Publications by authors named "Michael D Rosenblum"

78 Publications

Single-cell analyses identify circulating anti-tumor CD8 T cells and markers for their enrichment.

J Exp Med 2021 Apr;218(4)

Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA.

The ability to monitor anti-tumor CD8+ T cell responses in the blood has tremendous therapeutic potential. Here, we used paired single-cell RNA and TCR sequencing to detect and characterize "tumor-matching" (TM) CD8+ T cells in the blood of mice with MC38 tumors or melanoma patients using the TCR as a molecular barcode. TM cells showed increased activation compared with nonmatching T cells in blood and were less exhausted than matching cells in tumors. Importantly, PD-1, which has been used to identify putative circulating anti-tumor CD8+ T cells, showed poor sensitivity for identifying TM cells. By leveraging the transcriptome, we identified candidate cell surface markers for TM cells in mice and patients and validated NKG2D, CD39, and CX3CR1 in mice. These data show that the TCR can be used to identify tumor-relevant cells for characterization, reveal unique transcriptional properties of TM cells, and develop marker panels for tracking and analysis of these cells.
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http://dx.doi.org/10.1084/jem.20200920DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7933992PMC
April 2021

TGF-β: Deletion of TGF-β in T revisited.

Sci Immunol 2021 Feb;6(56)

Department of Dermatology, University of California San Francisco, San Francisco, CA, USA.

T-specific ablation of TGF-β clarifies the complicated role of T-derived TGF-β in immunosuppression in vivo.
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http://dx.doi.org/10.1126/sciimmunol.abg7284DOI Listing
February 2021

Histologic progression of acne inversa/hidradenitis suppurativa: Implications for future investigations and therapeutic intervention.

Exp Dermatol 2020 Dec 30. Epub 2020 Dec 30.

AbbVie Inc, North Chicago, IL, USA.

Since first recognized in 1839, the pathogenesis of acne inversa (AI) has undergone repeated revisions. Although there is agreement that AI involves occlusion of hair follicles with subsequent inflammation and the formation of tracts, the histologic progression of this disease still requires refinement. The objective of this study was to examine the histologic progression of AI based on the examination of a large cohort of punch biopsies and excisional samples that were examined first by hematoxylin and eosin staining. The most informative of these samples were step-sectioned and stained by immunohistochemistry for epithelial and inflammatory markers. Based on this examination, the following observations were made: 1) AI arises from the epithelium of the infundibulum of terminal and vellus hairs; 2) These form cysts and epithelial tendrils that extend into soft tissue; 3) Immunohistochemical staining demonstrates the epithelium of AI is disordered with infundibular and isthmic differentiation and de novo expression of stem cell markers; 4) The inflammatory response in AI is heterogeneous and largely due to cyst rupture. The conclusions of this investigation were that AI is an epithelial-driven disease caused by infiltrative, cyst forming tendrils and most of the inflammation is due to cyst rupture and release of cornified debris and bacteria. Cyst rupture often occurs below the depths of punch biopsy samples indicating their use for analysis may give an incomplete picture of the disease. Finally, our data suggest that unless therapies inhibit tendril development, it is unlikely they will cause prolonged treatment-induced remission in AI.
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http://dx.doi.org/10.1111/exd.14273DOI Listing
December 2020

Single-cell RNA sequencing of psoriatic skin identifies pathogenic Tc17 cell subsets and reveals distinctions between CD8 T cells in autoimmunity and cancer.

J Allergy Clin Immunol 2020 Dec 9. Epub 2020 Dec 9.

Department of Dermatology, University of California San Francisco, San Francisco, Calif. Electronic address:

Background: Psoriasis is an inflammatory, IL-17-driven skin disease in which autoantigen-induced CD8 T cells have been identified as pathogenic drivers.

Objective: Our study focused on comprehensively characterizing the phenotypic variation of CD8 T cells in psoriatic lesions.

Methods: We used single-cell RNA sequencing to compare CD8 T-cell transcriptomic heterogeneity between psoriatic and healthy skin.

Results: We identified 11 transcriptionally diverse CD8 T-cell subsets in psoriatic and healthy skin. Among several inflammatory subsets enriched in psoriatic skin, we observed 2 Tc17 cell subsets that were metabolically divergent, were developmentally related, and expressed CXCL13, which we found to be a biomarker of psoriasis severity and which achieved comparable or greater accuracy than IL17A in a support vector machine classifier of psoriasis and healthy transcriptomes. Despite high coinhibitory receptor expression in the Tc17 cell clusters, a comparison of these cells with melanoma-infiltrating CD8 T cells revealed upregulated cytokine, cytolytic, and metabolic transcriptional activity in the psoriatic cells that differed from an exhaustion program.

Conclusion: Using high-resolution single-cell profiling in tissue, we have uncovered the diverse landscape of CD8 T cells in psoriatic and healthy skin, including 2 nonexhausted Tc17 cell subsets associated with disease severity.
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http://dx.doi.org/10.1016/j.jaci.2020.11.028DOI Listing
December 2020

Developing Human Skin Contains Lymphocytes Demonstrating a Memory Signature.

Cell Rep Med 2020 Nov 3;1(8):100132. Epub 2020 Nov 3.

Department of Dermatology, University of California, San Francisco, San Francisco, CA 94143, USA.

Lymphocytes in barrier tissues play critical roles in host defense and homeostasis. These cells take up residence in tissues during defined developmental windows, when they may demonstrate distinct phenotypes and functions. Here, we utilized mass and flow cytometry to elucidate early features of human skin immunity. Although most conventional αβ T (Tconv) cells in fetal skin have a naive, proliferative phenotype, a subset of CD4 Tconv and CD8 cells demonstrate memory-like features and a propensity for interferon (IFN)γ production. Skin regulatory T cells dynamically accumulate over the second trimester in temporal and regional association with hair follicle development. These fetal skin regulatory T cells (Tregs) demonstrate an effector memory phenotype while differing from their adult counterparts in expression of key effector molecules. Thus, we identify features of prenatal skin lymphocytes that may have key implications for understanding antigen and allergen encounters and in infancy.
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http://dx.doi.org/10.1016/j.xcrm.2020.100132DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7691438PMC
November 2020

Cutting Edge: Tissue Antigen Expression Levels Fine-Tune T Cell Differentiation Decisions In Vivo.

J Immunol 2020 11 9;205(10):2577-2582. Epub 2020 Oct 9.

Department of Biosciences, University of Salzburg, 5020 Salzburg, Austria;

Immune homeostasis in peripheral tissues is, to a large degree, maintained by the differentiation and action of regulatory T cells (Treg) specific for tissue Ags. Using a novel mouse model, we have studied the differentiation of naive CD4 T cells into Foxp3 Treg in response to a cutaneous Ag (OVA). We found that expression of OVA resulted in fatal autoimmunity and in prevention of peripheral Treg generation. Inhibiting mTOR activity with rapamycin rescued the generation of Foxp3 T cells. When we varied the level of Ag expression to modulate TCR signaling, we found that low Ag concentrations promoted the generation of Foxp3 T cells, whereas high levels expanded effector T cells and caused severe autoimmunity. Our findings indicate that the expression level of tissue Ag is a key determinant of the balance between tissue-reactive effector and peripheral Foxp3 T cells, which determines the choice between tolerance and autoimmunity.
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http://dx.doi.org/10.4049/jimmunol.1901094DOI Listing
November 2020

Keepin' it regulatory: Foxp3 gets a BAFfling SWItch.

Sci Immunol 2020 09;5(51)

Department of Dermatology, University of California, San Francisco, CA 94143, USA.

A novel role for SWI/SNF complexes in tuning Foxp3 expression and activity in T.
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http://dx.doi.org/10.1126/sciimmunol.abe5124DOI Listing
September 2020

Contribution of plasma cells and B cells to hidradenitis suppurativa pathogenesis.

JCI Insight 2020 10 2;5(19). Epub 2020 Oct 2.

Division of Rheumatology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan, USA.

Hidradenitis suppurativa (HS) is a debilitating chronic inflammatory skin disease characterized by chronic abscess formation and development of multiple draining sinus tracts in the groin, axillae, and perineum. Using proteomic and transcriptomic approaches, we characterized the inflammatory responses in HS in depth, revealing immune responses centered on IFN-γ, IL-36, and TNF, with lesser contribution from IL-17A. We further identified B cells and plasma cells, with associated increases in immunoglobulin production and complement activation, as pivotal players in HS pathogenesis, with Bruton's tyrosine kinase (BTK) and spleen tyrosine kinase (SYK) pathway activation as a central signal transduction network in HS. These data provide preclinical evidence to accelerate the path toward clinical trials targeting BTK and SYK signaling in moderate-to-severe HS.
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http://dx.doi.org/10.1172/jci.insight.139930DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7566715PMC
October 2020

Immunopathogenesis of hidradenitis suppurativa and response to anti-TNF-α therapy.

JCI Insight 2020 10 2;5(19). Epub 2020 Oct 2.

Department of Dermatology, University of California at San Francisco (UCSF), San Francisco, California, USA.

Hidradenitis suppurativa (HS) is a highly prevalent, morbid inflammatory skin disease with limited treatment options. The major cell types and inflammatory pathways in skin of patients with HS are poorly understood, and which patients will respond to TNF-α blockade is currently unknown. We discovered that clinically and histologically healthy appearing skin (i.e., nonlesional skin) is dysfunctional in patients with HS with a relative loss of immune regulatory pathways. HS skin lesions were characterized by quantitative and qualitative dysfunction of type 2 conventional dendritic cells, relatively reduced regulatory T cells, an influx of memory B cells, and a plasma cell/plasmablast infiltrate predominantly in end-stage fibrotic skin. At the molecular level, there was a relative bias toward the IL-1 pathway and type 1 T cell responses when compared with both healthy skin and psoriatic patient skin. Anti-TNF-α therapy markedly attenuated B cell activation with minimal effect on other inflammatory pathways. Finally, we identified an immune activation signature in skin before anti-TNF-α treatment that correlated with subsequent lack of response to this modality. Our results reveal the fundamental immunopathogenesis of HS and provide a molecular foundation for future studies focused on stratifying patients based on likelihood of clinical response to TNF-α blockade.
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http://dx.doi.org/10.1172/jci.insight.139932DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7566733PMC
October 2020

Exhausted T cell signature predicts immunotherapy response in ER-positive breast cancer.

Nat Commun 2020 07 17;11(1):3584. Epub 2020 Jul 17.

Division of Hematology and Oncology, University of California, San Francisco, CA, USA.

Responses to immunotherapy are uncommon in estrogen receptor (ER)-positive breast cancer and to date, lack predictive markers. This randomized phase II study defines safety and response rate of epigenetic priming in ER-positive breast cancer patients treated with checkpoint inhibitors as primary endpoints. Secondary and exploratory endpoints included PD-L1 modulation and T-cell immune-signatures. 34 patients received vorinostat, tamoxifen and pembrolizumab with no excessive toxicity after progression on a median of five prior metastatic regimens. Objective response was 4% and clinical benefit rate (CR + PR + SD > 6 m) was 19%. T-cell exhaustion (CD8PD-1/CTLA-4) and treatment-induced depletion of regulatory T-cells (CD4 Foxp3/CTLA-4) was seen in tumor or blood in 5/5 patients with clinical benefit, but only in one non-responder. Tumor lymphocyte infiltration was 0.17%. Only two non-responders had PD-L1 expression >1%. This data defines a novel immune signature in PD-L1-negative ER-positive breast cancer patients who are more likely to benefit from immune-checkpoint and histone deacetylase inhibition (NCT02395627).
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http://dx.doi.org/10.1038/s41467-020-17414-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7367885PMC
July 2020

A novel humanized mouse model to study the function of human cutaneous memory T cells in vivo in human skin.

Sci Rep 2020 07 7;10(1):11164. Epub 2020 Jul 7.

Department of Biosciences, University of Salzburg, Salzburg, Austria.

Human skin contains a population of memory T cells that supports tissue homeostasis and provides protective immunity. The study of human memory T cells is often restricted to in vitro studies and to human PBMC serving as primary cell source. Because the tissue environment impacts the phenotype and function of memory T cells, it is crucial to study these cells within their tissue. Here we utilized immunodeficient NOD-scid IL2rγ (NSG) mice that carried in vivo-generated engineered human skin (ES). ES was generated from human keratinocytes and fibroblasts and was initially devoid of skin-resident immune cells. Upon adoptive transfer of human PBMC, this reductionist system allowed us to study human T cell recruitment from a circulating pool of T cells into non-inflamed human skin in vivo. Circulating human memory T cells preferentially infiltrated ES and showed diverse functional profiles of T cells found in fresh human skin. The chemokine and cytokine microenvironment of ES closely resembled that of non-inflamed human skin. Upon entering the ES T cells assumed a resident memory T cell-like phenotype in the absence of infection, and a proportion of these cutaneous T cells can be locally activated upon injection of monocyte derived dendritic cells (moDCs) that presented Candida albicans. Interestingly, we found that CD69 memory T cells produced higher levels of effector cytokines in response to Candida albicans, compared to CD69 T cells. Overall, this model has broad utility in many areas of human skin immunology research, including the study of immune-mediated skin diseases.
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http://dx.doi.org/10.1038/s41598-020-67430-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7341892PMC
July 2020

Layilin augments integrin activation to promote antitumor immunity.

J Exp Med 2020 09;217(9)

Department of Dermatology, University of California, San Francisco, San Francisco, CA.

Tumor-infiltrating CD8+ T cells mediate antitumor immune responses. However, the mechanisms by which T cells remain poised to kill cancer cells despite expressing high levels of inhibitory receptors are unknown. Here, we report that layilin, a C-type lectin domain-containing membrane glycoprotein, is selectively expressed on highly activated, clonally expanded, but phenotypically exhausted CD8+ T cells in human melanoma. Lineage-specific deletion of layilin on murine CD8+ T cells reduced their accumulation in tumors and increased tumor growth in vivo. Congruently, gene editing of LAYN in human CD8+ T cells reduced direct tumor cell killing ex vivo. On a molecular level, layilin colocalized with integrin αLβ2 (LFA-1) on T cells, and cross-linking layilin promoted the activated state of this integrin. Accordingly, LAYN deletion resulted in attenuated LFA-1-dependent cellular adhesion. Collectively, our results identify layilin as part of a molecular pathway in which exhausted or "dysfunctional" CD8+ T cells enhance cellular adhesiveness to maintain their cytotoxic potential.
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http://dx.doi.org/10.1084/jem.20192080DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7478725PMC
September 2020

Tumor Immune Profiling-Based Neoadjuvant Immunotherapy for Locally Advanced Melanoma.

Ann Surg Oncol 2020 Oct 2;27(11):4122-4130. Epub 2020 Jun 2.

Department of Medicine, Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, CA, USA.

Background: The frequency of "exhausted" or checkpoint-positive (PD-1CTLA-4) cytotoxic lymphocytes (Tex) in the tumor microenvironment is associated with response to anti-PD-1 therapy in metastatic melanoma. The current study determined whether pretreatment Tex cells in locally advanced melanoma predicted response to neoadjuvant anti-PD-1 blockade.

Methods: Pretreatment tumor samples from 17 patients with locally advanced melanoma underwent flow cytometric analysis of pretreatment Tex and regulatory T cell frequency. Patients who met the criteria for neoadjuvant checkpoint blockade were treated with either PD-1 monotherapy or PD-1/CTLA-4 combination therapy. Best overall response was evaluated by response evaluation criteria in solid tumors version 1.1, with recurrence-free survival (RFS) calculated by the Kaplan-Meier test. The incidence and severity of adverse events were tabulated by clinicians using the National Cancer Institute Common Terminology Criteria for Adverse Events version 4.

Results: Of the neoadjuvant treated patients, 10 received anti-PD-1 monotherapy and 7 received anti-CTLA-4/PD-1 combination therapy. Of these 17 patients, 12 achieved a complete response, 4 achieved partial responses, and 1 exhibited stable disease. Surgery was subsequently performed for 11 of the 17 patients, and 8 attained a complete pathologic response. Median RFS and overall survival (OS) were not reached. Immune-related adverse events comprised four grade 3 or 4 events, including pneumonitis, transaminitis, and anaphylaxis.

Conclusion: The results showed high rates of objective response, RFS, and OS for patients undergoing immune profile-directed neoadjuvant immunotherapy for locally advanced melanoma. Furthermore, the study showed that treatment stratification based upon Tex frequency can potentially limit the adverse events associated with combination immunotherapy. These data merit further investigation with a larger validation study.
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http://dx.doi.org/10.1245/s10434-020-08648-7DOI Listing
October 2020

Phase II Trial of IL-12 Plasmid Transfection and PD-1 Blockade in Immunologically Quiescent Melanoma.

Clin Cancer Res 2020 06 6;26(12):2827-2837. Epub 2020 May 6.

University of California, San Francisco, San Francisco, California.

Purpose: Tumors with low frequencies of checkpoint positive tumor-infiltrating lymphocytes (cpTIL) have a low likelihood of response to PD-1 blockade. We conducted a prospective multicenter phase II trial of intratumoral plasmid IL-12 (tavokinogene telseplasmid; "tavo") electroporation combined with pembrolizumab in patients with advanced melanoma with low frequencies of checkpoint positive cytotoxic lymphocytes (cpCTL).

Patients And Methods: Tavo was administered intratumorally days 1, 5, and 8 every 6 weeks while pembrolizumab (200 mg, i.v.) was administered every 3 weeks. The primary endpoint was objective response rate (ORR) by RECIST, secondary endpoints included duration of response, overall survival and progression-free survival. Toxicity was evaluated by the CTCAE v4. Extensive correlative analysis was done.

Results: The combination of tavo and pembrolizumab was well tolerated with adverse events similar to those previously reported with pembrolizumab alone. Patients had a 41% ORR ( = 22, RECIST 1.1) with 36% complete responses. Correlative analysis showed that the combination enhanced immune infiltration and sustained the IL-12/IFNγ feed-forward cycle, driving intratumoral cross-presenting dendritic cell subsets with increased TILs, emerging T cell receptor clones and, ultimately, systemic cellular immune responses.

Conclusions: The combination of tavo and pembrolizumab was associated with a higher than expected response rate in this poorly immunogenic population. No new or unexpected toxicities were observed. Correlative analysis showed T cell infiltration with enhanced immunity paralleling the clinical activity in low cpCTL tumors.
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http://dx.doi.org/10.1158/1078-0432.CCR-19-2217DOI Listing
June 2020

Regulatory T cells in skin injury: At the crossroads of tolerance and tissue repair.

Sci Immunol 2020 05;5(47)

Department of Dermatology, University of California, San Francisco, 513 Parnassus Avenue, San Francisco, CA 94143, USA.

Skin injury is a highly inflammatory process that is carefully regulated to mitigate tissue damage and allow for proper barrier repair. Regulatory T cells (T) are crucial coordinators of the immune response to injury in several organs. Here, we review the emerging role of T in facilitating skin repair after injury. We focus on recently discovered interactions between lymphocytes and nonhematopoietic cells during wound healing and discuss how these interactions are regulated both by "classical" suppressive mechanisms of T and by "nonclassical" reparative T functions.
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http://dx.doi.org/10.1126/sciimmunol.aaz9631DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7274208PMC
May 2020

Dermal Adipocyte Lipolysis and Myofibroblast Conversion Are Required for Efficient Skin Repair.

Cell Stem Cell 2020 06 16;26(6):880-895.e6. Epub 2020 Apr 16.

Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT 06511, USA; Department of Dermatology, Yale University, New Haven, CT 06511, USA. Electronic address:

Mature adipocytes store fatty acids and are a common component of tissue stroma. Adipocyte function in regulating bone marrow, skin, muscle, and mammary gland biology is emerging, but the role of adipocyte-derived lipids in tissue homeostasis and repair is poorly understood. Here, we identify an essential role for adipocyte lipolysis in regulating inflammation and repair after injury in skin. Genetic mouse studies revealed that dermal adipocytes are necessary to initiate inflammation after injury and promote subsequent repair. We find through histological, ultrastructural, lipidomic, and genetic experiments in mice that adipocytes adjacent to skin injury initiate lipid release necessary for macrophage inflammation. Tamoxifen-inducible genetic lineage tracing of mature adipocytes and single-cell RNA sequencing revealed that dermal adipocytes alter their fate and generate ECM-producing myofibroblasts within wounds. Thus, adipocytes regulate multiple aspects of repair and may be therapeutic for inflammatory diseases and defective wound healing associated with aging and diabetes.
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http://dx.doi.org/10.1016/j.stem.2020.03.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7853423PMC
June 2020

Sex drives T into fat.

Sci Immunol 2020 04;5(46)

Department of Dermatology, University of California at San Francisco (UCSF), San Francisco, CA 94143, USA. Email:

Androgens promote inflammation in visceral adipose tissue (VAT), leading to the expansion of a distinct IL-33 producing stromal population and recruitment of T.
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http://dx.doi.org/10.1126/sciimmunol.abb8620DOI Listing
April 2020

Regulatory T cells use arginase 2 to enhance their metabolic fitness in tissues.

JCI Insight 2019 12 19;4(24). Epub 2019 Dec 19.

Department of Dermatology.

Distinct subsets of Tregs reside in nonlymphoid tissues where they mediate unique functions. To interrogate the biology of tissue Tregs in human health and disease, we phenotypically and functionally compared healthy skin Tregs with those in peripheral blood, inflamed psoriatic skin, and metastatic melanoma. The mitochondrial enzyme, arginase 2 (ARG2), was preferentially expressed in Tregs in healthy skin, increased in Tregs in metastatic melanoma, and reduced in Tregs from psoriatic skin. ARG2 enhanced Treg suppressive capacity in vitro and conferred a selective advantage for accumulation in inflamed tissues in vivo. CRISPR-mediated deletion of this gene in primary human Tregs was sufficient to skew away from a tissue Treg transcriptional signature. Notably, the inhibition of ARG2 increased mTOR signaling, whereas the overexpression of this enzyme suppressed it. Taken together, our results suggest that Tregs express ARG2 in human tissues to both regulate inflammation and enhance their metabolic fitness.
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http://dx.doi.org/10.1172/jci.insight.129756DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6975275PMC
December 2019

An Injectable Cytokine Trap for Local Treatment of Autoimmune Disease.

Biomaterials 2020 02 12;230:119626. Epub 2019 Nov 12.

Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA, 94158, USA. Electronic address:

Systemic cytokine therapy is limited by toxicity due to activation of unwanted immune cells in off-target tissues. Injectable nanomaterials that interact with the immune system have potential to offer improved pharmacokinetics and cell specificity compared to systemic cytokine therapy by instead capturing and potentiating endogenous cytokine. Here we demonstrate the use of high aspect ratio polycaprolactone nanowires conjugated to cytokine-binding antibodies that assemble into porous matrices when injected into the subcutaneous space. Nanowires are well tolerated in vivo over several weeks, incite minimal foreign body response and resist clearance. Nanowires conjugated with JES6-1, an anti-interleukin-2 (IL-2) antibody, were designed to capture endogenous IL-2 and selectively activate tissue resident regulatory T cells (Tregs). Together these nanowire-antibody matrices were capable of sequestering endogenous IL-2 in the skin and were successful in rebalancing local immune compartments to a more suppressive, Treg-mediated phenotype in both wild type and transgenic murine autoimmune disease models.
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http://dx.doi.org/10.1016/j.biomaterials.2019.119626DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6930339PMC
February 2020

Regulatory T cells in skin are uniquely poised to suppress profibrotic immune responses.

Sci Immunol 2019 09;4(39)

Department of Dermatology, University of California, San Francisco, San Francisco, CA, USA.

At the center of fibrosing diseases is the aberrant activation of tissue fibroblasts. The cellular and molecular mechanisms of how the immune system augments fibroblast activation have been described; however, little is known about how the immune system controls fibroblast function in tissues. Here, we identify regulatory T cells (T) as important regulators of fibroblast activation in skin. Bulk cell and single-cell analysis of T in murine skin and lungs revealed that T in skin are transcriptionally distinct and skewed toward T helper 2 (T2) differentiation. When compared with T in lung, skin T preferentially expressed high levels of GATA3, the master T2 transcription factor. Genes regulated by GATA3 were highly enriched in skin "T2 T" subsets. In functional experiments, T depletion resulted in a preferential increase in T2 cytokine production in skin. Both acute depletion and chronic reduction of T resulted in spontaneous skin fibroblast activation, profibrotic gene expression, and dermal fibrosis, all of which were exacerbated in a bleomycin-induced murine model of skin sclerosis. Lineage-specific deletion of in T resulted in an exacerbation of T2 cytokine secretion that was preferential to skin, resulting in enhanced fibroblast activation and dermal fibrosis. Together, we demonstrate that T play a critical role in regulating fibroblast activation in skin and do so by expressing a unique tissue-restricted transcriptional program that is mediated, at least in part, by GATA3.
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http://dx.doi.org/10.1126/sciimmunol.aaw2910DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6848056PMC
September 2019

Human CD4CD103 cutaneous resident memory T cells are found in the circulation of healthy individuals.

Sci Immunol 2019 07;4(37)

Department of Biosciences, University of Salzburg, Salzburg, Austria.

Tissue-resident memory T cells (T) persist locally in nonlymphoid tissues where they provide frontline defense against recurring insults. T at barrier surfaces express the markers CD103 and/or CD69, which function to retain them in epithelial tissues. In humans, neither the long-term migratory behavior of T nor their ability to reenter the circulation and potentially migrate to distant tissue sites has been investigated. Using tissue explant cultures, we found that CD4CD69CD103 T in human skin can down-regulate CD69 and exit the tissue. In addition, we identified a skin-tropic CD4CD69CD103 population in human lymph and blood that is transcriptionally, functionally, and clonally related to the CD4CD69CD103 T population in the skin. Using a skin xenograft model, we confirmed that a fraction of the human cutaneous CD4CD103 T population can reenter circulation and migrate to secondary human skin sites where they reassume a T phenotype. Thus, our data challenge current concepts regarding the strict tissue compartmentalization of CD4 T cell memory in humans.
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http://dx.doi.org/10.1126/sciimmunol.aav8995DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7057121PMC
July 2019

CD27 Promotes CD4 Effector T Cell Survival in Response to Tissue Self-Antigen.

J Immunol 2019 08 17;203(3):639-646. Epub 2019 Jun 17.

Department of Dermatology, University of California, San Francisco, CA 94143;

Signaling through CD27 plays a role in T cell activation and memory. However, it is currently unknown how this costimulatory receptor influences CD4 effector T (Teff) cells in inflamed tissues. In the current study, we used a murine model of inducible self-antigen expression in the epidermis to elucidate the functional role of CD27 on autoreactive Teff cells. Expression of CD27 on Ag-specific Teff cells resulted in enhanced skin inflammation when compared with CD27-deficient Teff cells. CD27 signaling promoted the accumulation of IFN-γ and IL-2-producing T cells in skin draining lymph nodes in a cell-intrinsic fashion. Surprisingly, this costimulatory pathway had minimal effect on early T cell activation and proliferation. Instead, signaling through CD27 resulted in the progressive survival of Teff cells during the autoimmune response. Using BH3 profiling to assess mitochondrial cell priming, we found that CD27-deficient cells were equally as sensitive as CD27-sufficient cells to mitochondrial outer membrane polarization upon exposure to either BH3 activator or sensitizer peptides. In contrast, CD27-deficient Teff cells expressed higher levels of active caspase 8. Taken together, these results suggest that CD27 does not promote Teff cell survival by increasing expression of antiapoptotic BCL2 family members but instead acts by preferentially suppressing the cell-extrinsic apoptosis pathway, highlighting a previously unidentified role for CD27 in augmenting autoreactive Teff cell responses.
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http://dx.doi.org/10.4049/jimmunol.1900288DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6650327PMC
August 2019

Y'all comeback now.

Sci Immunol 2019 May;4(35)

Department of Dermatology, University of California, San Francisco, CA 94143, USA. Department of Pathology, University of California, San Francisco, CA 94143, USA. Email:

Keratinocytes regulate circulating CD8 T cell memory after cutaneous infection.
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http://dx.doi.org/10.1126/sciimmunol.aax8197DOI Listing
May 2019

Adaptive Immune Resistance Emerges from Tumor-Initiating Stem Cells.

Cell 2019 05 25;177(5):1172-1186.e14. Epub 2019 Apr 25.

Robin Chemers Neustein Laboratory of Mammalian Cell Biology and Development, Howard Hughes Medical Institute, The Rockefeller University, New York, NY 10065, USA. Electronic address:

Our bodies are equipped with powerful immune surveillance to clear cancerous cells as they emerge. How tumor-initiating stem cells (tSCs) that form and propagate cancers equip themselves to overcome this barrier remains poorly understood. To tackle this problem, we designed a skin cancer model for squamous cell carcinoma (SCC) that can be effectively challenged by adoptive cytotoxic T cell transfer (ACT)-based immunotherapy. Using single-cell RNA sequencing (RNA-seq) and lineage tracing, we found that transforming growth factor β (TGF-β)-responding tSCs are superior at resisting ACT and form the root of tumor relapse. Probing mechanism, we discovered that during malignancy, tSCs selectively acquire CD80, a surface ligand previously identified on immune cells. Moreover, upon engaging cytotoxic T lymphocyte antigen-4 (CTLA4), CD80-expressing tSCs directly dampen cytotoxic T cell activity. Conversely, upon CTLA4- or TGF-β-blocking immunotherapies or Cd80 ablation, tSCs become vulnerable, diminishing tumor relapse after ACT treatment. Our findings place tSCs at the crux of how immune checkpoint pathways are activated.
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http://dx.doi.org/10.1016/j.cell.2019.03.025DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6525024PMC
May 2019

Treg-Cell Control of a CXCL5-IL-17 Inflammatory Axis Promotes Hair-Follicle-Stem-Cell Differentiation During Skin-Barrier Repair.

Immunity 2019 03;50(3):655-667.e4

Department of Dermatology, University of California, San Francisco, CA, USA. Electronic address:

Restoration of barrier-tissue integrity after injury is dependent on the function of immune cells and stem cells (SCs) residing in the tissue. In response to skin injury, hair-follicle stem cells (HFSCs), normally poised for hair generation, are recruited to the site of injury and differentiate into cells that repair damaged epithelium. We used a SC fate-mapping approach to examine the contribution of regulatory T (Treg) cells to epidermal-barrier repair after injury. Depletion of Treg cells impaired skin-barrier regeneration and was associated with a Th17 inflammatory response and failed HFSC differentiation. In this setting, damaged epithelial cells preferentially expressed the neutrophil chemoattractant CXCL5, and blockade of CXCL5 or neutrophil depletion restored barrier function and SC differentiation after epidermal injury. Thus, Treg-cell regulation of localized inflammation enables HFSC differentiation and, thereby, skin-barrier regeneration, with implications for the maintenance and repair of other barrier tissues.
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http://dx.doi.org/10.1016/j.immuni.2019.02.013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6507428PMC
March 2019

Regulatory T cells in inflammatory skin disease: from mice to humans.

Int Immunol 2019 07;31(7):457-463

Department of Dermatology, Medical Sciences Building, University of California, San Francisco, CA, USA.

The skin is the largest organ in the body and one of the primary barriers to the environment. In order to optimally protect the host, the skin is home to numerous immune cell subsets that interact with each other and other non-immune cells to maintain organ integrity and function. Regulatory T cells (Tregs) are one of the largest immune cell subsets in skin. They play a critical role in regulating inflammation and facilitating organ repair. In doing so, they adopt unique and specialized tissue-specific functions. In this review, we compare and contrast the role of Tregs in cutaneous immune disorders from mice and humans, with a specific focus on scleroderma, alopecia areata, atopic dermatitis, cutaneous lupus erythematosus and psoriasis.
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http://dx.doi.org/10.1093/intimm/dxz020DOI Listing
July 2019

Inflammatory T cells maintain a healing disposition.

Sci Immunol 2019 01;4(31)

Department of Dermatology, University of California, San Francisco, CA 94143, USA. Email:

Commensal-specific T cells dually possess type-2 and type-17 effector potential, allowing plasticity in orchestrating tissue immunity.
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http://dx.doi.org/10.1126/sciimmunol.aav9723DOI Listing
January 2019

The TNFRSF members CD27 and OX40 coordinately limit T17 differentiation in regulatory T cells.

Sci Immunol 2018 12;3(30)

Department of Dermatology, University of California, San Francisco, CA 94143, USA.

Regulatory T cells (T) are closely related to T17 cells and use aspects of the T17-differentiation program for optimal immune regulation. In several chronic inflammatory human diseases, T express IL-17A, suggesting that dysregulation of T17-associated pathways in T may result in either loss of suppressive function and/or conversion into pathogenic cells. The pathways that regulate the T17 program in T are poorly understood. We have identified two TNF receptor superfamily (TNFRSF) members, CD27 and OX40, that are preferentially expressed by skin-resident T Both CD27 and OX40 signaling suppressed the expression of T17-associated genes from T in a cell-intrinsic manner in vitro and in vivo. However, only OX40 played a nonredundant role in promoting T accumulation. T that lacked both CD27 and OX40 were defective in controlling skin inflammation and expressed high levels of IL-17A, as well as the master T17 transcription factor, RORγt. Last, we found that CD27 expression was inversely correlated with T IL-17 production in skin of patients with psoriasis and hidradenitis suppurativa. Together, our results suggest that TNFRSF members play both redundant and distinct roles in regulating T plasticity in tissues.
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http://dx.doi.org/10.1126/sciimmunol.aau2042DOI Listing
December 2018

Tissue-specific contributions of to atopic dermatitis and mast cell-mediated histaminergic itch.

Proc Natl Acad Sci U S A 2018 12 21;115(51):E12091-E12100. Epub 2018 Nov 21.

Department of Physiology, University of California, San Francisco, CA 94143;

Atopic dermatitis (AD) is the most common skin disease in children. It is characterized by relapsing inflammation, skin-barrier defects, and intractable itch. However, the pathophysiology of itch in AD remains enigmatic. Here, we examine the contribution of , an orphan transmembrane protein linked to AD in both mice and humans. We show that is expressed by both keratinocytes and sensory neurons, but that loss of keratinocytic is sufficient to elicit robust scratching. mice demonstrate an accumulation of dermal mast cells, which are diminished following chronic treatment with cyclooxygenase inhibitors and an EP3 receptor antagonist. In mice, mast cell degranulation produces histaminergic itch in a histamine receptor 1/histamine receptor 4 (H4R/H1R)-dependent manner that may involve activation of TRPV1 afferents. TMEM79 has limited sequence homology to a family of microsomal glutathione transferases and confers protection from cellular accumulation of damaging reactive species, and may thus play a role in regulating oxidative stress. In any case, mechanistic insights from this model suggest that therapeutics targeting PGE and/or H1R/H4R histaminergic signaling pathways may represent useful avenues to treat -associated AD itch. Our findings suggest that individuals with mutations in develop AD due to the loss of protection from oxidative stress.
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http://dx.doi.org/10.1073/pnas.1814132115DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6305010PMC
December 2018

Adoptive Treg Cell Therapy in a Patient With Systemic Lupus Erythematosus.

Arthritis Rheumatol 2019 03 24;71(3):431-440. Epub 2019 Jan 24.

University of California, San Francisco.

Objective: Adoptive Treg cell therapy has great potential to treat autoimmune disease. Currently, very little is known about how these cells impact inflamed tissues. This study was undertaken to elucidate how autologous Treg cell therapy influences tissue inflammation in human autoimmune disease.

Methods: We describe a systemic lupus erythematosus (SLE) patient with active skin disease who received adoptive Treg therapy. We comprehensively quantified Treg cells and immune activation in peripheral blood and skin, with data obtained at multiple time points posttreatment.

Results: Deuterium tracking of infused Treg cells revealed the transient presence of cells in peripheral blood, accompanied by increased percentages of highly activated Treg cells in diseased skin. Flow cytometric analysis and whole transcriptome RNA sequencing revealed that Treg cell accumulation in skin was associated with a marked attenuation of the interferon-γ pathway and a reciprocal augmentation of the interleukin-17 (IL-17) pathway. This phenomenon was more pronounced in skin relative to peripheral blood. To validate these findings, we investigated Treg cell adoptive transfer of skin inflammation in a murine model and found that it also resulted in a pronounced skewing away from Th1 immunity and toward IL-17 production.

Conclusion: We report the first case of a patient with SLE treated with autologous adoptive Treg cell therapy. Taken together, our results suggest that this treatment leads to increased activated Treg cells in inflamed skin, with a dynamic shift from Th1 to Th17 responses.
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http://dx.doi.org/10.1002/art.40737DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6447289PMC
March 2019