T-cell tolerance and autoimmunity in transgenic models of central and peripheral tolerance

Experiments with transgenic mice expressing genes encoding both antigens in defined tissues and T-cell receptor genes of known specificities have enhanced our understanding of the mechanisms involved in the pathogenesis of autoimmune states. They have also shed light on the means by which potentially autoreactive cells may be prevented from exerting their autoaggressive potential. The value of the transgenic approach is that it can overcome the low frequency of peptide-specific T cells occurring in normal animals, and also provide a tissue-specific, cognate antigen that is absent in controls. These factors allow reactive T cells to be isolated or quantified by flow cytometry and their responses to antigen in vitro and in vivo be defined.     

The effect of bone marrow and thymus chimerism between non-obese diabetic (NOD) and NOD-E transgenic mice,on the expression and prevention of diabetes

The non-obese diabetic (NOD) mouse is an established animal model of the autoimmune disease, insulin-dependent diabetes mellitus (IDDM). The NOD-E mouse is a transgenic mouse which expresses the I-E molecule (absent in NOD mice). Expression of I-E protects these mice from both insulitis and IDDM.We have investigated the possible mechanisms of this protection by constructing bone marrow, and combined bone marrow and thymus chimeras between NOD and NOD-E mice. Our data suggest that thymic epithelium may play no direct role in either protection against, or promotion of, IDDM. Protection from diabetes is provided either by NOD-E donor bone marrow or NOD-E recipient non-thymic radioresistant cells. The means by which protection may be achieved in this system are discussed.     

Complete prevention of diabetes in transgenic NOD mice expressing I-E molecules.

Previously, we showed that transgenic expression of the MHC (major histocompatibility complex) class II I-E molecules prevented insulitis in non-obese diabetic (NOD) mice at the age of 19 weeks. To rule out the possibility that the I-E expression merely delays the onset of insulitis, we have further characterized the expression and function of the I-E molecule expressed in transgenic NOD mice and confirmed our previous observations. Northern blot analysis showed that the transgenic E alpha d gene was expressed in a pattern similar to the endogenous E alpha d gene in BALB/c mice. The newly expressed I-E molecules were recognized as an alloantigen by the T lymphocytes of normal NOD mice as shown by mixed lymphocyte reaction (MLR). Transgenic NOD mice were resistant to the treatment by cyclophosphamide, which effectively induces diabetes in normal NOD mice, and did not develop diabetes up to 40 weeks of age. On the basis of these findings, we discuss the role of I-E molecules in the prevention of diabetes in NOD mice.     

Control of central and peripheral tolerance by Aire

The negative selection of self-reactive thymocytes depends on the expression of tissue-specific antigens by medullary thymic epithelial cells. The autoimmune regulator (Aire) protein plays an important role in turning on these antigens, and the absence of even one Aire-induced tissue-specific antigen in the thymus can lead to autoimmunity in the antigen-expressing target organ. Recently, Aire protein has been detected in peripheral lymphoid organs, suggesting that peripheral Aire plays a complementary role here. In these peripheral sites, Aire was found to regulate the expression of a group of tissue-specific antigens that is distinct from those expressed in the thymus. Furthermore, transgenic antigen expression in extrathymic Aire-expressing cells (eTACs) can mediate deletional tolerance, but the immunological relevance of Aire-dependent, endogenous tissue-specific antigens remains to be determined.     

Mosaic theory revised: inflammation and salt play central roles in arterial hypertension

The mosaic theory of hypertension was advocated by Irvine Page ~80 years ago and suggested that hypertension resulted from the close interactions of different causes. Increasing evidence indicates that hypertension and hypertensive end-organ damage are not only mediated by the proposed mechanisms that result in hemodynamic injury. Inflammation plays an important role in the pathophysiology and contributes to the deleterious consequences of arterial hypertension. Sodium intake is indispensable for normal body function but can be detrimental when it exceeds dietary requirements. Recent data show that sodium levels also modulate the function of monocytes/macrophages, dendritic cells, and different T-cell subsets. Some of these effects are mediated by changes in the microbiome and metabolome due to high-salt intake. The purpose of this review is to propose a revised and extended version of the mosaic theory by summarizing and integrating recent advances in salt, immunity, and hypertension research. Salt and inflammation are placed in the middle of the mosaic because both factors influence each of the remaining pieces.     

Transplantation of retrovirally transduced bone marrow prevents autoimmune disease in aged mice by peripheral tolerance mechanisms

Transplantation of bone marrow (BM) engineered to express self-antigen has been shown to protect 100% of young mice from myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE), with thymic clonal deletion as a tolerance mechanism. Here, we asked whether aged mice can also be tolerised following transplantation with self-antigen-engineered BM and whether castration-induced thymus regrowth can enhance this outcomes. Then, 50% of aged mice were protected from EAE regardless of castration-induced thymus regrowth. EAE-free and diseased mice demonstrated MOG-specific lymphocyte proliferation and antibody production regardless of castration-induced thymus regrowth, consistent with lack of intrathymic deletion of self-antigen-reactive T cells. Although low chimerism levels ( < 4%) were observed, EAE-free mice showed significantly higher chimerism levels in lymphocytes in peripheral lymphoid organs compared with thymus. CD4⁺CD25⁺ regulatory T cells were elevated in lymph nodes of EAE-free mice. We conclude that transplantation of self-antigen expressing BM protects 50% of aged mice and castration-induced thymic regrowth had no effect on outcomes. Peripheral tolerance mechanisms are implicated since protection is associated with higher chimerism levels in peripheral T and B lymphocytes and with elevated regulatory T cells.     

Transplantation of retrovirally transduced bone marrow prevents autoimmune disease in aged mice by peripheral tolerance mechanisms

Transplantation of bone marrow (BM) engineered to express self-antigen has been shown to protect 100% of young mice from myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE), with thymic clonal deletion as a tolerance mechanism. Here, we asked whether aged mice can also be tolerised following transplantation with self-antigen-engineered BM and whether castration-induced thymus regrowth can enhance this outcomes. Then, 50% of aged mice were protected from EAE regardless of castration-induced thymus regrowth. EAE-free and diseased mice demonstrated MOG-specific lymphocyte proliferation and antibody production regardless of castration-induced thymus regrowth, consistent with lack of intrathymic deletion of self-antigen-reactive T cells. Although low chimerism levels (?相似文献   

B and T cell tolerance and autoimmunity in autoantibody transgenic mice

One hallmark of systemic lupus erythematosus (SLE) is the presence of autoantibodies directed at a diverse group of proteins of the U1/Sm small nuclear ribonucleoprotein particles (snRNP). Patients with SLE and murine models of this disease generate high titers of affinity mature, isotype-switched autoantibodies characteristic of T cell-dependent immune responses. In this investigation, we made use of anti-snRNP Ig transgenic mice (2-12 Tg) to track regulation of autoreactive B cells in normal and autoimmune-prone mice. Autoantibody studies demonstrated that the regulation of anti-snRNP B cells is intact in non-autoimmune Tg mice, but not in MRL-lpr/lpr mice. We further utilize autoreactive Tg B cells as antigen-presenting cells (APC) and individual snRNP peptides to assess the presence of autoreactive T cells in the repertoire of non-autoimmune and MRL-lpr/lpr mice. We found that Tg B cells can direct specific T cell tolerance in a non-autoimmune-prone (C57Bl/6) background, whereas the same autoantibody transgene in MRL-lpr/lpr mice drives T cell autoimmunity. Moreover, Tg B cell APC could stimulate autoreactive T cells from wild-type (non-Tg) C57Bl/6 mice, indicating a lack of tolerance induction in the absence of the autoantigenic-presenting B cells. Thus, we have defined dual roles for autoantigen-presenting B lymphocytes in stimulating self-reactive T cells that inhabit the normal repertoire or, under some conditions, providing tolerance signals.     

Immunological tolerance: new approaches using transgenic mice

Transgenic technology allows the introduction into the germline of an animal of a known gene coding for a normally foreign antigen, and by means of a specific promoter, the direction of its expression to specific tissues. The antigen is therefore synthesized by the animal as an authentic self molecule, at a particular stage in development, and in a particular site. In this review, J.F.A.P. Miller and colleagues discuss this radically new approach to the investigation of the mechanism of acquired immunological tolerance to self components.     

Mechanisms of hypoxic tolerance in presymptomatic APP23 transgenic mice

In the B6-Tg (ThylAPP)23Sdz (APP23tg) transgenic mouse model of Alzheimer's disease hypoxic tolerance is impaired prior to amyloid deposition. We therefore investigated mechanisms known to mediate resistance to hypoxic episodes in presymptomatic APP23tg and appropriate control strains. The mRNA expression levels in the hippocampus of adenosine receptor subtypes A1 and A3, estrogen receptors alpha and beta, progesterone receptor, and neuronal and endothelial nitric oxide synthase were investigated with semi-quantitative RT-PCR. Mice were pretreated in vivo with a low dose of 3-nitropropionate, an inhibitor of succinic dehydrogenase, known to mediate hypoxic tolerance within 1h. We found increased expression levels in presymptomatic, untreated APP23tg animals of adenosine A3 receptor mRNA and estrogen receptor alpha mRNA. In addition, we observed an increase in nNOS expression levels upon mild cellular hypoxia induced by 3-NP in transgenic but not in wild-type animals. We conclude that overexpression of human APP results in differential expression of receptors conferring hypoxic tolerance prior to amyloid deposition. Up-regulation of nNOS expression levels upon hypoxic challenge in APP23tg transgenic animals may therefore reflect a selective vulnerability in these animals even before amyloid deposition.     

Sex differences in central and peripheral mechanisms of fatigue in cyclists

We examined peripheral versus central contributions to fatigue in men and women during prolonged cycling using a peripheral nerve magnetic stimulation-based technique. 11 men (41 ± 3 years) and 9 women (38 ± 2 years) cycled for 2 h at ventilatory threshold with 5, 1-min sprints interspersed, followed by a 3-km time trial. Quadriceps strength testing was performed isometrically in a semi-reclined position pre- and post-cycling: (1) MVC; (2) MVC with superimposed 3-s magnetic stimulation to measure central activation ratio (CAR), a measure of central fatigue; (3) peripheral magnetic stimulation (PMS) alone of the femoral nerve in a 4-s pulse train, a measure of peripheral fatigue. Data were analyzed with mixed model ANOVA. When adjusted for body mass, men and women had similar strength (p = 0.876), and changes in MVC with time were similar between sexes, declining 22 % in men and 16 % in women (p = 0.360). CAR was similar between sexes and decreased 15 % (effect of time, p < 0.001). Changes in PMS-elicited force were different between sexes: only men lost stimulated strength (6.30 to 5.21 vs. 5.48 to 5.53 N kg^?1, interaction p = 0.036). Results clearly demonstrate that quadriceps fatigue after >2 h of cycling was of both central and peripheral origin in men but solely due to central mechanisms in women.     

Independent down-regulation of central and peripheral tumor necrosis factor production as a result of lipopolysaccharide tolerance in mice.

Lipopolysaccharide (LPS) induces a variety of central and peripheral effects that are largely mediated by cytokines, including tumor necrosis factor (TNF). Peripheral (intravenous [i.v.]) administration of LPS (2.5 micrograms per mouse) induced TNF levels in the serum and spleen but not in the brain, while central (intracerebroventricular [i.c.v.]) administration of LPS induced TNF production both in the brain and in the periphery. Mice challenged with LPS after LPS pretreatment (35 micrograms per mouse, intraperitoneally, as a single dose on day -3 or as a 4-day treatment on days -5 to -2) were unresponsive in terms of induction of serum TNF. When peripherally LPS-tolerant mice (where LPS pretreatment was given intraperitoneally) were challenged with an i.c.v. dose of LPS, brain (but not serum) TNF was still produced, meaning that the LPS-tolerant state was confined to the periphery. However, if LPS pretreatment was given i.c.v. (35 micrograms, as a single dose), the brain, like the periphery, became LPS tolerant in terms of TNF production. We investigated how tolerance to LPS affected two of its actions, decrease in food intake and induction of serum corticosterone (CS). After an i.v. challenge in peripherally LPS-tolerant mice, no decrease in food intake was observed, but this response was still elicited by an i.c.v. challenge. LPS tolerance reduced the CS response to i.v. and i.c.v. challenge. These results suggest that LPS-induced decrease in food intake might be a fully central effect, while the increase of serum CS might be due to both central and peripheral actions.     

Breakage of B cell tolerance and autoantibody production in anti-erythrocyte transgenic mice

In summary, there are two pathways for activation of peritoneal B-1 cells in HL mice: T cell-dependent and T cell-independent pathways. In both pathways, IL-10 is suggested to play an important role (Fig. 1). We have not yet known what type of cells secrete IL-10 by and whether other soluble factors are involved in each pathway. These questions are to be elucidated by further studies on HL mice.     

Evidence for peripheral mechanisms inducing tissue tolerance during ontogeny

Tissue grafts from a histoincompatible donor of the same developmental stage were introduced into an early chick embryo host in order to probe the immune response to the graft after birth, when the host has reached immune maturity. Limb buds from B4 or B12 chicken strains were grafted in situ on (B15 x B21)F1 recipients that were allowed to hatch. The grafted wing grew normally and was tolerated in a nearly perfect way during the host's lifetime, although reversible rejection crises severely affected the fundamentally healthy state of the grafted tissues. Skin grafts of the same major histocompatibility complex haplotype as the wing were performed on the adult wing-chimera and were permanently tolerated. In contrast, host peripheral blood lymphocytes maintained their capacity to proliferate against donor cells in the mixed lymphocyte reaction. These results, while showing that in vitro and in vivo tolerance are separable phenomena, suggest the existence of a peripheral mechanism inducing tolerance to self that complements the elimination of self-reactive clones by the thymus.     

Morphine tolerance and dependence in nociceptin/orphanin FQ transgenic knock-out mice

It has been hypothesized that morphine tolerance and dependence in mice following chronic exposure may reflect increased compensatory activity of antiopioid systems. The endogenous peptide nociceptin/orphanin FQ has been shown to have anti-opioid effects, for example antagonizing morphine analgesia. Moreover, chronic morphine administration increases synthesis of the peptide, and morphine tolerance and dependence can be attenuated or reversed by antagonists and agonists of the nociceptin/orphanin FQ receptor, respectively. The present study seeks to confirm a role for nociceptin/orphanin FQ in opioid tolerance and dependence by comparing morphine ED(50) values and naloxone-precipitated withdrawal jumping in mice homozygous (knock-out) and heterozygous for a null mutation of the Npnc1 gene encoding the nociceptin/orphanin FQ propeptide, and their wild type littermates, following chronic morphine exposure.Relative to morphine-naive control mice, significant rightward shifts in the morphine dose-response curve, resulting in increased morphine ED(50) values (approximately two to three-fold), was observed for all genotypes following three days of repeated systemic morphine injections. However, no differences between genotypes in the magnitude of tolerance were observed. In contrast, knock-out mice displayed significantly increased naloxone-precipitated withdrawal jumping relative to heterozygous and wild-type mice following implantation with a morphine pellet (25mg) for 72h.Use of nociception/orphaninFQ transgenic knock-out mice thus demonstrate the differential involvement of nociceptin/orphanin FQ in morphine tolerance and dependence.     

A key role for CCR7 in establishing central and peripheral tolerance

Early studies identified the CC-chemokine receptor (CCR)7 as an important homing molecule controlling the lymph node entry of naive T cells through high endothelial venules and of activated mature dendritic cells through afferent lymphatics. Consequently, these properties initially branded CCR7 as a central organizer of the primary immune response. However, recent studies have demonstrated that a variety of immune cells crucially rely on CCR7-directed migration not only for the induction of protective immunity but also for the establishment of immunological tolerance. In this review, we therefore highlight some of the recent advances in understanding the multiple roles of CCR7 in the induction and maintenance of central and peripheral tolerance.     

Antigen and lipopolysaccharide play synergistic roles in the effector phase of airway inflammation in mice

T helper (Th) cells play major roles in orchestrating asthmatic airway inflammation, but the molecular mechanisms controlling Th-cell recruitment to the airways remain incompletely defined. Innate immunity contributes importantly to the recruitment of effector T cells into sites of inflammation. To understand better the role of innate immune signals in the development of airway inflammation, we used a murine model in which lipopolysaccharide (LPS) contaminating the antigen is thought to trigger Toll-like receptor 4 (TLR4). To investigate the importance of the TLR4-signaling pathway in induction of lung inflammation, we compared recruitment of adoptively transferred ovalbumin-specific Th1 and Th2 cells in wild-type and TLR4 mutant (TLR4m) mice after intranasal or aerosol challenge. Intranasal challenge of TLR4m mice with ovalbumin resulted in decreased recruitment of Th1 and Th2 cells compared with that of wild-type mice. The numbers of Th1 and Th2 cells recruited to the airways of TLR4m mice were less profoundly reduced after aerosol ovalbumin challenge. Comparing the effects of altering the dose of ovalbumin with that of LPS suggested that both contribute to the magnitude of the response in wild-type mice. Our findings demonstrate the importance of both antigen and endotoxin acting in a synergistic manner in the development of airway inflammation.