Method of amplifying natural killer T cells

A natural killer and cell technology, applied in animal cells, vertebrate cells, cell culture active agents, etc., can solve problems such as difficult access to cellular immunotherapy

Inactive Publication Date: 2003-09-24
KIRIN BREWERY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, even from umbilical cord blood, it is difficult to obtain sufficient quantities of these cells for immunotherapy

Method used

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  • Method of amplifying natural killer T cells
  • Method of amplifying natural killer T cells
  • Method of amplifying natural killer T cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052]Example 1. Human Vα24 + Expansion of NKT cells

[0053] (1) Preparation of PBMCs and preparation of G-PBSCs

[0054] Buffy coats (crude PBMCs) prepared from 400 ml of whole blood from healthy individuals were provided by the Japanese Red Cross Blood Center. Cellular fractions containing PBSCs (crude G-PBSCs) were obtained from chemotherapy-treated patients who received G-CSF subcutaneously at a dose of 100-250 μg / person / day for 6-10 days after chemotherapy (Table 1 ), within 2-24 hours of the last G-CSF injection, with COBE Spectra TM Cell Separator (LAKE WOOD, CO USA 80215) to complete apheresis (Hematopoietic Stem Cells: Applications in Biology and Therapeutics. Levit DJ, Mertelsmann R (eds), Marcel Dekker, Inc., New York, 1995, 611- 630).

[0055] From the obtained crude PBMCs and crude G-PBSCs, mononuclear cell fractions (referred to as "PBMCs" and "G-PBSCs", respectively) were prepared using Lymphosepal density gradient medium (Immuno-Biological Laboratories Gun...

Embodiment 2

[0068] Example 2 Evaluation of Vα24 + Cytotoxic activity of NKT cells against human tumor cells

[0069] Purification of human Vα24 from day 12 cultured cells using the FACS Vantage cell sorting system + NKT cells, and the cytotoxic activity of these cells against tumor cells and the cytokine production activity of these cells were assessed.

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Abstract

Expansion of human Vα24+ natural killer T by culturing a monocyte fraction obtainable from human peripheral blood in the presence of a cytokine capable of proliferating and / or activating lymphocytes and α-glycosylceramide cells, the hematopoietic stem cells in the peripheral blood are mobilized by granulocyte-colony stimulating factor. The cell fraction containing human Vα24+ natural killer T cells expanded by this method can be used as an effective cancer therapeutic agent.

Description

technical field [0001] The present invention relates to a method for amplifying human Vα24 + Method for natural killer T (NKT) cells, and human Vα24 obtained by the method + NKT cells and a human Vα24-containing + Use of components of NKT cells. Background technique [0002] NKT cells are a unique subset of mature lymphocytes, which have both NK cell receptors and T cell receptors (Annu.Rev.Immunol., 15, 535-562, 1997; J.Exp.Med., 182 , 633-638, 1995). Murine NKT cells express NK1.1 and TCRαβ receptors and are particularly dense in bone marrow (J. Immunol., 145, 3209-3215, 1990) and liver (J. Exp. Med., 180, 699-704, 1994) . Such cells express a very limited TCR repertoire (J. Exp. Med., 180, 1097-1106, 1994; Int. Immunol., 7, 1157-1161, 1995), including a constant alpha chain. These results imply that the ligands of NKT cells are non-polymorphic and non-classical MHC class I molecules that exhibit a unique process of specific antigen presentation via a TAP (transporte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K39/00A61P35/00C12N5/02C12N5/0783
CPCA61K39/0011A61K2035/124A61K2039/5158A61P35/00C12N5/0646C12N2501/22C12N2501/23C12N5/0636
Inventor 若杉寻
Owner KIRIN BREWERY CO LTD
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