we wanted to examine the effects of anti-CD45RB antibody treatment on the course of an autoimmune disorder, experimental allergic encephalomyelitis (EAE), a Th1-mediated process. Mice immunized with myelin basic protein and treated with anti-CD45RB antibody did not develop EAE. Histologically, there was no evidence of lymphocytic infiltrates in the central nervous system. T cell proliferation and TNF-alpha production were significantly decreased in anti-CD45RB-treated mice. Furthermore, there was a significant reduction in the production of other Th1 cytokines including interferon-gamma and IL-2, but not IL-4 or IL-6. However, levels of a number of adhesion markers or markers of activation such as VLA-4 and LFA-1 on T cells were no different in treated versus control animals. Thus, anti-CD45RB can prevent EAE and appears to do so by altering T cell proliferation and cytokine production.
Assuntos
Anticorpos Monoclonais/uso terapêutico , Encefalomielite Autoimune Experimental/prevenção & controle , Antígenos Comuns de Leucócito/imunologia , Animais , Divisão Celular , Citocinas/biossíntese , Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/patologia , Encefalomielite Autoimune Experimental/terapia , Integrina alfa4beta1 , Integrinas/biossíntese , Antígeno-1 Associado à Função Linfocitária/biossíntese , Camundongos , Ratos , Receptores de Retorno de Linfócitos/biossíntese , Linfócitos T/imunologiaRESUMO
Systemic lupus erythematosus is characterized by polyclonal B cell activation, the production of autoantibodies, and often by renal disease. Previous studies demonstrated that unfractionated B cells from several strains of mice with lupus hyperproliferate in culture when stimulated with lipopolysaccharide (LPS) or anti-IgM. We wished to further examine proliferation of resting B cells from the BXSB mouse model of lupus and mice with the Yaa allele, when activated with a number of stimuli. Our work demonstrates that: (1) resting B cells from mice containing the Yaa allele hyperproliferated compared to that seen with B cells from mice lacking the Yaa allele, (2) this hyperproliferation occurred whether cells were stimulated with phorbol myristate acetate/ionomycin, LPS, anti-IgM, or CD40L cross-linking, (3) this hyperproliferation is specific to B and not T cells. Taken together these data suggest that one mechanism by which the Yaa allele contributes to the accelerated onset of lupus in BXSB male mice is through its influence on B cell activation.