Differential Leukocyte miRNA Responses Following Pan T Cell, Allorecognition and Allosecretome-Based Therapeutics Activation

Abstract Background Effective immunomodulation of T cell responses is critical in treating both autoimmune diseases and cancer. Our previous studies have demonstrated that secretomes derived from control or methoxypolyethylene glycol (mPEG) mixed lymphocyte alloactivation assays exerted potent immunomodulatory activity that was mediated by microRNAs (miRNA). In this study, the immunomodulatory effects of biomanufactured miRNA-based allo-secretome therapeutics (SYN, TA1, IA1 and IA2) were compared to Pan T cell activators (PHA and anti-CD3/CD28) and alloactivation (MHC-disparate donors; ± mPEG grafting). The differential effects of these activation strategies on resting PBMC were assessed via T cell differentiation and proliferation as well as the differential expression of multiple miRNA.Results Mitogen-induced PBMC proliferation (average of > 85%) significantly exceed that arising from either allostimulation (~ 30%) or the proinflammatory IA1 secretome product (~ 12%). Consequent to stimulation, the ratio of CD4 to CD8 cells of the resting PBMC (CD4:CD8; 1.7 ± 0.1) decreased in the Pan-T cell, allrecognition and IA1 activated cells (average of 1.1 ± 0.2; 1.2 ± 0.1 and 1.0 ± 0.1). These changes arose consequent to the expansion of both CD4+CD8+ and CD4-CD8- populations and the shrinkage of the CD4 subset relative to the expansion of the CD8 T cells. Most importantly, this study demonstrated that these activation strategies exert profoundly unique effects on the differential expression of miRNA within the treated PBMC and that these 'differential patterns of miRNA expression' are associated with significant differences in cellular differentiation and biological function.Conclusions These findings support the concept that the 'differential pattern of miRNA expression', not a change in a single miRNA, governs the biologic immune response in a 'lock and key' manner. The biomanufacturing of miRNA-enriched secretome biotherapeutics may be a successful approach for producing miRNA cocktails (e.g., TA1 and IA1) that replicate the normal biological 'lock and key' miRNA configuration necessary for the systemic treatment of autoimmune diseases (TA1) or enhancing the endogenous immune response to cancer (IA1).

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Differential Leukocyte miRNA Responses Following Pan T Cell, Allorecognition and Allosecretome-Based Therapeutics Activation

10.21203/rs.2.18715/v1 ◽

2019 ◽

Author(s):

Xining Yang ◽

Wendy M. Toyofuku ◽

Mark D. Scott

Keyword(s):

T Cell ◽

Autoimmune Diseases ◽

T Cell Activation ◽

Mirna Expression ◽

Cell Activation ◽

Expression Profiles ◽

Immunomodulatory Activity ◽

Mirna Expression Profiles

Abstract Background: Effective immunomodulation of T cell responses is critical in treating both autoimmune diseases and cancer. Our previous studies have demonstrated that nanoscale bioengineering of cell surfaces with methoxypolyethylene glycol (mPEG) induces a potent tolerogenic immunomodulatory effect. Moreover, secretomes derived from mPEG- or control mixed lymphocyte alloactivation assays also exerted potent immunomodulatory activity that was mediated by microRNAs (miRNA). In this study, the immunomodulatory effects of Pan T cell activators (PHA and anti-CD3/CD28), alloactivation (MHC-disparate donors; ± mPEG grafting) and biomanufactured miRNA-based allo-secretome therapeutics (SYN, TA1, IA1 and IA2) were examined on T cell proliferation, subset differentiation and leukocyte miRNA expression profiles of resting human PBMC. Results: In contrast to Pan T cell activation, allorecognition and the pro-inflammatory IA1 secretome product induced increasingly controlled proliferation of resting PBMC. The differential effects of the activation strategies were also apparent in T cell differentiation and the Teff:Treg ratio and in the miRNA expression profiles noted in the treated PBMC. In contrast, the mPEG-PBMC and TA1 secretome products inhibited alloproliferation. Importantly, the activation strategies exerted significantly different miRNA expression in the treated leukocytes that was associated with differences in proliferation and cellular differentiation. Conclusions: Immunomodulatory secretome-derived, miRNA-enriched, therapeutics can be reproducibly biomanufactured that will induce the specific bioregulatory events necessary to induce the differentiation of naïve T cells to produce a tolerogeneic (TA1) or inflammatory (IA1) response both in vitro and in vivo. The successful development and biomanufacturing of immunomodulatory, miRNA-enriched, secretome biotherapeutics may provide potent tools for the systemic treatment of autoimmune diseases or enhancing the endogenous immune response to cancer while reducing the potential adverse risks of more non-specific immunomodulatory approaches.

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IL-7 and IL-15 combined with strong TCR stimulation decrease Treg suppressive activity in healthy donors and patients with rheumatoid arthritis

10.22541/au.160760000.09053245/v1 ◽

2020 ◽

Author(s):

Daniil Shevyrev ◽

Valeriy Tereshchenko ◽

Alexey Sizikov ◽

Vladimir Kozlov

Keyword(s):

Rheumatoid Arthritis ◽

T Cell ◽

Autoimmune Diseases ◽

Homeostatic Proliferation ◽

Suppressive Activity ◽

Cd8 Cells ◽

Cell Clones ◽

Healthy Donors ◽

Tcr Signals ◽

And Function

Homeostatic proliferation (HP) is a physiological process to reconstitute the T-cell pool after lymphopenia with IL-7 and IL-15 being the key cytokines regulating the process. However, there is no evidence whether these cytokines influence the function of regulatory T cells (Tregs). Since lymphopenia often accompanies autoimmune diseases, we decided to study the proliferation rate and function of Tregs stimulated by IL-7 and IL-15 in patients with rheumatoid arthritis (RA) compared to healthy donors (HD). The study used peripheral blood from 14 RA patients and 18 HD. Proliferation of purified CD3CD4CD25CD127 cells was assessed by flow cytometry using CFSE. Tregs were stimulated by anti-CD3, IL-7, IL-15, IL-7, or IL-15 combined with anti-CD3, and by IL-2+anti-CD3, and their functional activity was evaluated in each case by CD4 and CD8 cells proliferation inhibition. The suppressive activity of peripheral Tregs did not differ between RA and HD; however, it significantly decreased when IL-7 or IL-15 were applied together with strong TCR stimulation with anti-CD3 antibodies. Herewith Treg proliferation caused by IL-7 and IL-15 was lower in RA than in HD. The revealed decrease in Treg suppressive activity can lead to the proliferation of potentially self-reactive T-cell clones, which can receive relatively strong TCR signals. This may be another explanation of why lymphopenia is associated with the development of autoimmune diseases. The revealed decrease of Treg proliferation under IL-7 and IL-15 may lead to a delay in Treg pool reconstitution in patients with rheumatoid arthritis.

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Introduction to Clinical Immunology: Overview of the Immune Response, Autoimmune Conditions, and Immunosuppressive Therapeutics for Rheumatic Diseases

10.2310/im.1328 ◽

2016 ◽

Author(s):

Steven K. Lundy ◽

Alison Gizinski ◽

David A. Fox

Keyword(s):

Immune Response ◽

Immune System ◽

T Cell ◽

Immune Responses ◽

Autoimmune Diseases ◽

Adaptive Immunity ◽

Cell Populations ◽

Hematopoietic Stem ◽

Innate And Adaptive Immunity ◽

Adaptive Immune

The immune system is a complex network of cells and mediators that must balance the task of protecting the host from invasive threats. From a clinical perspective, many diseases and conditions have an obvious link to improper functioning of the immune system, and insufficient immune responses can lead to uncontrolled acute and chronic infections. The immune system may also be important in tumor surveillance and control, cardiovascular disease, health complications related to obesity, neuromuscular diseases, depression, and dementia. Thus, a working knowledge of the role of immunity in disease processes is becoming increasingly important in almost all aspects of clinical practice. This review provides an overview of the immune response and discusses immune cell populations and major branches of immunity, compartmentalization and specialized immune niches, antigen recognition in innate and adaptive immunity, immune tolerance toward self antigens, inflammation and innate immune responses, adaptive immune responses and helper T (Th) cell subsets, components of the immune response that are important targets of treatment in autoimmune diseases, mechanisms of action of biologics used to treat autoimmune diseases and their approved uses, and mechanisms of other drugs commonly used in the treatment of autoimmune diseases. Figures show the development of erythrocytes, platelets, lymphocytes, and other immune system cells originating from hematopoietic stem cells that first reside in the fetal liver and later migrate to the bone marrow, antigen–major histocompatibility complex recognition by T cell receptor control of T cell survival and activation, and Th cells as central determinants of the adaptive immune response toward different stimuli. Tables list cell populations involved in innate and adaptive immunity, pattern recognition receptors with known ligands, autoantibody-mediated human diseases: examples of pathogenic mechanisms, selected Food and Drug Administration–approved autoimmune disease indications for biologics, and mechanism of action of biologics used to treat autoimmune diseases. This review contains 3 highly rendered figures, 5 tables, and 64 references.

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CMV Immunity Status in Older (>50 Years Old) Subjects after Non-Myeloablative or Reduced Intensity Regimen for Hematopoietic Cell Transplantation (HCT): A Comparison with a Younger Cohort after Ablative HCT.

Blood ◽

10.1182/blood.v106.11.3246.3246 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3246-3246

Author(s):

Ghislaine Gallez-Hawkins ◽

Lia Thao ◽

Simon F. Lacey2 ◽

Joybelle Martinez ◽

Anne E. Franck ◽

...

Keyword(s):

Immune Response ◽

Stem Cell ◽

T Cell ◽

Cell Transplantation ◽

Immune Reconstitution ◽

Cmv Infection ◽

Cd8 Cells ◽

Group 2 ◽

Reduced Intensity ◽

Group 1

Abstract Immunity declines with age as demonstrated by cell-mediated and humoral responses to alloantigens. The susceptibility of these elderly subjects to endogenous virus infection, such as human cytomegalovirus (HCMV) reactivation, is a particular concern during the process of hematopoietic stem cell transplantation (HCT) and immune reconstitution. In this report, the host contribution to stem cell engraftment and differentiation was evaluated by comparing the HCMV immune response in older subjects (> 50 y.o.) to a younger (< 50 y.o.) transplant population. This was a retrospective analysis of a subset of data collected prospectively and with IRB approval for characterization of the CMV immune response of allogeneic transplant patients. Within the dataset, two groups of patients were compared. Group 1 consisted of 10 patients >50 y.o. who had received reduced intensity or non-myeloablative conditioning regimen, and Group 2 consisted of 13 patients <50 y.o., most of whom had received a myeloablative regimen. Because 9 of 10 in Group 1 had had CMV reactivation, Group 2 was selected from the subset of younger patients with known post-transplant CMV infection. CMV infection was defined as either a positive CMV blood culture using shell vial assay or a positive CMV PCR on plasma. Subjects were assessed on days 40, 90, 120, 150, 180, and 360 post-HCT by CMV-specific tetramer-binding assay using CD8 cells, assays for intracellular INF-g response of CD4 and CD8 cells, and a T-cell receptor excision circle (TREC) assay. There were no significant differences observed in the CD4+/IFN-g+ cell responses to CMV antigen nor were the rates of activated CD4+/CD69+/IFN-g+ cells different between the groups. Group 1 was also characterized by a robust CD8+/IFN-g+ response to HLA-specific CMV peptides, and all subjects had ≥ 2cells/μl by day 150 post-HCT. The frequency of CMV tetramer positive cells (≥ 2cells/μl) was 50% in Group 1 by day 90 post-HCT and was not statistically different from Group 2. The T cell renewal in the thymus as measured by the TREC spanned over 0 -- 92 copies/μg of total cellular DNA in Group 1 and from 0 – 129 copies/μg in Group 2 during the first year post-HCT (n.s.). In conclusion, CMV immune reconstitution in older transplant subjects, who undergo a reduced intensity or non-myeloablative regimen, is robust and, in this small sampling, did not differ from that observed in a younger adult group.

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In VitroImmunomodulatory Activity of a Transition-State Analog Inhibitor of Human Purine Nucleoside Phosphorylase in Cutaneous Leishmaniasis

Journal of Immunology Research ◽

10.1155/2017/3062892 ◽

2017 ◽

Vol 2017 ◽

pp. 1-6 ◽

Cited By ~ 1

Author(s):

Natália Barbosa Carvalho ◽

Fernanda Ventin de Oliveira Prates ◽

Rafael de Castro da Silva ◽

Mayra Elizabeth Ferreira Dourado ◽

Camila Farias Amorim ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

Cutaneous Leishmaniasis ◽

T Cell Activation ◽

Purine Nucleoside Phosphorylase ◽

Purine Nucleoside ◽

Fourth Generation ◽

Immunomodulatory Activity ◽

Nucleoside Phosphorylase

Cutaneous leishmaniasis (CL) is the most common clinical form of American tegumentary leishmaniasis caused byLeishmania(Viannia)braziliensis. CL is associated with a strong Th1 immune response. This exacerbated inflammatory response is correlated with severity of disease and delays the healing time of the ulcer. The fourth-generation immucillin derivative (DI4G), a potent inhibitor of purine nucleoside phosphorylase, has been proposed as a promising agent in the treatment of diseases associated with T cell activation. Herein, we evaluated thein vitroimmunomodulatory activity of DI4G in cells of patients presenting with CL. Peripheral blood mononuclear cells (PBMC) from CL patients were stimulated with soluble leishmania antigen (SLA), in the presence or absence of DI4G, and IFN-γ, TNF, CXCL9, and CXCL10 levels were determined by ELISA. Lymphocyte proliferation in the presence or absence of DI4G was also evaluated, using flow cytometry. DI4G was able to decrease (p<0.05) IFN-γproduction but did not change the TNF, CXCL9, and CXCL10 levels. DI4G decreased (p<0.05) the lymphoproliferative response mediated by CD8+T cells, but not that by CD4+T cells. DI4G is able to attenuate the exaggerated immune response in CL, exhibiting immunomodulatory activity in IFN-γproduction and in CD8+T cell proliferation.

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Adoptive Transfer of Tracer-Alloreactive CD4+T Cell Receptor Transgenic T Cells Alters the Endogenous Immune Response to an Allograft

American Journal of Transplantation ◽

10.1111/ajt.13821 ◽

2016 ◽

Vol 16 (10) ◽

pp. 2842-2853 ◽

Cited By ~ 6

Author(s):

M. L. Miller ◽

J. Chen ◽

M. D. Daniels ◽

M. G. McKeague ◽

Y. Wang ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

T Cell Receptor ◽

Adoptive Transfer ◽

Cell Receptor ◽

Cd4 T Cell ◽

Endogenous Immune Response

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Brain Toxoplasmosis Comorbid with Autoimmune Disease: Complicated Immune Response And Case Demonstration

Canadian Journal of Neurological Sciences / Journal Canadien des Sciences Neurologiques ◽

10.1017/cjn.2021.164 ◽

2021 ◽

Vol 48 (s2) ◽

pp. S8-S8

Author(s):

Alice Graham ◽

Crystal Fong ◽

Asghar Naqvi ◽

Jian-Qiang Lu

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

Autoimmune Diseases ◽

Lupus Erythematosus ◽

Incidental Finding ◽

Cell Infiltration ◽

List Type ◽

Pathological Features ◽

T Cell Infiltration

Toxoplasmosis is an opportunistic infection caused by Toxoplasma gondii (TG), commonly involving the brain. Symptomatic clinical disease of TG infection is much more commonly associated with immunodeficiency; clinicopathological manifestations of brain toxoplasmosis are linked to individual immune responses including brain infiltration of T-cells that are thought to fight against toxoplasmosis. In patients with autoimmune diseases, immune status is typically characterized by T-cell infiltration and complicated mainly by immunosuppressant and/or immunomodulatory treatment. In this study, we demonstrate clinical and radiological features correlated with pathological features of brain toxoplasmosis at different disease stages in a patient with coexisting autoimmune diseases, including systemic lupus erythematosus and autoimmune hepatitis. The infiltration of CD8+ T-cells in toxoplasma immunostaining-positive acute lesions was greater than that in toxoplasma immunostaining-negative chronic lesions. We also review previously reported cases of brain toxoplasmosis with comorbid autoimmune diseases. Our present case and literature review suggest that brain toxoplasmosis in patients with autoimmune diseases may be asymptomatic unless disease complications occur; it may present as an incidental finding at postmortem examination of rapidly developed lesions. T-cell infiltration in patients with autoimmune diseases and coexisting toxoplasmosis may be at least partially reduced; ultimately, the roles of T-cell infiltration in brain toxoplasmosis deserve further investigation.Learning ObjectivesDiscuss complicated immune response to toxoplasmosis in patients with autoimmune diseases.Describe clinical, radiological, and pathological features of brain toxoplasmosis in patients with autoimmune diseases.

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Cytomegalovirus antigen-specific T Cell immune response in patients with autoimmune diseases under different cytomegalovirus infection status

10.21203/rs.3.rs-309275/v1 ◽

2021 ◽

Author(s):

Huimin Ma ◽

Yuting Tan ◽

Xiaoqing Liu ◽

Xiaochun Shi ◽

Wenjie Zheng ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

Autoimmune Diseases ◽

Cd4 T Cell ◽

Cell Immune Response ◽

Cmv Infection ◽

Infection Group ◽

Cell Counts ◽

Ifn Γ

Abstract Background: T-cell immunity is important for the control of cytomegalovirus (CMV) infection. The frequency of IFN-γ secreting T cells after stimulation with CMV-specific protein-1 (IE-1) and phosphoprotein 65 (pp65) antigen can help predict the risk of active CMV infection. Patients with autoimmune diseases have a high incidence of active CMV infection, but the CMV antigen-specific T cell immune response of this population is still blank in the world. This study aimed to use T-SPOT.CMV to investigate CMV antigen-specific T cell immune response in patients with autoimmune diseases under different CMV infection conditions.Methods: Patients with autoimmune diseases in the Peking Union Medical College Hospital from March, 2017 to October, 2020 were continuously selected. According to the definition, the subjects were divided into latent CMV infection group and active CMV infection group. T-SPOT.CMV was used to evaluate CMV antigen-specific T cell immune response under different CMV infection status, and the possible influential factors of CMV antigen-specific T cell immune response were further analyzed.Results: Fifty patients with latent CMV infection and fifty patients with active CMV infection were enrolled. After stimulated by immediate early IE-1 and pp65 antigen, the median frequency of IFN-γ secreting T cells in active CMV infection group were all significantly lower than that in latent CMV infection group (p<0.001), and in CMV disease group was significantly lower than that in latent CMV infection group (p<0.001). After stimulated by pp65, the median frequency of IFN-γ secreting T cells with CD4+ T cell counts < 200/ul was significantly lower than that of CD4+ T cell counts ≥ 200/ul (p=0.043), and those with CD8+ T cell counts < 250/ul was significantly lower than that of CD8+ T cell counts ≥ 250/ul (p=0.03). The frequency of IFN-γ secreting T cells stimulated by pp65 was significantly higher than IE-1.Conclusions: In patients with autoimmune diseases, the CMV antigen-specific T-cell immune response in patients with active CMV infection was significantly lower than that with latent CMV infection. IE-1 was considered as a more stable antigen with better effect than pp65. Lymphocyte, CD4+T cell and CD8+T cell count might affect CMV antigen-specific T cell immune response.

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Comparison of miRNA Expression Profiles between HIV-1 and HIV-2 Infected Monocyte-Derived Macrophages (MDMs) and Peripheral Blood Mononuclear Cells (PBMCs)

International Journal of Molecular Sciences ◽

10.3390/ijms21186970 ◽

2020 ◽

Vol 21 (18) ◽

pp. 6970

Author(s):

Santanu Biswas ◽

Emily Chen ◽

Mohan Haleyurgirisetty ◽

Sherwin Lee ◽

Indira Hewlett ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Signaling Pathways ◽

Differential Expression ◽

Signaling Pathway ◽

Mirna Expression ◽

Expression Profiles ◽

Mirna Expression Profile ◽

Differentially Expressed Mirnas ◽

Hiv 1

During the progression of HIV-1 infection, macrophage tropic HIV-1 that use the CCR5 co-receptor undergoes a change in co-receptor use to CXCR4 that is predominately T cell tropic. This change in co-receptor preference makes the virus able to infect T cells. HIV-2 is known to infect MDMs and T cells and is dual tropic. The aim of this study was to elucidate the differential expression profiles of host miRNAs and their role in cells infected with HIV-1/HIV-2. To achieve this goal, a comparative global miRNA expression profile was determined in human PBMCs and MDMs infected with HIV-1/HIV-2. Differentially expressed miRNAs were identified in HIV-1/HIV-2 infected PBMCs and MDMs using the next-generation sequencing (NGS) technique. A comparative global miRNA expression profile in infected MDMs and PBMCs with HIV-1 and HIV-2 identified differential expression of several host miRNAs. These differentially expressed miRNAs are likely to be involved in many signaling pathways, like the p53 signaling pathway, PI3K-Akt signaling pathways, MAPK signaling pathways, FoxO signaling pathway, and viral carcinogenesis. Thus, a comparative study of the differential expression of host miRNAs in MDMs and T cell in response to HIV-1 and HIV-2 infection will help us to identify unique biomarkers that can differentiate HIV-1 and HIV-2 infection.

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