title experimental studies of anti-tumor effect induced by ... · preparation of splenic...

Title Experimental Studies of Anti-tumor Effect Induced byMicrowave Tumor Coagulation

Author(s) NOGUCHI, HIROYUKI

Citation 日本外科宝函 (1984), 53(2): 324-337

Issue Date 1984-03-01

URL http://hdl.handle.net/2433/208772

Right

Type Departmental Bulletin Paper

Textversion publisher

Kyoto University

Arch Jpn Chir 53(2), 324～337, Marz, 1984

Experimental Studies of Anti-tumor E妊ectInduced by

Microwave Tumor Coagulation

HIROYUKI NOGUCHI

Department of Gastroenterological Surgery, Wakayama Medical College (Director: Prof. Dr. MASAHARU KATSUMI)

Received for Publication, Oct. 20, 1983.

Introduction

Microwave has been utilized for medical diathermy. Recently, a microwave tissue coagula-

tor (MTC), which is an apparatus used for coagulating tissues by converging the energy of

microwaves in as small an area as possible, was developed 27>_ One of the characteristics of MTC

is non-carbonizing effect on tissues. Since MTC is favorably compared with an electrocautery

and a laser with regard to hemostatic performance, it is useful for operation, particularly on

parenchymal organs such as the liver2s,29，制 andspleena1i, and an increasing number of surgeons

have become attentive to its clinical usefulness. In the treatment of bleeding or stenosis from

gastric, rectal or esophageal cancer, endoscopically guided microwave irradiation done to destroy

tumor tissues in a focal region assures us of a satisfactory resuJt1,u,a2>. Furthermore, microwave

coagulation therapy intended for tumor tissue destruction has begun to be attempted as one of

multidisciplinary treatments of unresectable malignancies involving the digestive tracts and liver

metastases.

In the present experiment, the anti-tumor effect induced by microwave coagulation therapy

was studied.

Materials and Methods

APPARATUS

The apparatus used for microwave coagulation therapy was composed of 1) the generator

which produced microwaves of 2,450 MHz in frequency and 12 cm in wavelength, 2) the coaxial

cable which transmitted microwaves so produced, and 3) the operating monopolar antenna of 10

mm in diameter at the base and 7.8 mm in length.

ANIMALS

Seven to nine-week-old male inbred BALB/c mice were offered for the experiments.

TUMORS

The tumors used in the位 perimentswe陀 asfollows; Meth A 18>, a transplantable fibrosarcoma

Key words: Microwave Coagulation Therapy, Anti-tumor Effect, Microwave Tumor Co唱gulation,Microwave・ immunotherapy. 索引語： 7 イクロ波凝周治療，抗腫蕩効果，マイクロ波腫湯凝固，マイクロ波免疫治療．Present Address: Wakayama Medical College 1 7 bancho Wakayama city, 640, Japan.

ANTI-TUMOR EFFECT INDUCED BY MICROWAVE TUMOR COAGULATION 325

induced in a BALB/c mouse by 3-methyl cholanthrene and Crocker tumor 1802•> detected in

albino mouse and generally known as Sarcoma 180 (S-180). Tumor cells were maintained by

serial passage in ascitic form in BALB/c mice, and were used after complete removing of red

blood cells43>.

EVALUATION OF ANTI-TUMOR EFFECT

Living tumor cells (1×105 cells) were injected into the abdominal cavity of treated or im-

muD;ized mice for challenge, and the anti-tumor effect was evaluated in consideration of the 90-day-

survival rate or survival period 17>.

PREPARATION OF MICRO羽TAVETREATED TUMOR CELLS

A dish of 3 cm in diameter holding 6 ml of a Meth A (3×107 cells) suspension in Roswell

Park Memorial Institute Medium 1640 (RPMI-1640) (Nissui Pharmaceutical Co., Tokyo, Japan)

or phosphate buffered saline (PBS) was buried in heat insulating material, and the monopolar

antenna was immersed in the suspension (Fig. 1.). Microwave-treatment was performed while

the antenna was being moved gently. The electric output was 20 watts. The degree of treat-

ment was assessed on the basis of the viability of Meth A microscopically determinanced by the

use of 0.5% trypan blue.

FRACTIONATION OF MICROWAVE-TREATED CELL SUSPENSION

The microwave-treated suspension was centrifuged at 1,000 rpm for 5 minutes. The re-

sultant supernatant was taken as a liquid fraction. The precipitate as a cellular fraction was

washed 3 times with RPMI-1640 or PBS, and was used after addition of an appropriate amount

of RPMI-1640 or PBS to regain the original volume.

PREPARATION OF SPLENIC LYMPHOCYTES

The mouse spleen excised under an a・septic condition was swollen by infusion of cooled

RPMI-1640, and the spleen was pecked scrupulously with dental tweezers to obtain splenic cells.

The suspension of those cells was aspirated into a syringe and poured out 3 to 4 times, so that

individual cells came apart4o>. Then, a suspension of splenic lymphocytes was prepared by use

ofCedarlane lympholyte-M cell separation medium.

Mono polar Antenna

6ml of Meth A Sarcoma (3×107cells) Suspension in RPMl-1640 or PBS

326 日外宝第53巻第2号（昭和59年3月）

TREATMENT WITH ANTI司Thy1, 2 ANTIBODY AND COMPLEMENT

The suspension of splenic lymphocytes, the antibody, and the complement were diluted

with RPMI-1640 containing 25 mM HEPES buffer (pH 7.3, GIBCO) and 0.3% bovine serum

albumine (Sigma Chemical Co., Ltd.). The diluted suspension of splenic lymphocytes (1×107

cells/ml) was incubated at 4°C for 60 minutes after addition of a 1/20 volume of anti-Thy 1,2

monoclonal antibody (Cedarlane Laboratories, Ltdふ Themixture was incubated at 37。Cfor

60 minutes, and was washed a time, and then was incubated at 37。Cfor 60 minutes after addition

of a 1/10 volume of Low

washed 3 times with PBS. Thereafter, dead cells were removed by the use of Cedarlane l戸n-

pholyte・Mcell separation medium.

WINN ASSAY

According to the method of WINNa4>, splenic lymphocytes were mixed with Meth A

(1×105 cells) in 0.2 ml PBS (lymphocyte: Meth A=23 : 1). The mixture solution was im-

mediately injected subcutaneously into the fta此 ofBALB/c mice. The weight of a tumor mass

was measured 14 days after the tumor cell transplantation. The anti-tumor activity was expressed

m terms of the percent growth inhibition of the tumor.

Results

1) ANTI-TUMOR EFFECT INDUCED BY MICROWAVE COAGULATION FOR

SOLID TUMOR11>

A solid tumor was produced by transplanting Meth A(4×106 cells) into the subcutis of a

mouse hind limb. Fourteen days after transplantation, microwave coagulation treatment (20

watts, 4×15 sec) was performed (microwave group) (Fig. 2.), or the tumor-bearing limb was

amputated (amputation group). These groups were compared with the control group (without

microwave coagulation or amputation) in the following items:

A) Anti-tumor effect 2 weeks later: The 90-day-survival rates of experimental animals

Coaxial Cable

Tumor

Microwave

Tissue

Coagulator

20 watts, 15 sec

Fig. 2. Microwave coagulation treatment (20 watts, 15 sec) for solid tumor (from four directions and four times. in total）’，


「ーT時間pl，叩tion(M帥 A,4×10「一一MicrowaveTreatment & Amputation

Survival I Rate I I 「一－Challenge (Meth A ' 1 x I 0' cells咽）

% I ;. ;.

100

50

・・・Mier。waveGroupホ n=19 - Amputation Gr。upホ n=22

Survival f'----Ttme

-2a -14 o io 4・0 孔 90Day

Fi邑.3. Anti-tumor effect at 2 weeks after microwave treatment for tumor and amputation of tumor bearing limb. *Survival rates were significant at pく0.05by x2-test as compared with control group.

were 32% and 27% in the microwave group and the amputation group, respectively. The former

rates was significantly higher than that (0%) in the control group (p< 0.05, by x2 test), but the

latter was not (Fig. 3).

B) Anti-tumor effect in the presence of many residual tumor cells (ex. peritoneal carcino-

matosis): The survival time of experimental animals (in days) was significantly longer in the

microwave group than that in the control group (p< 0.05, by Mann-Whitney test). By contraries,

there was no significant difference in the survival time between the amputation group and the

control (Fig. 4.).

2) TRANSPLANTING RATE OF MICROWAVE-TREATED TUMOR CELLS (safety

of those cells used for immunization)

「一一一一Transplantation(Meth A 4X 106 cells} Survival i 「ーChallenge(Meth A・Ix I os cells i.p.} Rate －｜「ーMicrowaveTreatment & Amputation

%1ι↓ 100

50

-7 0 20 40

ー－-Microwave Groupホ n=19-Amputation Group n=21

Fig. 4. Anti-tumor effect in the presence of many residual tumor cells (ex. peritoneal carcmomatos1s}. *Survival tim~ (in days) of microwave group was significant at Pく0.05by Mann-¥Vhitney test as compared with control group.


The suspension of microwave-treated Meth A(l×106 cells, 0.2 ml) was injected intraperitone-

ally, and the survival rate (as the transplanting rate) was determined 90 days after the injection.

In case the viability of injected Meth A was suppressed below 48% by microwave irradiation, all

mice concerned could survive more than 90 days (tumor transplanting rate: O°/o). This may be

construed as meaning that tumor cells (Meth A) whose viability is lower than 48°/o can be used

safely for immunization (Table 1).

The eff1巴ctof temperature on the viability of microwave-treated tumor c巴llswas studied.

When the temperature of the suspension of tumor cells (Meth A) rose to 50-60。Cunder microwave

irradiation, their viability was lowered sharply. When it was about 1 %, the suspension showed

its temperature of about 63°C (Fig. 5ふ

3) CANCEROCIDAL EFFECT OF INTRAPERITONEAL IMMUNIZATION WITH

MICROWAVE-TREATED TUMOR CELLS AND HEAT-DENATURALIZED

TUMOR CELLS

In Group 3-A, Meth A (1×10s cells, cell viability: 0.9°/o, RPMI・1640suspension: 0.2 ml,

temperature: about 63°C) treated with microwaves in a dish was infused intraperitoneally. In

Control 3・A,RPMI-1640 (0.2 ml) treated with microwaves for the same period of time was used

instead of Meth A. In Group 3-B, Meth A (1×106 cells, viability: 1 %, RPMI-1640 suspension:

0.2 ml, temperature: about 62°C) subjected to hot water bathing a test tube was infused. In

Control 3・B,RPMI-1640 (0.2 ml) subjected to hot water bathing was used.

In Group 3-A, the 90-day-survival rate was 27°/o and the survival time was significantly

longer than that in Control 3-A(p<0.01, by Mann-Whitney test). There was no significant

Table 1. Transplanting rate of microwave-treated tumor cells (safety of those cells used for immunization)

Viability of Number Survival Rate Tr町喝p陥nted 。f of Mice m町 e

Me廿、 A Mice サ、an90 Days （%】（%）

85 10 。81 11 。70 10 。66 11 。64 11 27

61 10 60

57 10 70

48 11 100

31 10 100

18 10 100

8 11 100


Viability 。fTi:mor cells

% 100

．．

．．．

80 ー． •I

60

40

20

Temperature of Suspension of Tum or

手与〒一－－.－ー Cells (M柑 A)io 4o so so℃

Fig. 5. Effect of temperature on viability of microwave treated tumor cells.

difference in th巴 survivaltime between Group 3-B and Control 3-B (Fig. 6).


FRACTIONS OF MICROWAVE-TREATED TUMOR CELL SUSPENSION

The microwave-treated Meth A suspension (cell viability: 0.9%) was separated into a cellular

fraction and a liquid fraction. These fractions were adjusted to the cell count of l×1oa and/or

the volume of 0.2 ml, and both or either of them were injected intraperitoneally. Immunization

was performed with both fractions in Group 4-A, with the liquid fraction alone in Group 4 B,

and with the cellular fraction alone in Group 4 C. On the other hand, two control groups were

provided, one called Control 4-A/4-B(control for Group 4 A or Group 4-B) and the other called

3・A Microwave Treatment Group 3・B Hot Water Bathing Group (Viability of Tumor Cells: 0.9 %) (Viability of Tum。rCells: 1.0%)

Survival Survival Rate Rate

% 100

% 100

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10 20 30 40 90 Day f 10 20 30 Day Challenge Challenge

Fig. 6. Cancerocidal effect of one intraperitoneal immunization with microwave-treated tumor cells and with tumor cells subjected to hot water bathing. On the 21st day after immunization, living tumor cells (1×105) were i吋ectedintraperitoneally (Challenge). *Survival time (in days) was significant at Pく0.01by Mann-Whitney test as compared with control group.


Control 4-C(control for Group 4-C). In Control 4・A/4-B,RPMI-1640 (0.2 ml) treated with

microwaves for the same period was used and, in Control 4-C, untreated RPMI・1640was used.

In Group 4-A, the survival time was signi五cantlylonger than that in Control 4-A/4-B

(p<0.01, by Mann-Whitney test). In Group B, no significant difference was observed by

Mann-Whitney test, but such a difference was detected by Student t-test (p< 0.05) or Kolmogorov司

Smimov test (p< 0.05). In Group 4-C, the survival time was not significantly different from that

in Control 4-C (Fig. 7ふ

5) TRANSPLANTING RATES OF INDIYIDUAL FRACTIONS OF MICROWAVE-

TREATED TUMOR CELL SUSPENSION

Both or either of the cellular fraction and the liquid fraction, which were adjusted to the cell

count of 1×106 and/or the liquid volume of 0.2 ml, were injected into the h此 subcutis. In the

mice given the liquid fraction alone (n=lO) or both fractions (n=lO), no tumor mass was formed.

In contrast, tumors were formed in 80% of the mice given the cellular fraction alone (n=lO).

Thus, the liquid fraction exerted an inhibitory effect on tumor growth.


LIQUID FRACTION OF MICROWAVE-TREATED TUMOR CELL SUSPENSION

The liquid fraction (0.2 ml) of a microwave-treated Meth A suspension in PBS (cell viability:

about 1 %) was intraperitoneally injected once a week and four times in total for immunization.

Survival Rate 4・ALiquid & Cellular Fraction

% 100

50

A

41

帥

PG

Con廿。14-A--- n=11

T Challenge

20 40 60 e

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4・BLiquid Fraction 4・CCellular Fraction Survival Rate

% 100

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ー－Gr。up•• 4－自

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SR

ー－Gr。up4C n=11

---c。n廿。14日 50n=ll

---c。ntrol4-C n=lO

Survival Time I ！「 SurvivalTime T 2o 4目0 60 Day T 20 40 60 Day

Challenge Challenge Fig. 7. Cane氾rocidaleffect of intraperitoneal immunization with fractions of the microwave-

treated tumゅrcell suspension. On the 21st day after immunization, living tumor回 lls(1×105 cells) wモreinjected intraperitoneally (Challenge). *Survival time (in days) was signi五cantat Pく0.01by Mann-Whitney test回 com-pared with contr百，Jgr百up.榊 Surviva_ltime (in days) was significant at pく0.05by Student t-test and Kolmo-gorov-Sm1rnov test as compared with cont叫 group.


Once week after the last inJection, immunized mice were challenged by living Meth A or S-180.

Control animals received PBS only.

The 90-day-survival rate after challenge with Meth A was 78°/o. The survival time after the

same challenge was significantly longer than that in the control group (p< 0.01, by Mann-Whitney

test). The survival time after challenge with S-180 was not significantly different from the

control value (Fig. 8.).

7) INDIRECT IMMUNOFLUORESCENCE TEST FOR DETECTION OF ANTI-

TUMOR CELL ANTIBODY IN SERUM9，叫42)

The liquid fraction (0.2 ml) of a microwave-treated Meth A suspension in PBS (cell viability:

about 1 %) was intraperitoneally injected once a week and four times in total for immunization.

Serum was obtained a week after the last injection and was subjected to heat inactivation at 56°C

for 30 minutes. The serum so inactivated was used as primary serum. Anti mouse-y-globulin

conjunction FITC (BEHRING INSTITUTE) served as secondary serum. Living Meth A

cells were employed as target cells.

The primary serum was examined for antibody to target cells by means of indirect immuno-

fluorescence using the primary serum diluted twice and the secondary serum diluted ten times.

As a result, no fluorescent cell was detected at all. In other words, it could not be demonstrated

anti-tumor antibody in the serum from those mice which acquired such a strong immunity against

tumors as in experiment 6.

8) ANTI-TUMOR IMMUNITY OF SPLENIC LYMPHOCYTES

The anti-tumor immunity, represented by growth inhibition, of splenic lymphocytes was

studied by Winn assay. In the immune lymphocyte group, the liquid fraction (0.2 ml) of a

microwave-treated Meth A suspension in PBS (cell viability: about 1 %) was intraperitoneally injected once a week and four times in total for immunization, and splenic lymphocytes were

Meth A Challenge Group S-180 Challenge Group

Survival Rate

% 100

Survival Rate

% 100

50

園園開ImmunizedGroup n=9

---Control G「oupn=9

50

＊

ーーーImmunizedGroup n=9

十

20

e

dzo n

e

i－

－

y咽

－

h

ひ－。’R白ぜ0

4 o

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し－－・・；・－－－，~val Time

20 30 40 Day

． ’ ’ I

－．．．，． ---c。ntrolGroup n=9

e

z－

町

8

au’

1・副岳、i

α

F

10 10

Cancerocidal effect of intraperitoneal immunization with the liquid fraction of microwave-treated tumor cell (Meth A) suspension. On the 7th day fter last injection (injection once a week and four times in total for immunization), living tumor cells (Meth A or S-180) were i吋ectedintra・ peritoneally (Challenge). 本Survivaltime was significant at p<0.01 by Mann-Whitney test as compared with control group.


o

no

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Z

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Lym凶、。cyte:Tum。rCell=23: 1

500 244土138n=10

400

300

207土72n=10

200

100 22±19 n=9 *

M士SD.

Control Immune Normal Treated Group Lymphocyte Lymphocyte Immune

Group Group Lymphocyte Group

Fig. 9. Anti-tumor immunity of splenic lymphocytes from immunized mice in which u’ere i吋ectedliquid fraction of a microwave-treated Meth A suspension once a week and four times in total for immunization. *Tumor weight of immune lymphocyte group was significant at p<0.01 by Stu・ dent t-test as compared with that of the other three groups.

obtained a week after the last injection. These lymphocytes and Meth A (1×105 cells) were

transplanted. In the normal lymphocyte group, PBS (0.2 ml) was used in place of the liquid

fraction. The other procedures were the same as those in the immune lymphocyte group. In

the treated immune lymphocyte group, immune lymphocytes prepared by the above mentioned

method were treated with anti-Thy 1,2 antibody and complement (cell viability after treatment:

68%). After removing dead lymphocytes, living cells were transplanted together with Meth A.

In the control group, only Meth A was transplanted.

Fourteen days a氏ertransplantation, the tumor weight was 22土19mg (mean土S.D.)in the

immune lymphocyte group, 207土72mg in the normal lymphocyte group, 244土138mg in the

treated immune lymphocyte group, and 327土220mg in the control group, respectively. There

was a significant difference in tumor weight between the immune lymphocyte group and each of

the other three groups (p<0.01, by Student t-test). No significant difference was recognized

among the normal lymphocyte group, treated immune lymphocyte group, and control group.

Namely, immune lymphocytes inhibited tumor growth, but this inhibitory activity was

impeded by treatment with anti-Thy 1, 2 antibody and complement. Normal lymphocytes (not

immunized) exerted no inhibitory effect (Fig. 9.).

Discussion

Microwave are electromagnetic waves ranging from 1to100 cm in wavelength and from 300

to 30,000 MHz in frequency呂町． The medical use of microwaves of 2,450土50MHz in frequency


and 12 cm in wavelength is permitted internationally. Biologically, microwaves are said to exert

a thermal, a specific thermal, and a non-thermal effect22,24，附. Although the thermal effect of

microwaves has been well established, there are still controversies about the specific and the non-

thermal effect4•24>. When microwaves are applied to living tissues, they mainly act upon watery

component, which is abundant in a living body, and produce dielectric heat, which is utilized in

surgery or for some other therapeutic purposes.

The MTC converges microwaves of 2,450 MHz in frequency and 12 cm in wavelength to a

small area to coagulate living tissues. In a canine experiment, liver resection using MTC was

reported12>. The changes observed on the liver stump immediately after resection may be under-

stood from the following descriptions: A carbonized layer was unseen and a vacuolated layer, a

coagulative necrosis layer, a liquefactive necrosis layer, and a healthy area, arranged in increasing

order of depth, were recognized. The depth from the vacuolated layer to the healthy area was

5 mm or more. In the healing process, the layer between the coagulative necrosis layer and the

healthy area, i.e., the liquefactive necrosis lay巴rcharged into a fibrotic layer of 3 mm in width凶．

When tumor cells were subjected to the microwave treatment in a dish, the cell membrane,

nucleus, and intracellular granules were maintained morphologically, even in case the cell vi-

ability was lost completely. In addition, the number of cells was not changed by microwave

irradiation even though irradiation time was extended considerably.

It was reported that when rabbit liv巴rtissues were irradiat巴dby microwaves (50 60 watts)

emitted from the monopolar antenna connected to MTC, the maximum temperature of tissues

3 mm away from the focus of irradiation was 65°C, though temperature measurement was not

possible in an area nearer to the focus27>. When the tumor cell suspension in a dish of 3 cm in

diameter was subjected to the microwave treatment (20 watts) by immersing the tip of the antenna

into the suspension, its temperature did not rise above 90°C irrespective of the treatment time.

The characteristic of microwave coagulation using MTC is that, though its thermal effect

on living tissues is comparatively mild, even the tissues far from the antenna are degenerated.

Many authors who described the anti-tumor effect of microwaves only reported general or

regional hyperthermia10,21,2a>. However, it has not been described yet whether or not microwaves

converged in a small area have an anti-tumor effect.

Cytotoxic T匂rmphocytes(CTL)5,1s,aa,a7,41) and natural killer (NK) cellss,13,3M1> immuno-

logically reveal句totoxicityto tumor cells. Furthermore, antibody dependent cell mediated

cytotoxicity (ADCC)37,39> and activated macrophage mediated cytotoxicity36•37•38> are known.

Complement exerts a cytolytic action44>, but it has a decisive weak point that the nuclear mem-

brane of tumor cells is free from this action. Thus, only cell-mediated cytotoxicity seems to be

promising.

In this study, microwave treated Meth A produced an anti-tumor effect, which was mainly

Thy 1+ cell dependent. ADCC was not considered to have a notable anti tumor effect, because

the antibody concerned was not detected in the serum. Among Thy 1 + cells, CTLs and NK

cells are recognized as those which bring about a strong anti-tumor effect. The Thy 1 antigen of

NK cells is weak. Thy 1 + NK cells are said to account for 35-50% of total NK cells7•14> and


approximately 3% of splenic lymphocytes2>. The loss of NK cell activity by one-time treatment

with anti-Thy 1, 2 antibody and complement is unlikely to be large.

The inhibition of tumor growth determined by Winn assay in syngeneic tumor bearing mice

well re但ectedthe immune resistance of the hosts. It has been described that the presence of

T-cells among donor lymphocytes is essential for growth inhibition 8>.

However, it is said that effector T-cells derived from the spleen are often immature, so that

they do not act directly on target cells8•11>. In this connection, the following two mechanisms have

been suggested: 1) Effector T-cells, which are memory T-cells, produce an anti-tumor effect

only after reaction with immune cells of a recipient8>. 2) Effector T-cells, which are immature

cytotoxic T-cells, are differentiated into mature CTLs after coming in contact with tumor cellsui.

In future, it will have to bεclarified, in details, how the final effector cells play a rule in strengthen-

ing immunological resistance to tumor cells with microwav巴treatment.

Generally speaking, the antigenicity of tumor associated antigen (T AA), which the immuno”

logical response of a host to tumor cells is based, is so weak that the host response is hardly induced.

It is desirable to enhance the antigenicity of tumor cells in order to take part in immunoreaction,

on the one hand, and increase the immunological offensive power of a host, on the other.

Meth A antigen in the liquid fract10n of a microwave treated Meth A cell suspension was

examined by the indirect immunofluorescence method. The serum, obtained from rabbits sensit四

ized by intravenous injection of the liquid fraction, was heat inactivated, and was subjected to

absorption treatment with the homogenates of BALB/c mouse spleen and liver tissues. This

serum was used as primary serum, and anti-rabbit－γ－globulin conjunction FITC(Hoechst) was

employed as secondary serum. Living Meth A cells and acetone fixed Meth A cells served as

target cells. As a result, both target cells showed ring-form fluorescence.

Meth A possess two kinds of T AA, one in the cytoplasm (tumor associated transplantation

antigen) and the other on the cell membrane (tumor specific surface antigen). These antigens

are different from each other in their antigenicitya,19,20>.

These results suggest that tumor associated antigen on the cell membrane might have been

detached by microwave treatment into the liquid fraction, which induced an anti-tumor effect.

It is true that when tumor cells are exposed to microwaves, antigen is released from them, but it is

still unclear what mechanism works to cause this phenomenon. At least, the possibility that it

may be due to the thermal effect of microwave treatment can be excluded, because hot water

bathing failed to induce an anti-tumor effect.

There is a possibility that antibody to the normal tissue or autoantibody may be produced by

microwave treatment, though no evidence supporting autoimmunoreaction was obtained in mice,

which underwent microwave coagulation of normal tissuesiη．

Conclusions

In the basic experiments using BALB/c mice and ascitic type tumor cells (Meth A, S-180),

the anti-tumor effect induced by microwave treatment with MTC was studied. The results are

summarized, as follows :


1) The anti-tumor effect induced by microwave coagulation treatment for solid tumor was

stronger than that of surgical resection.

2) In an in vitro system, microwave treatment exerted a potent anti-tumor effect without the

necessity of using an adjuvant.

3) This effect of microwave treatment was thought to be non-thermal effect.

4) It was suggested that this effect of microwave treatment was due to Thy 1 + cell (T-cell

and/or NK cell) dependent anti-tumor immunity and ADCC did not play an important

role in mediating this immunoreaction.

Acknowledgments

The author wishes to thank Prof. M.】（ATSUMIof the Dept. of Gastroenterological Surgery and Prof. H. MIYAMOTO of the Dept. of Microbiology, Wakayama Medical College, for their advice and help. The author is also indebted to my collaborators of the Dept.。fGastroenterological Surgery, Wakayama Medical College, for their kind encouragement given to me for so long.

References

1) Aoyama 0, Tabuse K, et al: A Case of Endoscopic Microwave Tissue Coagulation Therapy for Hemorrhagic Ulcer. Gastroenterological Endoscopy, in press. (in Japanese)

2) Cantor H, Kasai M, et al: Immunogenic Analysis of“Natural Killer" Activity in the Mouse. Immunological Rev 44: 3-12, 1979.

3) DuBois GC, Appella E, et al: lmmunogenic Properties of Soluble Cytosol Fractions of Meth A Sarcoma

Cells. Cancer Research 40: 4204-4208, 1980. 4) Epstein MA & Cook HF: The EFFECTS OF MICROWAVES ON THE ROUS NO. 1 FOWL SAR・

COMA VIRUS. Brit J Cancer 5: 244-252, 1951. 5) H出 himotoY, Nishimura T: The Mechanism of T-cell Mediated Cytotoxicity. Clin Immunol 13(2): 98-108,

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337

和文抄録

マイクロ波腫虜凝固による抗腫療効果に関する実験的研究

和歌山県立医科大学消化器外科（主任：勝見正治教授）

野口博志

マイクロ波は，従来より理学的療法として医療に用

いられてきたが，最近，マイクロ波のエネルギーをで

きるだけ小領域K集中させるととにより組織を凝固さ

せる装置（MicrowaveTissue Coagulator）が開発され

た．乙の装置を用い，実験的lζ腫蕩（MethA Sarcoma)

を処置し，抗腫湯効果lζ関して以下のような成績を得

た．

(1）固型腫蕩にマイクロ波凝固を行なうと，腫蕩組織

を外科的lζ切除して得られる抗腫蕩力より強い抗腫揚

力が得られた．

(2）腫蕩細胞を直接生体外で MicrowaveTissue Coa-

gulatorを用い処置すると，特に Adjuvantの必要な

しに，強い抗腫蕩力が獲得できた．

(3）腫場細胞を MicrowaveTissue Coagulatorで処置

して得られる抗腫揚力はマイクロ波のnon-thermal効

果によることが示唆された．

(4) 乙の抗腫蕩カは，主として， Thy1＋の細胞（T

細胞あるいは NK細胞）依存性の抗腫蕩免疫により生

じるものであり， ADCCは主要な働きをしていない

ととが示唆された．

title experimental studies of anti-tumor effect induced by ... · preparation of splenic...

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