The nuclear hormone receptor retinoic acid-related orphan receptor gamma t (RORγt) is a transcription factor (TF) specific to TH17 cells that produce interleukin (IL)-17 and have been implicated in a ...wide range of autoimmunity. Here, we developed a novel therapeutic strategy to modulate the functions of RORγt using cell-transducible form of transcription modulation domain of RORγt (tRORγt-TMD), which can be delivered effectively into the nucleus of cells and into the central nerve system (CNS). tRORγt-TMD specifically inhibited TH17-related cytokines induced by RORγt, thereby suppressing the differentiation of naïve T cells into TH17, but not into TH1, TH2, or Treg cells. tRORγt-TMD injected into experimental autoimmune encephalomyelitis (EAE) animal model can be delivered effectively in the splenic CD4⁺ T cells and spinal cord-infiltrating CD4⁺ T cells, and suppress the functions of TH17 cells. The clinical severity and incidence of EAE were ameliorated by tRORγt-TMD in preventive and therapeutic manner, and significant reduction of both infiltrating CD4⁺ IL-17⁺ T cells and inflammatory cells into the CNS was observed. As a result, the number of spinal cord demyelination was also reduced after tRORγt-TMD treatment. With the same proof of concept tTbet-TMD specifically blocking TH1 differentiation improved the clinical incidence of rheumatoid arthritis (RA). Therefore, tRORγt-TMD and tTbet-TMD can be novel therapeutic reagents with the natural specificity for the treatment of inflammatory diseases associated with TH17 or TH1. This strategy can be applied to treat various diseases where a specific transcription factor has a key role in pathogenesis.
Excessive expression of Tbet and IFNγ is evidence of systemic lupus erythematosus (SLE) in lupus patients. In this study, the nucleus-transducible form of Transcription Modulation Domain (TMD) of ...Tbet (ntTbet-TMD), which is a fusion protein between Protein Transduction Domain Hph-1 (Hph-1-PTD) and the TMD of Tbet comprising DNA binding domain and isotype-specific domain, was generated to inhibit Tbet-mediated transcription in the interactomic manner. ntTbet-TMD was effectively delivered into the nucleus of the cells and specifically inhibited Tbet-mediated transcription without influencing the differentiation of other T cell subsets and signaling events for T cell activation. The severity of nephritis was significantly reduced by ntTbet-TMD as effectively as methylprednisolone in lupus-prone mice. The number of Th1, Th2 or Th17 cells and the secretion of their cytokines substantially decreased in the spleen and kidney of lupus-prone mice by ntTbet-TMD treatment. In contrast to methylprednisolone, the marked increase of Treg cells and the secretion of their immunosuppressive cytokine were detected in the spleen of (NZB/NZW) F1 mice treated with ntTbet-TMD. Thus, ntTbet-TMD can improve nephritis in lupus-prone mice by modulating the overall proinflammatory microenvironment and rebalancing T cell subsets, leading to new immune therapeutics for Th1-mediated autoimmune diseases.
Early life environmental exposure, particularly during perinatal period, can have a life-long impact on organismal development and physiology. The biological rationale for this phenomenon is to ...promote physiological adaptations to the anticipated environment based on early life experience. However, perinatal exposure to adverse environments can also be associated with adult-onset disorders. Multiple environmental stressors induce glucocorticoids, which prompted us to investigate their role in developmental programming. Here, we report that perinatal glucocorticoid exposure had long-term consequences and resulted in diminished CD8 T cell response in adulthood and impaired control of tumor growth and bacterial infection. We found that perinatal glucocorticoid exposure resulted in persistent alteration of the hypothalamic-pituitary-adrenal (HPA) axis. Consequently, the level of the hormone in adults was significantly reduced, resulting in decreased CD8 T cell function. Our study thus demonstrates that perinatal stress can have long-term consequences on CD8 T cell immunity by altering HPA axis activity.
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•Early-life stress has a long-term effect on the immune system•Perinatal glucocorticoids exposure changes the set-point of the HPA axis•Reduced GR signaling decreases CD8 T cell function by altering chromatin at key loci•Perinatal glucocorticoids result in susceptibility to tumors and bacterial infection
Early life glucocorticoid exposure has long-term effects on immunity, including diminished CD8+ T cell responses in adulthood and impaired control of tumor growth as well as infection.
As sentinels of immune system, dendritic cells (DC) constantly sense environment and acquire antigens from the periphery in order to instigate adequate adaptive immune responses. DCs at the periphery ...acquire antigens by pattern recognition receptors (PRRs), process them into peptides to present to CD4 T cells via MHCII along with cytokine signals that play pivotal roles in skewing into specific types of T helper cells. However, this view does not rationalize how type 2 responses are activated in response to parasitic worms and allergens that are extensively diverse in their properties. Thus, it has been hypothesized that the initiation process for type 2 immune responses is mostly independent of PRRs but involves recognition of the consequences incurred by functional properties of type 2 stimuli such as proteolytic activity. Moreover, whether there are Th2-inducing cytokine signals derived from antigen-presenting DCs has been enigmatic for long time. A few cytokines and surface molecules have been suggested to contribute to the interaction of DC-T cells for Th2 induction yet it has been shown that they are likely involved in T cell activation in general rather than specific regulation of T helper cell differentiation. In our previous study, it was found that IRF4-dependent DC subset expressing PDL2 and CD301b specializes in promoting Th2 responses. I aimed to find a unique feature of this specific DC subset that enables it to induce Th2 activation. Interestingly, I found that high expression levels of ICAM1 on PDL2+ DC control its Th2-inducing function such that inhibition of ICAM1-mediated interaction with LFA1 was sufficient for Th2 differentiation in vitro. Indeed, when papain-sensitized mice were treated with recombinant ICAM1, Th2 responses in vivo were significantly reduced, indicating that ICAM1 engagement does regulate Th2 induction. In addition, it was observed that one of the possible mechanisms by which ICAM1 engagement can be circumvented for in vivo Th2 activation involves migration of Langerhans cells with high surface expressions of ICAM1. Also, this study showed that Th2 responses induced by blocking ICAM1 were dependent on OX40L and CD40–mediated signaling, and modulation of ICAM1 engagement may affect the clustering of OX40L on the immunologic synapse for efficient activation of subsequent signaling pathways.
•This study presents a novel strategy to modulate FoxP3 function directly.•The nt-FoxP3-subdomains are effectively delivered into the nucleus without toxicity.•The nt-FoxP3-FKH is intranuclear ...interactomic inhibitor of FoxP3.
Regulatory T cells (Treg cells) are crucial for the maintenance of immunological tolerance, and it has been reported that Treg cells are enriched within the tumor micro-environment for immune evasion due to their immunosuppressive functions. To inhibit Treg cells functions, FoxP3, a lineage-specific transcription factor responsible for the differentiation and functions of Treg cells, was functionally targeted by a nucleus-transducible (nt) form of various FoxP3 functional subdomains. These nt modified domains can be delivered into the nucleus effectively and work as interactomic inhibitors via disruption of the endogenous FoxP3-mediated transcription complex. Among these domains, nt-FoxP3-FKH (Forkhead DNA binding domain) is most effective at restoring NFAT activity suppressed by FoxP3, and inhibiting the binding of endogenous FKH-containing proteins to FKH DNA binding sequences without influencing the viability and activation of T cells. The suppressive functions of TGF-β-induced iTreg cells and thymus-derived tTreg cells were substantially blocked by nt-FoxP3-FKH, accompanied with down-regulation of CTLA-4 surface expression and IL-10 secretion of Treg cells. In addition, nt-FoxP3-FKH upregulated the expression of IL-2 and IFN-γ in Treg cells. Therefore, nt-FoxP3-FKH has the potential to be a novel therapeutic agent to modulate the immune-evasive tumor environment created by Treg cells without the need for genetic modifications.
Significance
T
H
17 cells are a subset of CD4
+
T helper cells that secrete the cytokine IL-17 and play a role in autoimmunity. RORγt is identified as a key transcription factor driving the T
H
17 ...differentiation. Sequence analysis indicated that transcription factor contains several conserved DNA-binding domain and isotype-specific domain that we termed transcription modulation domain (TMD). We designed a novel therapeutics, tRORγt-TMD, to deliver RORγt-TMD efficiently into the nucleus of the cells that regulates T
H
17 cell functions and T
H
17-mediated autoimmune diseases. With the same concept, tTbet-TMD also can regulate T
H
1 functions. In conclusion, tRORγt-TMD/tTbet-TMD can be novel and highly specific therapeutics for the treatment of T
H
17/T
H
1-mediated inflammatory disease and further allows us to discover new function of RORγt/Tbet in animals without genetic alteration.
The nuclear hormone receptor retinoic acid-related orphan receptor gamma t (RORγt) is a transcription factor (TF) specific to T
H
17 cells that produce interleukin (IL)-17 and have been implicated in a wide range of autoimmunity. Here, we developed a novel therapeutic strategy to modulate the functions of RORγt using cell-transducible form of transcription modulation domain of RORγt (tRORγt-TMD), which can be delivered effectively into the nucleus of cells and into the central nerve system (CNS). tRORγt-TMD specifically inhibited T
H
17-related cytokines induced by RORγt, thereby suppressing the differentiation of naïve T cells into T
H
17, but not into T
H
1, T
H
2, or T
reg
cells. tRORγt-TMD injected into experimental autoimmune encephalomyelitis (EAE) animal model can be delivered effectively in the splenic CD4
+
T cells and spinal cord-infiltrating CD4
+
T cells, and suppress the functions of T
H
17 cells. The clinical severity and incidence of EAE were ameliorated by tRORγt-TMD in preventive and therapeutic manner, and significant reduction of both infiltrating CD4
+
IL-17
+
T cells and inflammatory cells into the CNS was observed. As a result, the number of spinal cord demyelination was also reduced after tRORγt-TMD treatment. With the same proof of concept, tTbet-TMD specifically blocking T
H
1 differentiation improved the clinical incidence of rheumatoid arthritis (RA). Therefore, tRORγt-TMD and tTbet-TMD can be novel therapeutic reagents with the natural specificity for the treatment of inflammatory diseases associated with T
H
17 or T
H
1. This strategy can be applied to treat various diseases where a specific transcription factor has a key role in pathogenesis.
The nuclear hormone receptor retinoic acid-related orphan receptor gamma t (ROR...t) is a transcription factor (TF) specific to ... cells that produce interleukin (IL)-17 and have been implicated in ...a wide range of autoimmunity. Here, we developed a novel therapeutic strategy to modulate the functions of ROR...t using cell-transducible form of transcription modulation domain of ROR...t (tROR...t-TMD), which can be delivered effectively into the nucleus of cells and into the central nerve system (CNS). tROR...t-TMD specifically inhibited ...-related cytokines induced by ROR...t, thereby suppressing the differentiation of naive T cells into ..., but not into ..., ..., or Treg cells. tROR...t-TMD injected into experimental autoimmune encephalomyelitis (EAE) animal model can be delivered effectively in the splenic CD4+ T cells and spinal cord-infiltrating CD4+ T cells, and suppress the functions of ... cells. The clinical severity and incidence of EAE were ameliorated by tROR...t-TMD in preventive and therapeutic manner, and significant reduction of both infiltrating CD4+ IL-17+ T cells and inflammatory cells into the CNS was observed. As a result, the number of spinal cord demyelination was also reduced after tROR...t-TMD treatment. With the same proof of concept, tTbet-TMD specifically blocking ... differentiation improved the clinical incidence of rheumatoid arthritis (RA). Therefore, tROR...t-TMD and tTbet-TMD can be novel therapeutic reagents with the natural specificity for the treatment of inflammatory diseases associated with ... or ... This strategy can be applied to treat various diseases where a specific transcription factor has a key role in pathogenesis. (ProQuest: ... denotes formulae/symbols omitted.)
The nuclear hormone receptor retinoic acid-related orphan receptor gamma t (ROR...t) is a transcription factor (TF) specific to ... cells that produce interleukin (IL)-17 and have been implicated in ...a wide range of autoimmunity. Here, we developed a novel therapeutic strategy to modulate the functions of ROR...t using cell-transducible form of transcription modulation domain of ROR...t (tROR...t-TMD), which can be delivered effectively into the nucleus of cells and into the central nerve system (CNS). tROR...t-TMD specifically inhibited ...-related cytokines induced by ROR...t, thereby suppressing the differentiation of naive T cells into ..., but not into ..., ..., or Treg cells. tROR...t-TMD injected into experimental autoimmune encephalomyelitis (EAE) animal model can be delivered effectively in the splenic CD4+ T cells and spinal cord-infiltrating CD4+ T cells, and suppress the functions of ... cells. The clinical severity and incidence of EAE were ameliorated by tROR...t-TMD in preventive and therapeutic manner, and significant reduction of both infiltrating CD4+ IL-17+ T cells and inflammatory cells into the CNS was observed. As a result, the number of spinal cord demyelination was also reduced after tROR...t-TMD treatment. With the same proof of concept, tTbet-TMD specifically blocking ... differentiation improved the clinical incidence of rheumatoid arthritis (RA). Therefore, tROR...t-TMD and tTbet-TMD can be novel therapeutic reagents with the natural specificity for the treatment of inflammatory diseases associated with ... or ... This strategy can be applied to treat various diseases where a specific transcription factor has a key role in pathogenesis. (ProQuest: ... denotes formulae/symbols omitted.)
Abstract The accumulation of soluble oligomeric amyloid-β peptide (oAβ) proceeds the formation of senile plaques and contributes to synaptic and memory deficits in Alzheimer's disease (AD). The ...mechanism of mediating microglial oAβ clearance remains unclear and thought to occur via scavenger receptors (SRs) in microglia. SRs respond to their ligands in a subtype-specific manner. Therefore, we sought to identify the specific subtypes of SRs that mediate oAβ internalization and proteases that degrade oAβ species in naïve primary microglia. The component of oAβ species were characterized by western blot analysis, analytical ultracentrifugation analysis, and atomic force microscopy. The oAβ species remained soluble in the medium and microglial lysates during incubation at 37 °C. SR-A, but not CD36, mediated oAβ internalization in microglia as suggested by the use of subtype-specific neutralizing antibodies and small interfering RNAs (siRNAs). Immunoprecipitation analysis showed that oAβ interacted with SR-A on the plasma membrane. After internalization, over 40% of oAβ vesicles were trafficked toward lysosomes and degraded by cysteine proteases, including cathepsin B. The inhibitors of proteasome, neprilysin, matrix metalloproteinases, and insulin degrading enzyme failed to protect internalized oAβ from degradation. Our study suggests that SR-A and lysosomal cathepsin B are critical in microglial oAβ clearance, providing insight into how microglia are involved in the clearance of oAβ and their roles in the early stages of AD.
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