RÉSUMÉ
Most systemic autoimmune diseases occur more frequently in females than in males. This is particularly evident in Sjögren's syndrome, systemic lupus erythromatosis (SLE) and thyroid autoimmunity, where the ratio of females to males ranges from 20:1 to 8:1. Our understanding of the etiology of SLE implies important roles for genetics, environmental factors and sex hormones, but the relative significance of each remains unknown. Using the New Zealand hybrid mouse model system of SLE, we present here a new fetal liver chimera-based system in which we can segregate effects of immune system genes from that of sex hormones in vivo. We show that female hematopoietic cells express an intrinsic capacity to drive lupus-like disease in both male and female recipient mice, suggesting that this capacity is hormone independent. Particularly, only chimeric mice with a female hematopoietic system showed significantly increased numbers of germinal center B cells, memory B cells and plasma cells followed by a spontaneous loss of tolerance to nuclear components and hence elevated serum antinuclear autoantibodies. A protective effect of testosterone was noted with regard to disease onset, but not disease incidence. Thus, genetic factors encoded within the female hematopoietic system can effectively drive lupus-like disease even in male recipients.
Sujet(s)
Auto-immunité , Cellules souches hématopoïétiques , Hybridation génétique , Animaux , Autoanticorps/biosynthèse , Autoanticorps/immunologie , Maladies auto-immunes/étiologie , Sous-populations de lymphocytes B/immunologie , Sous-populations de lymphocytes B/métabolisme , Cellules de la moelle osseuse , Transplantation de moelle osseuse , Femelle , Foetus , Hormones sexuelles stéroïdiennes/métabolisme , Hépatocytes/métabolisme , Hépatocytes/transplantation , Interféron alpha/sang , Maladies du rein/étiologie , Activation des lymphocytes/immunologie , Mâle , Souris , Grossesse , Chimère obtenue par transplantationRÉSUMÉ
Several studies have described a role for type I interferons (IFNalphabeta) in the initiation and/or prolongation of autoimmune diseases. Most pronounced has been the association of disease activity with what is now known as 'the interferon signature' of gene expression in peripheral blood mononuclear cells from lupus patients. In correlation, studies have shown that inhibition of IFNalphabeta signaling abrogates disease in various mouse models of lupus. New Zealand black (NZB) and B6.Nba2 congenic mice spontaneously develop elevated levels of serum anti-nuclear autoantibodies (ANAs). Nevertheless, neither of these strains develop fatal renal disease. The female F1 offspring of NZB or B6.Nba2 crossed with New Zealand white (NZW) mice do, however, develop kidney disease. We have previously shown that increases in endogenous IFNalphabeta levels in (B6.Nba2 x NZW)F1 mice leads to accelerated development of renal disease in an IFNalphabeta-dependent manner. We now show that B6.Nba2 and (B6.Nba2 x NZW)F1 mice deficient for the IFNalphabeta-receptor fail to develop ANA and renal disease, although the mice have substantial immune complex deposition in the glomeruli. Thus, endogenous IFNalphabeta might influence disease by affecting B-cell activation and differentiation, as well as the kidneys' susceptibility to damage, the latter perhaps through induction of a local inflammatory milieu.
Sujet(s)
Prédisposition génétique à une maladie , Interféron alpha/métabolisme , Interféron bêta/métabolisme , Glomérulonéphrite lupique/immunologie , Récepteur à l'interféron alpha-bêta/génétique , Animaux , Lymphocytes B/immunologie , Femelle , Glomérulonéphrite lupique/génétique , Activation des lymphocytes , Souris , Souris congéniques , Transduction du signal/génétiqueRÉSUMÉ
The Bcl-2 family member Bax plays a critical role in apoptosis. In healthy resting cells, Bax resides in the cytoplasm and loosely attached to the mitochondrial membrane. Apoptotic stimuli induce Bax activation, which is characterized by translocation and multimerization on the mitochondrial membrane surface resulting in exposure of an amino terminal epitope recognized by the monoclonal antibody 6A7. To understand the structural changes that occur during Bax activation, we determined the crystal structure of a Bax peptide bound to the 6A7 Fab fragment to a resolution of 2.3 A. The structure reveals the conformation of the 6A7 peptide epitope on Bax in the activated form and elucidates the extensive structural changes that Bax must undergo during the conversion from its native to its activated conformation.
Sujet(s)
Fragments Fab d'immunoglobuline/composition chimique , Peptides/composition chimique , Conformation des protéines , Protéine Bax/composition chimique , Anticorps , Sites de fixation , Lignée cellulaire tumorale , Cristallisation , Épitopes/composition chimique , Épitopes/métabolisme , Humains , Fragments Fab d'immunoglobuline/métabolisme , Modèles moléculaires , Peptides/métabolisme , Liaison aux protéines , Structure tertiaire des protéines , Relation structure-activité , Protéine Bax/métabolismeRÉSUMÉ
Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown etiology. Associations between viral infections and the onset of SLE have been suggested, and recent studies have provided evidence that type I interferons (IFNalpha/beta) might play a role in the SLE disease process. Viruses and interferons have also been implicated in mouse models of SLE. We generated a model of accelerated proteinuria, in which lupus-prone mice were injected repeatedly with polyinosinic:polycytidylic acid (polyI:C), mimicking exposure to virus-derived double stranded RNA (dsRNA), leading to the production of IFNalpha/beta. PolyI:C-treated (B6.Nba2 x NZW)F1 and (B6 x NZW)F1 hybrid mice developed significantly increased levels of anti-dsDNA autoantibodies, characteristic of lupus. Most significantly, polyI:C-treated (B6.Nba2 x NZW)F1 mice, but not (B6 x NZW)F1 or parental strains, developed lupus-like nephritis in an accelerated fashion, which was dependent on IFNalpha/beta and associated with elevated deposition of total IgG, IgG2a and complement factor C3 in the glomerular capillary walls. These data suggest that reagents, which increase the levels of endogenous IFNalpha/beta (directly or indirectly), can accelerate the course of lupus-like nephritis, the development of which is dependent on the presence of both NZW- and Nba2-encoded genes.
Sujet(s)
Interféron alpha/biosynthèse , Interféron bêta/biosynthèse , Lupus érythémateux disséminé/génétique , Lupus érythémateux disséminé/immunologie , Poly I-C/pharmacologie , Animaux , Anticorps antinucléaires/sang , Modèles animaux de maladie humaine , Femelle , Prédisposition génétique à une maladie , Humains , Immunoglobuline G/métabolisme , Glomérule rénal/effets des médicaments et des substances chimiques , Glomérule rénal/immunologie , Glomérule rénal/anatomopathologie , Glomérulonéphrite lupique/étiologie , Glomérulonéphrite lupique/anatomopathologie , Souris , Protéinurie/étiologie , Transduction du signalRÉSUMÉ
The ratio of CD4 T cells to CD8 T cells (CD4:CD8 ratio) varies over two-fold between C57BL/6 and DBA/2 mice for both T cell precursors in the thymus and mature T cells in the periphery. Correlation analysis of the CD4:CD8 ratio in thymic precursors vs peripheral T cells in F2 and backcross mice found that thymic precursor ratios are inherited independently from those in the periphery, indicating that the CD4:CD8 ratios in these populations are affected by distinct genetic mechanisms. A genome scan of progeny in the phenotypic extremes identified three quantitative trait loci (QTLs). Trmq1 (for T cell ratio modifier QTL 1) was detected in the telomeric end of c6 (peak marker D6Mit15 at 74 cM) and had a maximum LOD score of 4.6. Trmq2, in the telomeric half of c2, peaked at D2MIT483 and had a maximum LOD score of 3.41. Both of these QTLs impacted the CD4:CD8 ratios in peripheral T cells and had no impact on variation in this ratio among thymic precursors. However, heterozygosity for the H2 complex was suggestively associated (LOD score of 2.43) with increases in CD4 T cells among T cell precursors in the thymus. All of these QTLs were affected by epistatic interactions, indicating that additional modifiers in the B6 and DBA/2 genomes modulate this phenotype.
Sujet(s)
Rapport CD4-CD8 , Lymphocytes T CD4+ , Lymphocytes T CD8+ , Liaison génétique , Animaux , Cartographie chromosomique , Croisements génétiques , Lod score , Noeuds lymphatiques/métabolisme , Souris , Souris de lignée C57BL , Souris de lignée DBA , Spécificité d'espèce , Thymus (glande)/métabolismeRÉSUMÉ
The production of stable cell lines is an important technique in cell biology, and it is often the rate-limiting step in studies involving the characterization of the function of novel genes or gene mutations. To facilitate this process, a novel family of retroviral vectors, the pE vector family, has been generated. The retroviral sequences in the pE vectors have been taken from the Moloney murine leukemia virus (MMLV) vector pMFG, which has been shown to express cDNA inserts more consistently and at higher levels than earlier generations of MMLV vectors. These vectors contain four different internal ribosome entry site-selectable markers, allowing high-efficiency selection of transductants expressing the desired cDNA. The pE vectors have an episomal design to allow long-term production of high-titer virus without the need for subcloning the producer line. Using a strategy of combinatorial infection followed by combinatorial drug selection, we demonstrate that the pE vectors can be used to generate stable, polyclonal cell lines expressing at least three novel cDNAs in less than 2 weeks. The use of these vectors will thus dramatically accelerate the production of complex stable cell lines.
Sujet(s)
Techniques de culture cellulaire/méthodes , Vecteurs génétiques/génétique , Virus de la leucémie murine de Moloney/génétique , Transduction génétique/méthodes , Animaux , Lignée cellulaire , Cytométrie en flux , Souris , Virus de la leucémie murine de Moloney/physiologie , Plasmides/génétiqueRÉSUMÉ
Previous work has established a role for CD40-mediated signals in eliciting helper-dependent CD8(+) T cell responses. Here we investigated the effects of in vivo CD40 stimulation on the survival and function of tumor-specific CD8(+) T cells in a mouse melanoma model system. We found that agonistic anti-CD40 antibody treatment alone of tumor-bearing mice accelerated the deletion of tumor-antigen-specific T cells. However, long-term survival and function of tumor-antigen-specific T cells could be achieved when viral immunization with tumor antigen and anti-CD40 treatment were combined. This rescue of CD8(+) T cells could not be easily replicated by inflammatory or antigen-specific stimuli alone, demonstrating the specificity of signals that regulate the deletion or survival of tumor-specific T cells. These results demonstrate that opposing effects can be elicited by CD40 stimulation in vivo and suggest the need for caution in using this treatment for cancer patients.
Sujet(s)
Antigènes CD40/immunologie , Lymphocytes T CD8+/immunologie , Mélanome expérimental/immunologie , Animaux , Antigènes néoplasiques/immunologie , Femelle , Expression des gènes , Déplétion lymphocytaire , Souris , Souris de lignée C57BL , Transplantation tumorale/immunologie , Ovalbumine/génétique , Ovalbumine/immunologie , Lymphocytes T/immunologie , Cellules cancéreuses en culture , Vaccination , Virus de la vaccineRÉSUMÉ
Autoimmune disease is controlled by genetic and environmental factors. Both of these affect susceptibility to autoimmunity at three levels: the overall reactivity of the immune system, the specific antigen and its presentation, and the target issue.
Sujet(s)
Maladies auto-immunes/étiologie , Antigènes/immunologie , Maladies auto-immunes/génétique , Maladies auto-immunes/immunologie , Cartographie chromosomique , Réactions croisées , Humains , Modèles biologiquesSujet(s)
Antigènes d'histocompatibilité/métabolisme , Complexe majeur d'histocompatibilité , Récepteur lymphocytaire T antigène, alpha-bêta/immunologie , Sous-populations de lymphocytes T/immunologie , Sous-populations de lymphocytes T/métabolisme , Allergie et immunologie/tendances , Séquence d'acides aminés , Animaux , Séquence nucléotidique , Différenciation cellulaire/génétique , Différenciation cellulaire/immunologie , Antigènes d'histocompatibilité/génétique , Humains , Complexe majeur d'histocompatibilité/génétique , Récepteur lymphocytaire T antigène, alpha-bêta/génétique , Sociétés médicales , Sous-populations de lymphocytes T/cytologie , États-UnisRÉSUMÉ
IE/DR MHC class II molecules have an extensive H-bonding network under the bound peptide. In IE(k), two alpha chain acidic amino acids in the core of this network were mutated to amides. At low pH, the mutant molecule exchanged peptide much more rapidly than the wild-type. The crystal structure of the mutant IE(k) revealed the loss of a single buried water molecule and a reorganization of the predicted H-bonding network. We suggest that these mutations enhance the transition of MHC class II to an open conformation at low pH allowing the bound peptide to escape. In wild-type IE(k), the need to protonate these amino acids also may be a bottleneck in the return to a closed conformation after peptide binding.
Sujet(s)
Présentation d'antigène/immunologie , Antigènes d'histocompatibilité de classe II/génétique , Antigènes d'histocompatibilité de classe II/immunologie , Peptides/immunologie , Séquence d'acides aminés , Animaux , Antigènes d'histocompatibilité de classe II/composition chimique , Cinétique , Souris , Modèles moléculaires , Données de séquences moléculaires , Mutagenèse , Structure secondaire des protéinesRÉSUMÉ
Injection of soluble protein antigen into animals causes abortive proliferation of the responding T cells. Immunological adjuvants boost T cell responses at least in part by increasing the survival of activated T cells during and after the initial proliferative phase of their clonal expansion. To understand how adjuvants promote T cell survival, we used gene microarrays to analyze gene expression in T cells activated either with antigen alone or in the presence of two different adjuvants. Among the genes whose expression was increased by both adjuvants was the IkappaB family member Bcl-3. Retroviral infection experiments showed that expression of Bcl-3 increased survival of activated T cells in vitro and in vivo. Adjuvants may therefore improve survival of activated T cells via induction of Bcl-3.
Sujet(s)
Adjuvants immunologiques , Activation des lymphocytes , Protéines proto-oncogènes/biosynthèse , Lymphocytes T/immunologie , Animaux , Protéine-3 du lymphome à cellules B , Mort cellulaire , Femelle , Analyse de profil d'expression de gènes , Lipopolysaccharides/immunologie , Souris , Souris de lignée BALB C , Souris de lignée C57BL , Données de séquences moléculaires , Séquençage par oligonucléotides en batterie , Lymphocytes T/cytologie , Facteurs de transcription , Virus de la vaccine/immunologieRÉSUMÉ
Older humans and mice frequently contain very large clones of CD8(+) T cells. In mice these cells are phenotypically very similar to memory CD8(+) T cells. Like memory CD8(+) T cells, most members of the clones are in continuous slow division, apparently independently of Ag stimulation. Proliferation of the CD8(+) clonal T cells is inhibited in mice treated with Ab to the IL-2R beta-chain that blocks signaling by either IL-2 or IL-15. However, inhibition of IL-2 increases the numbers of dividing clonal cells. Therefore, like normal memory CD8(+) T cells, expansion of the clones is driven by IL-15 and inhibited by IL-2 and is probably limited by the amounts of IL-15 and IL-2 present in the host. Control by these two cytokines may account for the fact that, although the clones can be very large, they do not overwhelm or kill their hosts. Nevertheless the clonal cells compete successfully with normal memory CD8(+) T cells for growth. Perhaps the clonal cells use IL-15 more effectively or are more resistant to the inhibitory effects of IL-2. Thus they might affect the immune response of their hosts by competing for factors that stimulate and inhibit normal CD8(+) memory T cells.
Sujet(s)
Vieillissement/immunologie , Lymphocytes T CD8+/cytologie , Lymphocytes T CD8+/immunologie , Cytokines/physiologie , Animaux , Antigènes de surface/analyse , Division cellulaire/immunologie , Séparation cellulaire , Survie cellulaire/immunologie , Clones cellulaires/cytologie , Clones cellulaires/immunologie , Inhibiteurs de croissance/physiologie , Antigènes d'histocompatibilité de classe I/physiologie , Mémoire immunologique , Interleukine-15/physiologie , Interleukine-2/physiologie , Souris , Souris de lignée C57BL , Souris de lignée DBA , Souris knockout , Sous-populations de lymphocytes T/cytologie , Sous-populations de lymphocytes T/immunologie , Facteurs tempsRÉSUMÉ
In order to track hematopoetic cells of all lineages unambiguously at all stages of development, we have developed C57BL/6 mice that express a transgene coding for green fluorescent protein (GFP) under control of the human ubiquitin C promoter. These mice, called UBI-GFP/BL6, express GFP in all tissues examined, with high levels of GFP expression observed in hematopoetic cells. UBI-GFP/BL6 mice are unique in that B cells, T cells, and dendritic cells have distinct levels of GFP fluorescence. In cell transfer experiments, leukocytes from UBI-GFP/BL6 mice are readily identified by FACS or fluorescence microscopy. We demonstrate that transplanted UBI-GFP/BL6 dendritic cells are easily identified in secondary lymphoid tissues. Direct interactions between individual dendritic cells and multiple naïve CD8+ T cells are observed in lymph nodes within 12 h of cell transfer and require loading of the dendritic cells with the appropriate peptide antigen. Dendritic cells undergo specific morphologic changes following interactions with antigen-specific T cells.
Sujet(s)
Présentation d'antigène , Antigènes/immunologie , Lymphocytes T CD8+/immunologie , Cellules dendritiques/immunologie , Colorants fluorescents/analyse , Protéines luminescentes/analyse , Noeuds lymphatiques/immunologie , Transfert adoptif , Animaux , Communication cellulaire , Lignage cellulaire , Mouvement cellulaire , Chimiotaxie des leucocytes , Régulation de l'expression des gènes , Gènes rapporteurs , Protéines à fluorescence verte , Noeuds lymphatiques/cytologie , Mâle , Souris , Souris de lignée C57BL , Souris transgéniques , Spécificité d'organe , Régions promotrices (génétique) , Chimère post-radique , Protéines de fusion recombinantes/analyse , Rate/cytologie , Rate/immunologie , Ubiquitine C/génétiqueRÉSUMÉ
These studies tested whether antigenic competition between T cells occurs. We generated CD8(+) T cell responses in H-2(b) mice against the dominant ovalbumin epitope SIINFEKL (ova8) and subdominant epitope KRVVFDKL, using either vaccinia virus expressing ovalbumin (VV-ova) or peptide-pulsed dendritic cells. CD8(+) T cell responses were visualized by major histocompatibility complex class I-peptide tetrameric molecules. Transfer of transgenic T cells with high affinity for ova8 (OT1 T cells) completely inhibited the response of host antigen-specific T cells to either antigen, demonstrating that T cells can directly compete with each other for response to antigen. OT1 cells also inhibited CD8(+) T cell responses to an unrelated peptide, SIYRYGGL, providing it was presented on the same dendritic cells as ova8. These inhibitions were not due to a more rapid clearance of virus or antigen-presenting cells (APCs) by the OT1 cells. Rather, the inhibition was caused by competition for antigen and antigen-bearing cells, since it could be overcome by the injection of large numbers of antigen-pulsed dendritic cells. These results imply that common properties of T cell responses, such as epitope dominance and secondary response affinity maturation, are the result of competitive interactions between antigen-bearing APC and T cell subsets.
Sujet(s)
Cellules présentatrices d'antigène/immunologie , Lymphocytes T CD8+/immunologie , Lymphocytes T/immunologie , Séquence d'acides aminés , Animaux , Communication cellulaire , Cellules cultivées , Cellules dendritiques/immunologie , Épitopes/immunologie , Femelle , Mâle , Souris , Souris de lignée C57BL , Lignées consanguines de souris , Souris transgéniques , Ovalbumine/immunologie , Fragments peptidiques/immunologie , Récepteurs aux antigènes des cellules T/génétique , Récepteurs aux antigènes des cellules T/immunologieRÉSUMÉ
We characterized antigen-specific CD4(+) T cells in six patients with treatment-resistant Lyme arthritis, using an HLA-DRB1*0401 major histocompatibility complex (MHC) class II tetramer covalently loaded with OspA(164-175), an immunodominant epitope of Borrelia burgdorferi. Direct analysis of OspA-tetramer binding CD4(+) cells in patients expressing the HLA-DRB1*0401 allele revealed frequencies of between <0.005 and 0.1% in peripheral blood (n = 6), and between <0.005 and 3.1% in synovial fluid (n = 3). OspA-tetramer(+)CD4(+) cells were directly cloned at 1 cell per well and expanded by mitogen and IL-2 on allogeneic feeder cells. As measured by [(3)H]thymidine incorporation, 95% of 168 T cell clones from synovial fluid binding the OspA-tetramer were antigen-reactive. Clones generated from peripheral blood revealed a different pattern of responsiveness when compared with clones generated from synovial fluid, as measured by proliferation, IFN-gamma, and IL-13 secretion. These clones, selected on the basis of their peptide binding, also responded to whole protein, but with a different cytokine profile. Our studies demonstrate that MHC class II tetramers can be used in humans to directly identify, isolate, and characterize antigen-reactive T cells from an inflammatory compartment.
Sujet(s)
Groupe Borrelia burgdorferi/immunologie , Lymphocytes T CD4+/immunologie , Antigènes HLA-DR/immunologie , Lipoprotéines , Antigènes de surface/immunologie , Protéines de la membrane externe bactérienne/immunologie , Vaccins antibactériens , Biopolymères , Clones cellulaires , Cytokines/immunologie , Relation dose-réponse (immunologie) , Antigènes HLA-DR/administration et posologie , Chaines HLA-DRB1 , Humains , Techniques in vitro , Maladie de Lyme/immunologie , Vaccins contre la maladie de Lyme/immunologieRÉSUMÉ
Memory T cells maintain their numbers for long periods after antigen exposure. Here we show that CD8+ T cells of memory phenotype divide slowly in animals. This division requires interleukin-15 and is markedly increased by inhibition of interleukin-2 (IL-2). Therefore, the numbers of CD8+ memory T cells in animals are controlled by a balance between IL-15 and IL-2.
Sujet(s)
Lymphocytes T CD8+/cytologie , Lymphocytes T CD8+/immunologie , Mémoire immunologique , Interleukine-15/physiologie , Interleukine-2/physiologie , Animaux , Anticorps , Lymphocytes T CD8+/transplantation , Mort cellulaire , Division cellulaire , Homéostasie , Antigènes CD44/analyse , Interleukine-15/immunologie , Interleukine-2/immunologie , Interleukine-7/immunologie , Interleukine-7/physiologie , Numération des lymphocytes , Souris , Souris de lignée C57BL , Phénotype , Récepteurs à l'interleukine-15 , Récepteurs à l'interleukine-2/analyse , Récepteurs à l'interleukine-2/immunologie , Récepteurs à l'interleukine-7/immunologie , Récepteurs à l'interleukine-7/physiologie , Organismes exempts d'organismes pathogènes spécifiquesRÉSUMÉ
The cytokines interleukin (IL)-2, IL-4, IL-6, IL-7, and IL-15 have all previously been shown to inhibit resting T cell death in vitro. We have found a difference in the response of T cells to IL-6, depending on the activation status of the cells. IL-6 inhibited the death of naive T cells, but had no effect on the death of either superantigen-activated T cells, or T cells bearing memory markers. This was true even when the resting and activated T cells were isolated from the same animal; thus, the determining factor for IL-6 insensitivity was the activation status or activation history of the cell, and not the milieu in the animal from which the cells were isolated. Activated T cells expressed lower levels of IL-6 receptors on their surfaces, yet there were sufficient levels of receptors for signaling, as we observed similar levels of signal transducer and activator of transcription (Stat)3 phosphorylation in resting and activated T cells treated with IL-6. However, there was profound inhibition of IL-6-induced Stat1 phosphorylation in activated T cells compared with resting T cells. These data suggest that there is activation-induced inhibition of IL-6 receptor signaling in T cells. This inhibition appears to be specific for some but not all of the IL-6-mediated signaling cascades in these cells.
Sujet(s)
Interleukine-6/antagonistes et inhibiteurs , Interleukine-6/physiologie , Activation des lymphocytes , Protéines de répression , Transduction du signal/immunologie , Sous-populations de lymphocytes T/cytologie , Sous-populations de lymphocytes T/immunologie , Facteur de transcription AP-1/antagonistes et inhibiteurs , Facteur de transcription AP-1/physiologie , Animaux , Protéines de transport/biosynthèse , Mort cellulaire/immunologie , Survie cellulaire/immunologie , Cellules cultivées , Cytokines/physiologie , Protéines de liaison à l'ADN/antagonistes et inhibiteurs , Protéines de liaison à l'ADN/métabolisme , Relation dose-réponse (immunologie) , Régulation négative/immunologie , Femelle , Mémoire immunologique , Immunophénotypage , Interphase/immunologie , Souris , Souris de lignée BALB C , Souris de lignée C57BL , Souris transgéniques , Phosphorylation , Récepteurs à l'interleukine-6/antagonistes et inhibiteurs , Récepteurs à l'interleukine-6/biosynthèse , Facteur de transcription STAT-1 , Facteur de transcription STAT-3 , Protéine-1 suppressive de la signalisation des cytokines , Protéines SOCS , Sous-populations de lymphocytes T/métabolisme , Transactivateurs/antagonistes et inhibiteurs , Transactivateurs/métabolismeRÉSUMÉ
Recent advances in gene array technology and isolation of lymphocytes now allow comprehensive analysis of gene expression in many different types of T cells. So far only a few sets of results have been published. However it is already clear that these analyses provide accurate measurements of gene expression in T cells. This technology offers the first opportunity to examine global and subtle changes in gene expression in response to specific stimuli.
Sujet(s)
Analyse de profil d'expression de gènes , Génome , Activation des lymphocytes/génétique , Séquençage par oligonucléotides en batterie , Lymphocytes T/immunologie , Animaux , Séparation cellulaire , Bases de données factuelles , Étiquettes de séquences exprimées , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Température élevée , Humains , Cellules Jurkat , Activation des lymphocytes/effets des médicaments et des substances chimiques , Souris , Esters de phorbol/pharmacologie , ARN messager/analyse , ARN messager/biosynthèse , ARN messager/génétique , Lymphocytes T/effets des médicaments et des substances chimiques , Lymphocytes T/métabolismeRÉSUMÉ
Considerable evidence indicates that CD4(+) T cells are important in the pathogenesis of rheumatoid arthritis (RA), but the antigens recognized by these T cells in the joints of patients remain unclear. Previous studies have suggested that type II collagen (CII) and human cartilage gp39 (HCgp39) are among the most likely synovial antigens to be involved in T cell stimulation in RA. Furthermore, experiments have defined dominant peptide determinants of these antigens when presented by HLA-DR4, the most important RA-associated HLA type. We used fluorescent, soluble peptide-DR4 complexes (tetramers) to detect synovial CD4(+) T cells reactive with CII and HCgp39 in DR4(+) patients. The CII-DR4 complex bound in a specific manner to CII peptide-reactive T cell hybridomas, but did not stain a detectable fraction of synovial CD4(+) cells. A background percentage of positive cells (<0.2%) was not greater in DR4 (DRB1*0401) patients compared with those without this disease-associated allele. Similar results were obtained with the gp39-DR4 complex for nearly all RA patients. In a small subset of DR4(+) patients, however, the percentage of synovial CD4(+) cells binding this complex was above background and could not be attributed to nonspecific binding. These studies demonstrate the potential for peptide-MHC class II tetramers to be used to track antigen-specific T cells in human autoimmune diseases. Together, the results also suggest that the major oligoclonal CD4(+) T cell expansions present in RA joints are not specific for the dominant CII and HCgp39 determinants.