悪性リンパ腫の免疫組織化学 各種リンパ腫マーカーと抗原賦活処理

悪性リンパ腫の免疫組織化学染色
悪性リンパ腫の免疫組織化学
各種リンパ球マーカーと抗原賦活処理
DakoCyotmation, CO, LTD
BioMedical Science Department
谷 洋一
悪性リンパ腫の免疫組織化学染色
Lymphoma ?
LCA
Cytokeratin S100 protein
Lymphoma
+*
-
-
Carcinoma
-
+
-/+
Melanoma
-
-
+
Sarcoma
-
-
-/+
LCA-negative lymphoma:
Anaplastic large cell lymphoma and lymphoblastic lymphoma
悪性リンパ腫の免疫組織化学染色
Pre pre B cell
Pre B cell
Precursor cells; antigen
independent
TdT
CD79a
CD19
CD20, CD22
Cytoplasmic CD22
CD10
Bcl-6
CD138
Immature
B cell
Mature
B cell
Virgin B cells; antigen
independent
Activate
d B cell
Plasma
cell
Germinal center & post GC
cells; antigen dependent
悪性リンパ腫の免疫組織化学染色
B Cell lineage markers
CD20 ( L26)
- Positive in B cell lymphomas
- Negative in plasmacytomas and plasmablastic lymphomas
- B cell lymphoblastic lymphoma may be negative
- Heat-induced epitope retrieval causes undesirable nuclear
staining.
CD79a (JCB117)
- Positive in B cell lymphomas (97%)
- Positive in plasmacytomas (50%)
- Positive in B cell lymphoblastic lymphomas
- May be positive in some T cell lymphoblastic lymphoma/
leukemias (6%)
悪性リンパ腫の免疫組織化学染色
Prothymocyte
Subcapsular
thymocyte
Cortical
thymocyte
Medullary
thymocyte
Precursor cells
Peripheral
T cell
Mature T cells
TdT
CD7
CD2
CD3
Cytoplasmic
Surface
CD4
CD4, CD8
CD1
CD8
悪性リンパ腫の免疫組織化学染色
T Cell lineage markers
CD3 (Cytoplasmic)
(Polyclonal Ab against CD3ε,monoclonal Ab, PS1, F7.2.38)
-Highly sensitive and specific for T lineage
-Positive in T / NK cell lymphomas (90% >)
CD45RO (UCHL1)
-Some T cell lymphomas are CD3 -/CD45RO+
- May be positive in myeloid cells and their tumors
- Heat-induced epitope retrieval causes undesirable nuclear staining
悪性リンパ腫の免疫組織化学染色
Immunophenotyping of B or T cell lymphomas
Antibody
Precursor
B cell
Mature
B cell
Precursor
T cell
Mature
T cell
CD20
CD79a
-(-+)
+
+
+
-(-+)
-
CD3
CD45RO
CD7
CD1a
-(-+)
-
-
+
-(-+)
+
+
+
+
+
-
+
-
+
-
TdT
Target Retrieval Methods for Major Primary Antibodies
悪性リンパ腫の免疫組織化学染色
CD20
CD79a
bcl-2
Cyclin D1
min)
CD5
(L26)
( JCB117)
(124)
(DCS-6)
no treatment
CB or TE buffer
TE buffer
TE buffer
(4C7)
TE buffer
(95℃, 20 min)
CD3
CD3
(Polyclonal)
(PC3/188A)
TE buffer
(95℃, 20 min)
CD45RO (UCHL1)
CD7
(CBC.37)
CD4
(IF6)
CD8
(C8/144B)
CD56
(1B6)
Granzyme B (GrB-7)
no treatment
S1700
TE buffer
TE buffer
TE buffer
TE buffer
(95℃,
(95℃,
(95℃,
(95℃,
(95℃,
CD15
CD30
Fascin
min)
ALK-1
(C3D-1)
(Ber-H2)
(55K-2)
no treatment
TE buffer
CB or TE buffer
(95℃, 20 min)
(95℃, 20 – 40
(ALK1)
TE buffer
(95℃, 20 min)
(95℃, 20 min)
(95℃, 20 – 40
20
20
20
20
20
min)
min)
min)
min)
min)
(95℃, 20 – 40 min)
CB: Citrate
Buffer(pH6.0)
TE: Tris-EDTA(pH9.0)
S1700: DAKO S1700
悪性リンパ腫の免疫組織化学染色
CD20(L26) and CD45RO(UCHL1)
Heat-induced epitope retrieval causes undesirable nuclear staining
CD20
CD45RO
CD20
CD45RO
悪性リンパ腫の免疫組織化学染色
Mantle Cell lymphoma
Cyclin D1
TRIS-EDTA (pH9.0) , 95℃ 20 min
悪性リンパ腫の免疫組織化学染色
NK/T cell lymphoma, cytotoxic phenotype
CD8
CD56
TRIS-EDTA (pH9.0) , 95℃ 20 min
Granzyme B
悪性リンパ腫の免疫組織化学染色
Anaplastic Large Cell Lymphoma (ALCL)
ALK1
TRIS-EDTA (pH9.0) , 95℃ 20 min
悪性リンパ腫の免疫組織化学染色
Hodgkin lymphoma
CD30
Fascin
TRIS-EDTA (pH9.0) , 95℃ 20 min