scholarly journals OP0023 GENERATION OF PT101, A HIGHLY SELECTIVE IL-2 MUTEIN FOR TREATMENT OF AUTOIMMUNE DISEASES

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 13.1-13
Author(s):  
J. Visweswaraiah ◽  
E. Sampson ◽  
P. Petaipimol ◽  
A. Monsef ◽  
K. Kis-Toth ◽  
...  
Keyword(s):  
Clinical Trial ◽  
T Cells ◽  
Natural Killer Cells ◽  
Autoimmune Diseases ◽  
Natural Killer ◽  
Cytokine Production ◽  
Immune Cell ◽  
Cell Types ◽  
Killer Cells

Background:Regulatory T cells (Tregs) play a critical role in immune homeostasis and are dysfunctional in many autoimmune diseases. Interleukin 2 (IL-2) drives the proliferation and function of Tregs. via the heterotrimeric IL-2 receptor (CD25/CD122/CD132). As a result, CD25 loss-of-function in mice is associated with Treg deficiency and widespread autoimmunity. Low dose IL-2 is being evaluated for treatment of autoimmune diseases and has been shown to expand Tregs, however it has a narrow selectivity window before activating conventional T cells and natural killer cells. To enhance IL-2 selectivity and improve its therapeutic utility for activating and expanding Tregs, mutations can be introduced that reduce CD122/CD132 affinity thus creating a dependency on CD25 binding for signaling through CD122/CD132 upon IL-2 facilitated CD25/CD122/CD132 trimer formation.Objectives:To generate a highly selective IL-2 mutein that activates and expands Tregs selectively that can be used for treatment of autoimmune diseasesMethods:Using a structure guided approach, we introduced mutations in IL-2 that significantly decreased CD122 binding affinity in addition to other mutations that increased CD25 binding affinity. Finally, we explored additional mutations, format, orientation, and linker lengths to generate the most potent, selective molecule with drug-like manufacturability. These structure activity relationship efforts culminated in the generation of PT101, a mutant IL-2 Fc fusion that is selective in activating and expanding Tregs in vitro and in vivo.Results:PT101 selectively induced STAT5 phosphorylation in human and cynomolgus monkey Tregs in vitro. In humanized NOD-scid IL-2Rg-null (NSG) mice and cynomolgus monkeys, administration of PT101 dose-dependently and selectively expanded Tregs without significant effects on other immune cell types, and without eliciting proinflammatory cytokine production. The Tregs from PT101-dosed humanized mice have increased expression of FOXP3 and CD25, suggesting enhanced function and stability. In a Phase 1a single ascending dose clinical trial, PT101 was well-tolerated and selectively expanded total Tregs, with a mean maximal increase of up to 3.6-fold over baseline in healthy volunteers. There was no evidence of expansion of natural killer cells nor pro-inflammatory conventional T cells at any of the doses studied.Conclusion:PT101 selectively activated and expanded Tregs without significant effects on other immune cell types, and without eliciting proinflammatory cytokine production. These Tregs have enhanced function and stability as seen by increase in expression of FOXP3 and CD25 in these cells. PT101 maintained selectivity in Phase 1 a clinical trial with no evidence of expansion of natural killer cells nor pro-inflammatory conventional T at any dose studied. A Phase 1b/2a clinical trial in patients with ulcerative colitis and a Phase 2 clinical trial in patients with systemic lupus erythematosus are planned to further evaluate PT101.Disclosure of Interests:None declared

scholarly journals Single-Cell Sequencing of Mouse Heart Immune Infiltrate in Pressure Overload–Driven Heart Failure Reveals Extent of Immune Activation

Circulation ◽  
2019 ◽  
Vol 140 (25) ◽  
pp. 2089-2107 ◽  
Author(s):  
Elisa Martini ◽  
Paolo Kunderfranco ◽  
Clelia Peano ◽  
Pierluigi Carullo ◽  
Marco Cremonesi ◽  
...  
Keyword(s):  
Heart Failure ◽  
T Cells ◽  
Mast Cells ◽  
Natural Killer Cells ◽  
Regulatory T Cells ◽  
Natural Killer ◽  
Immune Activation ◽  
Immune Cell ◽  
Pressure Overload ◽  
Cell Types

Background: Inflammation is a key component of cardiac disease, with macrophages and T lymphocytes mediating essential roles in the progression to heart failure. Nonetheless, little insight exists on other immune subsets involved in the cardiotoxic response. Methods: Here, we used single-cell RNA sequencing to map the cardiac immune composition in the standard murine nonischemic, pressure-overload heart failure model. By focusing our analysis on CD45 + cells, we obtained a higher resolution identification of the immune cell subsets in the heart, at early and late stages of disease and in controls. We then integrated our findings using multiparameter flow cytometry, immunohistochemistry, and tissue clarification immunofluorescence in mouse and human. Results: We found that most major immune cell subpopulations, including macrophages, B cells, T cells and regulatory T cells, dendritic cells, Natural Killer cells, neutrophils, and mast cells are present in both healthy and diseased hearts. Most cell subsets are found within the myocardium, whereas mast cells are found also in the epicardium. Upon induction of pressure overload, immune activation occurs across the entire range of immune cell types. Activation led to upregulation of key subset-specific molecules, such as oncostatin M in proinflammatory macrophages and PD-1 in regulatory T cells, that may help explain clinical findings such as the refractivity of patients with heart failure to anti–tumor necrosis factor therapy and cardiac toxicity during anti–PD-1 cancer immunotherapy, respectively. Conclusions: Despite the absence of infectious agents or an autoimmune trigger, induction of disease leads to immune activation that involves far more cell types than previously thought, including neutrophils, B cells, Natural Killer cells, and mast cells. This opens up the field of cardioimmunology to further investigation by using toolkits that have already been developed to study the aforementioned immune subsets. The subset-specific molecules that mediate their activation may thus become useful targets for the diagnostics or therapy of heart failure.

scholarly journals The endometrial immune environment of women with endometriosis

2019 ◽  
Vol 25 (5) ◽  
pp. 565-592 ◽  
Author(s):  
Júlia Vallvé-Juanico ◽  
Sahar Houshdaran ◽  
Linda C Giudice
Keyword(s):  
T Cells ◽  
Immune System ◽  
Natural Killer Cells ◽  
Natural Killer ◽  
Immune Cells ◽  
Immune Cell ◽  
Reproductive Age ◽  
Cycle Phase ◽  
Cell Populations ◽  
Killer Cells

Abstract BACKGROUND Endometriosis, a common oestrogen-dependent inflammatory disorder in women of reproductive age, is characterized by endometrial-like tissue outside its normal location in the uterus, which causes pelvic scarring, pain and infertility. While its pathogenesis is poorly understood, the immune system (systemically and locally in endometrium, pelvic endometriotic lesions and peritoneal fluid) is believed to play a central role in its aetiology, pathophysiology and associated morbidities of pain, infertility and poor pregnancy outcomes. However, immune cell populations within the endometrium of women with the disease have had incomplete phenotyping, thereby limiting insight into their roles in this disorder. OBJECTIVE AND RATIONALE The objective herein was to determine reproducible and consistent findings regarding specific immune cell populations and their abundance, steroid hormone responsiveness, functionality, activation states, and markers, locally and systemically in women with and without endometriosis. SEARCH METHODS A comprehensive English language PubMed, Medline and Google Scholar search was conducted with key search terms that included endometriosis, inflammation, human eutopic/ectopic endometrium, immune cells, immune population, immune system, macrophages, dendritic cells (DC), natural killer cells, mast cells, eosinophils, neutrophils, B cells and T cells. OUTCOMES In women with endometriosis compared to those without endometriosis, some endometrial immune cells display similar cycle-phase variation, whereas macrophages (Mø), immature DC and regulatory T cells behave differently. A pro-inflammatory Mø1 phenotype versus anti-inflammatory Mø2 phenotype predominates and natural killer cells display abnormal activity in endometrium of women with the disease. Conflicting data largely derive from small studies, variably defined hormonal milieu and different experimental approaches and technologies. WIDER IMPLICATIONS Phenotyping immune cell subtypes is essential to determine the role of the endometrial immune niche in pregnancy and endometrial homeostasis normally and in women with poor reproductive history and can facilitate development of innovative diagnostics and therapeutics for associated symptoms and compromised reproductive outcomes.

In vitro identification of human pro-B cells that give rise to macrophages, natural killer cells, and T cells

Blood ◽  
2003 ◽  
Vol 101 (11) ◽  
pp. 4313-4321 ◽  
Author(s):  
Damien Reynaud ◽  
Nathalie Lefort ◽  
Elodie Manie ◽  
Laure Coulombel ◽  
Yves Levy
Keyword(s):  
T Cells ◽  
Natural Killer Cells ◽  
B Cell ◽  
Natural Killer ◽  
Functional Characterization ◽  
Killer Cells ◽  
Interleukin 7 ◽  
Human Cd34 ◽  
Rag 1

Abstract In this study we report the molecular and functional characterization of very early interleukin 7 receptor α (IL-7Rα)+-CD79a+CD19– B-cell progenitors, produced by human CD34+CD19–CD10– cord blood cells grown in the presence of stromal cells and cytokines. Purified IL-7Rα+CD79a+CD19– cells transcribed the B-lymphoid specific genes E2A, EBF, TdT, Rag-1, had initiated DJH rearrangements, but almost lacked Pax-5 mRNA. When exposed to appropriate environmental conditions, these cells repressed B-cell genes and completely differentiated into CD14+ macrophages, CD56+ natural killer cells, and CD4high T cells. Retention of the DJH rearranged genes in both CD14+ and CD56+ cells unambiguously demonstrates that early B-cell genes, expressed prior to Pax-5, can be activated in a multipotent human progenitor cell whose final fate, including in non-B lineages, is determined by external signals.

scholarly journals Lymphocytes and Trogocytosis-Mediated Signaling

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1478
Author(s):  
Jim Reed ◽  
Madison Reichelt ◽  
Scott A. Wetzel
Keyword(s):  
T Cells ◽  
Natural Killer Cells ◽  
Natural Killer ◽  
Intracellular Signaling ◽  
Immune Cell ◽  
Nk Cell ◽  
Γδ T Cells ◽  
Killer Cells ◽  
The Individual ◽  
The Impact

Trogocytosis is the intercellular transfer of membrane and membrane-associated molecules. This underappreciated process has been described in a variety of biological settings including neuronal remodeling, fertilization, viral and bacterial spread, and cancer, but has been most widely studied in cells of the immune system. Trogocytosis is performed by multiple immune cell types, including basophils, macrophages, dendritic cells, neutrophils, natural killer cells, B cells, γδ T cells, and CD4+ and CD8+ αβ T cells. Although not expressed endogenously, the presence of trogocytosed molecules on cells has the potential to significantly impact an immune response and the biology of the individual trogocytosis-positive cell. Many studies have focused on the ability of the trogocytosis-positive cells to interact with other immune cells and modulate the function of responders. Less understood and arguably equally important is the impact of these molecules on the individual trogocytosis-positive cell. Molecules that have been reported to be trogocytosed by cells include cognate ligands for receptors on the individual cell, such as activating NK cell ligands and MHC:peptide. These trogocytosed molecules have been shown to interact with receptors on the trogocytosis-positive cell and mediate intracellular signaling. In this review, we discuss the impact of this trogocytosis-mediated signaling on the biology of the individual trogocytosis-positive cell by focusing on natural killer cells and CD4+ T lymphocytes.

An In Vitro Coculture Model to Study Cytokine Profiles of Natural Killer Cells During Maternal Immune Cell-Trophoblast Interactions

2006 ◽  
Vol 13 (3) ◽  
pp. 196-202 ◽  
Author(s):  
Evangelos Ntrivalas ◽  
Joanne Kwak-Kim ◽  
Kenneth Beaman ◽  
Harilaos Mantouvalos ◽  
Alice Gilman-Sachs
Keyword(s):  
Natural Killer Cells ◽  
Natural Killer ◽  
Immune Cell ◽  
Killer Cells ◽  
Cytokine Profiles ◽  
Coculture Model

scholarly journals Effect of combined contraceptive pill on immune cell of ovarian endometriotic tissue

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Wanwisa Waiyaput ◽  
Keerati Wattanakamolchai ◽  
Yada Tingthanatikul ◽  
Srithean Lertvikool ◽  
Siriluk Tantanavipas ◽  
...  
Keyword(s):  
T Cells ◽  
Natural Killer Cells ◽  
Regulatory T Cells ◽  
Natural Killer ◽  
Cyst Wall ◽  
Immune Cell ◽  
Ethinyl Estradiol ◽  
Untreated Group ◽  
Killer Cells ◽  
Endometriotic Cyst

Abstract Background Dysregulation of immune response is associated with development of endometriosis. The study aim was to evaluate effect of combined oral contraceptive pills (COCs) consisting of ethinyl estradiol (EE) and desogestrel on the expression of macrophage, natural killer cells, and regulatory T cells of ovarian endometriotic cysts. Methods Endometriotic cyst wall tissues were collected from women with endometriosis who were treated (n = 22) with COCs (one table per day of EE 0.03 mg and desogestrel 0.15 mg administered for 28 to 35 days before surgery) or untreated (n = 22). The tissues were collected from endometriotic cyst wall during laparoscopic or laparotomy ovarian cystectomy. Immunohistochemistry for anti-CD68, anti-CD56, and anti-forkhead–winged helix transcription factor (FoxP3), a marker for macrophages, natural killer cells, and regulatory T cells, respectively, were investigated. Results The median (interquartile range [IQR]) number of anti-CD68 positive cells in the COC group was significantly lower than in the untreated group (12.7; 4.9–19.3) versus 45.7 (26.0–70.7), p < 0.001). Tissue infiltration of anti-CD56 positive cells in endometriotic cyst was significantly higher after the treatment when compared with tissue from untreated group (42.9, 27.4–68.9 versus 25.3 (14.1–37.3; p = 0.009). The number of regulatory T cells was also significantly increased in the COC group (6.3, 2.8–15.5) versus 0 (0–1.8; p < 0.001). Conclusions The effects of COC, containing EE 0.30 mg with desogestrel 0.15 mg, on the immune system was demonstrated by a significant decrease in the number of macrophages and an increase in natural killer and regulatory T cells.

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