Amphotericin B both inhibits and enhances T-cell proliferation: Inhibitory effect is mediated through H2O2 production via cyclooxygenase pathway by macrophages

Abstract Abstract 1916 Proteinase 3 (P3), a serine protease constitutively expressed in primary granules and on the membrane of some resting granulocytes, is the target of T cell-mediated autoimmunity in Wegener's granulomatosis (WG) and of anti-leukemia immunity mediated by PR1 (VLQELNVTV)-specific cytotoxic T lymphocytes (PR1-CTL). We have shown that soluble P3 is increased by 5-fold in sera from acute myeloid leukemia (AML) patients compared to healthy controls, and soluble P3 mediates enzyme-independent inhibition of T-cell proliferation. Moreover, tumor-associated neutrophils (TANs) are associated with a poor prognosis in a number of cancers including renal cell carcinoma and lung cancer, and P3 is also over-expressed in a variety of AML and chronic myeloid leukemia (CML). Therefore, we hypothesized that membrane-bound P3 (mP3) may similarly regulate adaptive immunity by suppressing T-cell proliferation. To study this, T cells from healthy donors were labeled with the membrane dye CFSE and stimulated with anti-CD3 and anti-CD28 in the presence or absence of mP3-expressing PMNs for five days. The percentage of proliferating cells was determined by flow cytometry. Proliferation of autologous CD8+ and CD4+ T cells was significantly inhibited by 75% and 72%, respectively, when PMNs were co-incubated with lymphocytes at a ratio of 3:1, and by > 90% at 5:1. This cell contact-dependent inhibitory effect was limited to PMN since PBMCs added to lymphocytes in place of PMNs at 5:1 had no effect on T cell proliferation. To determine whether the inhibitory effect was specifically mediated by mP3, we FAC-sorted CD177+PMNs and CD177−PMNs to obtain highly purified (>98%) mP3+ and mP3− PMNs, respectively, because CD177 and mP3 are co-expressed on same subset of resting PMNs. At a ratio of 3:1 (CD177+PMNs or CD177−PMNs to lymphocytes), mP3+PMNs mediated > 75% growth inhibition of both CD8+ and CD4+ T cells compared to < 55% inhibition by mP3−PMNs (p<0.05). Furthermore, the inhibitory effect of mP3+PMNs on T cell proliferation was blocked (< 10% inhibition of proliferation) by anti-P3 but not by isotype control mAb. The inhibitory effect of mP3 was enzyme-independent because Elafin or α1-anti-trypsin did not affect inhibition by mP3+PMNs. In addition, mP3-mediated inhibition was fully reversible as T cells proliferated normally with anti-CD3/anti-CD28 stimulation after PMNs were removed from co-culture. Similarly, mP3+AML blasts inhibited autologous CD8+ and CD4+ T cell proliferation by 50% and 30%, respectively, at a 2:1 ratio of AML blasts: lymphocytes. Interestingly, bone marrow myeloid derived suppressor cells (MDSC) from leukemia patients express significantly higher mP3 (79.4±5.23% (mean±SEM, n=7)), compared to 22.4±11.55% mP3 on MDSC from healthy donors (p= 0.0007, n=3). Taken together, these data support an important new function of membrane-bound P3 on innate immune cells and leukemia in controlling adaptive T cell immunity. These findings suggest a novel mechanism whereby neutrophils could promote tumor growth in vivo through contact-mediated suppression of tumor-infiltrating lymphocytes by mP3 or by soluble P3 secreted by activated TANs in cancer and myeloid leukemia. Thus, targeting P3 with anti-P3 antibodies may be explored as a novel therapeutic strategy for leukemia and other cancers. Disclosures: No relevant conflicts of interest to declare.

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Interaction of leukocyte integrins with ligand is necessary but not sufficient for function.

The Journal of Cell Biology ◽

10.1083/jcb.116.6.1527 ◽

1992 ◽

Vol 116 (6) ◽

pp. 1527-1535 ◽

Cited By ~ 90

Author(s):

I Dransfield ◽

C Cabañas ◽

J Barrett ◽

N Hogg

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Killer Cell ◽

Inhibitory Effect ◽

T Cell Proliferation ◽

Lymphocyte Function ◽

Receptor Ligand ◽

Leukocyte Integrins ◽

Cell Assays ◽

Ligand Interactions

The leukocyte integrins (CD11/CD18 or beta 2-type integrins) are expressed exclusively on leukocytes and participate in many adhesion-dependent functions (Arnaout, M.A. 1990. Blood. 75:1037-1050; Springer, T. A. 1990. Nature. (Lond.) 346:425-434; Dustin, M. L., and T. S. Springer. 1991. Annu. Rev. Immunol. 9:27-66). The avidity of leukocyte integrin binding to their ligands or counter-receptors is dependent upon response to intracellular signals (Wright, S. D., and B. C. Meyer. 1986. J. Immunol. 136:1759-1764; Dustin, M. A., and T. S. Springer. 1989. Nature (Lond.). 341:619-624). We have investigated the effects of a novel mAb (mAb 24) which defines a leukocyte integrin alpha subunit epitope that is Mg(2+)-dependent and may be used as a "reporter" of the activation state of these receptors (Dransfield, I., and N. Hogg. 1989. EMBO (Eur. Mol. Biol. Organ) J. 8:3759-3765; Dransfield, I., A.-M. Buckle, and N. Hogg. 1990. Immunol. Rev. 114:29-44; Dransfield, I., C. Cabañas, A. Craig, and N. Hogg. 1992. J. Cell Biol.) Data is presented to show that this mAb inhibits monocyte-dependent, antigen-specific T cell proliferation and IL-2-activated natural killer cell assays which are both dependent on lymphocyte function-associated antigen-1 (LFA-1), and complement receptor type 3 (CR3)-mediated neutrophil chemotaxis to f-Met-Leu-Phe. This inhibitory effect is not caused by the prevention of receptor/ligand binding because LFA-1/ICAM-1, LFA-1/ICAM-2,3 and CR3/iC3b interactions are, under activating conditions, promoted rather than blocked by mAb 24. As it does not interfere with mitogen-stimulated T cell proliferation, it is unlikely that mAb 24 transduces a "negative" or antiproliferative signal to the T cells to which it is bound. Using a model system of transient activation of LFA-1, we have found that mAb 24 prevents "deadhesion" of receptor/ligand pairs, possibly locking leukocyte integrins in an "active" conformation. It is speculated that inhibition of leukocyte integrin function by this mAb reflects the necessity for dynamic leukocyte integrin/ligand interactions.

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Specific Inhibitory Action of Anisodamine against a Staphylococcal Superantigenic Toxin, Toxic Shock Syndrome Toxin 1 (TSST-1), Leading to Down-Regulation of Cytokine Production and Blocking of TSST-1 Toxicity in Mice

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.12.3.399-408.2005 ◽

2005 ◽

Vol 12 (3) ◽

pp. 399-408 ◽

Cited By ~ 8

Author(s):

Saori Nakagawa ◽

Koji Kushiya ◽

Ikue Taneike ◽

Ken'ichi Imanishi ◽

Takehiko Uchiyama ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

Proinflammatory Cytokines ◽

Toxic Shock Syndrome ◽

Inhibitory Effect ◽

Herbal Extract ◽

T Cell Proliferation ◽

Toxic Shock ◽

Toxic Shock Syndrome Toxin

ABSTRACT Toxic shock syndrome toxin 1 (TSST-1), produced by Staphylococcus aureus (including methicillin-resistant S. aureus), is a superantigenic toxin responsible for toxic shock syndrome as well as neonatal TSS-like exanthematous disease. TSST-1 exhibits its deleterious effects by leading to the abnormal proliferation of, e.g., Vβ2+ T cells and overproduction of proinflammatory cytokines. In the present study we examined the inhibitory effect of a Chinese herbal extract, anisodamine, on TSST-1 using human peripheral blood mononuclear cells (PBMCs). Anisodamine inhibited the production of proinflammatory cytokines better than interleukin-10 (an anti-inflammatory cytokine). The inhibitory effect of anisodamine was greater than that of any tropane alkaloid examined. Anisodamine acted directly on both monocytes and T cells in human PBMCs, and the effect was confirmed at the transcriptional level. Inhibition of NF-κB activation was also demonstrated. In contrast, no significant inhibition of Vβ2+ T-cell proliferation was observed. In mice injected with TSST-1, anisodamine treatment significantly decreased serum proinflammatory cytokine levels and prevented TSST-1-induced death. These results suggest that anisodamine specifically acts against the production of cytokines (inflammatory cytokines in particular) and not against Vβ2+ T-cell proliferation and that anisodamine may have a beneficial effect on TSST-1-associated disease.

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A procedure for in vitro evaluation of the immunosuppressive effect of mesenchymal stem cells on activated T cell proliferation

10.21203/rs.3.rs-63604/v1 ◽

2020 ◽

Author(s):

Catalina Marinescu ◽

Bogdan Preda ◽

Alexandrina Burlacu

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Inhibitory Effect ◽

Immunosuppressive Effect ◽

T Cell Proliferation ◽

Immunomodulatory Capacity ◽

Activated T Cell ◽

Dose Dependent

Abstract Background: Mesenchymal stem/stromal cells (MSC) represent adult cells with multipotent capacity. Besides their capacity to differentiate into multiple lineages in vitro and in vivo, increasing evidence points towards the immunomodulatory capacity of these cells, as an important feature for their therapeutic power. Although not included in the minimal criteria established by the International Society for Cellular Therapy as a defining MSC attribute, demonstration of the immunomodulatory capacity of MSC can be useful for the characterization of these cells before being considered MSC. Here we present a simple and reliable protocol by which the immunosuppressive effect of MSC can be evaluated in vitro. It is based on the measuring of the proliferation of activated T cells cultured in direct contact with irradiated MSC.Results: Our results showed that MSC have a dose-dependent inhibitory effect on activated T cell proliferation, which can be quantified as a percentage of maximum proliferation. Our data shows that batch-to-batch variability can be determined within one or multiple experiments, by extracting the area under curve of T cell proliferation plotted against the absolute number of MSC in co-culture.Conclusions: The validation of the immunomodulatory capacity of MSC could be added to the characterization of the cells before being used in various MSC-based approaches to treat immunological diseases. Our results showed that MSC have a dose-dependent inhibitory effect on activated T cell proliferation. The immunosuppressive properties of MSC vary between batches, but not between different passages of the same batch.

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Sphingosine 1-phosphate is a novel inhibitor of T-cell proliferation

Blood ◽

10.1182/blood-2002-09-2962 ◽

2003 ◽

Vol 101 (12) ◽

pp. 4909-4915 ◽

Cited By ~ 69

Author(s):

Yixin Jin ◽

Eirunn Knudsen ◽

Ling Wang ◽

Yenan Bryceson ◽

Bassam Damaj ◽

...

Keyword(s):

Cell Proliferation ◽

T Cells ◽

Dendritic Cells ◽

T Cell ◽

Interleukin 2 ◽

Inhibitory Effect ◽

Interferon Γ ◽

T Cell Proliferation ◽

Inhibitory Molecule ◽

Sphingosine 1 Phosphate

Abstract Sphingosine 1-phosphate (S1P) is a pleiotropic lysosphingophospholipid stored and secreted by platelets. Using reverse transcription–polymerase chain reaction and flow cytometric analyses, we determined the expression of S1P receptors (S1P1, S1P3, S1P4, and S1P5) in peripheral blood T cells. T cells were induced to proliferate in the presence of phorbol 12-myristate 13-acetate (PMA) plus ionomycin, anti-CD3 plus anti-CD28, and allogeneic immature or mature dendritic cells. This activity was inhibited by the addition of S1P. Enhanced T-cell proliferation was observed when these cells were stimulated with the same stimuli, but were incubated in serum-free media (SFM). Addition of S1P to SFM inhibited the stimulation of T cells induced by T-cell stimuli, suggesting that S1P is an important inhibitory molecule present in the serum. T-cell proliferation was also inhibited by the addition of dihydrosphingosine 1-phosphate (DHS1P), sphingosine, and ceramide; however, the latter 2 sphingolipids required higher concentrations than S1P. Pretreatment of T cells with pertussis toxin (PTX) blocked the inhibitory effect of S1P on activation with PMA plus ionomycin, but not on activation with anti-CD3 plus anti-CD28. This is corroborated with the down-regulation of S1P1 in T cells stimulated with anti-CD3 plus anti-CD28. Similarly, PTX did not affect the inhibitory effect of S1P on T-cell proliferation when dendritic cells were used as stimuli. Further, S1P or DHS1P but not ceramide or sphingosine enhanced rather than decreased secretion of interleukin 2 and interferon γ by T cells stimulated with anti-CD3 plus anti-CD28. These results show differential effects of S1P on polyclonal T-cell proliferation and cytokine secretion.

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Interaction between the CD45 antigen and phytohemagglutinin. Inhibitory effect on the lectininduced T cell proliferation by anti-CD45 monoclonal antibody

European Journal of Immunology ◽

10.1002/eji.1830171012 ◽

1987 ◽

Vol 17 (10) ◽

pp. 1461-1466 ◽

Cited By ~ 43

Author(s):

Carmelo Bernabeu ◽

Ana C. Carrera ◽

Manuel O. De Landázuri ◽

Francisco Sánchez-Madrid

Keyword(s):

Monoclonal Antibody ◽

Cell Proliferation ◽

T Cell ◽

Inhibitory Effect ◽

T Cell Proliferation

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A procedure for in vitro evaluation of the immunosuppressive effect of mouse mesenchymal stem cells on activated T cell proliferation

Stem Cell Research & Therapy ◽

10.1186/s13287-021-02344-3 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Catalina-Iolanda Marinescu ◽

Mihai Bogdan Preda ◽

Alexandrina Burlacu

Keyword(s):

Cell Proliferation ◽

T Cell ◽

Inhibitory Effect ◽

Immunosuppressive Effect ◽

T Cell Proliferation ◽

Immunomodulatory Capacity ◽

Activated T Cell ◽

Dose Dependent

Abstract Background Mesenchymal stem/stromal cells (MSC) represent adult cells with multipotent capacity. Besides their capacity to differentiate into multiple lineages in vitro and in vivo, increasing evidence points towards the immunomodulatory capacity of these cells, as an important feature for their therapeutic power. Although not included in the minimal criteria established by the International Society for Cellular Therapy as a defining MSC attribute, demonstration of the immunomodulatory capacity of MSC can be useful for the characterization of these cells before being considered MSC. Methods Here we present a simple and reliable protocol by which the immunosuppressive effect of mouse bone marrow-derived MSC can be evaluated in vitro. It is based on the measuring of the proliferation of activated T cells cultured in direct contact with irradiated MSC. Results Our results showed that mouse MSC have a dose-dependent inhibitory effect on activated T cell proliferation, which can be quantified as a percentage of maximum proliferation. Our data shows that batch-to-batch variability can be determined within one or multiple experiments, by extracting the area under curve of T cell proliferation plotted against the absolute number of MSC in co-culture. Conclusions The validation of the immunosupressive capacity of MSC could be added to the characterization of the cells before being used in various MSC-based approaches to treat immunological diseases. Our results showed that mouse MSC have a dose-dependent inhibitory effect on activated T cell proliferation. The immunosuppressive properties of MSC vary between batches, but not between different passages of the same batch.

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