Analysis of the adpative immune response-dependence of the PRKCG cistromes in human Jurkat cells and CD4+ lymphocytes

Gradients of soluble molecules coordinate cellular communication in a diverse range of multicellular systems. Chemokine-driven chemotaxis is a key orchestrator of cell movement during organ development, immune response and cancer progression. Chemotaxis assays capable of examining cell responses to different chemokines in the context of various extracellular matrices will be crucial to characterize directed cell motion in conditions which mimic whole tissue conditions. Here, a microfluidic device which can generate different chemokine patterns in flow-free gradient chambers while controlling surface extracellular matrix (ECM) to study chemotaxis either at the population level or at the single cell level with high resolution imaging is presented. The device is produced by combining additive manufacturing (AM) and soft lithography. Generation of concentration gradients in the device were simulated and experimentally validated. Then, stable gradients were applied to modulate chemotaxis and chemokinetic response of Jurkat cells as a model for T lymphocyte motility. Live imaging of the gradient chambers allowed to track and quantify Jurkat cell migration patterns. Using this system, it has been found that the strength of the chemotactic response of Jurkat cells to CXCL12 gradient was reduced by increasing surface fibronectin in a dose-dependent manner. The chemotaxis of the Jurkat cells was also found to be governed not only by the CXCL12 gradient but also by the average CXCL12 concentration. Distinct migratory behaviors in response to chemokine gradients in different contexts may be physiologically relevant for shaping the host immune response and may serve to optimize the targeting and accumulation of immune cells to the inflammation site. Our approach demonstrates the feasibility of using a flow-free gradient chamber for evaluating cross-regulation of cell motility by multiple factors in different biologic processes.

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Human Cellular Immune Response to the Saliva of Phlebotomus papatasi Is Mediated by IL-10-Producing CD8+ T Cells and Th1-Polarized CD4+ Lymphocytes

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0001345 ◽

2011 ◽

Vol 5 (10) ◽

pp. e1345 ◽

Cited By ~ 23

Author(s):

Maha Abdeladhim ◽

Mélika Ben Ahmed ◽

Soumaya Marzouki ◽

Nadia Belhadj Hmida ◽

Thouraya Boussoffara ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

Cd8 T Cells ◽

Cellular Immune Response ◽

Phlebotomus Papatasi ◽

Cellular Immune ◽

Cd4 Lymphocytes

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Gastrin-CCK-B type receptors on human T lymphoblastoid Jurkat cells

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.1995.268.3.g522 ◽

1995 ◽

Vol 268 (3) ◽

pp. G522-G529 ◽

Cited By ~ 3

Author(s):

J. Dornand ◽

S. Roche ◽

F. Michel ◽

J. P. Bali ◽

S. Cabane ◽

...

Keyword(s):

Immune Response ◽

Binding Sites ◽

Gastrointestinal Hormones ◽

Jurkat Cells ◽

Affinity Binding ◽

Single Class ◽

High Affinity Binding ◽

High Affinity ◽

Inositol 1,4,5 Trisphosphate ◽

Specific Receptors

The presence of specific receptors for gastrointestinal hormones on T cells and their involvement in the immune response are still matters of debate. We reported the effects of gastrin-cholecystokinin (CCK)-related peptides on J.RT3-T3.5 Jurkat cells. A single class of high-affinity binding sites (dissociation constant approximately 0.1 nM) for gastrin and CCK-8 was evident on these cells. These peptides dose-dependently induced a transient increase in intracellular Ca2+ concentration ([Ca2+]i), which was independent of extracellular Ca2-. L-365,260 was 150- to 300-fold more potent than L-364,718 to inhibit radiolabeled ligand binding or peptide-stimulated [Ca2+]i increase, confirming the gastrin-CCK-B nature of the receptor. Gastrin caused a rise in inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] level within 5 s of stimulation. Finally, gastrin increased interleukin (IL)-2 secretion in J.RT3-T3.5 cells. We conclude that 1) J.RT3-T3.5 cells possess "gastrin-CCK-B type" receptors coupled to phospholipase C activation, Ins(1,4,5)P3 formation, and Ca2+ release from intracellular Ca2+ pools, and 2) these receptors could be involved in the regulation of IL-2 production.

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Low Ozone Concentrations Affect the Structural and Functional Features of Jurkat T Cells

Processes ◽

10.3390/pr9061030 ◽

2021 ◽

Vol 9 (6) ◽

pp. 1030

Author(s):

Enrica Cappellozza ◽

Manuela Costanzo ◽

Laura Calderan ◽

Mirco Galiè ◽

Osvaldo Angelini ◽

...

Keyword(s):

Immune Response ◽

Blood Cells ◽

Inflammatory Responses ◽

Antioxidant Response ◽

Jurkat Cells ◽

Therapeutic Effects ◽

Basic Study ◽

Human T Lymphocyte ◽

Functional Features

Autohemotherapy is the most used method to administer O2-O3 systemically. It consists in exposing a limited amount of blood to a gaseous O2-O3 and reinfusing it, thus activating a cascade of biochemical pathways involving plasma and blood cells that gives rise to antioxidant and anti-inflammatory responses. The therapeutic effects strictly depend on the O3 dose; it is therefore necessary to understand the relationship between the O3 concentration and the effects on blood cells involved in antioxidant and immune response. Here we performed a basic study on the effects of the low O3 concentrations used for autohemotherapy on the structural and functional features of the human T-lymphocyte-derived Jurkat cells. Ultrastructural, biomolecular, and bioanalytic techniques were used. Our findings showed that 10, 20, and 30 µg O3 concentrations were able to trigger Nrf2-induced antioxidant response and increase IL-2 secretion. However, viability and proliferation tests as well as ultrastructural observations revealed stress signs after treatment with 20 and 30 µg O3, thus designating 10 µg O3 as the optimal concentration in combining cell safety and efficient antioxidant and immune response in our in vitro system. These data offer novel evidence of the fine regulatory role played by the oxidative stress level in the hormetic response of T lymphocytes to O2-O3 administration.

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Development of a PML-RARα-IL-2 Recombinant Plasmid DNA for APL.

Blood ◽

10.1182/blood.v110.11.4881.4881 ◽

2007 ◽

Vol 110 (11) ◽

pp. 4881-4881

Author(s):

Yubing Zhou ◽

Gang Hu ◽

Dongzhi Cen ◽

Lijian Yang ◽

Shaohua Chen ◽

...

Keyword(s):

Immune Response ◽

Dna Vaccine ◽

Recombinant Plasmid ◽

Dna Vaccines ◽

Gene Segment ◽

Interleukin 2 ◽

A549 Cells ◽

Jurkat Cells ◽

Active Immunotherapy ◽

Nb4 Cells

Abstract Specific active immunotherapy with DNA vaccines is one of the best approaches to eradicate minimal residual disease in cancer patients, when appropriate tumor-specific antigens are identified for this disease. However, little data are regarding the DNA vaccines in acute leukemia, recent reports has shown that a human PML-RARα breakpoint-drived DNA vaccine can prevent APL in mice model, but the host immune response were weakly, due to the weak immunogenicity. In order to improve the effect of DNA vaccine for acute promyelocytic leukemia (APL) therapy, we develop a vector coexpressing PML-RARα gene fused to human Interleukin- 2 (hIL-2) gene. PML-RARα fusion gene segment and the full-lenght hIL-2 gene were amplified from NB4 cells or Jurkat cells. Both PCR products were cloned into PIRES plasmid respectively to construct a recombinant plasmid PML-RARα-IRES-hIL-2. The recombinant plasmids were then transfected into K562 or A549 cells respectively. The expression of the PML-RARα/hIL-2 mRNA and protein in transfected cells were identified by RT-PCR, dot blotting, ELISA and Western-Blot respectively. By in vivo assays, BALB/c mice were vaccinated at 6–8 week of age with a total of 200 μg DNA in normal saline, injected into two sites in the quadriceps muscles on day 0, 7 and 21. The plasmid containing the same sequence of PML-RARα gene and blank plasmid served as controls. Two weeks after the final DNA boost, both PML-RARα/hIL-2 mRNA and protein, serum INF-γ and anti-NB4 cells specific cytotoxicity of splenocytes following 7 days of stimulation in vitro with freeze thawing NB4 cells and recombinant human IL-2 were assessed by ELISA and LDH assays. The results showed that the sequence of the fragments inserted in multi-clone site (MCS) A and MCS B of PIRES plasmid were absolutely correct by double restriction enzyme cutting analysis (Sal I/Not I) and sequence analysis, the PML-RARα/hIL-2 mRNA and protein could be identified in transfected K562 or A549 cells and in mice quadriceps muscles. The level of serum INF-γ and cytotoxicity of splenocytes against NB4 cells from immunized mice was significant increased than that from control groups. Our results indicated that the recombinant plasmid expressing PML-RARα and hIL-2 was successfully constructed, which can induce more effective immune response and anti-APL cells effect in animal models. The data suggest that effective vaccination approaches should be possible against APL. It could be farther used in the research as PML-RARα DNA vaccine for APL.

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Features of the lymphocytic microenvironment in metastatic uveal melanoma

Уральский медицинский журнал ◽

10.52420/2071-5943-2021-20-2-36-42 ◽

2021 ◽

Vol 20 (2) ◽

pp. 36-42

Author(s):

A. Y. Shamanova ◽

E. L. Kazachkov ◽

I. Е. Panova ◽

A. V. Vazhenin ◽

Т. N. Shamaeva ◽

...

Keyword(s):

Immune Response ◽

Uveal Melanoma ◽

Antitumor Effect ◽

Primary Melanoma ◽

Distant Metastases ◽

Melanoma Cells ◽

The Body ◽

Immune Reactivity ◽

Tumor Metastases ◽

Cd4 Lymphocytes

Introduction. Uveal melanoma is a malignant neoplasm of the vascular tract of the eye. Prevention of metastasis of this tumor is one of the main tasks in order to increase the rates of relapse-free survival of patients. Despite the pronounced immunosuppressive activity of uveal melanoma cells, its lymphocytic microenvironment exerts its antitumor effect.Aim of the study. Compare the lymphocytic microenvironment of primary uveal melanomas and distant metastases (to the liver).Мaterials and methods. The tissue material of choroid melanoma after enucleation and the material of tumor metastases for the period 2013-2018 were studied. An immunohistochemical study was performed using CD8, CD4, and CD56 markers for the qualitative and quantitative assessment of lymphocytes in the tumor stroma.Results. Differences were found in the lymphocytic infiltration of the uveal melanoma stroma and its distant metastases. A statistically significantly greater representation of CD4, CD8-lymphocytes and CD56 cells in tumor metastases than in primary melanoma tissue samples, with CD4-lymphocytes predominant. A direct high-strength correlation was registered between the number of CD4-lymphocytes and CD8-lymphocytes.Discussion. Malignant cells actively modify their cellular and stromal-vascular environment, ensuring their active growth and reproduction. The question of the immune reactivity of the surrounding cells in relation to uveal melanoma remains debatable. According to our data, which is consistent with a number of other studies, uveal melanoma cells do not completely evade the body's immune response. Thus, the determination of possible points of antitumor exposure can be based on a detailed study of the microenvironment of uveal melanoma. Conclusions. The pronounced lymphocytic infiltrate found in uveal melanoma metastases in comparison with the primary tumor indicates an active immune response of the body to the tumor. These results of our study confirm the importance of further studying the immune-mediated antitumor effect on uveal melanoma and the need to investigate possible approaches to immunotherapy.

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