1  5908 123 TARGETED DE-METHYLATION OF THE FOXP3-TSDR IS SUFFICIENT TO INDUCE PHYSIOLOGICAL FOXP3 EXPRESSION BUT NOT A FUNCTIONAL TREG PHENOTYPE. CD4+ REGULATORY T CELLS (TREGS) ARE KEY MEDIATORS OF IMMUNOLOGICAL TOLERANCE AND PROMISING EFFECTOR CELLS FOR IMMUNO-SUPPRESSIVE ADOPTIVE CELLULAR THERAPY TO FIGHT AUTOIMMUNITY AND CHRONIC INFLAMMATION. THEIR FUNCTIONAL STABILITY IS CRITICAL FOR THEIR CLINICAL UTILITY AND HAS BEEN CORRELATED TO THE DEMETHYLATED STATE OF THE TSDR/CNS2 ENHANCER ELEMENT IN THE TREG LINEAGE TRANSCRIPTION FACTOR FOXP3. HOWEVER, PROOF FOR A CAUSAL CONTRIBUTION OF THE TSDR DE-METHYLATION TO FOXP3 STABILITY AND TREG INDUCTION IS SO FAR LACKING. WE HERE ESTABLISHED A POWERFUL TRANSIENT-TRANSFECTION CRISPR-CAS9-BASED EPIGENETIC EDITING METHOD FOR THE SELECTIVE DE-METHYLATION OF THE TSDR WITHIN THE ENDOGENOUS CHROMATIN ENVIRONMENT OF A LIVING CELL. THE INDUCED DE-METHYLATED STATE WAS STABLE OVER WEEKS IN CLONAL T CELL PROLIFERATION CULTURES EVEN AFTER EXPRESSION OF THE EDITING COMPLEX HAD CEASED. EPIGENETIC EDITING OF THE TSDR RESULTED IN FOXP3 EXPRESSION, EVEN IN ITS PHYSIOLOGICAL ISOFORM DISTRIBUTION, PROVING A CAUSAL ROLE FOR THE DE-METHYLATED TSDR IN FOXP3 REGULATION. HOWEVER, SUCCESSFUL FOXP3 INDUCTION WAS NOT ASSOCIATED WITH A SWITCH TOWARDS A FUNCTIONAL TREG PHENOTYPE, IN CONTRAST TO WHAT HAS BEEN REPORTED FROM FOXP3 OVEREXPRESSION APPROACHES. THUS, TSDR DE-METHYLATION IS REQUIRED, BUT NOT SUFFICIENT FOR A STABLE TREG PHENOTYPE INDUCTION. THEREFORE, TARGETED DEMETHYLATION OF THE TSDR MAY BE A CRITICAL ADDITION TO PUBLISHED IN VITRO TREG INDUCTION PROTOCOLS WHICH SO FAR LACK FOXP3 STABILITY.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
2  3454  39 HYPOMETHYLATION AT THE REGULATORY T CELL-SPECIFIC DEMETHYLATED REGION IN CD25HI T CELLS IS DECOUPLED FROM FOXP3 EXPRESSION AT THE INFLAMED SITE IN CHILDHOOD ARTHRITIS. THE MAINTENANCE OF FOXP3 EXPRESSION IN CD25(HI) REGULATORY T CELLS (TREGS) IS CRUCIAL TO THE CONTROL OF INFLAMMATION AND ESSENTIAL FOR SUCCESSFUL TREG TRANSFER THERAPIES. COEXPRESSION OF CD25 AND FOXP3 IN COMBINATION WITH A HYPOMETHYLATED REGION WITHIN THE FOXP3 GENE, CALLED THE TREG-SPECIFIC DEMETHYLATED REGION (TSDR), IS CONSIDERED THE HALLMARK OF STABLE TREGS. THE TSDR IS AN EPIGENETIC MOTIF THAT IS IMPORTANT FOR STABLE FOXP3 EXPRESSION AND IS USED AS A BIOMARKER TO MEASURE TREG LINEAGE COMMITMENT. IN THIS STUDY, WE REPORT THAT, UNLIKE IN PERIPHERAL BLOOD, CD4(+) T CELL EXPRESSION OF CD25 AND FOXP3 IS FREQUENTLY DISSOCIATED AT THE INFLAMED SITE IN PATIENTS WITH JUVENILE IDIOPATHIC ARTHRITIS, WHICH LED US TO QUESTION THE STABILITY OF HUMAN TREGS IN CHRONIC INFLAMMATORY ENVIRONMENTS. WE DESCRIBE A NOVEL CD4(+)CD127(LO)CD25(HI) HUMAN T CELL POPULATION THAT EXHIBITS EXTENSIVE TSDR AND PROMOTER DEMETHYLATION IN THE ABSENCE OF STABLE FOXP3 EXPRESSION. THIS POPULATION EXPRESSES HIGH LEVELS OF CTLA-4 AND CAN SUPPRESS T CONVENTIONAL CELL PROLIFERATION IN VITRO. THESE DATA COLLECTIVELY SUGGEST THAT THIS POPULATION MAY REPRESENT A CHRONICALLY ACTIVATED FOXP3(LO) TREG POPULATION. WE SHOW THAT THESE CELLS HAVE DEFECTS IN IL-2 SIGNALING AND REDUCED EXPRESSION OF A DEUBIQUITINASE IMPORTANT FOR FOXP3 STABILITY. CLINICALLY, THE PROPORTIONS OF THESE CELLS WITHIN THE CD25(HI) T CELL SUBSET ARE INCREASED IN PATIENTS WITH THE MORE SEVERE COURSES OF DISEASE. OUR STUDY DEMONSTRATES, THEREFORE, THAT HYPOMETHYLATION AT THE TSDR CAN BE DECOUPLED FROM FOXP3 EXPRESSION IN HUMAN T CELLS AND THAT ENVIRONMENT-SPECIFIC BREAKDOWN IN FOXP3 STABILITY MAY COMPROMISE THE RESOLUTION OF INFLAMMATION IN JUVENILE IDIOPATHIC ARTHRITIS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
3  1061  43 CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS IS CHARACTERIZED BY AN UPREGULATION OF PERIPHERAL T-REGULATORY CELL POLARIZED TOWARDS T-BET AND TIGIT. BACKGROUND: NON-INFECTIOUS UVEITIS CAN CAUSE CHRONIC RELAPSING AND REMITTING OCULAR INFLAMMATION, WHICH MAY REQUIRE HIGH DOSE SYSTEMIC IMMUNOSUPPRESSION TO PREVENT SEVERE SIGHT LOSS. IT HAS BEEN CLASSICALLY DESCRIBED AS AN AUTOIMMUNE DISEASE, MEDIATED BY PRO-INFLAMMATORY TH1 AND TH17 T-CELL SUBSETS. STUDIES SUGGEST THAT NATURAL IMMUNOSUPPRESSIVE CD4(+)CD25(+)FOXP3(+) T-REGULATORY CELLS (TREGS) ARE INVOLVED IN RESOLUTION OF INFLAMMATION AND MAY BE INVOLVED IN THE MAINTENANCE OF CLINICAL REMISSION. OBJECTIVE: TO INVESTIGATE WHETHER THERE IS A PERIPHERAL BLOOD IMMUNOREGULATORY PHENOTYPE ASSOCIATED WITH CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS BY COMPARING PERIPHERAL BLOOD LEVELS OF TREG, TH1, AND TH17, AND ASSOCIATED DNA METHYLATION AND CYTOKINE LEVELS IN PATIENTS WITH ACTIVE UVEITIC DISEASE, CONTROL SUBJECTS AND PATIENTS (WITH PREVIOUSLY ACTIVE DISEASE) IN CLINICAL REMISSION INDUCED BY IMMUNOSUPPRESSIVE DRUGS. METHODS: ISOLATED PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMC) FROM PERIPHERAL BLOOD SAMPLES FROM PROSPECTIVELY RECRUITED SUBJECTS WERE ANALYZED BY FLOW CYTOMETRY FOR CD3, CD4, FOXP3, TIGIT, T-BET, AND RELATED ORPHAN RECEPTOR GAMMAT. EPIGENETIC DNA METHYLATION LEVELS OF FOXP3 TREG-SPECIFIC DEMETHYLATED REGION (TSDR), FOXP3 PROMOTER, TBX21, RORC2, AND TIGIT LOCI WERE DETERMINED IN CRYOPRESERVED PBMC USING A NEXT-GENERATION SEQUENCING APPROACH. RELATED CYTOKINES WERE MEASURED IN BLOOD SERA. FUNCTIONAL SUPPRESSIVE CAPACITY OF TREG WAS ASSESSED USING T-CELL PROLIFERATION ASSAYS. RESULTS: FIFTY PATIENTS WITH UVEITIS (INTERMEDIATE, POSTERIOR, AND PANUVEITIS) AND 10 CONTROL SUBJECTS WERE RECRUITED. THE FREQUENCY OF CD4(+)CD25(+)FOXP3(+) TREG, TIGIT(+) TREG, AND T-BET(+) TREG AND THE RATIO OF TREG TO TH1 WERE SIGNIFICANTLY HIGHER IN REMISSION PATIENTS COMPARED WITH PATIENTS WITH ACTIVE UVEITIC DISEASE; AND TIGIT(+) TREGS WERE A SIGNIFICANT PREDICTOR OF CLINICAL REMISSION. TREG FROM PATIENTS IN CLINICAL REMISSION DEMONSTRATED A HIGH LEVEL OF IN VITRO SUPPRESSIVE FUNCTION COMPARED WITH TREG FROM CONTROL SUBJECTS AND FROM PATIENTS WITH UNTREATED ACTIVE DISEASE. PBMC FROM PATIENTS IN CLINICAL REMISSION HAD SIGNIFICANTLY LOWER METHYLATION LEVELS AT THE FOXP3 TSDR, FOXP3 PROMOTER, AND TIGIT LOCI AND HIGHER LEVELS AT RORC LOCI THAN THOSE WITH ACTIVE DISEASE. CLINICAL REMISSION WAS ALSO ASSOCIATED WITH SIGNIFICANTLY HIGHER SERUM LEVELS OF TRANSFORMING GROWTH FACTOR BETA AND IL-10, WHICH POSITIVELY CORRELATED WITH TREG LEVELS, AND LOWER SERUM LEVELS OF IFNGAMMA, IL-17A, AND IL-22 COMPARED WITH PATIENTS WITH ACTIVE DISEASE. CONCLUSION: CLINICAL REMISSION OF SIGHT-THREATENING NON-INFECTIOUS UVEITIS HAS AN IMMUNOREGULATORY PHENOTYPE CHARACTERIZED BY UPREGULATION OF PERIPHERAL TREG, POLARIZED TOWARD T-BET AND TIGIT. THESE FINDINGS MAY ASSIST WITH INDIVIDUALIZED THERAPY OF UVEITIS, BY INFORMING WHETHER DRUG THERAPY HAS INDUCED PHENOTYPICALLY STABLE TREG ASSOCIATED WITH LONG-TERM CLINICAL REMISSION.	2018	

4  3373  32 HISTONE MODULATION BLOCKS TREG-INDUCED FOXP3 BINDING TO THE IL-2 PROMOTER OF VIRUS-SPECIFIC CD8(+) T CELLS FROM FELINE IMMUNODEFICIENCY VIRUS-INFECTED CATS. CD8(+) T CELLS ARE CRITICAL FOR CONTROLLING HIV INFECTION. DURING THE CHRONIC PHASE OF LENTIVIRAL INFECTION, CD8(+) T CELLS LOSE THEIR PROLIFERATIVE CAPACITY AND EXHIBIT IMPAIRED ANTIVIRAL FUNCTION. THIS LOSS OF CD8(+) T CELL FUNCTION IS DUE, IN PART, TO CD4(+)CD25(+) T REGULATORY (TREG) CELL-MEDIATED SUPPRESSION. OUR RESEARCH GROUP HAS DEMONSTRATED THAT LENTIVIRUS-ACTIVATED CD4(+)CD25(+) TREG CELLS INDUCE THE REPRESSIVE TRANSCRIPTION FACTOR FORKHEAD BOX P3 (FOXP3) IN AUTOLOGOUS CD8(+) T CELLS FOLLOWING CO-CULTURE. WE HAVE RECENTLY REPORTED THAT TREG-INDUCED FOXP3 BINDS THE INTERLEUKIN-2 (IL-2), INTERFERON-GAMMA (IFN- GAMMA), AND TUMOR NECROSIS FACTOR-ALPHA (TNF-ALPHA) PROMOTERS IN VIRUS-SPECIFIC CD8(+) T CELLS. THESE DATA SUGGEST AN IMPORTANT ROLE OF FOXP3-MEDIATED CD8(+) T CELL DYSFUNCTION IN LENTIVIRAL INFECTION. TO ELUCIDATE THE MECHANISM OF THIS SUPPRESSION, WE PREVIOUSLY REPORTED THAT DECREASED METHYLATION FACILITATES FOXP3 BINDING IN MITOGEN-ACTIVATED CD8(+) T CELLS FROM FELINE IMMUNODEFICIENCY VIRUS (FIV)-INFECTED CATS. WE DEMONSTRATED THE REDUCED BINDING OF FOXP3 TO THE IL-2 PROMOTER BY INCREASING METHYLATION OF CD8(+) T CELLS. IN THE STUDIES PRESENTED HERE, WE ASK IF ANOTHER FORM OF EPIGENETIC MODULATION MIGHT ALLEVIATE FOXP3-MEDIATED SUPPRESSION IN CD8(+) T CELLS. WE HYPOTHESIZED THAT DECREASING HISTONE ACETYLATION IN VIRUS-SPECIFIC CD8(+) T CELLS WOULD DECREASE TREG-INDUCED FOXP3 BINDING TO THE IL-2 PROMOTER. INDEED, USING ANACARDIC ACID (AA), A KNOWN HISTONE ACETYL TRANSFERASE (HAT) INHIBITOR, WE DEMONSTRATE A REDUCTION IN FOXP3 BINDING TO THE IL-2 PROMOTER IN VIRUS-SPECIFIC CD8(+) T CELLS CO-CULTURED WITH AUTOLOGOUS TREG CELLS. THESE DATA IDENTIFY A NOVEL MECHANISM OF FOXP3-MEDIATED CD8(+) T CELL DYSFUNCTION DURING LENTIVIRAL INFECTION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
5  3861  29 ISOLATION OF HUMAN ANTIGEN-SPECIFIC REGULATORY T CELLS WITH HIGH SUPPRESSIVE FUNCTION. ADOPTIVE TRANSFER OF REGULATORY T (TREG) CELLS COULD BE AN ALTERNATIVE TO CHRONIC IMMUNOSUPPRESSION FOR PREVENTION OF ALLOGENEIC GRAFT REJECTION. WHILE POLYSPECIFIC TREG CELLS CAN PREVENT IMMUNE RESPONSES UNDER LYMPHOPENIC CONDITIONS, AG-SPECIFIC TREG CELLS ARE NEEDED TO TREAT AUTOIMMUNITY AND GRAFT REJECTION. YET, RELIABLE MARKERS FOR AG-SPECIFIC TREG CELLS ARE MISSING. WE REPORT THAT LATENCY-ASSOCIATED PEPTIDE (LAP) AND GLYCOPROTEIN A REPETITIONS PREDOMINANT (GARP) CAN IDENTIFY HUMAN AG-SPECIFIC TREG CELLS. IN ADDITION, WE SHOW THAT THE DEPLETION OF CD154(+) CELLS FROM LAP(+) OR GARP(+) TREG CELLS INCREASES THE TREG-CELL PURITY TO OVER 90%, AS ASSESSED BY EPIGENETIC ANALYSIS. THESE AG-SPECIFIC TREG CELLS CAN BE ISOLATED MAGNETICALLY AND MIGHT CONTRIBUTE TO THE DEVELOPMENT OF GMP-BASED PROTOCOLS. IN ADDITION, AG-SPECIFIC TREG CELLS ARE FUNCTIONALLY FAR SUPERIOR TO CD4(+) CD25(HIGH) OR CD4(+) CD25(HIGH) CD127(LOW) TREG CELLS IN VITRO AND IN PREVENTING STRONG ALLOREACTIONS IN HUMANIZED MICE. THEY COULD, THEREFORE, HAVE A HIGH THERAPEUTIC POTENTIAL FOR THE CONTROL OF ALLOIMMUNE, AUTOIMMUNE, AND ALLERGIC IMMUNE RESPONSES IN PATIENTS.	2014	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                           
6  3662  40 INDUCTION OF STABLE HUMAN FOXP3(+) TREGS BY A PARASITE-DERIVED TGF-BETA MIMIC. IMMUNE HOMEOSTASIS IN THE INTESTINE IS TIGHTLY CONTROLLED BY FOXP3(+) REGULATORY T CELLS (TREGS), DEFECTS OF WHICH ARE LINKED TO THE DEVELOPMENT OF CHRONIC CONDITIONS, SUCH AS INFLAMMATORY BOWEL DISEASE (IBD). AS A MECHANISM OF IMMUNE EVASION, SEVERAL SPECIES OF INTESTINAL PARASITES BOOST TREG ACTIVITY. THE PARASITE HELIGMOSOMOIDES POLYGYRUS IS KNOWN TO SECRETE A MOLECULE (HP-TGM) THAT MIMICS THE ABILITY OF TGF-BETA TO INDUCE FOXP3 EXPRESSION IN CD4(+) T CELLS. THE STUDY AIMED TO INVESTIGATE WHETHER HP-TGM COULD INDUCE HUMAN FOXP3(+) TREGS AS A POTENTIAL THERAPEUTIC APPROACH FOR INFLAMMATORY DISEASES. CD4(+) T CELLS FROM HEALTHY VOLUNTEERS WERE EXPANDED IN THE PRESENCE OF HP-TGM OR TGF-BETA. TREG INDUCTION WAS MEASURED BY FLOW CYTOMETRIC DETECTION OF FOXP3 AND OTHER TREG MARKERS, SUCH AS CD25 AND CTLA-4. EPIGENETIC CHANGES WERE DETECTED USING CHIP-SEQ AND PYROSEQUENCING OF FOXP3. TREG PHENOTYPE STABILITY WAS ASSESSED FOLLOWING INFLAMMATORY CYTOKINE CHALLENGE AND TREG FUNCTION WAS EVALUATED BY CELLULAR CO-CULTURE SUPPRESSION ASSAYS AND CYTOMETRIC BEAD ARRAYS FOR SECRETED CYTOKINES. HP-TGM EFFICIENTLY INDUCED FOXP3 EXPRESSION (> 60%), IN ADDITION TO CD25 AND CTLA-4, AND CAUSED EPIGENETIC MODIFICATION OF THE FOXP3 LOCUS TO A GREATER EXTENT THAN TGF-BETA. HP-TGM-INDUCED TREGS HAD SUPERIOR SUPPRESSIVE FUNCTION COMPARED WITH TGF-BETA-INDUCED TREGS, AND RETAINED THEIR PHENOTYPE FOLLOWING EXPOSURE TO INFLAMMATORY CYTOKINES. FURTHERMORE, HP-TGM INDUCED A TREG-LIKE PHENOTYPE IN IN VIVO DIFFERENTIATED TH1 AND TH17 CELLS, INDICATING ITS POTENTIAL TO RE-PROGRAM MEMORY CELLS TO ENHANCE IMMUNE TOLERANCE. THESE DATA INDICATE HP-TGM HAS POTENTIAL TO BE USED TO GENERATE STABLE HUMAN FOXP3(+) TREGS TO TREAT IBD AND OTHER INFLAMMATORY DISEASES.	2021	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
7  6071  35 THE DNA METHYLATION INHIBITOR 5-AZACYTIDINE INCREASES REGULATORY T CELLS AND ALLEVIATES AIRWAY INFLAMMATION IN OVALBUMIN-SENSITIZED MICE. BACKGROUND: ASTHMA IS CHARACTERIZED AS A CHRONIC INFLAMMATORY DISORDER OF THE AIRWAYS ASSOCIATED WITH AN ENHANCED TH2 RESPONSE TO INHALED ALLERGENS. CD4+ T REGULATORY (TREG) CELLS ARE CONTROLLED BY THE MASTER TRANSCRIPTION FACTOR FOXP3 AND STRICTLY MAINTAIN PERIPHERAL IMMUNOTOLERANCE. EPIGENETIC REGULATION OF FOXP3 BY DNA METHYLTRANSFERASE INHIBITORS, SUCH AS 5-AZACYTIDINE (AZA), CAN GENERATE A STEADY SUPPLY OF FUNCTIONAL TREG CELLS. THEREFORE, WE PROPOSE THAT AZA CAN AUGMENT TREG CELLS IN VIVO TO PREVENT THE PATHOGENESIS OF ASTHMA. METHODS: BALB/C MICE WERE SENSITIZED WITH CHICKEN OVALBUMIN (OVA) AND TREATED WITH DIFFERENT DOSES OF AZA. AIRWAY HYPERRESPONSIVENESS TO METHACHOLINE, EOSINOPHILIA IN BRONCHOALVEOLAR LAVAGE FLUID, CIRCULATING TITERS OF OVA-SPECIFIC IGG1 AND IGE, AND STIMULATING LEVELS OF TH2 CYTOKINES FROM SPLENOCYTES WERE THEN DETERMINED. CELLULAR POPULATIONS WERE EXAMINED BY FLOW CYTOMETRY. PC61 ANTIBODY, WHICH DEPLETES CD25+ CELLS, WAS USED TO VERIFY THE ROLE OF CD25+ CELLS IN AZA-INDUCED TOLERANCE. RESULTS: ADMINISTRATION OF AZA TO OVA-SENSITIZED MICE DIMINISHED AIRWAY HYPERREACTIVITY, PULMONARY EOSINOPHILIA, LEVELS OF OVA-SPECIFIC IGG1 AND IGE IN SERUM, AND SECRETION OF TH2 CYTOKINES FROM OVA-STIMULATED SPLENOCYTES IN A DOSE-DEPENDENT MANNER. PERCENTAGES OF CD25+ AND FOXP3+ CELLS IN THE CD4+ CELL POPULATION WERE NOTABLY INCREASED IN AZA-TREATED MICE COMPARED TO SENSITIZED CONTROL MICE. FURTHERMORE, THE MAJOR SYMPTOMS OF ASTHMA WERE EXACERBATED BY DEPLETING CD25+ CELLS IN AZA-TREATED MICE. CONCLUSIONS: AZA MAY HAVE APPLICATIONS AS A NOVEL CLINICAL STRATEGY TO INCREASE THE PRODUCTION OF TREG CELLS IN ORDER TO MODULATE THE AIRWAY INFLAMMATION ASSOCIATED WITH ASTHMA.	2013	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                        
8  6020  39 THE ATTENUATION OF RENAL FIBROSIS BY HISTONE DEACETYLASE INHIBITORS IS ASSOCIATED WITH THE PLASTICITY OF FOXP3(+)IL-17(+) T CELLS. BACKGROUND: THE HISTONE DEACETYLASE (HDAC) INHIBITOR, WHICH HAS POTENTIAL EFFECTS ON EPIGENETIC MODIFICATIONS, HAD BEEN REPORTED TO ATTENUATE RENAL FIBROSIS. CD4(+) FORKHEAD BOX P3 (FOXP3)(+) T REGULATORY (TREG) CELLS MAY BE CONVERTED TO INFLAMMATION-ASSOCIATED T HELPER 17 CELLS (TH17) WITH TISSUE FIBROSIS PROPERTIES. THE ASSOCIATION BETWEEN FOXP3(+)IL-17(+) T CELLS AND THE ATTENUATION OF RENAL FIBROSIS BY THE HDAC INHIBITOR IS NOT CLEAR. METHODS: THIS STUDY EVALUATED THE ROLES OF THE HDAC INHIBITOR, TREG CELLS AND THEIR DIFFERENTIATION INTO TH17 CELLS, WHICH AGGRAVATE CHRONIC INFLAMMATION AND RENAL FIBROSIS IN A UNILATERAL URETERAL OBSTRUCTION (UUO) MOUSE MODEL. THE STUDY GROUPS INCLUDED CONTROL AND UUO MICE THAT WERE MONITORED FOR 7, 14 OR 21 DAYS. RESULTS: JUXTAGLOMERULAR (JG) HYPERPLASIA, ANGIOTENSIN II TYPE 1 RECEPTOR (AT1R) EXPRESSION AND LYMPHOCYTE INFILTRATION WERE OBSERVED IN RENAL TISSUES AFTER UUO BUT WERE DECREASED AFTER TRICHOSTATIN A (TSA) TREATMENT, A HDAC INHIBITOR. THE NUMBER OF CD4(+)FOXP3(+) T CELLS INCREASED PROGRESSIVELY, ALONG WITH THE NUMBER OF FOXP3(+)INTERLEUKIN (IL)-17(+) T CELLS, AFTER 14 DAYS, AND THEIR NUMBERS THEN PROGRESSIVELY DECREASED WITH INCREASING CD4(+)IL-17(+) T CELL NUMBERS, AS DEMONSTRATED BY DOUBLE IMMUNOHISTOCHEMISTRY. PROGRESSIVE RENAL FIBROSIS WAS ASSOCIATED WITH THE LOSS OF CD4(+)FOXP3(+)IL-17(+) T CELLS IN SPLENIC SINGLE-CELL SUSPENSIONS. FOXP3(+)IL-17(+) T CELLS EXPRESSED TGF-BETA1 BOTH IN VITRO AND IN VIVO, AND TGF-BETA1 EXPRESSION WAS SIGNIFICANTLY KNOCKDOWN BY IL-17 SIRNA IN VITRO. THESE CELLS WERE FOUND TO PLAY A ROLE IN CONVERTING TREGS INTO IL-17- AND TGF-BETA1-PRODUCING CELLS. CONCLUSIONS: TSA TREATMENT DECREASED JG HYPERPLASIA, THE PERCENTAGE OF FOXP3(+)IL-17(+) CELLS AND THE DEGREE OF FIBROSIS, SUGGESTING THAT THERAPEUTIC BENEFITS MAY RESULT FROM EPIGENETIC MODIFICATIONS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                         
9  5049  35 PHARMACOLOGICAL INHIBITION OF RORC2 ENHANCES HUMAN TH17-TREG STABILITY AND FUNCTION. INFLAMMATORY BOWEL DISEASES (IBD) ARE CHRONIC CONDITIONS THAT RESULT FROM UNCONTROLLED INTESTINAL INFLAMMATION. PATHOGENIC TH17 CELLS, CHARACTERIZED BY PRODUCTION OF IL-17A IN THE ABSENCE OF IL-10, ARE THOUGHT TO CONTRIBUTE TO THIS INFLAMMATION, BUT IN HUMANS, ANTIBODY-MEDIATED BLOCKADE OF IL-17A IS AN INEFFECTIVE IBD THERAPY WHEREAS IL-23 BLOCKADE IS EFFECTIVE. HERE, WE INVESTIGATED THE EFFECTS OF PHARMACOLOGICAL INHIBITION OF RORC2, THE TH17 CELL LINEAGE-DEFINING TRANSCRIPTION FACTOR, ON IN VIVO-DIFFERENTIATED HUMAN TH17 CELLS AND TH17-LIKE TREGS (TH17-TREGS). BMS-336, A SMALL MOLECULE RORC2 INVERSE AGONIST, INHIBITED EXPRESSION OF RORC2-REGULATED GENES IN PERIPHERAL TH17 CELLS (CD4(+) CD25(-) CD127(+) CXCR3(-) CCR4(+) CCR6(+) ) IN A DOSE-DEPENDENT MANNER, WITH SIMILAR INHIBITORY EFFECTS ON LAMINAR PROPRIA MONONUCLEAR CELLS FROM IBD AND NON-IBD SUBJECTS. EXPOSURE OF PERIPHERAL TH17-TREGS (CD4(+) CD25(HI) CD127(LO) CXCR3(-) CCR4(+) CCR6(+) ) TO BMS-336 ALSO INHIBITED IL-17A PRODUCTION AND PREVENTED INFLAMMATORY CYTOKINE-INDUCED DESTABILIZATION, AS EVIDENCED BY PRESERVED FOXP3 EXPRESSION AND EPIGENETIC STATUS OF THE TREG-SPECIFIC DEMETHYLATION REGION. IN PARALLEL, RORC2 INHIBITION INCREASED THE PRODUCTION OF IL-10 IN TH17-TREGS, RESULTING IN ENHANCED SUPPRESSION OF INFLAMMATORY CYTOKINES FROM MYELOID CELLS. THUS, VIA ITS ABILITY TO SIMULTANEOUSLY INHIBIT TH17 CELLS AND ENHANCE THE STABILITY AND FUNCTION OF TH17-TREGS, PHARMACOLOGICAL INHIBITION OF RORC2 IS A PROMISING APPROACH TO SUPPRESS INFLAMMATION AND PROMOTE IMMUNE REGULATION IN IBD.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                             
10   407  35 ANALYSIS OF FOXP3+ REGULATORY T CELLS THAT DISPLAY APPARENT VIRAL ANTIGEN SPECIFICITY DURING CHRONIC HEPATITIS C VIRUS INFECTION. WE REPORTED PREVIOUSLY THAT A PROPORTION OF NATURAL CD25(+) CELLS ISOLATED FROM THE PBMC OF HCV PATIENTS CAN FURTHER UPREGULATE CD25 EXPRESSION IN RESPONSE TO HCV PEPTIDE STIMULATION IN VITRO, AND PROPOSED THAT VIRUS-SPECIFIC REGULATORY T CELLS (TREG) WERE PRIMED AND EXPANDED DURING THE DISEASE. HERE WE DESCRIBE EPIGENETIC ANALYSIS OF THE FOXP3 LOCUS IN HCV-RESPONSIVE NATURAL CD25(+) CELLS AND SHOW THAT THESE CELLS ARE NOT ACTIVATED CONVENTIONAL T CELLS EXPRESSING FOXP3, BUT HARD-WIRED TREG WITH A STABLE FOXP3 PHENOTYPE AND FUNCTION. OF APPROXIMATELY 46,000 GENES ANALYZED IN GENOME WIDE TRANSCRIPTION PROFILING, ABOUT 1% WERE DIFFERENTIALLY EXPRESSED BETWEEN HCV-RESPONSIVE TREG, HCV-NON-RESPONSIVE NATURAL CD25(+) CELLS AND CONVENTIONAL T CELLS. EXPRESSION PROFILES, INCLUDING CELL DEATH, ACTIVATION, PROLIFERATION AND TRANSCRIPTIONAL REGULATION, SUGGEST A SURVIVAL ADVANTAGE OF HCV-RESPONSIVE TREG OVER THE OTHER CELL POPULATIONS. SINCE NO TREG-SPECIFIC ACTIVATION MARKER IS KNOWN, WE TESTED 97 NS3-DERIVED PEPTIDES FOR THEIR ABILITY TO ELICIT CD25 RESPONSE (ASSUMING IT IS A SURROGATE MARKER), ACCOMPANIED BY HIGH RESOLUTION HLA TYPING OF THE PATIENTS. SOME REACTIVE PEPTIDES OVERLAPPED WITH PREVIOUSLY DESCRIBED EFFECTOR T CELL EPITOPES. OUR DATA OFFERS NEW INSIGHTS INTO HCV IMMUNE EVASION AND TOLERANCE, AND HIGHLIGHTS THE NON-SELF SPECIFIC NATURE OF TREG DURING INFECTION.	2009	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
11   556  47 AZACYTIDINE TREATMENT INHIBITS THE PROGRESSION OF HERPES STROMAL KERATITIS BY ENHANCING REGULATORY T CELL FUNCTION. OCULAR INFECTION WITH HERPES SIMPLEX VIRUS 1 (HSV-1) SETS OFF AN INFLAMMATORY REACTION IN THE CORNEA WHICH LEADS TO BOTH VIRUS CLEARANCE AND CHRONIC LESIONS THAT ARE ORCHESTRATED BY CD4 T CELLS. APPROACHES THAT ENHANCE THE FUNCTION OF REGULATORY T CELLS (TREG) AND DAMPEN EFFECTOR T CELLS CAN BE EFFECTIVE TO LIMIT STROMAL KERATITIS (SK) LESION SEVERITY. IN THIS REPORT, WE EXPLORE THE NOVEL APPROACH OF INHIBITING DNA METHYLTRANSFERASE ACTIVITY USING 5-AZACYTIDINE (AZA; A CYTOSINE ANALOG) TO LIMIT HSV-1-INDUCED OCULAR LESIONS. WE SHOW THAT THERAPY BEGUN AFTER INFECTION WHEN VIRUS WAS NO LONGER ACTIVELY REPLICATING RESULTED IN A PRONOUNCED REDUCTION IN LESION SEVERITY, WITH MARKEDLY DIMINISHED NUMBERS OF T CELLS AND NONLYMPHOID INFLAMMATORY CELLS, ALONG WITH REDUCED CYTOKINE MEDIATORS. THE REMAINING INFLAMMATORY REACTIONS HAD A CHANGE IN THE RATIO OF CD4 FOXP3(+) TREG TO EFFECTOR TH1 CD4 T CELLS IN OCULAR LESIONS AND LYMPHOID TISSUES, WITH TREG BECOMING PREDOMINANT OVER THE EFFECTORS. IN ADDITION, COMPARED TO THOSE FROM CONTROL MICE, TREG FROM AZA-TREATED MICE SHOWED MORE SUPPRESSOR ACTIVITY IN VITRO AND EXPRESSED HIGHER LEVELS OF ACTIVATION MOLECULES. ADDITIONALLY, CELLS INDUCED IN VITRO IN THE PRESENCE OF AZA SHOWED EPIGENETIC DIFFERENCES IN THE TREG-SPECIFIC DEMETHYLATED REGION (TSDR) OF FOXP3 AND WERE MORE STABLE WHEN EXPOSED TO INFLAMMATORY CYTOKINES. OUR RESULTS SHOW THAT THERAPY WITH AZA IS AN EFFECTIVE MEANS OF CONTROLLING A VIRUS-INDUCED INFLAMMATORY REACTION AND MAY ACT MAINLY BY THE EFFECTS ON TREG.IMPORTANCE HSV-1 INFECTION HAS BEEN SHOWN TO INITIATE AN INFLAMMATORY REACTION IN THE CORNEA THAT LEADS TO TISSUE DAMAGE AND LOSS OF VISION. THE INFLAMMATORY REACTION IS ORCHESTRATED BY GAMMA INTERFERON (IFN-GAMMA)-SECRETING TH1 CELLS, AND REGULATORY T CELLS PLAY A PROTECTIVE ROLE. HENCE, NOVEL THERAPEUTICS THAT CAN REBALANCE THE RATIO OF REGULATORY T CELLS TO EFFECTORS ARE A RELEVANT ISSUE. THIS STUDY OPENS UP A NEW AVENUE IN TREATING HSV-INDUCED SK LESIONS BY INCREASING THE STABILITY AND FUNCTION OF REGULATORY T CELLS USING THE DNA METHYLTRANSFERASE INHIBITOR 5-AZACYTIDINE (AZA). AZA INCREASED THE FUNCTION OF REGULATORY T CELLS, LEADING TO ENHANCED SUPPRESSIVE ACTIVITY AND DIMINISHED LESIONS. HENCE, THERAPY WITH AZA, WHICH ACTS MAINLY BY ITS EFFECTS ON TREG, CAN BE AN EFFECTIVE MEANS TO CONTROL VIRUS-INDUCED INFLAMMATORY LESIONS.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
12  4967  32 PATHOLOGICAL CONDITIONS RE-SHAPE PHYSIOLOGICAL TREGS INTO PATHOLOGICAL TREGS. CD4(+)FOXP3(+) REGULATORY T CELLS (TREGS) ARE A SUBSET OF CD4 T CELLS THAT PLAY AN ESSENTIAL ROLE IN MAINTAINING PERIPHERAL IMMUNE TOLERANCE, CONTROLLING ACUTE AND CHRONIC INFLAMMATION, ALLERGY, AUTOIMMUNE DISEASES, AND ANTI-CANCER IMMUNE RESPONSES. OVER THE PAST 20 YEARS, SIGNIFICANT PROGRESS HAS BEEN MADE SINCE TREGS WERE FIRST CHARACTERIZED IN 1995. MANY CONCEPTS AND PRINCIPLES REGARDING TREGS GENERATION, PHENOTYPIC FEATURES, SUBSETS (TTREGS, PTREGS, ITREGS, AND ITREG35), TISSUE SPECIFICITY (CENTRAL TREGS, EFFECTOR TREGS, AND TISSUE RESIDENT TREGS), HOMEOSTASIS (HIGHLY DYNAMIC AND APOPTOTIC), REGULATION OF TREGS BY RECEPTORS FOR PAMPS AND DAMPS, TREG PLASTICITY (RE-DIFFERENTIATION TO OTHER CD4 T HELPER CELL SUBSETS, TH1, TH2, TFH AND TH17), AND EPIGENETIC REGULATION OF TREGS PHENOTYPES AND FUNCTIONS HAVE BEEN INNOVATED. IN THIS CONCISE REVIEW, WE WANT TO BRIEFLY ANALYZE THESE EIGHT NEW PROGRESSES IN THE STUDY OF TREGS. WE HAVE ALSO PROPOSED FOR THE FIRST TIME A NOVEL CONCEPT THAT "PHYSIOLOGICAL TREGS" HAVE BEEN RE-SHAPED INTO "PATHOLOGICAL TREGS" IN VARIOUS PATHOLOGICAL ENVIRONMENTS. CONTINUING OF THE IMPROVEMENT IN OUR UNDERSTANDING ON THIS IMPORTANT CELLULAR COMPONENT ABOUT THE IMMUNE TOLERANCE AND IMMUNE SUPPRESSION, WOULD LEAD TO THE FUTURE DEVELOPMENT OF NOVEL THERAPEUTICS APPROACHES FOR ACUTE AND CHRONIC INFLAMMATORY DISEASES, ALLERGY, ALLOGENEIC TRANSPLANTATION-RELATED IMMUNITY, SEPSIS, AUTOIMMUNE DISEASES, AND CANCERS.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
13   362  31 AMBIENT AIR POLLUTION IMPAIRS REGULATORY T-CELL FUNCTION IN ASTHMA. BACKGROUND: ASTHMA IS THE MOST FREQUENT CHRONIC DISEASE IN CHILDREN, AND CHILDREN ARE AT HIGH RISK FOR ADVERSE HEALTH CONSEQUENCES ASSOCIATED WITH AMBIENT AIR POLLUTION (AAP) EXPOSURE. REGULATORY T (TREG) CELLS ARE SUPPRESSORS OF IMMUNE RESPONSES INVOLVED IN ASTHMA PATHOGENESIS. TREG-CELL IMPAIRMENT IS ASSOCIATED WITH INCREASED DNA METHYLATION OF FORKHEAD BOX TRANSCRIPTION FACTOR 3 (FOXP3), A KEY TRANSCRIPTION FACTOR IN TREG-CELL ACTIVITY. BECAUSE AAP EXPOSURE CAN INDUCE EPIGENETIC CHANGES, WE HYPOTHESIZED THAT TREG-CELL FUNCTION WOULD BE IMPAIRED BY AAP, ALLOWING AMPLIFICATION OF AN INFLAMMATORY RESPONSE. OBJECTIVES: TO ASSESS WHETHER EXPOSURE TO AAP LED TO HYPERMETHYLATION OF THE FOXP3 GENE, CAUSING IMPAIRED TREG-CELL SUPPRESSION AND WORSENED ASTHMA SYMPTOM SCORES. METHODS: CHILDREN WITH AND WITHOUT ASTHMA FROM FRESNO, CALIF (HIGH POLLUTION, FRESNO ASTHMA GROUP [FA], N = 71, AND FRESNO NON ASTHMATIC GROUP, N = 30, RESPECTIVELY), AND FROM STANFORD, CALIF (LOW POLLUTION, STANFORD ASTHMA GROUP, N = 40, AND STANFORD NON ASTHMATIC GROUP, N = 40), WERE ENROLLED IN A CROSS-SECTIONAL STUDY. PERIPHERAL BLOOD TREG CELLS WERE USED IN FUNCTIONAL AND EPIGENETIC STUDIES. ASTHMA OUTCOMES WERE ASSESSED BY GLOBAL INITIATIVE IN ASTHMA SCORE. RESULTS: FRESNO ASTHMA GROUP TREG-CELL SUPPRESSION WAS IMPAIRED AND FA TREG-CELL CHEMOTAXIS WERE REDUCED COMPARED WITH OTHER GROUPS (P </= .05). TREG-CELL DYSFUNCTION WAS ASSOCIATED WITH MORE PRONOUNCED DECREASES IN ASTHMA GLOBAL INITIATIVE IN ASTHMA SCORE IN FA VERSUS THE STANFORD ASTHMA GROUP. FOXP3 WAS DECREASED IN FA COMPARED WITH THE FRESNO NON ASTHMATIC GROUP (P </= .05). FA ALSO CONTAINED SIGNIFICANTLY HIGHER LEVELS OF METHYLATION AT THE FOXP3 LOCUS (P </= .05). CONCLUSION: INCREASED EXPOSURE TO AAP IS ASSOCIATED WITH HYPERMETHYLATION OF THE FOXP3 LOCUS, IMPAIRING TREG-CELL FUNCTION AND INCREASING ASTHMA MORBIDITY. AAP COULD PLAY A ROLE IN MEDIATING EPIGENETIC CHANGES IN TREG CELLS, WHICH MAY WORSEN ASTHMA BY AN IMMUNE MECHANISM.	2010	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                 
14  1994  27 EPIGENETIC AND GENE EXPRESSION ALTERATIONS OF FOXP3 IN THE T CELLS OF EAE MOUSE MODEL OF MULTIPLE SCLEROSIS. MULTIPLE SCLEROSIS (MS) IS A CHRONIC AUTOIMMUNE DISEASE WITH DEMYELINATION AND NEURODEGENERATION OF THE CENTRAL NERVOUS SYSTEM. IT HAS BEEN SHOWN THAT THE REGULATORY T (TREG) CELLS ARE RESPONSIBLE FOR MAINTAINING TOLERANCE TO SELF-ANTIGENS AND CAN SUPPRESS THE AUTOIMMUNE PROCESS IN SEVERAL ANIMAL MODELS SUCH AS EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE), A MOUSE MODEL OF MS. RECENT BASIC STUDIES HAVE DEMONSTRATED THAT FORKHEAD BOX P (FOXP3) AND BTB DOMAIN AND CNC HOMOLOG 2 (BACH2) ARE THE MASTER TRANSCRIPTION FACTORS OF THESE CELLS PLAYING A PIVOTAL ROLE IN THE POLARIZATION OF NAIVE T CELLS INTO TREG CELLS. IN THE CURRENT STUDY, THE EXPRESSION OF FOXP3 AND BACH2 GENES AND FOXP3 PROMOTER METHYLATION WERE EVALUATED IN T CELLS OF THE EAE-INDUCED MICE. THE RESULTS OF THIS STUDY SHOWED A PROMINENT AND SIGNIFICANT HYPERMETHYLATION OF THE FOXP3 GENE PROMOTER IN THE EAE-INDUCED MICE COMPARED TO THE SHAM AND CONTROL GROUPS. THE EXPRESSION OF FOXP3 AND BACH2 GENES WAS SIGNIFICANTLY DECREASED IN THE EAE GROUP IN COMPARISON WITH THE SHAM AND CONTROL GROUPS. THIS STUDY SUGGESTS THAT THE EPIGENETIC MODIFICATION OF FOXP3 GENE IS INVOLVED IN THE PATHOGENESIS OF EAE AND THIS COULD BE IMPORTANT IN THERAPY IN AN APPROPRIATE AND LOGICAL STATEMENT.	2017	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
15  5431  35 REGULATORY T CELLS: PATHOPHYSIOLOGICAL ROLES AND CLINICAL APPLICATIONS. INFLAMMATION AND IMMUNE RESPONSES AFTER TISSUE INJURY PLAY PIVOTAL ROLES IN THE RESOLUTION OF INFLAMMATION, TISSUE RECOVERY, FIBROSIS, AND REMODELING. REGULATORY T CELLS (TREGS) ARE RESPONSIBLE FOR IMMUNE TOLERANCE AND ARE USUALLY ACTIVATED IN SECONDARY LYMPHATIC TISSUES. ACTIVATED TREGS SUBSEQUENTLY REGULATE EFFECTOR T CELL AND DENDRITIC CELL ACTIVATION. FOR CLINICAL APPLICATIONS SUCH AS THE SUPPRESSION OF BOTH AUTOIMMUNE DISEASES AND THE REJECTION OF TRANSPLANTED ORGANS, METHODS TO GENERATE STABILIZED ANTIGEN-SPECIFIC TREGS ARE REQUIRED. FOR THIS PURPOSE, TRANSCRIPTIONAL AND EPIGENETIC REGULATION OF FOXP3 EXPRESSION HAS BEEN INVESTIGATED. IN ADDITION TO CONVENTIONAL TREGS, THERE ARE SOME TREGS THAT RESIDE IN TISSUES AND ARE CALLED TISSUE TREGS. TISSUE TREGS EXHIBIT TISSUE-SPECIFIC FUNCTIONS THAT CONTRIBUTE TO THE MAINTENANCE OF TISSUE HOMEOSTASIS AND REPAIR. SUCH TISSUE TREGS COULD ALSO BE USEFUL FOR TREG-BASED CELL THERAPY. WE RECENTLY DISCOVERED BRAIN TREGS THAT ACCUMULATE IN THE BRAIN DURING THE CHRONIC PHASE OF ISCHEMIC BRAIN INJURY. BRAIN TREGS RESEMBLE OTHER TISSUE TREGS, BUT ARE UNIQUE IN EXPRESSING NEURAL CELL-SPECIFIC GENES SUCH AS THE SEROTONIN RECEPTOR (HTR7); CONSEQUENTLY, BRAIN TREGS RESPOND TO SEROTONIN. HERE, WE DESCRIBE OUR EXPERIENCES IN THE USE OF TREGS TO SUPPRESS GRAFT-VERSUS-HOST DISEASE AND TO PROMOTE NEURAL RECOVERY AFTER STROKE.	2020	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     
16  3998  33 LOSS OF EPIGENETIC MODIFICATION DRIVEN BY THE FOXP3 TRANSCRIPTION FACTOR LEADS TO REGULATORY T CELL INSUFFICIENCY. REGULATORY T (TREG) CELLS, DRIVEN BY THE FOXP3 TRANSCRIPTION FACTOR, ARE RESPONSIBLE FOR LIMITING AUTOIMMUNITY AND CHRONIC INFLAMMATION. WE SHOWED THAT A WELL-CHARACTERIZED FOXP3(GFP) REPORTER MOUSE, WHICH EXPRESSES AN N-TERMINAL GFP-FOXP3 FUSION PROTEIN, IS A HYPOMORPH THAT CAUSES PROFOUNDLY ACCELERATED AUTOIMMUNE DIABETES ON A NOD BACKGROUND. ALTHOUGH NATURAL TREG CELL DEVELOPMENT AND IN VITRO FUNCTION ARE NOT MARKEDLY ALTERED IN FOXP3(GFP) NOD AND C57BL/6 MICE, TREG CELL FUNCTION IN INFLAMMATORY ENVIRONMENTS WAS PERTURBED AND TGF-BETA-INDUCED TREG CELL DEVELOPMENT WAS REDUCED. FOXP3(GFP) WAS UNABLE TO INTERACT WITH THE HISTONE ACETYLTRANSFERASE TIP60, THE HISTONE DEACETYLASE HDAC7, AND THE IKAROS FAMILY ZINC FINGER 4, EOS, WHICH LED TO REDUCED FOXP3 ACETYLATION AND ENHANCED K48-LINKED POLYUBIQUITYLATION. COLLECTIVELY THIS RESULTS IN AN ALTERED TRANSCRIPTIONAL LANDSCAPE AND REDUCED FOXP3-MEDIATED GENE REPRESSION, NOTABLY AT THE HALLMARK IL-2 PROMOTER. LOSS OF CONTROLLED FOXP3-DRIVEN EPIGENETIC MODIFICATION LEADS TO TREG CELL INSUFFICIENCY THAT ENABLES AUTOIMMUNITY IN SUSCEPTIBLE ENVIRONMENTS.	2012	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                   
17  2392  28 EPIGENETIC REPRESSION OF INTERLEUKIN 2 EXPRESSION IN SENESCENT CD4+ T CELLS DURING CHRONIC HIV TYPE 1 INFECTION. THE MOLECULAR MECHANISMS FOR IL2 GENE-SPECIFIC DYSREGULATION DURING CHRONIC HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 (HIV-1) INFECTION ARE UNKNOWN. HERE, WE INVESTIGATED THE ROLE OF DNA METHYLATION IN SUPPRESSING INTERLEUKIN 2 (IL-2) EXPRESSION IN MEMORY CD4(+) T CELLS DURING CHRONIC HIV-1 INFECTION. WE OBSERVED THAT CPG SITES IN THE IL2 PROMOTER OF CD4(+) T CELLS WERE FULLY METHYLATED IN NAIVE CD4(+) T CELLS AND SIGNIFICANTLY DEMETHYLATED IN THE MEMORY POPULATIONS. INTERESTINGLY, WE FOUND THAT THE MEMORY CELLS THAT HAD A TERMINALLY DIFFERENTIATED PHENOTYPE AND EXPRESSED CD57 HAD INCREASED IL2 PROMOTER METHYLATION RELATIVE TO LESS DIFFERENTIATED MEMORY CELLS IN HEALTHY INDIVIDUALS. IMPORTANTLY, EARLY EFFECTOR MEMORY SUBSETS FROM HIV-1-INFECTED SUBJECTS EXPRESSED HIGH LEVELS OF CD57 AND WERE HIGHLY METHYLATED AT THE IL2 LOCUS. FURTHERMORE, THE INCREASED CD57 EXPRESSION ON MEMORY CD4(+) T CELLS WAS INVERSELY CORRELATED WITH IL-2 PRODUCTION. THESE DATA SUGGEST THAT DNA METHYLATION AT THE IL2 LOCUS IN CD4(+) T CELLS IS COUPLED TO IMMUNOSENESCENCE AND PLAYS A CRITICAL ROLE IN THE BROAD DYSFUNCTION THAT OCCURS IN POLYCLONAL T CELLS DURING HIV-1 INFECTION.	2015	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  
18  5509  38 RHPLD2 INHIBITS AIRWAY INFLAMMATION IN AN ASTHMATIC MURINE MODEL THROUGH INDUCTION OF STABLE CD25(+) FOXP3(+) TREGS. OUR PREVIOUS STUDIES HAVE SHOWN THAT RECOMBINANT HUMAN PHOSPHOLIPASE D2 (RHPLD2) PLAYS A MODULATOR ROLE ON NF-KAPPAB AND PKC SIGNALING PATHWAYS. IT ALSO INHIBITS IL-5-INDUCED INFLAMMATORY RESPONSE IN CHRONIC ASTHMATIC GUINEA PIGS. ADDITIONALLY, INCREASING EVIDENCE ALSO HAS REVEALED THAT THE ADOPTIVE TRANSFER OF INDUCED REGULATORY T CELLS (TREGS) MAY BE A THERAPEUTIC SOLUTION TO AIRWAY ALLERGIC DISEASES. TO INVESTIGATE THE EPIGENETIC, TRANSCRIPTOMIC AND PHENOTYPIC VARIABILITY OF TREG POPULATION IN AN OVALBUMIN (OVA)-INDUCED AIRWAY INFLAMMATION MODEL DERIVED FROM THE INDUCTION OF RHPLD2, OVA-INDUCED ASTHMATIC MURINE MODEL IS USED IN THIS STUDY. THE LUNG INFLAMMATION, EOSINOPHIL INFILTRATION, THE DIFFERENTIATION AND PROLIFERATION OF T HELPER CELLS AND THE AMPLIFICATION OF TREGS WERE EXAMINED IN THIS MOUSE MODEL WITH AND WITHOUT RHPLD2 INDUCTION. OUR DATA SHOWED THAT RHPLD2 ADMINISTRATION IN ASTHMATIC MICE SIGNIFICANTLY INCREASES CD4(+)CD25(+) FOXP3(+) TREG CELL NUMBERS AND ALLEVIATES LUNG INFLAMMATION. THE ADDITION OF RHPLD2 IN VITRO ENHANCED THE DEMETHYLATION OF TREG-SPECIFICDEMETHYLATED REGION (TSDR) IN ITREGS, SUGGESTING THAT RHPLD2 PROTEIN MAY BE INVOLVED IN IMPROVING THE QUALITY AND QUANTITY OF TREG CELLS THAT EVENTUALLY SIGNIFICANTLY REDUCES LUNG INFLAMMATION IN ASTHMATIC MURINE MODEL. THESE RESULTS SUGGEST THAT RHPLD2 COULD HAVE A CLINICAL IMPACT TREATING PATIENTS WITH ALLERGIC AIRWAY INFLAMMATION VIA PROMOTING AND STABILIZING ITREG DIFFERENTIATION AND FUNCTION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
19  3953  27 LOCUS-SPECIFIC REVERSIBLE DNA METHYLATION REGULATES TRANSIENT IL-10 EXPRESSION IN TH1 CELLS. IL-10 IS A PLEIOTROPIC CYTOKINE WITH MULTIFACETED FUNCTIONS IN ESTABLISHING IMMUNE HOMEOSTASIS. ALTHOUGH EXPRESSED BY TH1 AND TH2 CELLS, CONVENTIONAL TH1 CELLS PRODUCE MARGINAL LEVELS OF IL-10 COMPARED WITH THEIR TH2 COUNTERPARTS. IN THIS STUDY, WE INVESTIGATED THE EPIGENETIC MECHANISMS OF IL-10 GENE EXPRESSION IN TH1 CELLS. BIOINFORMATICS EMBOSS CPG PLOT ANALYSIS AND BISULFITE PYROSEQUENCING REVEALED THREE CPG DNA METHYLATION SITES IN THE IL-10 GENE LOCUS. PROGRESSIVE DNA METHYLATION AT ALL OF THE CPG REGIONS OF INTEREST (ROIS) ESTABLISHED A REPRESSIVE PROGRAM OF IL-10 GENE EXPRESSION IN TH1 CELLS. INTERESTINGLY, TH1 CELLS TREATED WITH IL-12 AND IL-27 CYTOKINES, THEREBY MIMICKING A CHRONIC INFLAMMATORY CONDITION IN VIVO, DISPLAYED A SIGNIFICANT INCREASE IN IL-10 PRODUCTION THAT WAS ACCOMPANIED BY SELECTIVE DNA DEMETHYLATION AT ROI 3 LOCATED IN INTRON 3. IL-10-PRODUCING T CELLS ISOLATED FROM LYMPHOCYTIC CHORIOMENINGITIS VIRUS-INFECTED MICE ALSO SHOWED ENHANCED DNA DEMETHYLATION AT ROI 3. BINDING OF STAT1 AND STAT3 TO DEMETHYLATED ROI 3 ENHANCED IL-10 EXPRESSION IN AN IL-12/IL-27-DEPENDENT MANNER. ACCORDINGLY, CD4(+) T CELLS ISOLATED FROM STAT1- OR STAT3-KNOCKOUT MICE WERE SIGNIFICANTLY DEFECTIVE IN IL-10 PRODUCTION. OUR DATA SUGGEST THAT, ALTHOUGH STABLY MAINTAINED DNA METHYLATION AT THE PROMOTER MAY REPRESS IL-10 EXPRESSION IN TH1 CELLS, LOCUS-SPECIFIC REVERSIBLE DNA DEMETHYLATION MAY SERVE AS A THRESHOLD PLATFORM TO CONTROL TRANSIENT IL-10 GENE EXPRESSION.	2018	
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      
20  5223  31 PRIMARY MURINE CD4+ T CELLS FAIL TO ACQUIRE THE ABILITY TO PRODUCE EFFECTOR CYTOKINES WHEN ACTIVE RAS IS PRESENT DURING TH1/TH2 DIFFERENTIATION. CONSTITUTIVE RAS SIGNALING HAS BEEN SHOWN TO AUGMENT IL-2 PRODUCTION, REVERSE ANERGY, AND FUNCTIONALLY REPLACE MANY ASPECTS OF CD28 CO-STIMULATION IN CD4+ T CELLS. THESE DATA RAISE THE POSSIBILITY THAT INTRODUCTION OF ACTIVE RAS INTO PRIMARY T CELLS MIGHT RESULT IN IMPROVED FUNCTIONALITY IN PATHOLOGIC SITUATIONS OF T CELL DYSFUNCTION, SUCH AS CANCER OR CHRONIC VIRAL INFECTION. TO TEST THE BIOLOGIC EFFECTS OF ACTIVE RAS IN PRIMARY T CELLS, CD4+ T CELLS FROM COXSACKIE-ADENOVIRUS RECEPTOR TRANSGENIC MICE WERE TRANSDUCED WITH AN ADENOVIRUS ENCODING ACTIVE RAS. AS EXPECTED, ACTIVE RAS AUGMENTED IL-2 PRODUCTION IN NAIVE CD4+ T CELLS. HOWEVER, WHEN CELLS WERE CULTURED FOR 4 DAYS UNDER CONDITIONS TO PROMOTE EFFECTOR CELL DIFFERENTIATION, ACTIVE RAS INHIBITED THE ABILITY OF CD4+ T CELLS TO ACQUIRE A TH1 OR TH2 EFFECTOR CYTOKINE PROFILE. THIS DIFFERENTIATION DEFECT WAS NOT DUE TO DEFICIENT STAT4 OR STAT6 ACTIVATION BY IL-12 OR IL-4, RESPECTIVELY, NOR WAS IT ASSOCIATED WITH DEFICIENT INDUCTION OF T-BET AND GATA-3 EXPRESSION. IMPAIRED EFFECTOR CYTOKINE PRODUCTION IN ACTIVE RAS-TRANSDUCED CELLS WAS ASSOCIATED WITH DEFICIENT DEMETHYLATION OF THE IL-4 GENE LOCUS. OUR RESULTS INDICATE THAT, DESPITE AUGMENTING ACUTE ACTIVATION OF NAIVE T CELLS, CONSTITUTIVE RAS SIGNALING INHIBITS THE ABILITY OF CD4+ T CELLS TO PROPERLY DIFFERENTIATE INTO TH1/TH2 EFFECTOR CYTOKINE-PRODUCING CELLS, IN PART BY INTERFERING WITH EPIGENETIC MODIFICATION OF EFFECTOR GENE LOCI. ALTERNATIVE STRATEGIES TO POTENTIATE RAS PATHWAY SIGNALING IN T CELLS IN A MORE REGULATED FASHION SHOULD BE CONSIDERED AS A THERAPEUTIC APPROACH TO IMPROVE IMMUNE RESPONSES IN VIVO.	2014