JPH06125996A - Monocyte activation acceleration method by fluctuating magnetic field - Google Patents
Monocyte activation acceleration method by fluctuating magnetic fieldInfo
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- JPH06125996A JPH06125996A JP4276232A JP27623292A JPH06125996A JP H06125996 A JPH06125996 A JP H06125996A JP 4276232 A JP4276232 A JP 4276232A JP 27623292 A JP27623292 A JP 27623292A JP H06125996 A JPH06125996 A JP H06125996A
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- magnetic field
- monocytes
- ifn
- fluctuating magnetic
- gamma
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Abstract
Description
【０００１】[0001]
【産業上の利用分野】本発明は、変動磁場を使用して、
単球を活性化させる方法に関する。さらに詳しくは、単
球をＩＦＮ−γと接触させ、同時に変動磁場をかけて、
該単球を活性化させる方法に関する。FIELD OF THE INVENTION The present invention uses a varying magnetic field to
It relates to a method of activating monocytes. More specifically, the monocytes are brought into contact with IFN-γ, and a varying magnetic field is applied at the same time,
It relates to a method of activating the monocytes.
【０００２】[0002]
【従来の技術】ＩＦＮ−γは、マイトジェンの刺激によ
り、リンパ球が産生し、分泌される低分子量の糖タンパ
ク質である。ＩＦＮ−γは、ウイルス感染に対してなん
らかの防御効果を発揮することや免疫機構に関与してい
ることが知られている。2. Description of the Related Art IFN-γ is a low molecular weight glycoprotein produced and secreted by lymphocytes upon stimulation with mitogen. It is known that IFN-γ exerts some protective effect against viral infection and is involved in the immune mechanism.
【０００３】現在、ウイルス感染の治療あるいは癌の治
療のために、ＩＦＮ−γを患者に直接投与することや患
者から採取した細胞を活性化することが行われている。
しかし、ＩＦＮ−γは、副作用が強いため、十分な治療
効果を得るために多量に投与したり使用したりすること
が困難な場合が多い。さらに、個体によってはＩＦＮ−
γに感受性が低い場合もある。従って、より少量のＩＦ
Ｎ−γで確実に細胞の活性化を行わせる手段が望まれて
いる。Currently, in order to treat viral infection or cancer, IFN-γ is directly administered to a patient or cells collected from the patient are activated.
However, since IFN-γ has strong side effects, it is often difficult to administer or use a large amount of IFN-γ in order to obtain a sufficient therapeutic effect. Furthermore, depending on the individual, IFN-
It may be less sensitive to γ. Therefore less IF
A means for reliably activating cells with N-γ is desired.
【０００４】[0004]
【発明が解決しようとする課題】本発明は、上記の欠点
を解決しようとするもので、その目的は、変動磁場を用
いることによって、少量のＩＦＮ−γによる刺激によっ
て単球の活性化が確実に行われるような方法を提供する
ことにある。DISCLOSURE OF THE INVENTION The present invention is intended to solve the above-mentioned drawbacks, and its purpose is to ensure activation of monocytes by stimulation with a small amount of IFN-γ by using a variable magnetic field. The purpose is to provide a method such as
【０００５】[0005]
【課題を解決するための手段】発明者らは、単球をＩＦ
Ｎ−γで活性化させる際に、同時に変動磁場をかけるこ
とにより、従来では作用しないような少量のＩＦＮ−γ
で細胞が活性化されること、および従来と同量のＩＦＮ
−γで、単球の活性化が大幅に促進されることを見い出
し、本発明を完成するに至った。SUMMARY OF THE INVENTION The inventors
By applying a fluctuating magnetic field at the same time when activating with N-γ, a small amount of IFN-γ that does not work conventionally is obtained.
Activation of cells and the same amount of IFN as before
It was found that -γ significantly promotes the activation of monocytes, and completed the present invention.
【０００６】本発明の単球を活性化させる方法は、
（ｉ）該単球をＩＦＮ−γと接触させる工程、および
（ｉｉ）該単球に変動磁場をかける工程、を包含する。The method of activating monocytes of the present invention comprises:
(I) contacting the monocytes with IFN-γ, and (ii) applying a varying magnetic field to the monocytes.
【０００７】好ましい実施態様においては、上記単球
は、ヒトの末梢血由来である。[0007] In a preferred embodiment, the monocytes are derived from human peripheral blood.
【０００８】好ましい実施態様においては、上記変動磁
場はパルス磁場である。In a preferred embodiment, the varying magnetic field is a pulsed magnetic field.
【０００９】好ましい実施態様においては、上記変動磁
場は、周波数０．１〜１００００Ｈｚ、平均磁場強度１
×１０-6〜１Ｔ、および最大磁場強度１×１０-5〜１０
Ｔである。In a preferred embodiment, the fluctuating magnetic field has a frequency of 0.1 to 10000 Hz and an average magnetic field strength of 1
× 10 -6 to 1 T, and maximum magnetic field strength 1 × 10 -5 to 10
T.
【００１０】本発明によって活性化が促進される単球
は、任意の哺乳類動物に由来する単球であり、好ましく
はヒト末梢血由来である。このような単球は、例えば、
生体の血液を採取し、この血液から単離精製することに
よって得られる。細胞の単離精製法としては、例えば、
密度勾配遠心分離法などの公知の任意の方法が用いられ
得る。The monocytes whose activation is promoted by the present invention are monocytes derived from any mammal, preferably human peripheral blood. Such monocytes are, for example,
It is obtained by collecting blood of a living body and isolating and purifying it from this blood. As a cell isolation and purification method, for example,
Any known method such as a density gradient centrifugation method can be used.
【００１１】本発明で、「単球が活性化」されるとは、
単球が、生理活性物質をより多く分泌するようになるこ
と、細菌や異物に対する細胞障害活性を有するようにな
ること、抗腫瘍活性を有するようになることなどをい
う。具体的には、単球の活性化は、例えば、細胞内カル
シウムの増加あるいは細胞のグルコース消費量の増加に
より測定され得る。In the present invention, "monocyte activation" means that
It means that monocytes secrete more physiologically active substances, have cytotoxic activity against bacteria and foreign substances, and have antitumor activity. Specifically, monocyte activation can be measured, for example, by an increase in intracellular calcium or an increase in cellular glucose consumption.
【００１２】本発明に用いられるＩＦＮ−γの由来は特
に限定されず、任意の生物に由来する天然のＩＦＮ−
γ、および遺伝子操作技術によって得られるＩＦＮ−γ
のいずれであってもよい。ＩＦＮ−γは、適当な緩衝液
に溶解させるかあるいは担体に固定して用いられ得る。
ＩＦＮ−γを単球に接触させるには、任意のあらゆる方
法が用いられ得る。例えば、ＩＦＮ−γを緩衝液に溶解
させて用いる場合、その溶液と単離精製して得られた単
球を含む緩衝液とを混合し、適当にインキュベートさせ
る方法を用い得る。さらに担体に固定した単球に、ＩＦ
Ｎ−γ溶液を接触させることもできる。ＩＦＮ−γを適
当な手段を用いて担体に固定して使用する場合、例えば
該ＩＦＮ−γ固定担体を、単球を含む溶液に接触させる
方法を用い得る。本発明で用いる場合、ＩＦＮ−γの使
用量は、ＩＦＮ−γ溶液単独で単球の活性化を行う場合
よりも少量でよく、好ましくは、細胞１〜２×１０6個
／ｍｌを含む培養液あるいは体液に０．１ＩＵ／ｍｌ〜
３００ＩＵ／ｍｌの割合で使用する。The origin of IFN-γ used in the present invention is not particularly limited, and natural IFN-derived from any organism is used.
γ, and IFN-γ obtained by genetic engineering techniques
Either of them may be used. IFN-γ can be used by dissolving it in an appropriate buffer or fixing it on a carrier.
Any method of contacting IFN-γ with monocytes can be used. For example, when IFN-γ is dissolved in a buffer and used, a method may be used in which the solution is mixed with a buffer containing monocytes obtained by isolation and purification, and the mixture is appropriately incubated. In addition, monocytes fixed on a carrier
It is also possible to contact the N-γ solution. When IFN-γ is immobilized on a carrier using an appropriate means and used, for example, a method of contacting the IFN-γ immobilized carrier with a solution containing monocytes can be used. When used in the present invention, the amount of IFN-γ used may be smaller than that in the case of activating monocytes with an IFN-γ solution alone, and preferably a culture containing 1 to 2 × 10 6 cells / ml. 0.1 IU / ml for liquid or body fluid
Use at a rate of 300 IU / ml.
【００１３】本発明に用いられる変動磁場には、例えば
交流磁場、パルス磁場があり、特にパルス磁場のような
磁場が速く変化する変動磁場が好ましい。変動磁場は、
周波数０．１〜１００００Ｈｚ、平均磁場強度１×１０
-6〜１Ｔ、最大磁場強度１×１０-5〜１０Ｔであること
が好ましい。The fluctuating magnetic field used in the present invention includes, for example, an alternating magnetic field and a pulse magnetic field, and a fluctuating magnetic field such as a pulse magnetic field in which the magnetic field changes rapidly is preferable. The fluctuating magnetic field is
Frequency 0.1-10000Hz, average magnetic field strength 1x10
It is preferable that the magnetic field intensity is −6 to 1 T and the maximum magnetic field strength is 1 × 10 −5 to 10 T.
【００１４】本発明の単球の活性化法においては、さら
に必要に応じて、細胞活性促進剤を単球に作用させ得
る。好ましくはこの細胞活性促進剤はリポ多糖（ＬＰ
Ｓ）である。In the method for activating monocytes of the present invention, a cell activity promoter may be allowed to act on monocytes, if necessary. Preferably, the cell activity promoter is lipopolysaccharide (LP
S).
【００１５】本発明の単球の活性化法は、具体的には、
例えば以下のような方法である。The method for activating monocytes of the present invention is specifically as follows.
For example, the following method is used.
【００１６】まず、変動磁場発生装置として、例えば、
適当な直径を有するヘルムホルツコイル、変動磁場の波
形を制御するファンクションジェネレーターおよび該ジ
ェネレーターの出力を増幅して電流を発生させ得るパワ
ーサプライからなる装置を準備する。発生させる磁場
は、例えば周波数５０〜１００Ｈｚ、ＤＵＴＹ４０〜６
０％、平均磁場強度２．２ｍＴ、最大磁場強度４．４ｍ
Ｔのパルス磁場で有り得る。First, as a variable magnetic field generator, for example,
A Helmholtz coil having an appropriate diameter, a function generator for controlling the waveform of the fluctuating magnetic field, and a power supply capable of amplifying the output of the generator to generate a current are prepared. The magnetic field to be generated is, for example, frequency 50 to 100 Hz, DUTY 40 to 6
0%, average magnetic field strength 2.2 mT, maximum magnetic field strength 4.4 m
It can be a pulsed magnetic field of T 2.
【００１７】活性化される単球は、例えばヒトの末梢血
から得られる。この血液を採取し、密度勾配遠心法など
の方法によって目的の単球が単離精製される。単離精製
した単球は、適当な緩衝液あるいは培養液（例えば１０
％ＦＢＳ−ＲＰＭＩ１６４０培地）中に浮遊させるか、
またはプレートに付着させて培養液に浸しておく。Activated monocytes are obtained, for example, from human peripheral blood. This blood is collected, and the target monocytes are isolated and purified by a method such as density gradient centrifugation. The isolated and purified monocytes may be used in an appropriate buffer or culture medium (for example, 10
% FBS-RPMI1640 medium), or
Alternatively, it is attached to the plate and immersed in the culture solution.
【００１８】一方、ＩＦＮ−γは、例えば上記単離精製
した単球を含む緩衝液あるいは培養液（例えば１０％Ｆ
ＢＳ−ＲＰＭＩ１６４０培地）に溶解させて、適当な濃
度に調製しておく。上記のように調製された単球を含む
液とＩＦＮ−γの溶液とを適当な温度で、約６０秒〜１
０時間、接触させると同時に、該溶液に上記変動磁場発
生装置を用いて、変動磁場を与える。このことにより単
球が活性化される。単球が活性化されたことは、細胞内
のカルシウム濃度が増加することによって確認され得
る。On the other hand, IFN-γ is, for example, a buffer solution or culture solution (for example, 10% F) containing the isolated and purified monocytes.
BS-RPMI1640 medium) to prepare an appropriate concentration. The solution containing monocytes prepared as described above and the solution of IFN-γ are heated at a suitable temperature for about 60 seconds to 1 second.
Simultaneously with contact for 0 hours, a fluctuating magnetic field is applied to the solution using the fluctuating magnetic field generator. This activates monocytes. Monocyte activation can be confirmed by an increase in intracellular calcium concentration.
【００１９】次に、この培養細胞から、ＩＦＮ−γを取
り除く。単球がプレートに付着している場合、培養液を
取り除くことによってＩＦＮ−γを簡単に取り除くこと
が可能である。ＩＦＮ−γ除去の後、必要であれば、Ｌ
ＰＳを０．１ｎｇ／ｍｌ〜１０ｍｇ／ｍｌの濃度となる
ように加えて、単球と接触させる。接触させる時間は、
好ましくは１〜１００時間、より好ましくは２４〜８０
時間である。ＬＰＳとの接触によって単球の抗腫瘍活性
が上昇する。この活性は、単球のグルコース消費量を測
定することによって検知され得る。グルコース消費量
は、当業者に公知の方法で測定される。Next, IFN-γ is removed from the cultured cells. If monocytes are attached to the plate, IFN-γ can be easily removed by removing the culture medium. After removal of IFN-γ, if necessary, L
PS is added to a concentration of 0.1 ng / ml to 10 mg / ml and brought into contact with monocytes. The contact time is
Preferably 1 to 100 hours, more preferably 24 to 80 hours
It's time. Contact with LPS increases the antitumor activity of monocytes. This activity can be detected by measuring the glucose consumption of monocytes. Glucose consumption is measured by methods known to those skilled in the art.
【００２０】[0020]
【実施例】以下、本発明を実施例に基づいて説明する
が、本発明はこれに限定されるものではない。EXAMPLES The present invention will be described below based on examples, but the present invention is not limited thereto.
【００２１】（実施例１）ヒトの新鮮な末梢血から、密
度勾配遠心法により単球を単離精製した。この単球を５
％ヒトＡＢ型血清−ＲＰＭＩ−１６４０中に細胞濃度１
×１０7個／ｍｌとなるように調製し、ウェル中に０．
５ｍｌずつ分注し、３７℃、５％ＣＯ2条件下で３０分
間インキュベートした。その後ウェルを温ハンクス溶液
でピペッティングして非付着細胞を取り除き、１ｍＭFu
raII溶液（FuraII-AM（同仁化学研究所製）２０μｌとP
luronic F-127 ４μｌを加えたものをハンクス溶液で
２ｍｌとする：１０μＭFuraII-AM）を０．５ｍｌ加
え、３７℃、５％ＣＯ2で３０分間インキュベートし
た。ウェル中のFuraII溶液を０．４ｍｌ取り除き、代わ
りにハンクス溶液を０．４ｍｌ加え（FuraII-AM濃度を
２μｌにする）、さらに３７℃、５％ＣＯ2の条件で３
０分間インキュベートした。インキュベート後、温ハン
クス溶液で３回洗浄し、ウェル中にHanks'を最終的に
０．５ｍｌになるように加えてこれを測定用の試料とし
た。このようにして調製した細胞試料を３７℃に温度設
定した細胞内カルシウムイオン濃度測定装置（CASALS：
オリンパス社製）の測定台上におき、ハンクス溶液を
０．２ｍｌ取り除いておいた。細胞試料に、３７℃に保
温しておいたＩＦＮ−γを１２５ＩＵ／ｍｌの濃度で含
むハンクス溶液０．２ｍｌをウェルに加えるのと同時に
垂直方向から９度の角度を持った、周波数１００Ｈｚ、
最大磁場強度４．４ｍＴ、平均磁場強度２．２ｍＴ、Ｄ
ＵＴＹ５０％のパルス磁場を与えた。この時のパルス磁
場は、直径２２ｃｍのヘルムホルツコイル、変動磁場の
波形を制御するファンクションジェネレーターおよびジ
ェネレーターの出力を増幅して電流を発生させることの
できるパワーサプライからなる装置によって発生させ
た。ヘルムホルツコイル内部では、磁場強度が上下２％
以内のほぼ一様なパルス磁場が得られた。ＩＦＮ−γお
よび磁場を与えた直後からの細胞内カルシウムイオン濃
度の変化をCASALSによって観察した。(Example 1) Monocytes were isolated and purified from fresh human peripheral blood by density gradient centrifugation. 5 this monocyte
% Human Serum AB Serum-RPMI-1640 with a cell concentration of 1
× 10 7 cells / ml, and 0.
5 ml aliquots were dispensed and incubated at 37 ° C. under 5% CO 2 for 30 minutes. The wells are then pipetted with warm Hanks solution to remove non-adherent cells and 1 mM Fu
raII solution (FuraII-AM (Dojindo Laboratories) 20 μl and P
4 μl of luronic F-127 was made up to 2 ml with Hanks solution: 0.5 ml of 10 μM FuraII-AM) was added and incubated for 30 minutes at 37 ° C., 5% CO 2 . Remove 0.4 ml of FuraII solution from the well, add 0.4 ml of Hank's solution instead (make the concentration of FuraII-AM 2 μl), and add 3% under the condition of 37 ° C. and 5% CO 2.
Incubated for 0 minutes. After the incubation, the plate was washed with a warm Hanks solution three times, and Hanks' was added to the well so that the final volume was 0.5 ml, and this was used as a sample for measurement. Cellular calcium ion concentration measuring device (CASALS:
It was placed on a measuring table (manufactured by Olympus) and 0.2 ml of the Hanks' solution was removed. To the cell sample, 0.2 ml of Hank's solution containing IFN-γ kept at 37 ° C. at a concentration of 125 IU / ml was added to the wells, and at the same time, an angle of 9 ° from the vertical direction, a frequency of 100 Hz,
Maximum magnetic field strength 4.4mT, average magnetic field strength 2.2mT, D
A pulse magnetic field of 50% UTY was applied. The pulsed magnetic field at this time was generated by a device including a Helmholtz coil having a diameter of 22 cm, a function generator controlling the waveform of the fluctuating magnetic field, and a power supply capable of amplifying the output of the generator to generate a current. Inside the Helmholtz coil, the magnetic field strength is up and down 2%.
A nearly uniform pulsed magnetic field within is obtained. Changes in intracellular calcium ion concentration immediately after applying IFN-γ and a magnetic field were observed by CASALS.
【００２２】比較の為に変動磁場を与えなかったこと以
外は、上記と同様にして、調製し、同様の操作を施した
単球の細胞内カルシウムイオン濃度変化を測定した。代
表的な細胞（４つ）の中のカルシウム濃度変化を図１お
よび図２に示す。丸印、菱形、三角、および点線は、そ
れぞれ別々の細胞を表す。図１は変動磁場を与えなかっ
た場合の結果を示し、図２は変動磁場を与えた場合の結
果を示している。横軸の時間は、ＩＦＮを与えてからの
時間を表している。この結果から、変動磁場を与えると
細胞内へのカルシウム流入が時間的に早く起こり、流入
量も増えることが判る。For the purpose of comparison, changes in intracellular calcium ion concentration of monocytes prepared and subjected to similar operations were measured in the same manner as above, except that a varying magnetic field was not applied. Changes in calcium concentration in representative cells (4) are shown in FIGS. 1 and 2. Circles, diamonds, triangles, and dotted lines represent separate cells. FIG. 1 shows the results when no fluctuating magnetic field was applied, and FIG. 2 shows the results when a fluctuating magnetic field was applied. The time on the horizontal axis represents the time after giving IFN. From this result, it is understood that when a fluctuating magnetic field is applied, calcium inflow into cells occurs earlier in time and the inflow amount increases.
【００２３】（実施例２）ヒト末梢血から、密度勾配遠
心法により分離精製した単球を、培地中に２×１０6個
／ｍｌになるように加えた。この培地を４つの培養プレ
ートに４００μｌずつ分注し、３７℃、５％ＣＯ2の条
件下で、２時間インキュベートして細胞をウェルに付着
させ、単球の標品を調製した。ヒト組換えＩＦＮ−γ
（Boehringer-Mannheim社製）をまず、１０％ＦＢＳ−
ＲＰＭＩ１６４０に溶解させ、それぞれのプレートに２
５IU/ml、５０IU/ml、１００IU/mlおよび２００IU/mlに
なるように加え、すべてのプレートに水平方向のパルス
磁場を与えながら約９時間培養した。この時のパルス磁
場は、直径２２ｃｍのヘルムホルツコイル、変動磁場の
波形を制御するファンクションジェネレーターおよびジ
ェネレーターの出力を増幅して電流を発生させることの
できるパワーサプライからなる装置によって発生させ
た、周波数５０Ｈｚ、ＤＵＴＹ５０％のパルス磁場（最
大磁場強度４．４ｍＴ、平均磁場強度２．２ｍＴ）であ
った。ヘルムホルツコイル内部では、磁場強度が上下２
％以内のほぼ一様な変動磁場が得られた。Example 2 Monocytes separated and purified from human peripheral blood by density gradient centrifugation were added to the medium at 2 × 10 6 cells / ml. 400 μl of this medium was dispensed into each of four culture plates and incubated at 37 ° C. under 5% CO 2 for 2 hours to allow cells to adhere to the wells to prepare a monocyte preparation. Human recombinant IFN-γ
(Boehringer-Mannheim), first, 10% FBS-
Dissolve in RPMI1640 and add 2 to each plate.
In addition to 5 IU / ml, 50 IU / ml, 100 IU / ml and 200 IU / ml, all plates were cultured for about 9 hours while applying a horizontal pulsed magnetic field. The pulsed magnetic field at this time was generated by a device including a Helmholtz coil having a diameter of 22 cm, a function generator for controlling the waveform of the fluctuating magnetic field, and a power supply capable of amplifying the output of the generator to generate a current, a frequency of 50 Hz, The pulse magnetic field was 50% DUTY (maximum magnetic field strength 4.4 mT, average magnetic field strength 2.2 mT). Inside the Helmholtz coil, the magnetic field strength is above and below 2
An almost uniform fluctuating magnetic field within% was obtained.
【００２４】培養が終了した後、ＩＦＮ−γを含む培養
液を取り除き、代わりにＬＰＳを含有する緩衝液をすべ
てのプレートに５０ｎｇ／ｍｌの濃度になるように加え
て、３７℃、５％ＣＯ2条件下で約６０時間あるいは約
８０時間培養した。６０時間あるいは８０時間の培養
後、それぞれの培養液中のグルコース濃度をユニキット
グルコースＥ（中外製薬社製）で測定し、間接的に抗腫
瘍活性を評価した。活性化インデックスは、以下の式に
より算出される。After the culture was completed, the culture solution containing IFN-γ was removed, and instead, a buffer solution containing LPS was added to all the plates to a concentration of 50 ng / ml, and the mixture was incubated at 37 ° C. with 5% CO 2. The cells were cultured under the two conditions for about 60 hours or about 80 hours. After culturing for 60 hours or 80 hours, the glucose concentration in each culture solution was measured by Unikit Glucose E (manufactured by Chugai Pharmaceutical Co., Ltd.) to indirectly evaluate the antitumor activity. The activation index is calculated by the following formula.
【００２５】[0025]
【化１】 [Chemical 1]
【００２６】なお、コントロール中の残留グルコース量
は、ＩＦＮ−γおよびＬＰＳを加えずに同じ工程を経た
細胞の培養液中のグルコース量のことである。The amount of residual glucose in the control is the amount of glucose in the culture solution of cells that have undergone the same steps without adding IFN-γ and LPS.
【００２７】比較のために、変動磁場を与えなかったこ
と以外は上記と同様にして調製し、同様の操作を施した
単球のグルコース消費量も測定した。結果を表１に示
す。For comparison, the glucose consumption of monocytes prepared in the same manner as above except that no variable magnetic field was applied and subjected to the same operation was also measured. The results are shown in Table 1.
【００２８】[0028]
【表１】 [Table 1]
【００２９】この結果から、ＩＦＮ−γの濃度が２５IU
/mlのときに、変動磁場による細胞活性化の促進が顕著
であることが判る。From this result, the concentration of IFN-γ was 25 IU.
It can be seen that the promotion of cell activation by the fluctuating magnetic field is remarkable when the amount is / ml.
【００３０】（実施例３）実施例１の実験を別のヒトか
ら得られた細胞（末梢血由来）についても同様に行っ
た。ただし、ＩＦＮ−γの濃度は、５０ＩＵ／ｍｌのみ
で、細胞濃度は、４×１０6個／ｍｌおよび２×１０6個
／ｍｌの２種類で行った。結果を表２に示す。Example 3 The experiment of Example 1 was similarly performed on cells (derived from peripheral blood) obtained from another human. However, the concentration of IFN-γ was 50 IU / ml only, and the cell concentration was 2 × 10 6 cells / ml and 2 × 10 6 cells / ml. The results are shown in Table 2.
【００３１】[0031]
【表２】 [Table 2]
【００３２】この末梢血は、ＩＦＮ−γのみの刺激に対
しては、感受性がなかったが、変動磁場を与えることに
よって、活性化された。This peripheral blood was not sensitive to stimulation with IFN-γ alone, but was activated by applying a fluctuating magnetic field.
【００３３】[0033]
【発明の効果】本発明によれば、変動磁場を使用して、
ＩＦＮ−γと接触した単球の活性化を促進させる方法が
提供される。本発明の方法では、ＩＦＮ−γの濃度が低
い場合でも、単球が十分に活性化され、さらに、ＩＦＮ
−γのみでは、十分に活性化されない単球でも活性化さ
せることができる。According to the present invention, a variable magnetic field is used,
Methods for promoting activation of monocytes contacted with IFN-γ are provided. According to the method of the present invention, monocytes are sufficiently activated even when the concentration of IFN-γ is low, and
-Γ alone can activate monocytes that are not sufficiently activated.
【図１】ＩＦＮ−γのみで活性化させた、ヒト末梢血由
来の単球のカルシウムイオンの取り込みの経時的変化を
示すグラフである。FIG. 1 is a graph showing the time course of calcium ion uptake by human peripheral blood-derived monocytes activated with IFN-γ alone.
【図２】ＩＦＮ−γおよび変動磁場で活性化させた、ヒ
ト末梢血由来の単球のカルシウムイオンの取り込みの経
時的変化を示すグラフである。FIG. 2 is a graph showing the time course of calcium ion uptake by human peripheral blood-derived monocytes activated with IFN-γ and a varying magnetic field.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 見藤 歩 北海道江別市野幌住吉町７−５ (72)発明者 宮崎 浩 北海道札幌市北区北20条西７丁目20登別荘 18号 (72)発明者 関口 定美 北海道札幌市南区真駒内上町５丁目６−３ (72)発明者 高橋 恒夫 北海道札幌市中央区宮の森１条６丁目４− 20−504 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Ayumi Mito 7-5 Nopporo, Sumiyoshi-cho, Ebetsu-shi, Hokkaido (72) Inventor Hiroshi Miyazaki 7-20-20 Kita-ku Nishi, Kita-ku, Sapporo No. 18 Noboribetsu Villa (72) Inventor Satomi Sekiguchi 5-6-3, Makomanaikamimachi, Minami-ku, Sapporo, Hokkaido (72) Inventor Tsuneo Takahashi 1-6-6, Miyanomori, Chuo-ku, Sapporo, Hokkaido 4-20-504
Claims (4)
って、（ｉ）該単球をＩＦＮ−γと接触させる工程、お
よび（ｉｉ）該単球に変動磁場をかける工程、を包含す
る、方法。1. A method for activating a mammal-derived monocyte, which comprises the steps of (i) contacting the monocyte with IFN-γ, and (ii) applying a varying magnetic field to the monocyte. how to.
求項１に記載の方法。2. The method according to claim 1, wherein the monocytes are derived from human peripheral blood.
項１に記載の方法。3. The method of claim 1, wherein the varying magnetic field is a pulsed magnetic field.
０Ｈｚ、平均磁場強度１×１０-6〜１Ｔ、および最大磁
場強度１×１０-5〜１０Ｔである、請求項１に記載の方
法。4. The fluctuating magnetic field has a frequency of 0.1 to 1000.
The method according to claim 1, wherein the magnetic field strength is 0 Hz, the average magnetic field strength is 1 × 10 −6 to 1 T, and the maximum magnetic field strength is 1 × 10 −5 to 10 T.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4276232A JPH06125996A (en) | 1992-10-14 | 1992-10-14 | Monocyte activation acceleration method by fluctuating magnetic field |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4276232A JPH06125996A (en) | 1992-10-14 | 1992-10-14 | Monocyte activation acceleration method by fluctuating magnetic field |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH06125996A true JPH06125996A (en) | 1994-05-10 |
Family
ID=17566541
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4276232A Pending JPH06125996A (en) | 1992-10-14 | 1992-10-14 | Monocyte activation acceleration method by fluctuating magnetic field |
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| Country | Link |
|---|---|
| JP (1) | JPH06125996A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005071057A1 (en) * | 2004-01-22 | 2005-08-04 | Japan Science And Technology Agency | Magnetic or electric field stimulating device and method for promoting, restraining, or obstructing growth and function of living cell or living tissue using the magnetic or electric field stimulating device |
| JP2008093453A (en) * | 1997-02-27 | 2008-04-24 | Catholic Univ Of America | Protection of living system from adverse effect of stress |
-
1992
- 1992-10-14 JP JP4276232A patent/JPH06125996A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008093453A (en) * | 1997-02-27 | 2008-04-24 | Catholic Univ Of America | Protection of living system from adverse effect of stress |
| WO2005071057A1 (en) * | 2004-01-22 | 2005-08-04 | Japan Science And Technology Agency | Magnetic or electric field stimulating device and method for promoting, restraining, or obstructing growth and function of living cell or living tissue using the magnetic or electric field stimulating device |
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