免疫学治疗 基因治疗

O
O
O
O
2'
(dGTP) OH
H
抑制DNA的合成
TNF gene :
Downregulation(下调) of MDR gene expression
Tumor cell chmosensitization
As the base: combining gene therapy with chemotherapy
gamma c, to cells of the
immune system. This treatment appeared very
successful, restoring
immune function to most of the children who received it.
1999,9,17，18岁(8,16)美 国青年Jesse Gelsinger:鸟氨 酸氨甲酰基转移酶（OTC） (-)遗传性疾病,而在美国宾 夕法尼亚州大学人类基因治 疗中心接受基因治疗时不幸 死亡，成为被报道的首例死 于基因治疗中的患者。
大剂量TNF注射造成恶液质的小鼠模型
在动物实验中TNF对于小鼠是强力抗肿瘤剂，鼠可耐受400g/kg体 重, TNF用于临床，当剂量达到8g/kg体重时即产生明显的副作用。
Attack tumor
specific
TIL
Tumor cell
TNF transfection
TIL
TNF
Producing
cells in the patient.
Strategies for Delivering Therapeutic Transgenes into Patients
(a) target cells are removed from the patient,
(b) target cells are selected for,
（常染色体隐性遗传） 引起免疫缺陷
腺苷 脱氧腺苷
dAMP
腺苷脱氨酶 (adenosine deaminase ) ADA
dADP
dATP
抑制核苷酸还原酶，阻碍DNA合成
T、B 细胞增殖障碍
S C I D 重度联合免疫缺陷症（ evere ombined mmuno eficiency ）
monocyte
TNF/ HSV-tk
TNF/ HSV-tk
transfection Tumor cell
Expression of TNF / HSV-tk
Injection of GCV
（二）免疫基因治疗
对细胞裂解性T细胞(CTL)的直接活化或通过其活化调节物的 间接活化是很多基因治疗的研究重心。
采用未修饰的TIL细胞治疗肿瘤，TIL细胞有专门攻击肿瘤 的特性，导入TNF基因的TIL细胞可望在肿瘤组织中安家，在 局部产生高浓度TNF以杀伤肿瘤而不带来严重的副作用。
(e) transplanted to the patient.
ex-vivo Schematic of
gene therapy protocol
The first case for gene therapy in the world is SCID
AMP
IMP
腺苷 ADA 肌苷
ADA(腺苷脱氨酶）基因 突变
Gene therapeutic vectors which transfer a cytotoxic mechanism to tumor cells
offer a new route of cancer therapy.
In the 1990s, late researchers tested a gene therapy treatment that would restore the function of a crucial gene,
TNF
TNF
Homing in tumor
Tumor infiltrating lymphocyte
（三）血管生成的抑制
Angiogenesis
VEGF promotes Tumor Angiogenesis
转染
突变信号传导缺陷性 VEGF受体基因
内皮细胞
抑制肿瘤的血管形成
大鼠和大鼠模型肿瘤系统中已获得成功
(3.8X1013)但只有1%的病 毒到达靶器官肝脏，而绝大 部分的病毒进入其它器官与 组织，从而引发了强烈的系 统性炎症反应。
一、癌症的基因治疗 （一）化学基因疗法
化学疗法
化疗出现前后肿瘤治疗水平的变化
100
80
有
效 率
60
40
20
0
化疗出现前 加化疗后
肾 绒 尤 瘤巴 胞视 白急 蕈 横 氏晚 细弥 睾转 骨
transduction (转导) refers to the
introduction of a transgene into a cell
through a viral vector system.
**Gene transfer can be performed by transfection
or transduction of target cells in vitro and then
二、自身免疫病的基因治疗
自身免疫性胰岛损伤
免疫复合物在 SLE 患者皮下沉积
I-型糖尿病患者血清中的自身抗体 SLE 患者血清中的抗中心粒抗体
Models of autoimmune diseases
The experimental models :
experimental auto-allergic encephalitis (脑炎), experimental thyroiditis (甲状腺炎) adjuvant induced arthritis (关节炎) Experimental antoimmune encephalomyelitis (脑脊 髓膜炎) (EAE)
Germ Line Therapy 生殖细胞治疗
Somatic Gene Therapy 体细胞治疗
Introduction of a nucleic acid or target
gene (transgene) directly into cells is
referred to as transfection(转 染).
母癌文
细
氏
胞
瘤
瘤
基 瘤网 血性 样 纹 病期 胞散 丸移 肉
特 淋
膜 母
病淋 巴
霉 菌 病
肌 肉 瘤
何 杰
淋巴瘤性组
癌性 胚
瘤
巴细 性
金织胎
化疗药是肿瘤治疗史上的不朽里程碑
过去治疗生存率（％） 现在治疗生存率（％）
几种肿瘤治疗效果的今昔（五年生存率）
90
生
80
存
70
率
60
50
40
30
20
10
0
骨肉瘤
乳腺癌
上颌窦癌 肾母细胞瘤
MDR的分子机制
肿瘤的MDR（multiple drug resistance）
自杀基因 (suicide gene) :
编码对肿瘤细胞有害的酶类(+相应的原药 )
单纯疱疹病毒胸苷激酶（herpes simplex thymidine
kinase, HSV-tk）基因
TNF:
O O P O CH2
O
O
N
6
N1
NH2 N
N
Guanine(G)
O
6
N1
OH OH N C C CH
O CH3
NH2 N
N
OH O
2'
OH
H
O
6
N1
OH OH N C C CH
O CH3
O
O
O
NH2 N
N
O P O P O P O CH2
OH O
O
O
O
2'
O
OH
H
N
N
O
O
O
NH2 N
N
O P O P O P O CH2
O N1 6
NH2
N
OH
HO CH2
O
OH
OH
N C C CH
O CH3
N
ganciclovir
OH
H
9-(1,3-二羟-2-丙氧甲基)鸟苷 (ganciclovir,GCV)
O
6
N1
OH OH N C C CH
O CH3
NH2 N
N
HO CH2
OH O
2'
OH
H
9-(1,3-二羟-2-丙氧甲基)鸟苷 (ganciclovir,GCV)
drug converting enzymes and then treatment with
systemic administration of the respective nontoxic
pro-drug.
geneexpression
Prodrug (nontoxic)
. HSV-tk (单纯疱疹病毒胸苷激酶) gene:
Naturally occurring models :
hemolytic anemia in NZB mice systemic lupus erythematosus(狼疮) in NZB/NZW (BW), BXSB and MRL mice diabetes in obese mice
分子外科
The idea is simple; the practice is not.
As you might guess from the low success rates of vector transfection, this is a dicey procedure, and the techniques are still crude, at best.
A new gene is injected into an adenovirus vector, which is used to introduce the modified DNA into a human cell. If the treatment is successful, the new gene will make a functional protein.
ADA
T cell
ADA
ADA
从1990年转移ADA基因到现在的大部分基因治疗临床试验都是先从病人体内获得某种 细胞（例如T淋巴细胞），进行培养，在体外完成基因转移后，筛选成功转移的细胞扩 增培养，然后重新输入患者体内。这种效果较为可靠，称其为体外（ex vivo）基因治疗。
cfdr
1994年美国科学家利用经过修饰的腺病毒为载体，成功地 将治疗遗传性囊性纤维化病的正常基因cfdr 转入患者肺组织 中。这种直接往人体组织细胞中转移基因的治病方法叫做体 内（in vivo）基因治疗。
（四）基因传递和基因导向技术
inherent advantages (优势): 1 integrating (整合) the therapeutic gene into the chromosomal DNA of a targetwenku.baidu.comcell,
2 deliver the therapeutic gene to large numbers of target cells.
administration of the modified cells (修饰的细
胞) to an animal or patient.
**In vivo gene transfer is accomplished by direct transfection or transduction of target
(c) transduced with a recombinant retrovirus coding the gene of interest
(d) expanded to obtain a therapeutically useful number of transduced number of cells, and
免疫治疗 Immunotherapy
第一节 基因治疗 (gene therapy)
Gene therapy can be broadly defined as the transfer of genetic material into a cell to transiently (短暂) or
permanently(永久) alter the cellular phenotype(表型).
Coding herpes simplex thymidine kinase
ganciclovir acyclovir bromovinyldeoxyuridine
HSV-tk
Phosphorylates 磷酸化
Monophosphates (一磷酸)
the anti-herpetic nucleoside analogues
能下调多种耐药性基因（mdr）的表达，从而使肿瘤 细胞发生化学致敏作用（chemosensitization）。这 一作用作为癌症的基因治疗和化学治疗联合疗法的基 础。
(1) suicide gene therapy :
Enzyme/Pro-drug Therapy:
delivery to tumor cells of genes encoding pro-