(精品) 分子病理学技术进展及临床应用课件

Centrocyte T cell Macrophage FDC
Proliferation zone
Selected for apoptosis
On-going mutation
Update to B-lymphocyte development model in 2000’s
B cell differentiation by immuno-markers
That is the beginning of our story today.
Understanding of Lymphocyte Development
1950s-2010
1. structure and function of lymphoid tissue
Germenal Center(GC)：structure，function，transformation
激素
(3) 癌基因蛋白的应用 (4)对肿瘤增生程度的评价 ki-67,PCNA
(5)微小病灶的发现
微小癌，微小病灶（如羊水栓塞） (6)在肿瘤分期上的意 义 (7)指导肿瘤的治疗 (8)免疫性疾病的辅助诊断 (9)病原微生物的检测
2．常用免疫组织化学方法： A、一步法 B、二步法 C、三步法 D、多步法 间接法 金银法 PAP和BigBee APAAP ABC法 EnVision法
For post-graduate program
Clinical pathology-ZR-201509
一、临床分子病理学常用方法
免疫组织化学 基因克隆性重排的检测 FISH及C G H 等位基因不平衡分析 杂合性缺失（ L O H ） 的检测 微卫星D N A 不稳定性（MSI）的检测
B cleaved cell
B non-cleaved cell
T lymphocyte
T lmmunocyte
Memory cell Plasma cell
Paradigm of Lymphocyte transform
by morphology and immunotyping considering molecular events
Hilus
Medullary sinuses Efferent lymphatic vessels
Could you recognize the following cellular elements in lymph node?
T-zone
Germinal center Mantle zone Marginal zone
For post-graduate program
Clinical pathology-ZR-
For post-graduate program
Clinical pathology-ZR-2014
2. 免疫组织化学的应用范围及优点：
1.
应用范围：
(1) 提高病理诊断准确性
(2)对疾病的预后和治疗的意义
APC DCC 5p 16q
RAS
Rb/P53
21q 13q14/17p13
Suppressgene oncogene oncogene
Suppressgene
Adenoma Malignant
Carcinoma Metastasis
Basic understanding of molecular carcinogenesis
The contemporary understanding of B and T cell differentiation 2009-2010
Understanding of the lymphocyte differentiation and transformation by multiple approaches inl. morphology,
Understanding based on 70’s of last Century
Only by morphology
Lymphocyte transform in center of Follicle Interfollicle zone
FDC B lymphocyte
B lmmunocyte
immunotyping and molecular means.
Understanding of general carcinogenesis from different patterns of cell & molecular
biology
From colonorectal carcinoma, we understand the multi-step of carcinogenesis but linearized.
Understanding of B cell differentiation until 2005 by H Stein
Bone Marrow
T cell-rich zone
Germinal Center
Precursor B Lymphoblast
Somatic Hypermutation/ Affinity Maturation
Even more, the molecules in the related cells are the key, especially involved in the development and differentiation of lymphoid & hematopoietic cells
V(D)J recombination
Naive B cell
Intact BCR
Antigen
BCL2 ↓ BCL6 ↑
B B
B
B DC
BT
Increased Affinity
B
Decreased Affinity
Class Switching IGM → IgG, IgA
DC
T
B
B
BCL6 ↓ BCL2 ↑ IRF4 ↑ B
BT法（CSA：Catalyzed signal amplification） 1:50、1:200、1:500、1:500、1:1000、1:5000、1:5000, 1:106
（一）免疫组织化学及其应用
1. 免疫组织化学的相关理论和技术 1. 免疫组织化学的工作原理
已知的特异性抗体或抗原能特异性结合
通过化学反应使标记于结合后的特异性抗体上的显示剂，如酶， 金属离子、同位素等，显示一定的信号（如：颜色）
借助显微镜、荧光显微镜或电子显微镜观察其颜色变化，从而在 抗原抗体结合部位确定组织、细胞结构
In this program we have a central focus: how to dig new biomarkers related with human diseases for both diagnosis, prognosis and clinical treatment
Immunohistochemistry Molecular Pathology Modern Immunopathology Genetic/hereditary pathology Pathology related with cell-communication Some branching of modern clinical pathologies
More than 100 years ago (1830), British physician Thomas Hodgkin discovered a type of Lymphoma with his naked eyes.
In the time, obviously Thomas Hodgkin did never know R-S cells in his cases before the microscope available in clinic.
Pathology in Personalized Medicine
Technical platforms in post-genomic era: High-throughput Gene Expression array Protein and antibody array Proteomics Epigenomic
分子病理学技术进展及临床应用
内容:
一、临床分子病理学方法1 二、当前分子病理学技术在临床的实际应用 三、分子病理学临床应用存在的问题及对策思考
来自临床的问题
Contemporary Understanding of Carcinogenesis by molecular biology
as our introduction
Modern Pathology:
1. Morphology 2. Phenotyping of cell 3. Genotyping of cell 4. Function of cells & molecules
Level of organ-cell by eyes & microscope Sub-cellular level Proteins, Nuclear acids and their reciprocal reactions
Lennert’s Presumption based on 80’s of last Century
Mwk.baidu.comntle zone
Germinal center Dark zone Light zone
Mantle zone Marginal zone
Centroblast Blast Immuoblast Macrophage FDC
分子医学视野下的结直肠癌: 一个病? 两个病? 三个病?
What we are facing on in Clinical practicing?
Classical Pathology: by morphology with cell-
level resolution as Golden Standard more then 100 years.
Apoptosis
No BCR
Apoptosis
B Centroblasts
Centrocytes
?
Post-Germinal Center
Plasma cell
Memory B cell
IgHV clonal rearrangement somatic hypermutation!
What happen to T cell since it has been less mentioned
Thymus-depending zone
Capsular and afferent lymphatics
Reticulum supporting lymphatic tissue
Cortex
Subcapsular sinus
Cortex
Follicle
GC
Medulla
Trabeculae Medullary cords
B cell differentiation with anatom ical sites of various stages
Could you recognize cellular elements as follows in lymph node?
T-zone
Germinal center Mantle zone Marginal zone
„…morphology is like a grand summary of all genetic events in a cell.“
Juan Rosai in : Sue Armstrong : A Matter of Life and Death. Dundee University Press -2008
While at present time, for us, what can be down with our naked eyes rather then with microscope or modern molecular techniques?
The cellular elements in lymphoid & hematopoietic tumors are the key to understand and diagnose their diseases.