EFFECT OF INTERLEUKINS 2, 7, 15 ON THE PROLIFERATION OF NATURAL KILLERS IN VITRO

Introduction. The actual task of modern adoptive cancer immunotherapy is the selection of the optimal composition of cytokines for ex vivo stimulation of immunocompetent cells for subsequent administration to oncological patients.The purpose of the study was to compare the effect of interleukin (IL) 2, 7, 15 and their combinations on the proliferation of natural killer cells in breast cancer (BC) patients in vitro.Materials and methods. The research was conducted on natural killer cells isolated by magnetic separation from mononuclear cells of peripheral blood of ten patients with locally advanced BC (stage II). After separation, the cells were cultured at a concentration of 2.5 × 105 cells / ml for 10 days in RPMI 1640 medium supplemented with cytokines at a concentration of 40 ng / ml each in five experimental variants: IL‑2; IL‑7; IL‑15; IL‑7 / IL‑15; IL‑2 / IL‑7 / IL‑15. On the 10th day of cultivation, the phenotype of cells and the cell cycle were studied by flow cytometry. For immunophenotyping of cells, we used monoclonal antibodies to antigens: CD3, CD16 / 56, CD45, CD4, CD19, and CD8. For cell cycle study cells were stained with propidium iodide.Results. On the final 10th day of cultivation the number of living cells expressed as percentage of the seeding numbers were significantly different from control (45.9 %) in samples IL‑2 (86.8 %) and IL‑7 / IL‑15 (85.6 %), IL‑15 (76.4 %), IL‑2 / IL‑7 / IL‑15 (75.8 %). The proportion of natural killers (CD16+CD56+) significantly differed from the control (18.2 %) in samples IL‑2 (45.6 %), IL‑15 (39.9 %), IL‑7 / IL‑15 (36.2 %), IL‑2 / IL‑7 / IL‑15 (35.9 %). The propor‑ tion of natural killer T cells (CD3+ / CD16+CD56+) significantly differed from the control (0.4 %) in samples incu‑ bated with IL‑2 (2.06 %), IL‑15 (2.2 %), IL‑7 (0.9 %), IL‑7 / IL‑15 (1.26 %), IL‑2 / IL‑7 / IL‑15 (2.46 %). All experimental tests also showed a significant increase in the proportion of cells in the S‑phase and increase in the proliferation index (G2 / M + S).Conclusion. The maximum stimulation of the proliferation of natural killer cells isolated from the blood of patients with BC in vitro was obtained by stimulation with IL‑15 alone and in combinations with γc‑cytokines.

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Enhancement of in vitro beta-thalassemic and normal hematopoiesis by a noncytotoxic monoclonal antibody, 9.1C3: evidence for negative regulation of hematopoiesis by monocytes and natural killer cells

Blood ◽

10.1182/blood.v72.4.1124.bloodjournal7241124 ◽

1988 ◽

Vol 72 (4) ◽

pp. 1124-1133

Author(s):

G Kannourakis ◽

GR Johnson ◽

CG Begley ◽

JA Werkmeister ◽

GF Burns

Keyword(s):

Monoclonal Antibody ◽

Bone Marrow ◽

Natural Killer Cells ◽

Natural Killer ◽

Colony Formation ◽

Mononuclear Cells ◽

Regulatory Control ◽

Beta Thalassemia ◽

Killer Cells

The enhancement of in vitro human hematopoiesis by the addition of a noncytotoxic monoclonal antibody, 9.1C3, is described. Enhancement of all aspects of in vitro hematopoiesis was observed on addition of 9.1C3 antibody to cultures of mononuclear cells from normal bone marrow, cord blood, and peripheral blood from beta-thalassemia major patients. In cultures with no exogenous colony-stimulating factor (CSF), the addition of 9.1C3 resulted in a two- to eightfold increase in nonerythroid colony formation. Similarly, for cultures maximally stimulated with CSF, the addition of 9.1C3 antibody resulted in a one- to fourfold increase in colony formation. These effects were abrogated by the removal of either adherent, Leu-M3+ or Leu-7+ cells. Colony- forming cells were shown to be present among the 9.1C3-negative cells when mononuclear cells were sorted by flow cytometry. Media conditioned in the presence of 9.1C3 and mononuclear cells were able to enhance colony formation in vitro for normal nonadherent bone marrow cells beyond that achieved with supramaximal amounts of human placental- conditioned medium and erythropoietin. The data suggest that natural killer cells interact with monocytes to exert a negative regulatory control on in vitro granulopoiesis and erythropoiesis. Consequently, the number of progenitor and multipotential cells in cultures of unfractionated cell populations may be greatly underestimated.

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Ethanol and Natural Killer Cells. II. Stimulation of Human Natural Killer Activity by Ethanol In Vitro

Alcoholism Clinical and Experimental Research ◽

10.1111/j.1530-0277.1997.tb04241.x ◽

1997 ◽

Vol 21 (6) ◽

pp. 981-987 ◽

Cited By ~ 11

Author(s):

Feng Li ◽

Robert T. Cook ◽

Carole Alber ◽

Wendy Rasmussen ◽

Jack T. Stapleton ◽

...

Keyword(s):

Natural Killer Cells ◽

Natural Killer ◽

Natural Killer Activity ◽

Killer Cells ◽

Killer Activity ◽

Stimulation Of

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Prolactin Induces IL-2 Associated TRAIL Expression on Natural Killer Cells from Chronic Hepatitis C PatientsIn vivoandIn vitro

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530319666181206125545 ◽

2019 ◽

Vol 19 (7) ◽

pp. 975-984

Author(s):

Maria L.H. Medel ◽

Gabriela G. Reyes ◽

Luz M. Porras ◽

Arturo R. Bernal ◽

Jesús S. Luna ◽

...

Keyword(s):

Viral Load ◽

Natural Killer Cells ◽

Natural Killer ◽

Mononuclear Cells ◽

Killer Cells ◽

Trail Expression ◽

Viral Loads ◽

Nkg2d Expression ◽

Ifn Γ

Background:Natural killer cells (NKC) are a major component of the innate immune response to HCV, mediating their effects through TRAIL and IFN-γ. However, their function is diminished in chronic HCV patients (HCVp). Prolactin is an immunomodulatory hormone capable of activating NKC.Objective:The study aims to explore if hyperprolactinemia can activate NKC in HCVp.Methods:We treated twelve chronic HCVp (confidence level =95%, power =80%) for 15 days with Levosulpiride plus Cimetidine to induce mild hyperprolactinemia. Before and after treatment, we determined TRAIL and NKG2D expression on peripheral blood NKC, along with cytokine profiles, viral loads and liver function. We also evaluated in vitro effects of prolactin and/or IL-2 on NKC TRAIL or NKG2D expression and IFN-γ levels on cultured blood mononuclear cells from 8 HCVp and 7 healthy controls.Results:The treatment induced mild hyperprolactinemia and increased TRAIL expression on NKC as well as the secretion of IL-1ra, IL-2, PDGF and IFN-γ. Viral loads decreased in six HCVp. IL-2 and TRAIL together explained the viral load decrease. In vitro, prolactin plus IL-2 synergized to increase TRAIL and NKG2D expression on NKC from HCVp but not in controls.Conclusion:Levosulpiride/Cimetidine treatment induced mild hyperprolactinaemia that was associated with NKC activation and Th1-type cytokine profile. Also, an increase in TRAIL and IL-2 was associated with viral load decrease. This treatment could potentially be used to reactivate NKC in HCVp.

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Enhancement of in vitro beta-thalassemic and normal hematopoiesis by a noncytotoxic monoclonal antibody, 9.1C3: evidence for negative regulation of hematopoiesis by monocytes and natural killer cells

Blood ◽

10.1182/blood.v72.4.1124.1124 ◽

1988 ◽

Vol 72 (4) ◽

pp. 1124-1133 ◽

Cited By ~ 3

Author(s):

G Kannourakis ◽

GR Johnson ◽

CG Begley ◽

JA Werkmeister ◽

GF Burns

Keyword(s):

Monoclonal Antibody ◽

Bone Marrow ◽

Natural Killer Cells ◽

Natural Killer ◽

Colony Formation ◽

Mononuclear Cells ◽

Regulatory Control ◽

Beta Thalassemia ◽

Killer Cells

Abstract The enhancement of in vitro human hematopoiesis by the addition of a noncytotoxic monoclonal antibody, 9.1C3, is described. Enhancement of all aspects of in vitro hematopoiesis was observed on addition of 9.1C3 antibody to cultures of mononuclear cells from normal bone marrow, cord blood, and peripheral blood from beta-thalassemia major patients. In cultures with no exogenous colony-stimulating factor (CSF), the addition of 9.1C3 resulted in a two- to eightfold increase in nonerythroid colony formation. Similarly, for cultures maximally stimulated with CSF, the addition of 9.1C3 antibody resulted in a one- to fourfold increase in colony formation. These effects were abrogated by the removal of either adherent, Leu-M3+ or Leu-7+ cells. Colony- forming cells were shown to be present among the 9.1C3-negative cells when mononuclear cells were sorted by flow cytometry. Media conditioned in the presence of 9.1C3 and mononuclear cells were able to enhance colony formation in vitro for normal nonadherent bone marrow cells beyond that achieved with supramaximal amounts of human placental- conditioned medium and erythropoietin. The data suggest that natural killer cells interact with monocytes to exert a negative regulatory control on in vitro granulopoiesis and erythropoiesis. Consequently, the number of progenitor and multipotential cells in cultures of unfractionated cell populations may be greatly underestimated.

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