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Drug and method for proliferating natural killer cells

a technology of natural killer cells and proliferating cells, which is applied in the direction of transferrins, drug compositions, immunodeficiency patients and mice that are deficient in the function of natural killers, can solve the problems that mice deficient in natural killer function are very easily infected by specific viruses, and achieve convenient collection, easy analysis of nk activity, and enhanced nk activity

Inactive Publication Date: 2007-05-24
MORINAGA MILK IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0020] When the inventors of the present invention examined a method for screening for a food or drink-derived factor that enhances the NK activity, they administered lactoferrin, which is a food (milk)-derived factor, to an animal for a predetermined period, then administered a Toll-like receptor ligand with specific timing during the administration period, and found that the proportion of NK cells among the intraperitoneal cells markedly increased in the peritoneal cavity of the animal. Further, they found that since it became possible to induce NK cells in the peritoneal cavity by the aforementioned method, the NK activity could be easily analyzed and NK cells could be conveniently collected. Thus, they accomplished the present invention.

Problems solved by technology

That is, even if T lymphocytes and B lymphocytes are normal, immunodeficient patients and mice that are deficient in the natural killer function are very easily infected by specific viruses.
However, since all of these tissues contain NK cells only in low proportions, technical skills and suitable equipment are required to secure NK cells necessary for experiments.
However, the technique disclosed in Patent document 1 is a method for proliferating human NK cells by mixed culture of peripheral blood mononuclear cells and cells of human fibroblast Wilms' tumor line HFWT, and any method for collecting NK cells proliferated in vivo and screening for a factor that enhances the natural killer activity are not disclosed in this document.
However, Patent document 3 does not describe the effect of the modified human lactoferrin for actually stimulating NK cells, and its basis has been unclear.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0065] In this example, a test was performed in order to examine the NK cell proliferating effect of lactoferrin.

(1) Test Method

[0066] Forty five 7-week old C57BL / 6 mice (purchased from Charles River Laboratories Japan, Inc.) were divided into 5 groups, each consisting of 9 animals, acclimatized and fed for 1 week. A solution of bovine lactoferrin (Morinaga Milk Industry Co., Ltd.) in physiological saline (Otsuka Pharmaceutical Co., Ltd.) was administered everyday to the animals of each group for 7 days in an amount of 0 (control sample), 30, 100, 300 or 1000 mg / kg body weight by using an oral sonde. After completion of the administration, the animals were left for 1 day, and the spleen was collected from each mouse, and a suspension of spleen cells was prepared. The number of cells in the prepared cell suspension was adjusted to 2×106 cells per specimen. The aforementioned cell suspension was added with 10 μl per specimen of anti-mouse CD16 / CD32 monoclonal antibody (Fc Block™, B...

example 2

[0069] In this example, a test was performed in order to examine the NK cell proliferating and inducing effect of lactoferrin and a Toll-like receptor ligand.

(1) Test Method

[0070] In the same manner as in Example 1, 9 mice per group, 63 mice in total, were acclimatized and fed for 1 week. Among the groups, 3 groups were assigned lactoferrin administration groups (LF groups, dose of lactoferrin: 300 mg / kg body weight / day), and the remaining 4 groups were assigned control groups (physiological saline was administered). An administration schedule was designed, in which each sample (lactoferrin or physiological saline) was administered everyday for 7 days to mice in all the 7 groups by using an oral sonde, then nothing was administered for 1 day, and the 8th day was assumed as the day of completion of the administration. Among the 3 lactoferrin administration groups and 3 groups of the control groups, groups of which mice were intraperitoneally administered with 100 μg of polyIC (Pol...

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Abstract

NK cells are obtained by administering a Toll-like receptor ligand such as polyinosinic-polycytidylic acid into the peritoneal cavity of an animal to which lactoferrin has been administered to proliferate NK cells in the peritoneal cavity and collecting NK cells from the peritoneal cavity.

Description

TECHNICAL FIELD [0001] The present invention relates to a drug and method for proliferating natural killer cells, a method for producing natural killer cells and a method for screening for a substance having a natural killer cell proliferating action. The present invention is useful in the fields of medicine, food and so forth. BACKGROUND ART [0002] Natural killer (NK) cells are a group of cells having a cytotoxic activity against various tumor cells, virus infected cells and cells having different major histocompatibility antigens, and their activation and inhibition are regulated primarily by receptors that recognize self and nonself. [0003] NK cells are considered to have important functions as one type of lymphocytes involved in natural immunity. Against viral infection, in particular, they play a very important role in immune responses in early infection until acquired immunity is established by T lymphocytes and B lymphocytes. [0004] That is, even if T lymphocytes and B lympho...

Claims

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Application Information

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IPC IPC(8): A61K35/14A61K48/00A61K38/40A61K38/17C12N5/07C12N5/0783C12N5/09C12Q1/04G01N33/15G01N33/50
CPCA61K31/295A61K31/787A61K38/40A61K45/06A61K2300/00A61P31/12A61P35/00A61P37/02A61P37/04A61P43/00A61P7/00A61K38/17C07K14/19
Inventor KUHARA, TETSUYAITOH, TAKEHITO
Owner MORINAGA MILK IND CO LTD
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