1 3716 115 INHIBITING INFLAMMATION WITH MYELOID CELL-SPECIFIC NANOBIOLOGICS PROMOTES ORGAN TRANSPLANT ACCEPTANCE. INDUCING GRAFT ACCEPTANCE WITHOUT CHRONIC IMMUNOSUPPRESSION REMAINS AN ELUSIVE GOAL IN ORGAN TRANSPLANTATION. USING AN EXPERIMENTAL TRANSPLANTATION MOUSE MODEL, WE DEMONSTRATE THAT LOCAL MACROPHAGE ACTIVATION THROUGH DECTIN-1 AND TOLL-LIKE RECEPTOR 4 (TLR4) DRIVES TRAINED IMMUNITY-ASSOCIATED CYTOKINE PRODUCTION DURING ALLOGRAFT REJECTION. WE CONDUCTED NANOIMMUNOTHERAPEUTIC STUDIES AND FOUND THAT A SHORT-TERM MTOR-SPECIFIC HIGH-DENSITY LIPOPROTEIN (HDL) NANOBIOLOGIC TREATMENT (MTORI-HDL) AVERTED MACROPHAGE AEROBIC GLYCOLYSIS AND THE EPIGENETIC MODIFICATIONS UNDERLYING INFLAMMATORY CYTOKINE PRODUCTION. THE RESULTING REGULATORY MACROPHAGES PREVENTED ALLOREACTIVE CD8(+) T CELL-MEDIATED IMMUNITY AND PROMOTED TOLEROGENIC CD4(+) REGULATORY T (TREG) CELL EXPANSION. TO ENHANCE THERAPEUTIC EFFICACY, WE COMPLEMENTED THE MTORI-HDL TREATMENT WITH A CD40-TRAF6-SPECIFIC NANOBIOLOGIC (TRAF6I-HDL) THAT INHIBITS CO-STIMULATION. THIS SYNERGISTIC NANOIMMUNOTHERAPY RESULTED IN INDEFINITE ALLOGRAFT SURVIVAL. TOGETHER, WE SHOW THAT HDL-BASED NANOIMMUNOTHERAPY CAN BE EMPLOYED TO CONTROL MACROPHAGE FUNCTION IN VIVO. OUR STRATEGY, FOCUSED ON PREVENTING INFLAMMATORY INNATE IMMUNE RESPONSES, PROVIDES A FRAMEWORK FOR DEVELOPING TARGETED THERAPIES THAT PROMOTE IMMUNOLOGICAL TOLERANCE. 2018 2 5619 20 SCALING UP PRENATAL NUTRITION COULD REDUCE THE GLOBAL BURDEN OF NONCOMMUNICABLE DISEASES IN THE NEXT GENERATION: A MODELING ANALYSIS. BACKGROUND: NUTRITIONAL CONDITIONS DURING PREGNANCY MAY INFLUENCE THE EPIGENETIC DEVELOPMENT OF AN INDIVIDUAL AND CONSEQUENTLY THEIR LATER-LIFE RISK OF NONCOMMUNICABLE DISEASE (NCD). IMPROVING NUTRITION FOR PREGNANT FEMALES MAY THEREFORE SERVE THE DUAL PURPOSE OF DIRECTLY IMPROVING PREGNANCY OUTCOMES AND PREVENTING NCDS IN THE NEXT GENERATION. OBJECTIVES: WE ESTIMATED THE IMPACT OF PRENATAL SUPPLEMENTATION WITH IRON AND FOLIC ACID (IFA), MULTIPLE MICRONUTRIENTS (MMS), OR CALCIUM AT 50%, 75%, OR 90% COVERAGE ON FUTURE NCDS BY AGE AND SEX IN 2015. METHODS: WE USED SECONDARY DATA SOURCES FROM 132 COUNTRIES TO QUANTIFY THE CASES OF DIABETES AND HYPERTENSION AND THE DEATHS FROM SELECTED NCDS THAT COULD BE AVERTED OR DELAYED BY SCALING UP PRENATAL MICRONUTRIENT SUPPLEMENTATION. RESULTS: GLOBALLY, >51,000 NCD DEATHS, 6 MILLION CASES OF HYPERTENSION, AND 3 MILLION CASES OF DIABETES COULD BE PREVENTED PER OFFSPRING BIRTH COHORT IF MOTHERS WERE PRENATALLY SUPPLEMENTED WITH MMS AT 90% COVERAGE. FOR IFA THESE NUMBERS WOULD BE ROUGHLY HALF. CALCIUM SUPPLEMENTATION AT 90% COULD DELAY 51,000 DEATHS PER BIRTH COHORT. OUR MODEL SUGGESTS THAT SUBSTANTIAL NUMBERS OF NCD DEATHS AND CASES OF HYPERTENSION AND DIABETES COULD BE PREVENTED IN FUTURE GENERATIONS BY SCALING UP MICRONUTRIENT SUPPLEMENTATION FOR MOTHERS DURING PREGNANCY. CONCLUSIONS: HIGHLIGHTING THE ADDITIONAL BENEFITS OF PROVEN NUTRITION INTERVENTIONS IS CRITICAL IN ENSURING ADEQUATE AND SUSTAINED INVESTMENTS, AND PROGRAMMATIC INTEGRATION. AS THE DOUBLE BURDEN OF DISEASE CONTINUES TO GROW, POPULATION-WIDE EFFORTS TO SCALE UP MICRONUTRIENT SUPPLEMENTATION TO PREGNANT FEMALES COULD HELP PREVENT BOTH UNDERNUTRITION AND CHRONIC DISEASE. 2022 3 3861 30 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 4 3373 28 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 5 987 24 CHRONIC SCHISTOSOMIASIS DURING PREGNANCY EPIGENETICALLY REPROGRAMS T-CELL DIFFERENTIATION IN OFFSPRING OF INFECTED MOTHERS. SCHISTOSOMIASIS IS A NONTRANSPLACENTAL HELMINTH INFECTION. CHRONIC INFECTION DURING PREGNANCY SUPPRESSES ALLERGIC AIRWAY RESPONSES IN OFFSPRING. WE ADDRESSED THE QUESTION WHETHER IN UTERO EXPOSURE TO CHRONIC SCHISTOSOME INFECTION (REG PHASE) IN MICE AFFECTS B-CELL AND T-CELL DEVELOPMENT. THEREFORE, WE FOCUSED OUR ANALYSES ON T-CELL DIFFERENTIATION CAPACITY INDUCED BY EPIGENETIC CHANGES IN PROMOTER REGIONS OF SIGNATURE CYTOKINES IN OFFSPRING. HERE, WE SHOW THAT NAIVE T CELLS FROM OFFSPRING OF SCHISTOSOME INFECTED FEMALE MICE HAD A STRONG CAPACITY TO DIFFERENTIATE INTO T(H) 1 CELLS, WHEREAS T(H) 2 DIFFERENTIATION WAS IMPAIRED. IN ACCORDANCE, REDUCED LEVELS OF HISTONE ACETYLATION OF THE IL-4 PROMOTER REGIONS WERE OBSERVED IN NAIVE T CELLS. TO CONCLUDE, OUR MOUSE MODEL REVEALED DISTINCT EPIGENETIC CHANGES WITHIN THE NAIVE T-CELL COMPARTMENT AFFECTING T(H) 2 AND T(H) 1 CELL DIFFERENTIATION IN OFFSPRING OF MOTHERS WITH CHRONIC HELMINTH INFECTION. THESE FINDINGS COULD EVENTUALLY HELP UNDERSTAND HOW HELMINTHS ALTER T-CELL DRIVEN IMMUNE RESPONSES INDUCED BY ALLERGENS, BACTERIAL OR VIRAL INFECTIONS, AS WELL AS VACCINES. 2017 6 1253 25 CURRENT PROBLEMS AND FUTURE DIRECTIONS OF TRANSFUSION-INDUCED ALLOIMMUNIZATION: SUMMARY OF AN NHLBI WORKING GROUP. IN APRIL 2010, A WORKING GROUP SPONSORED BY THE NATIONAL HEART, LUNG, AND BLOOD INSTITUTE WAS ASSEMBLED TO IDENTIFY RESEARCH STRATEGIES TO IMPROVE OUR UNDERSTANDING OF ALLOIMMUNIZATION CAUSED BY THE TRANSFUSION OF ALLOGENEIC BLOOD COMPONENTS AND TO EVALUATE POTENTIAL APPROACHES TO BOTH REDUCE ITS OCCURRENCE AND MANAGE ITS EFFECTS. SIGNIFICANT SEQUELAE OF ALLOIMMUNIZATION WERE DISCUSSED AND IDENTIFIED, INCLUDING DIFFICULTIES IN MAINTAINING CHRONIC TRANSFUSION OF RED BLOOD CELLS AND PLATELETS, HEMOLYTIC DISEASE OF THE NEWBORN, NEONATAL ALLOIMMUNE THROMBOCYTOPENIA, AND REJECTION OF TRANSPLANTED CELLS AND TISSUES. THE DISCUSSIONS RESULTED IN A CONSENSUS THAT IDENTIFIED KEY AREAS OF FUTURE RESEARCH AND DEVELOPMENTAL AREAS, INCLUDING GENETIC AND EPIGENETIC RECIPIENT FACTORS THAT REGULATE ALLOIMMUNIZATION, BIOCHEMICAL SPECIFICS OF TRANSFUSED PRODUCTS THAT AFFECT ALLOIMMUNIZATION, AND NOVEL TECHNOLOGIES FOR HIGH-THROUGHPUT GENOTYPING TO FACILITATE EXTENSIVE AND EFFICIENT ANTIGEN MATCHING BETWEEN DONOR AND RECIPIENT. ADDITIONAL AREAS OF IMPORTANCE INCLUDED ANALYSIS OF UNAPPRECIATED MEDICAL SEQUELAE OF ALLOIMMUNIZATION, SUCH AS CELLULAR IMMUNITY AND ITS EFFECT UPON TRANSPLANT AND AUTOIMMUNITY. IN ADDITION, SUPPORT FOR RESEARCH INFRASTRUCTURE WAS DISCUSSED, WITH AN EMPHASIS ON ENCOURAGING COLLABORATION AND SYNERGY OF ANIMAL MODELS BIOLOGY AND HUMAN CLINICAL RESEARCH. FINALLY, TRAINING FUTURE INVESTIGATORS WAS IDENTIFIED AS AN AREA OF IMPORTANCE. IN AGGREGATE, THIS COMMUNICATION PROVIDES A SYNOPSIS OF THE OPINIONS OF THE WORKING GROUP ON THE ABOVE ISSUES AND PRESENTS BOTH A LIST OF SUGGESTED PRIORITIES AND THE RATIONALE FOR THE TOPICS OF FOCUS. THE AREAS OF RESEARCH IDENTIFIED IN THIS REPORT REPRESENT POTENTIAL FERTILE GROUND FOR THE MEDICAL ADVANCEMENT OF PREVENTING AND MANAGING ALLOIMMUNIZATION IN ITS DIFFERENT FORMS AND MITIGATING THE CLINICAL PROBLEMS IT PRESENTS TO MULTIPLE PATIENT POPULATIONS. 2011 7 6789 22 [CURRENT STATUS OF ATL RESEARCH: EFFORTS FOR PREVENTION AND PRECISION MEDICINE FOR ATL]. THE INTRODUCTION OF NEW AGENTS AND HEMATOPOIETIC STEM CELL TRANSPLANTATION INTO THE TREATMENT OF ATL HAS ACTIVATED ITS CLINICAL RESEARCH. HOWEVER, THE PROGNOSIS OF ATL REMAINS POOR COMPARED WITH THOSE OF OTHER LEUKEMIAS AND LYMPHOMAS. THUS, SEEMINGLY WE HAVE TO RECONSIDER A NEW STRATEGY OF ATL THERAPY BASED ON ITS UNIQUE CHARACTERISTICS. HTLV-1 INFECTION OF T CELLS RESULTS IN CLONAL PROLIFERATION OF INFECTED CELLS THAT ACCUMULATE GENETIC AND EPIGENETIC ABNORMALITIES BEFORE THE ONSET OF ATL. THEREFORE, THE TREATMENT STRATEGY SHOULD INCLUDE THE PREVENTION OF HTLV-1 INFECTION AND ATL DEVELOPMENT IN ADDITION TO PRECISION MEDICINE BASED ON THE STRATIFICATION OF ATL CASES BY BIOMARKERS THAT DISCRIMINATE CLINICAL STAGES OF ATL. I SUMMARIZE HERE THE RECENT PROGRESS IN ATL RESEARCH FOCUSING ON THE BIOMOLECULAR ABNORMALITIES THAT LEAD TO CLONAL EXPANSION AND MALIGNANT TRANSFORMATION OF HTLV-1-INFECTED T CELLS. APPARENTLY, ONE OF THE BASES FOR THE PREVENTION OF ATL IS TO ESTABLISH A DISEASE ENTITY OF "CHRONIC ACTIVE HTLV-1 INFECTION" THAT DEFINES HIGH-RISK CARRIERS FOR ATL DEVELOPMENT AND ENABLES PREVENTIVE INTERVENTION. 2017 8 4760 27 NOVEL TREATMENTS OF ADULT T CELL LEUKEMIA LYMPHOMA. ADULT T CELL LEUKEMIA-LYMPHOMA (ATL) IS AN AGGRESSIVE MALIGNANCY SECONDARY TO CHRONIC INFECTION WITH THE HUMAN T CELL LEUKEMIA VIRUS TYPE I (HTLV-I) RETROVIRUS. ATL CARRIES A DISMAL PROGNOSIS. ATL CLASSIFIES INTO FOUR SUBTYPES (ACUTE, LYMPHOMA, CHRONIC, AND SMOLDERING) WHICH DISPLAY DIFFERENT CLINICAL FEATURES, PROGNOSIS AND RESPONSE TO THERAPY, HENCE REQUIRING DIFFERENT CLINICAL MANAGEMENT. SMOLDERING AND CHRONIC SUBTYPES RESPOND WELL TO ANTIRETROVIRAL THERAPY USING THE COMBINATION OF ZIDOVUDINE (AZT) AND INTERFERON-ALPHA (IFN) WITH A SIGNIFICANT PROLONGATION OF SURVIVAL. CONVERSELY, THE WATCH AND WAIT STRATEGY OR CHEMOTHERAPY FOR THESE INDOLENT SUBTYPES ALLIES WITH A POOR LONG-TERM OUTCOME. ACUTE ATL IS ASSOCIATED WITH CHEMO-RESISTANCE AND DISMAL PROGNOSIS. LYMPHOMA SUBTYPES RESPOND BETTER TO INTENSIVE CHEMOTHERAPY BUT SURVIVAL REMAINS POOR. ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION (HSCT) RESULTS IN LONG-TERM SURVIVAL IN ROUGHLY ONE THIRD OF TRANSPLANTED PATIENTS BUT ONLY A SMALL PERCENTAGE OF PATIENTS CAN MAKE IT TO TRANSPLANT. OVERALL, CURRENT TREATMENTS OF AGGRESSIVE ATL ARE NOT SATISFACTORY. PROGNOSIS OF REFRACTORY OR RELAPSED PATIENTS IS DISMAL WITH SOME ENCOURAGING RESULTS WHEN USING LENALIDOMIDE OR MOGAMULIZUMAB. TO OVERCOME RESISTANCE AND PREVENT RELAPSE, PRECLINICAL OR PILOT CLINICAL STUDIES USING TARGETED THERAPIES SUCH AS ARSENIC/IFN, MONOCLONAL ANTIBODIES, EPIGENETIC THERAPIES ARE PROMISING BUT WARRANT FURTHER CLINICAL INVESTIGATION. ANTI-ATL VACCINES INCLUDING TAX PEPTIDE-PULSED DENDRITIC CELLS, INDUCED TAX-SPECIFIC CTL RESPONSES IN ATL PATIENTS. FINALLY, BASED ON THE PROGRESS IN UNDERSTANDING THE PATHOPHYSIOLOGY OF ATL, AND THE RISK-ADAPTED TREATMENT APPROACHES TO DIFFERENT ATL SUBTYPES, TREATMENT STRATEGIES OF ATL SHOULD TAKE INTO ACCOUNT THE HOST IMMUNE RESPONSES AND THE HOST MICROENVIRONMENT INCLUDING HTLV-1 INFECTED NON-MALIGNANT CELLS. HEREIN, WE WILL PROVIDE A SUMMARY OF NOVEL TREATMENTS OF ATL IN VITRO, IN VIVO, AND IN EARLY CLINICAL TRIALS. 2020 9 2242 27 EPIGENETIC MODULATION OF CD8(+) T CELL FUNCTION IN LENTIVIRUS INFECTIONS: A REVIEW. CD8(+) T CELLS ARE CRITICAL FOR CONTROLLING VIREMIA DURING HUMAN IMMUNODEFICIENCY VIRUS (HIV) INFECTION. THESE CELLS PRODUCE CYTOLYTIC FACTORS AND ANTIVIRAL CYTOKINES THAT ELIMINATE VIRALLY- INFECTED CELLS. DURING THE CHRONIC PHASE OF HIV INFECTION, CD8(+) T CELLS PROGRESSIVELY LOSE THEIR PROLIFERATIVE CAPACITY AND ANTIVIRAL FUNCTIONS. THESE DYSFUNCTIONAL CELLS ARE UNABLE TO CLEAR THE PRODUCTIVELY INFECTED AND REACTIVATED CELLS, REPRESENTING A ROADBLOCK IN HIV CURE. THEREFORE, MECHANISMS TO UNDERSTAND CD8(+) T CELL DYSFUNCTION AND STRATEGIES TO BOOST CD8(+) T CELL FUNCTION NEED TO BE INVESTIGATED. USING THE FELINE IMMUNODEFICIENCY VIRUS (FIV) MODEL FOR LENTIVIRAL PERSISTENCE, WE HAVE DEMONSTRATED THAT CD8(+) T CELLS EXHIBIT EPIGENETIC CHANGES SUCH AS DNA DEMETHYLATION DURING THE COURSE OF INFECTION AS COMPARED TO UNINFECTED CATS. WE HAVE ALSO DEMONSTRATED THAT LENTIVIRUS-ACTIVATED CD4(+)CD25(+) T REGULATORY CELLS INDUCE FORKHEAD BOX P3 (FOXP3) EXPRESSION IN VIRUS-SPECIFIC CD8(+) T CELL TARGETS, WHICH BINDS THE INTERLEUKIN (IL)-2, TUMOR NECROSIS FACTOR (TNF)-Α, AND INTERFERON (IFN)-Γ PROMOTERS IN THESE CD8(+) T CELLS. FINALLY, WE HAVE REPORTED THAT EPIGENETIC MODULATION REDUCES FOXP3 BINDING TO THESE PROMOTER REGIONS. THIS REVIEW COMPARES AND CONTRASTS OUR CURRENT UNDERSTANDING OF CD8(+) T CELL EPIGENETICS AND MECHANISMS OF LYMPHOCYTE SUPPRESSION DURING THE COURSE OF LENTIVIRAL INFECTION FOR TWO ANIMAL MODELS, FIV AND SIMIAN IMMUNODEFICIENCY VIRUS (SIV). 2018 10 787 18 CELLS OF ADULT T-CELL LEUKEMIA EVADE HTLV-1 TAX/NF-KAPPAB HYPERACTIVATION-INDUCED SENESCENCE. HUMAN T-CELL LEUKEMIA VIRUS TYPE 1 (HTLV-1) IS THE ETIOLOGICAL AGENT OF ADULT T-CELL LEUKEMIA/LYMPHOMA (ATL). THE HTLV-1 VIRAL TRANS-ACTIVATOR/ONCOPROTEIN TAX IS A MAJOR DRIVER OF ATL, YET IT INDUCES RAPID P21(CIP1/WAF1) (P21)- AND P27(KIP1)-MEDIATED CELLULAR SENESCENCE THROUGH CONSTITUTIVE ACTIVATION (HYPERACTIVATION) OF NF-KAPPAB. ALTHOUGH CONSTITUTIVE NF-KAPPAB ACTIVATION IS A COMMON FEATURE OF T/B-CELL LEUKEMIA/LYMPHOMA, INCLUDING ATL, IT IS NOT KNOWN HOW ATL CELLS MAINTAIN CHRONIC NF-KAPPAB ACTIVATION WITHOUT UNDERGOING SENESCENCE. HERE, WE DEMONSTRATE THAT, IN CONTRAST TO HTLV-1(-) T-CELL LINES, ATL CELL LINES NO LONGER UNDERGO TAX-INDUCED SENESCENCE. ALTHOUGH TAX(+) AND TAX(-) ATL CELL LINES SHOWED SIGNATURES OF CONSTITUTIVE NF-KAPPAB ACTIVATION, THEIR ABILITY TO PROGRESS THROUGH THE CELL CYCLE WAS UNAFFECTED. IN SOME CASES, ATL CELL LINES CONTINUED TO PROLIFERATE DESPITE SIGNIFICANT UPREGULATION OF P21; ADDITIONALLY, MANY CELL LINES DISPLAYED ALTERED EXPRESSION OF G1 AND G1/S CYCLINS, PARTICULARLY OVEREXPRESSION OF CYCLIN D2. WE PROPOSE THAT, DURING THE COURSE OF ATL DEVELOPMENT, LEUKEMIA CELLS ACQUIRE GENETIC/EPIGENETIC CHANGES THAT CAN MITIGATE THE SENESCENCE RESPONSE TRIGGERED BY NF-KAPPAB HYPERACTIVATION. RESTORING THE NF-KAPPAB-INDUCED SENESCENCE RESPONSE WOULD LIKELY HELP TO CONTROL THE DEVELOPMENT AND PROGRESSION OF ATL AND SIMILAR LYMPHOID MALIGNANCIES. 2019 11 3454 29 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 12 5900 31 T-CELL EXHAUSTION IN ORGAN TRANSPLANTATION. EXHAUSTION OF T CELLS OCCURS IN RESPONSE TO LONG-TERM EXPOSURE TO SELF AND FOREIGN ANTIGENS. IT LIMITS T CELL CAPACITY TO PROLIFERATE AND PRODUCE CYTOKINES, LEADING TO AN IMPAIRED ABILITY TO CLEAR CHRONIC INFECTIONS OR ERADICATE TUMORS. T-CELL EXHAUSTION IS ASSOCIATED WITH A SPECIFIC TRANSCRIPTIONAL, EPIGENETIC, AND METABOLIC PROGRAM AND CHARACTERISTIC CELL SURFACE MARKERS' EXPRESSION. RECENT STUDIES HAVE BEGUN TO ELUCIDATE THE ROLE OF T-CELL EXHAUSTION IN TRANSPLANT. HIGHER LEVELS OF EXHAUSTED T CELLS HAVE BEEN ASSOCIATED WITH BETTER GRAFT FUNCTION IN KIDNEY TRANSPLANT RECIPIENTS. IN CONTRAST, REINVIGORATING EXHAUSTED T CELLS BY IMMUNE CHECKPOINT BLOCKADE THERAPIES, WHILE PROMOTING TUMOR CLEARANCE, INCREASES THE RISK OF ACUTE REJECTION. LYMPHOCYTE DEPLETION AND HIGH ALLOANTIGEN LOAD HAVE BEEN IDENTIFIED AS MAJOR DRIVERS OF T-CELL EXHAUSTION. THIS COULD ACCOUNT, AT LEAST IN PART, FOR THE REDUCED RATES OF ACUTE REJECTION IN ORGAN TRANSPLANT RECIPIENTS INDUCED WITH THYMOGLOBULIN AND FOR THE PRO-TOLEROGENIC EFFECTS OF A LARGE ORGAN SUCH AS THE LIVER. AMONG THE DRUGS THAT ARE WIDELY USED FOR MAINTENANCE IMMUNOSUPPRESSION, CALCINEURIN INHIBITORS HAVE A CONTRASTING INHIBITORY EFFECT ON EXHAUSTION OF T CELLS, WHILE THE INFLUENCE OF MTOR INHIBITORS IS STILL UNCLEAR. HARNESSING OR ENCOURAGING THE NATURAL PROCESSES OF EXHAUSTION MAY PROVIDE A NOVEL STRATEGY TO PROMOTE GRAFT SURVIVAL AND TRANSPLANTATION TOLERANCE. 2022 13 923 21 CHRONIC INFECTION DRIVES DNMT3A-LOSS-OF-FUNCTION CLONAL HEMATOPOIESIS VIA IFNGAMMA SIGNALING. AGE-RELATED CLONAL HEMATOPOIESIS (CH) IS A RISK FACTOR FOR MALIGNANCY, CARDIOVASCULAR DISEASE, AND ALL-CAUSE MORTALITY. SOMATIC MUTATIONS IN DNMT3A ARE DRIVERS OF CH, BUT DECADES MAY ELAPSE BETWEEN THE ACQUISITION OF A MUTATION AND CH, SUGGESTING THAT ENVIRONMENTAL FACTORS CONTRIBUTE TO CLONAL EXPANSION. WE TESTED WHETHER INFECTION PROVIDES SELECTIVE PRESSURE FAVORING THE EXPANSION OF DNMT3A MUTANT HEMATOPOIETIC STEM CELLS (HSCS) IN MOUSE CHIMERAS. WE CREATED DNMT3A-MOSAIC MICE BY TRANSPLANTING DNMT3A(-/-) AND WT HSCS INTO WT MICE AND OBSERVED THE SUBSTANTIAL EXPANSION OF DNMT3A(-/-) HSCS DURING CHRONIC MYCOBACTERIAL INFECTION. INJECTION OF RECOMBINANT IFNGAMMA ALONE WAS SUFFICIENT TO PHENOCOPY CH BY DNMT3A(-/-) HSCS UPON INFECTION. TRANSCRIPTIONAL AND EPIGENETIC PROFILING AND FUNCTIONAL STUDIES INDICATE REDUCED DIFFERENTIATION ASSOCIATED WITH WIDESPREAD METHYLATION ALTERATIONS, AND REDUCED SECONDARY STRESS-INDUCED APOPTOSIS ACCOUNTS FOR DNMT3A(-/-) CLONAL EXPANSION DURING INFECTION. DNMT3A MUTANT HUMAN HSCS SIMILARLY EXHIBIT DEFECTIVE IFNGAMMA-INDUCED DIFFERENTIATION. WE THUS DEMONSTRATE THAT IFNGAMMA SIGNALING INDUCED DURING CHRONIC INFECTION CAN DRIVE DNMT3A-LOSS-OF-FUNCTION CH. 2021 14 2421 20 EPIGENETIC SIGNATURE OF PD-1+ TCF1+ CD8 T CELLS THAT ACT AS RESOURCE CELLS DURING CHRONIC VIRAL INFECTION AND RESPOND TO PD-1 BLOCKADE. WE HAVE RECENTLY DEFINED A NOVEL POPULATION OF PD-1 (PROGRAMMED CELL DEATH 1)+ TCF1 (T CELL FACTOR 1)+ VIRUS-SPECIFIC CD8 T CELLS THAT FUNCTION AS RESOURCE CELLS DURING CHRONIC LCMV INFECTION AND PROVIDE THE PROLIFERATIVE BURST SEEN AFTER PD-1 BLOCKADE. SUCH CD8 T CELLS HAVE BEEN FOUND IN OTHER CHRONIC INFECTIONS AND ALSO IN CANCER IN MICE AND HUMANS. THESE CD8 T CELLS EXHIBIT STEM-LIKE PROPERTIES UNDERGOING SELF-RENEWAL AND ALSO DIFFERENTIATING INTO THE TERMINALLY EXHAUSTED CD8 T CELLS. HERE WE COMPARED THE EPIGENETIC SIGNATURE OF STEM-LIKE CD8 T CELLS WITH EXHAUSTED CD8 T CELLS. ATAC-SEQ ANALYSIS SHOWED THAT STEM-LIKE CD8 T CELLS HAD A UNIQUE SIGNATURE IMPLICATING ACTIVITY OF HMG (TCF) AND RHD (NF-KAPPAB) TRANSCRIPTION FACTOR FAMILY MEMBERS IN CONTRAST TO HIGHER ACCESSIBILITY TO ETS AND RUNX MOTIFS IN EXHAUSTED CD8 T CELLS. IN ADDITION, REGULATORY REGIONS OF THE TRANSCRIPTION FACTORS TCF7 AND ID3 WERE MORE ACCESSIBLE IN STEM-LIKE CELLS WHEREAS PRDM1 AND ID2 WERE MORE ACCESSIBLE IN EXHAUSTED CD8 T CELLS. WE ALSO COMPARED THE EPIGENETIC SIGNATURES OF THE 2 CD8 T CELL SUBSETS FROM CHRONICALLY INFECTED MICE WITH EFFECTOR AND MEMORY CD8 T CELLS GENERATED AFTER AN ACUTE LCMV INFECTION. BOTH CD8 T CELL SUBSETS GENERATED DURING CHRONIC INFECTION WERE STRIKINGLY DIFFERENT FROM CD8 T CELL SUBSETS FROM ACUTE INFECTION. INTERESTINGLY, THE STEM-LIKE CD8 T CELL SUBSET FROM CHRONIC INFECTION, DESPITE SHARING KEY FUNCTIONAL PROPERTIES WITH MEMORY CD8 T CELLS, HAD A VERY DISTINCT EPIGENETIC PROGRAM. THESE RESULTS SHOW THAT THE CHRONIC STEM-LIKE CD8 T CELL PROGRAM REPRESENTS A SPECIFIC ADAPTATION OF THE T CELL RESPONSE TO PERSISTENT ANTIGENIC STIMULATION. 2019 15 5283 19 PROPHYLACTIC OR PREEMPTIVE LOW-DOSE AZACITIDINE AND DONOR LYMPHOCYTE INFUSION TO PREVENT DISEASE RELAPSE FOLLOWING ALLOGENEIC TRANSPLANTATION IN PATIENTS WITH HIGH-RISK ACUTE MYELOGENOUS LEUKEMIA OR MYELODYSPLASTIC SYNDROME. BECAUSE OF THE PERSISTENTLY HIGH RATES OF RELAPSE OF PATIENTS WITH HIGH-RISK ACUTE MYELOGENOUS LEUKEMIA (AML) AND MYELODYSPLASTIC SYNDROME (MDS) FOLLOWING ALLOGENEIC HEMATOPOIETIC STEM CELL TRANSPLANTATION (ALLO-HSCT), POST-TRANSPLANTATION MAINTENANCE THERAPY HAS BEEN PROPOSED. WE PREVIOUSLY INITIATED A PHASE II TRIAL IN WHICH EPIGENETIC THERAPY WAS COMBINED WITH IMMUNOTHERAPY IN AN ATTEMPT TO REDUCE DISEASE RELAPSE. IN THAT STUDY, LOW-DOSE AZACITIDINE (AZA) AND ESCALATING DOSES OF DONOR LYMPHOCYTE INFUSION (DLI) WERE GIVEN AS POST-ALLO-HSCT MAINTENANCE TREATMENT. IN THE PRESENT STUDY, WE RETROSPECTIVELY ANALYZE A LARGER COHORT OF PATIENTS RECEIVING POST-TRANSPLANTATION MAINTENANCE THERAPY AND PROVIDE UPDATES ON SOME PATIENTS OF THE EARLIER STUDY. THE OBJECTIVES OF THE PRESENT STUDY WERE TO ANALYZE THE CUMULATIVE INCIDENCE OF RELAPSE (CIR), OVERALL SURVIVAL (OS), AND PROGRESSION-FREE SURVIVAL (PFS) AND THE INCIDENCE OF ACUTE AND CHRONIC GRAFT-VERSUS-HOST DISEASE (GVHD) OF PATIENTS WITH HIGH-RISK AML OR MDS RECEIVING POST-TRANSPLANTATION MAINTENANCE TREATMENT WITH AZA WITH OR WITHOUT DLI. WE RETROSPECTIVELY ANALYZED 77 PATIENTS (54 WITH AML, 23 WITH MDS) CONSIDERED AT HIGH RISK BASED ON EITHER THEIR GENOMIC OR CLINICAL STATUS AT TRANSPLANTATION. FOLLOWING ALLOGENEIC TRANSPLANTATION, THEY RECEIVED AT LEAST 1 CYCLE OF PROPHYLACTIC OR PREEMPTIVE LOW-DOSE AZA WITH OR WITHOUT ESCALATING DOSES OF DLI TO PREVENT DISEASE RELAPSE. ALMOST ONE-HALF OF THE PATIENTS (47%) WERE ABLE TO RECEIVE THE FULL 12 CYCLES OF SCHEDULED AZA, AND A MAJORITY (79%) RECEIVED AT LEAST 1 DLI. WITH A MEDIAN FOLLOW-UP OF 24 MONTHS, 19 PATIENTS (25%; 16 WITH AML, 3 WITH MDS) RELAPSED, AT A MEDIAN OF 9.8 MONTHS (RANGE, 4 TO 58.6 MONTHS), GIVING A 22% CIR AT 24 MONTHS. OS AND PFS AT 24 MONTHS WERE 70.8% AND 68.3%, RESPECTIVELY. THE CUMULATIVE INCIDENCES OF GRADE II-IV ACUTE GVHD AND CHRONIC GVHD WERE 27.4% AND 45%, RESPECTIVELY. ONLY A MINORITY OF PATIENTS (11%) REQUIRED DELAYED ADMINISTRATION OF AZA. THESE FINDINGS CONFIRM THAT AZA-DLI MAINTENANCE IS BOTH TOLERABLE AND EFFECTIVE IN REDUCING THE RISK OF POST-TRANSPLANTATION RELAPSE. 2021 16 5147 31 POTENTIAL ROLES OF LONG NONCODING RNAS AS THERAPEUTIC TARGETS IN ORGAN TRANSPLANTATION. ORGAN TRANSPLANTATION IS THE MOST PREFERRED TREATMENT OPTION FOR END-STAGE ORGAN DISEASES; HOWEVER, ALLOGRAFT REJECTION IS THE MAJOR HURDLE IN SUCCESSFUL LONG-TERM TRANSPLANT SURVIVAL. IN SPITE OF DEVELOPING BETTER HLA MATCHING AND MORE EFFECTIVE IMMUNOSUPPRESSIVE REGIMEN, ONE-YEAR GRAFT SURVIVAL HAS BEEN INCREASED BY NEARLY 90% AND THE INCIDENCE OF ACUTE REJECTION BY ONE-YEAR POST-TRANSPLANTATION HAS BEEN DECREASED BY 12.2% IN THE LAST DECADES, CHRONIC ALLOGRAFT REJECTION HAS REMAINED AS ONE OF THE MAJOR OBSTACLES TO THE LONG-LASTING SURVIVAL OF THE TRANSPLANTED ALLOGRAFT. THEREFORE, SEEMINGLY PREVENTING THE ALLOGRAFT REJECTION AND INDUCING IMMUNOLOGICAL TOLERANCE AGAINST TRANSPLANTED ALLOGRAFTS IS ONE OF THE PRIMARY GOALS IN TRANSPLANTATION RESEARCH TO ENABLE LONG-LASTING GRAFT SURVIVAL. VARIOUS MECHANISMS SUCH AS LONG NONCODING RNAS (LNCRNAS) HAVE BEEN PROPOSED THAT INDUCE IMMUNE TOLERANCE BY MODULATING THE GENE EXPRESSION AND REGULATING INNATE AND ADAPTIVE IMMUNE RESPONSES DURING TRANSPLANTATION. BESIDES, BECAUSE OF INVOLVEMENT IN REGULATING EPIGENETIC, TRANSCRIPTIONAL, AND POST-TRANSLATIONAL MECHANISMS, LNCRNAS COULD AFFECT ALLOGRAFT STATUS. THEREFORE, THESE MOLECULES COULD BE CONSIDERED AS THE POTENTIAL TARGETS FOR PREDICTION, PROGNOSIS, DIAGNOSIS, AND TREATMENT OF GRAFT REJECTION. IT IS SUGGESTED THAT THE NONINVASIVE PREDICTIVE BIOMARKERS HOLD PROMISE TO OVERCOME THE CURRENT LIMITATIONS OF CONVENTIONAL TISSUE BIOPSY IN THE DIAGNOSIS OF REJECTION. HENCE, THIS REVIEW AIMS TO PROVIDE A COMPREHENSIVE OVERVIEW OF LNCRNAS AND THEIR FUNCTION TO FACILITATE DIAGNOSIS, PROGNOSIS, AND PREDICTION OF THE RISK OF GRAFT REJECTION, AND THE SUGGESTIVE THERAPEUTIC CHOICES AFTER TRANSPLANTATION. 2022 17 5015 22 PERSISTENCE OF SELF-REACTIVE CD8+ T CELLS IN THE CNS REQUIRES TOX-DEPENDENT CHROMATIN REMODELING. SELF-REACTIVE CD8(+) T CELLS ARE IMPORTANT MEDIATORS OF PROGRESSIVE TISSUE DAMAGE IN AUTOIMMUNE DISEASES, BUT THE MOLECULAR PROGRAM UNDERLYING THESE CELLS' FUNCTIONAL ADAPTATION IS UNCLEAR. HERE WE CHARACTERIZE THE TRANSCRIPTIONAL AND EPIGENETIC LANDSCAPE OF SELF-REACTIVE CD8(+) T CELLS IN A MOUSE MODEL OF PROTRACTED CENTRAL NERVOUS SYSTEM (CNS) AUTOIMMUNITY AND COMPARE IT TO POPULATIONS OF CNS-RESIDENT MEMORY CD8(+) T CELLS EMERGING FROM ACUTE VIRAL INFECTION. WE FIND THAT AUTOIMMUNE CD8(+) T CELLS PERSISTING AT SITES OF SELF-ANTIGEN EXHIBIT CHARACTERISTIC TRANSCRIPTIONAL REGULATION TOGETHER WITH DISTINCT EPIGENETIC REMODELING. THIS SELF-REACTIVE CD8(+) T CELL FATE DEPENDS ON THE TRANSCRIPTIONAL REGULATION BY THE DNA-BINDING HMG-BOX PROTEIN TOX WHICH REMODELS MORE THAN 400 GENOMIC REGIONS INCLUDING LOCI SUCH AS TCF7, WHICH IS CENTRAL TO STEMNESS OF CD8(+) T CELLS. CONTINUOUS EXPOSURE TO CNS SELF-ANTIGEN SUSTAINS TOX LEVELS IN SELF-REACTIVE CD8(+) T CELLS, WHEREAS GENETIC ABLATION OF TOX IN CD8(+) T CELLS RESULTS IN SHORTENED PERSISTENCE OF SELF-REACTIVE CD8(+) T CELLS IN THE INFLAMED CNS. OUR STUDY ESTABLISHES AND CHARACTERIZES THE GENETIC DIFFERENTIATION PROGRAM ENABLING CHRONIC T CELL-DRIVEN IMMUNOPATHOLOGY IN CNS AUTOIMMUNITY. 2021 18 5908 22 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 19 6071 28 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 20 5704 22 SINGLE-CELL RNA-SEQ REVEALS TOX AS A KEY REGULATOR OF CD8(+) T CELL PERSISTENCE IN CHRONIC INFECTION. PROGENITOR-LIKE CD8(+) T CELLS MEDIATE LONG-TERM IMMUNITY TO CHRONIC INFECTION AND CANCER AND RESPOND POTENTLY TO IMMUNE CHECKPOINT BLOCKADE. THESE CELLS SHARE TRANSCRIPTIONAL REGULATORS WITH MEMORY PRECURSOR CELLS, INCLUDING T CELL-SPECIFIC TRANSCRIPTION FACTOR 1 (TCF1), BUT IT IS UNCLEAR WHETHER THEY ADOPT DISTINCT PROGRAMS TO ADAPT TO THE IMMUNOSUPPRESSIVE ENVIRONMENT. BY COMPARING THE SINGLE-CELL TRANSCRIPTOMES AND EPIGENETIC PROFILES OF CD8(+) T CELLS RESPONDING TO ACUTE AND CHRONIC VIRAL INFECTIONS, WE FOUND THAT PROGENITOR-LIKE CD8(+) T CELLS BECAME DISTINCT FROM MEMORY PRECURSOR CELLS BEFORE THE PEAK OF THE T CELL RESPONSE. WE DISCOVERED A COEXPRESSION GENE MODULE CONTAINING TOX THAT EXHIBITED HIGHER TRANSCRIPTIONAL ACTIVITY ASSOCIATED WITH MORE ABUNDANT ACTIVE HISTONE MARKS IN PROGENITOR-LIKE CELLS THAN MEMORY PRECURSOR CELLS. MOREOVER, THYMOCYTE SELECTION-ASSOCIATED HIGH MOBILITY GROUP BOX PROTEIN TOX (TOX) PROMOTED THE PERSISTENCE OF ANTIVIRAL CD8(+) T CELLS AND WAS REQUIRED FOR THE PROGRAMMING OF PROGENITOR-LIKE CD8(+) T CELLS. THUS, LONG-TERM CD8(+) T CELL IMMUNITY TO CHRONIC VIRAL INFECTION REQUIRES UNIQUE TRANSCRIPTIONAL AND EPIGENETIC PROGRAMS ASSOCIATED WITH THE TRANSCRIPTION FACTOR TOX. 2019