Immunostimulating Effect of Palmiboshinwhan (baweibushenwan) Extracts in Methotrexate-induced Immunosuppressed SD rats.

(1)

서 론

(innate immunity)

(adaptive immunity) ,

, ,

(specific immunity)

. (humoral im-

munity) (cell-mediated immunity)

1-4).

5)

. , , ,

6). ,

5).

八味補腎丸誘發

免疫機能低下影響

강석봉 Original Article

Immunostimulating Effect of Palmiboshinwhan (baweibushenwan) Extracts in Methotrexate-induced Immunosuppressed SD rats.

Seok-Bong Kang

Department of Oriental Medicine, Daegu Haay University, Daegu Korea

Objectives: The purpose of this study was to verify the effect of Palmiboshinwhan (PMBSW) in methotrexate (MTX)-induced immunosuppressed SD rats.

Methods: The test articles were once a day dosed for 14 days by gastric gavage from 2 days after last MTX-dosing, and changes in body weight, spleen weight and total blood leukocyte numbers were observed with total lymphocyte numbers, B and T lymphocyte percentages, CD3+CD4+, CD3+/CD8+, CD4+/CD8+ T lymphocyte percentages in the blood and spleen, the serum interleukin (IL)-2 levels and the productivity of IL-2 of splenic cells.

Result & Conclusion: It is concluded that PMBSW has relatively good immunostimulating effect in the MTX-induced immunosuppressed SD rats. Theefficient dosage was considered above 500mg/kg. In addition, it is considered that the immunostimulating effect of PMBSW was mediated to both the B and T lymphocytes. The more favorable effects were detected in T lymphocytes rather than B lymphocytes, and PMBSW showedrelatively good stimulating potential against CD4+ T lymphocytes but not any stimulating effect against CD8+ T lymphocytes in the present study.

Key Words : Palmiboshinwhan (baweibushenwan), methotrexate, immunosuppression

2008 4 16 2008 5 7

: , 165

(Tel : +82-53-770-2111, Fax : +82-53-770-2082, E-mail : [email protected])

(2)

6).

,

, , , , ,

8 ,

7).

.

8), ⁹⁾, ¹⁰⁾

, , ,

,

.

, methotr-

exate( MTX) SD rat

1000, 500 250mg/kg

2 ,

,

, B , T , CD3+

CD4+ T , CD3+CD8+ T , CD4 +/CD8+ T Interleukin (IL)-2

.

재료 및 방법 1. 실험동물

180g Sprague-Dawley

(6wks old upon receipt, Charles River,

Japan) ( , )

10 ,

, 3 (1000, 500

250mg/kg ) 5 ,

12 60 .

(20-25 ) (30-35%)

, 11-12

/hr, 12hr/day .

,

. 3

polycarbonate .

2. 실험군의 구분

MTX

MTX 5

45%

6 ,

6 .

, MTX ,

MTX 1000,

500 250mg/kg ( P1, P2 P3)

5 (Table 1).

Group ID Treatment Dosage MTX-induced

immunosuppression

Sham D.W. 0mg/kg 0mg/kg

Control D.W. 0mg/kg 2mg/kg, 4 times

P1 PMBSW Extract 1000mg/kg 2mg/kg, 4 times

PMBSW extract: Palmiboshinwhan extract; D.W.: Distilled water; PMBSW Extracts were dissolved in D.W. and dosed by gastric gavage at 10ml/kg levels and MTX was dosed by gastric gavage at 5ml/kg levels using saline as vehicle. PMBSW and vehicle were dosed for 14 days from 2 days after last dosing of MTX. MTX was dosed once a day for 4 days (Total 4 times).

Table 1.

(3)

3. 八味補腎丸의 조성 및 추출 1)

,

13)

1 Table 2 .

2)

5 (5531.25g)

5000ml

rotary vacuum evaporator(N-N type; LAB Camp, Dajeon, Korea)

programmable freeze dryer(PVTFD10A; ilShin Lab., Seoul, Korea)

1 236.41g, 1182.05g

( 21.37%)

.

4. MTX에 의한 면역억제 유발

MTX

(Sigma, USA) 2mg/kg 5ml/

kg 1 4

, MTX

. MTX 5 ,

45%

.

5. 실험물질의 투여

1000, 500 250mg/kg

10ml/kg 1

14 .

.

MTX 6 14 .

6. 체중 및 증체량의 측정

MTX 1 , MTX

, MTX 5 , ,

1, 7, 14 ,

,

18 .

MTX

.

Herbs Scientific Name Dose (g)

熟地黃 REHMANNIAE RADIX PREPARAT 300.00

兎絲子 CUSCUTAE SEMEN 300.00

當歸身 ANGELICAE GIGANTIS RADIX 131.25

肉蓉 CISTANCHES HERBA 187.50

山茱萸 CORNI FRUCTUS 93.75

黃柏 PHELLODENDRI CORTEX 37.50

知母 ANEMARRHENAE RHIZOMA 37.50

破古紙 PSORALEAE FRUCTUS 18.75

Total 8 types 1106.25g

Table 2.

(4)

Body weight gains during dosing (Day 0 - Day 14)

= Body weight at sacrifice - Body weight at initial dosing

Body weight gains during induction periods (MTX 0 - MTX 5)

= Body weight at 5 days after MTX-dosing - Body weight at MTX dosing

7. 비장 중량의 측정

, g

.

. Relative weight (%) =

[(Absolute spleen weight / Body weight at sacrifice) × 100]

8. 채혈

MTX 5 ethyl

ether 1ml

EDTA(ethylene diamine tetraa- ccetic acid dipotassium salt)

, 5ml 1ml EDTA

. IL-2

4ml

1 3,000rpm 10

(serum) .

9. 혈액 중 총백혈구 수의 계산

, MTX 5

.

pippet

1×10³/1mm³ .

Türk solution .

. Difference of total WBC numbers during dosing periods

= Numbers at 5 days after last MTX-dosing - Numbers at sacrifice

10. 비장 부유액의 준비

70% alcohol

, 4 RPMI-1640(Gibco

BRL, NY, USA) 2 , RPMI-

1640 Petri dish

homogenize

. (mesh No.

100: Sigma, USA)

RPMI-1640 1 , HBSS(Hanks Balanced Salt Solution,

Gibco BRL, USA) 2 .

hypotonic shock

, 10 HBSS 2

RPMI-1640

10% FBS(fetal bovine serum; Gibco BRL, USA)

.

11. 말초혈액 및 비장내의 림프구수 측정

EDTA

PBS Limphoprep

(1.077±0.0001g/ml, Nycomed Pharma As, Oslo,

Norway) 30 3000rpm

,

(5)

.

PBS 3000rpm 10 2

RPMI-1640 ,

, Trypan blue

exclusion .

12. 말초혈액 및 비장내의 B 세포율과 T 세포율 측정

EDTA tube

12×75 2 100 .

FITC Anti-Rat CD3 Monoclonal Antibody

(DiaSorin, MN, USA) 0.1 PE

Anti-Rat CD45R/B220 Monoclonal Antibody(Sigma,

USA) 0.5 Vortex mixer

30 Lysing solution(FASS

Lysing solution, Becton Dickinson, USA) 2

, 15 .

Lysis 1000rpm, 5

2 washing

solution(PBS) 1000rpm, 5

. 500 PBS

Vortex mixer Flow cytometer (Becton Dickinson, USA) .

13. 말초혈액 및 비장내의 CD3+CD4+ T 세포율 측정

EDTA tube

12×75 2 100 .

Anti-Rat CD4 Monoclonal Antibody(Sigma, USA)

0.5 Vortex mixer

30 Lysing solution(FACS Lysing solution, Becton Dickinson, USA) 2

15 . Lysis

100rpm, 5

2 washing solution

(PBS) 1000rpm, 5

. 500 PBS

Vortex mixer Flow cytometer(Becton

Dickinson, USA) .

14. 말초혈액 및 비장내의 CD3+CD8+ T 세포율 측정

EDTA tube

12×75 2 100 .

Anti-Rat CD8 Monoclonal Antibody(Sigma, USA)

0.5 Vortex mixer

30 Lysing solution(FACS Lysing solution, Becton Dickinson, USA) 2

15 . Lysis

100rpm, 5

2 washing solution

(PBS) 1000rpm, 5

. 500 PBS

Vortex mixer Flow cytometer(Becton

Dickinson, USA) .

15. 말초혈액 및 비장내의 CD4+/CD8+ T 세포 율 측정

CD3+CD4+ T

CD3+CD8+T , CD4

+/CD8+ T .

CD4+/CD8+ T lymphocytes (Ratio)

= (CD3+CD4+ T lymphocytes / CD3+CD8+ T lymphocytes)

(6)

16. 말초혈액내 IL-2 함량 및 비장세포의 IL-2 생산량의 측정

(serum) IL-2 mouse

IL-2 ELISA Kit Intertest-2X kit(Endogen, USA) 450nm

IL-2

. FBS 10%

RPMI-1640 1×10⁶ cell/ml

, concanavalin-A(Sigma, USA)

100 /ml 37 , 5% CO2

incubator 24 IL-2

mouse IL-2 ELISA Kit Intertest-2X kit(Endogen, USA) 450nm

IL-2 . 96 well plate

well 10

36 40 ,

well buffer 4

plate paper towel

well biotinylated polyclonal anti-mouse IL-2(Endogen, USA) 100

37 40 . well

buffer 4 plate paper towel

well streptoavidin-peroxidase(Sigma, USA)

100 37

25 . well

buffer 4 plate

towel well substrate mix

(Endogen, USA) 100

10 .

well 100 ELISA

450nm .

standard curve

IL-2 .

17. 통계처리

Mann-Whitney Wilcoxon's Rank Sum(MW) test

, SPSS

for Windows(Release 10.1.2, SPSS Inc., USA) .

% changes ,

% changes .

% Changes vs Sham

= [(Data of Sham -Data of Control)/Data of Sham×100]

% Changes vs Control

= [(Data of Control -Data of tested group)/

Data of Control]×100]

결 과

1. 체중 및 증체량의 변화

1)

MTX 1 , MTX , MTX 5

, , 1, 7, 13

MTX 5

. P1 13

(p<0.01 p<0.05) . P2

7 ,

. P3

13

(7)

, (Table 3).

2)

MTX MTX

,

(Table 4).

MTX

, P1 (p<0.05)

, P2, P3

, (Table 4).

2. 비장 중량의 변화

P1, P2

(p<0.01 p<0.05) , P3

, (Table 5).

P1

(p<0.05) , P2,

P3

Group ID¹⁾ Sham Control P1 P2 P3

Day-1 185.67 ± 6.98 186.00 ± 10.81 186.17 ± 9.83 187.33 ± 8.61 186.50 ± 7.26 MTX 0^a) 188.83 ± 5.95 189.17 ± 12.11 186.83 ± 9.79 188.17 ± 6.46 187.17 ± 9.75 MTX 5 241.17 ± 34.10 167.50 ± 13.02^** 165.50 ± 7.71^** 166.67 ± 9.35^** 166.50 ± 8.76^**

Day 0^2,b) 219.97 ± 34.14 147.67 ± 11.43^** 145.50 ± 9.40^** 145.67 ± 10.39^** 149.17 ± 9.37^**

Day 1 241.83 ± 38.51 147.00 ± 13.83^** 148.50 ± 9.89^** 145.33 ± 9.61^** 146.50 ± 10.56^**

Day 7 271.67 ± 33.22 144.00 ± 12.33^* 157.17 ± 13.44^** 147.33 ± 8.33^** 144.67 ± 10.35^**

Day 13 284.83 ± 29.65 145.33 ± 7.89^** 168.50 ± 9.79^**,## 150.67 ± 6.86^** 149.17 ± 4.88^**

Day 14^2,c) 258.17 ± 28.37 131.67 ± 8.50^** 149.00 ± 11.54^**,# 137.17 ± 7.88^** 136.50 ± 4.64^**

Mean ± S.D., g; 1) Group ID was listed in Table 1; 2) Overnight fasted; a) At initial MTX dosing; b) At initial Palmiboshinwhan extract dosing; c) At sacrifice; ** p<0.01 compared to that of Sham (MW test); * p<0.05 compared to that of Sham (MW test); ## p<0.01 compared to that of Control (MW test); # p<0.05 compared to that of Control (MW test).

Table 3.

Group ID¹⁾ Periods of body weight gains

MTX 0 - 5 Changes²⁾ Day 0 - 14 Changes

Sham 52.33 ± 30.67 38.50 ± 13.98

Control -21.67 ± 2.88^** -141.40% -16.00 ± 13.94^** -141.56%

P1 -21.33 ± 5.32^** 2.54% 3.50 ± 10.84^**,# 122.88%

P2 -21.50 ± 7.84^** 1.77% -8.50 ± 4.85^** 47.88%

P3 -20.67 ± 1.88^** 5.62% -12.67 ± 7.50 21.83%

Mean ± S.D., g; 1) Group ID was listed in Table 1; 2) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); # p<0.05 compared to that of Control (MW test).

Table 4.

(8)

, (Table 5).

3. 말초 혈액 중 총 백혈구 수의 변화

MTX 5

(Table 6).

P1

(p<0.05) , P2, P3

, (Table 6).

P1

Group ID¹⁾ Absolute weight (g) Changes³⁾ Relative weight (%)²⁾ Changes

Sham 0.721±0.087 0.283±0.052

Control 0.210±0.069^** -70.84% 0.159±0.049^* -43.88%

P1 0.337±0.056^**,## 60.35% 0.228±0.047^# 43.51%

P2 0.279±0.037^**,# 32.91% 0.203±0.021^* 27.89%

P3 0.238±0.052^** 13.01% 0.175±0.042^** 9.84%

Mean ± S.D. (n=6); 1) Group ID was listed in Table 1; 2) Relative weight (%) = [(Absolute spleen weight / Body weight at sacrifice) ×100];

3) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); * p<0.05 compared to that of Sham (MW test); ## p<0.01 compared to that of Control (MW test); # p<0.05 compared to that of Control (MW test).

Table 5.

Group ID¹⁾ At MTX 5 (× 10³cells/mm³) Changes²⁾ At Sacrifice (× 10³cells/mm³) Changes

Sham 8.70±1.37 8.81±1.64

Control 4.73±0.95^** -45.56% 3.88±0.42^** -55.91%

P1 4.78±0.91^** 0.95% 5.67±0.83^**,# 45.97%

P2 4.79±0.77^** 1.09% 4.59±1.17^** 18.28%

P3 4.71±0.84^** -0.46% 4.32±0.80^** 11.29%

Mean ± S.D. (n=6); 1) Group ID was listed in Table 1; 2) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); # p<0.05 compared to that of Control (MW test).

Table 6.

Group ID²⁾ At MTX 5 (× 103cells/mm³) Changes³⁾

Sham 0.11 ± 0.72

Control -0.85 ± 1.01 -850.00%

P1 0.89 ± 1.40^# 205.71%

P2 -0.19 ± 0.65 78.45%

P3 -0.39 ± 0.88 55.12%

Mean ± S.D. (n=6); 1) Difference of total WBC numbers during dosing periods = Numbers at sacrifice - Numbers at 2 days after last MTX-dosing; 2) Group ID was listed in Table 1; 3) Changes vs Sham or Control; # p<0.05 compared to that of Control (MW test).

Table 7.

(9)

(p<0.05) , P2, P3 , (Table 7).

4. 말초 혈액 및 비장내 임파구 수의 변화

P1,

P2 (p<

0.01 p<0.05) .

P3

, (Table 8).

P1 (p<0.01) , P2, P3

, (Table 8).

5. 말초 혈액 및 비장내 B 임파구율의 변화

B

(Table 9).

B P1, P2

(p<0.01

p<0.05) , P3

, (Table 9).

6. 말초 혈액 및 비장내 T 임파구율의 변화

T P1

(p<0.05) . P2, P3

, (Table 10).

Group ID¹⁾ Blood (× 10⁵cells/ml) Changes²⁾ Spleen (× 10⁶cells/ml) Changes

Sham 75.75±4.12 64.87±9.74

Control 43.88±4.19^** -42.07% 30.92±4.72^** -52.33%

P1 55.74±4.10^**,## 27.03% 41.24±4.80^**,## 33.36%

P2 52.43±5.93^**,# 19.49% 34.89±7.84^** 12.83%

P3 47.44±4.84^** 8.10% 33.04±2.84^** 6.85%

Mean ± S.D. (n=6); 1) Group ID was listed in Table 1; 2) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); # p<0.01 compared to that of Control (MW test); ## p<0.05 compared to that of Control (MW test).

Table 8.

Group ID¹⁾ Blood (%) Changes²⁾ Spleen (%) Changes

Sham 46.91±3.15 41.99±2.43

Control 39.63±2.86^** -15.53% 35.71±1.74^** -14.95%

P1 41.10±3.25^* 3.71% 42.23±4.54^## 18.26%

P2 38.45±3.92^** -2.98% 39.69±3.25^**,# 11.16%

P3 39.46±3.24^** -0.42% 37.38±2.63^* 4.68%

Mean ± S.D. (n=6); 1) Group ID was listed in Table 1; 2) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); * p<0.05 compared to that of Sham (MW test); ## p<0.01 compared to that of Control (MW test); # p<0.05 compared to that of Control (MW test).

Table 9.

(10)

T P1 (p<0.01)

T , P2, P3

, (Table 10).

7. 말초 혈액 및 비장내 CD3+CD4+ T 임파구율 의 변화

CD3+CD4+ T P1

(p<0.05) . P2, P3

, (Table 11).

CD3+CD4+ T P1, P2

(p<0.01 p<0.05)

, P3

, (Table 11).

8. 말초 혈액 및 비장내 CD3+CD8+ T 임파구율 의 변화

CD3+CD8+ T MTX

,

(Table 12).

CD3+CD8+ T MTX

,

(Table 12).

Sham 51.63±5.04 50.65±4.56

Control 43.19±3.83^* -16.34% 39.13±4.11^** -22.74%

P1 50.83±4.76^# 17.70% 46.76±2.73^## 19.48%

P2 50.25±7.29 16.35% 43.39±3.24^* 10.88%

P3 45.84±4.52^* 6.13% 41.16±4.05^** 5.19%

Table 10.

Sham 42.65±3.99 40.34±6.41

Control 31.41±4.24^** -26.36% 27.51±3.13^** -31.81%

P1 38.33±2.52^# 22.06% 35.47±6.07^# 28.95%

P2 34.44±4.24^** 9.66% 31.52±1.08^**,## 14.58%

P3 33.81±4.94^* 7.64% 28.92±4.05^** 5.14%

Table 11.

(11)

9. 말초 혈액 및 비장내 CD4+/CD8+ T 임파구 율의 변화

CD4+/CD8+ T P1, P2

(p<0.01 p<0.05) . P3

, (Table 13).

CD4+/CD8+ T

, (Table 13).

10. 말초 혈액 IL-2 함량 및 비장세포의 IL-2 생 산량의 변화

IL-2

, (Table 14).

Sham 16.52±0.73 20.32±1.41

Control 17.33±1.42 4.92% 19.67±2.41 -3.20%

P1 16.49±0.97 -4.87% 20.13±3.09 2.33%

P2 17.49±0.71 0.93% 19.38±1.87 -1.48%

P3 16.46±1.41 -5.05% 19.81±2.82 0.72%

Mean ± S.D. (n=6); 1) Group ID was listed in Table 1; 2) Changes vs Sham or Control.

Table 12.

Group ID²⁾ Blood (Ratio) Changes³⁾ Spleen (Ratio) Changes

Sham 2.59±0.34 1.98±0.25

Control 1.82±0.23^** -29.98% 1.43±0.32^* -27.99%

P1 2.33±0.20^## 28.43% 1.79±0.35 25.22%

P2 1.98±0.29^**,# 8.83% 1.64±0.15^* 14.76%

P3 2.06±0.31^* 13.66% 1.49±0.31^* 4.51%

Mean ± S.D. (n=6); 1) CD4+/CD8+ T lymphocytes (Ratio) = (CD3+CD4+ T lymphocytes / CD3+CD8+ T lymphocytes); 3) Group ID was listed in Table 1; 2) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); * p<0.05 compared to that of Sham (MW test); ## p<0.01 compared to that of Control (MW test); # p<0.05 compared to that of Control (MW test).

Table 13.

Group ID¹⁾ Blood levels (pg/ml) Changes²⁾ Spleen cell productivity (pg/ml) Changes

Sham 5.09±1.08 33.95±4.02

Control 2.16±0.76^** -57.51% 19.18±2.34^** -43.50%

P1 3.24±0.63^** 49.61% 25.40±4.64^*,# 32.40%

P2 2.54±0.43^** 17.33% 22.78±3.68^*,# 18.74%

P3 2.37±0.79^** 9.71% 20.43±1.35^** 6.53%

Mean ± S.D. (n=6); 1) Group ID was listed in Table 1; 2) Changes vs Sham or Control; ** p<0.01 compared to that of Sham (MW test); * p<0.05 compared to that of Sham (MW test); # p<0.05 compared to that of Control (MW test).

Table 14.

(12)

IL-2 P1, P2 (p<0.05) , P3

, (Table 14).

고 찰

(innate immunity)

(adaptive immunity) .

, ,

(humoral immunity) (cell-mediated immunity)

1-4).

B

, T

T cytokine

2-3,11)

.

T T (helper

T lymphocyte) T (cytotoxic T lymphocyte) ²⁾.

B

,

B B (BCR: B

cell receptor) ⁴⁾.

BCR B

(plasma cell)

BCR

2). T

. T

T (TCR: T cell receptor)

2). TCR

T , TCR

BCR

(MHC: Major Histo-compatibility Complex)

4).

T TCR

(CD4 CD8) , T

T (TH:

Helper T Cell) T (TC: cytotoxic T cell) ^4,11).

T (TH) CD4

MHC 2

cytokine . cytokine B

, T ,

3).

T CD8

MHC 1

11). Cytokine

(Interleukin), ,

(Interferon), (Lymphokine) ¹¹⁾.

IL-2 CD4 T cell NK cell

LAK(Lymphokine activated killer) cell

3-4).

(13)

. ,

5).

6). , , ,

6)

7) .

, ,

, , , ,

8 ^6,7).

,

, ,

,

, ,

12).

MTX (antimetabolites)

(dihydrofolate reductase inhibitor) , ,

,

, , ^13-14).

MTX ,

,

15-17)

.

8),

9), ¹⁰⁾, ¹⁸⁾ , ,

, ,

.

, MTX

SD rat 1000, 500 250mg/kg 2

,

, , B

(ratio of B lymphocytes), T ,

CD3+CD4+ T , CD3+CD8+ T ,

CD4+/CD8+ T T cytokine

Interleukin (IL)-2 .

MTX

MTX 5 .

P1 13

(p<0.01 p<0.05)

, P2 P3

,

, , ,

. MTX

16).

, P1, P2

(p<0.01 p<0.05) ,

P3

. P1

(p<0.05) ,

P2, P3 .

.

MTX ,

17) MTX

.

P1

(p<0.05) , P2, P3

. ,

P1, P2 P1 .

(14)

.

MTX B

, MTX

19).

B

, P1, P2, P3

P1, P2

(p<0.01 p<0.05) , P3

. B

T ,

.

MTX T

20-22)

,

T P1

. P2, P3 . T

.

CD3+CD4+ T MTX

23),

CD3+CD4

+ T P1

(p<0.05) . P2,

P3 .

CD3+CD4+ T P1, P2

(p<0.01 p<

0.05) , P3

.

T ,

.

MTX CD8+ T

24). CD3

+CD8+ T MTX

.

T T

.

MTX CD4+/CD8+ T

25) ,

P1, P2 CD4+/

CD8+ T P3

. CD4+/CD8+ T

.

CD8+ CD4+

.

MTX IL-2 T

21) IL-2

26).

P1, P2 IL-2

, P3 .

P1, P2, P3

.

MTX IL-2

B , T

.

MTX

(15)

. 500 250mg/kg

, ,

500mg/kg .

B T

,

B T

, T CD4+

, CD8+

.

요 약

, MTX SD

rat 1000, 500 250mg

/kg( P1, P2, P3 ) 2 ,

, , B

, T , CD3+CD4+ T , CD3 +CD8+ T , CD4+/CD8+ T

T cytokine Interleukin (IL)-2

, .

1.

P1 13

, P1

. 2.

P1

. 3.

P1

. 4.

P1

, P2

.

5. B

P1, P2 B

.

6. T

P1

T .

7. CD3+CD4+ T

P1

CD3+CD4+ T , P2

.

8. CD3+CD8+ T

MTX , MTX CD3+CD8+ T

.

9. CD4+/CD8+ T

P1, P2

CD4+/CD8+ T .

10. IL-2 IL-2

P1, P2

IL-2 .

500mg/kg .

B

T , T

CD4+

.

(16)

참고문헌

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1996:20-8.

2. 29 . . : .

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8. .

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. : . 2005.

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