20090114-1 Lymph Node Intro (1)

8/4/2019 20090114-1 Lymph Node Intro (1)

1/31

Lymph Node Normal Morphology

Cortex

Primary Follicle

Secondary Follicle

Mantle Zone

Paracortex

Medulla

Sinuses

8/4/2019 20090114-1 Lymph Node Intro (1)

2/31

Cortex

Primary B-Cell Follicles

Nodules of small lymphocytes

Lack germinal centers

Secondary B-Cell Follicles

Result of stimulation

Germinal Centers

Mantle zone

8/4/2019 20090114-1 Lymph Node Intro (1)

3/31

Germinal Centers

Pale zone

toward antigen entry

small cleaved cells / centrocytes

follicular dendritic cells

Dark zone

toward paracortex

large lymphoid cells / centroblasts

tingible body macrophages

8/4/2019 20090114-1 Lymph Node Intro (1)

4/31

Mantle Zone

polarized toward antigen entry

express bcl-2 protein

8/4/2019 20090114-1 Lymph Node Intro (1)

5/31

Paracortex

rich in T cells

CD4:CD8 ratio variable

interdigitating dendritic cell

S-100 positive

irregular vesicular nuclei

high endothelial venules

postcapillary vessel

cuboidal epithelium

8/4/2019 20090114-1 Lymph Node Intro (1)

6/31

Medullary areas

B cells predominate especially plasma cells

histiocytes

8/4/2019 20090114-1 Lymph Node Intro (1)

7/31

Handling the Fresh Specimen

Surgeon should excise the largest and most

abnormal node

Tissue for histology

Touch imprints

Fresh / frozen tissue for immunologicstudies

Sterile portion for cytogenetics

8/4/2019 20090114-1 Lymph Node Intro (1)

8/31

Frozen Section

Diagnostic frozen section should be

discouraged

Use frozen to assess adequacy or triage

tissue

8/4/2019 20090114-1 Lymph Node Intro (1)

9/31

Freezing for Immunologic

Studies

Liquid nitrogen or isopentane / dry ice mix

is best

Thin sections (

8/4/2019 20090114-1 Lymph Node Intro (1)

10/31

Fixation

Node sliced in 2-3 mm intervals

One metal based fixative (B5, Zenkers, zinc

sulfate)

One neutral buffered formaldehyde

(formalin)

8/4/2019 20090114-1 Lymph Node Intro (1)

11/31

Processing

Single most important factor for optimal

histology is section thickness

Sections should be one cell layer thick

8/4/2019 20090114-1 Lymph Node Intro (1)

12/31

Routine Stains

H&E

Giemsa - highlight nuclear features,

cytoplasmic granules and plasmacytoid

features

PAS - highlights mucin and glycogen,

immunoglobulin inclusions and blood

vessels

Methyl-green pyronin - highlights

plasmacytoid features

8/4/2019 20090114-1 Lymph Node Intro (1)

13/31

Common Errors in Fixation and

Processing

Drying of specimen - dark edge artifact;

autolysis if prolonged

Section >3 mm thick - soft unfixed core;

center cells show ballooning and are pale

Overfixation in B5 - brittle tissue; decreased

nuclear staining

Inadequate dehydration - numerous cracks

(dry earth look)

8/4/2019 20090114-1 Lymph Node Intro (1)

14/31

Common Errors in Fixation and

Processing

Paraffin too hot - muddy staining with poor

detail

Improper sectioning - Venetian-blind effect;

poor cytologic detail

Section drying too hot - bubbled nuclei and

antigen loss

8/4/2019 20090114-1 Lymph Node Intro (1)

15/31

Antibodies Employed in Paraffin

Tissue Sections

Antibody /

CD Anti en

Normal Reactivity Comments

LCA

CD45RB

Lymphocytes,

myeloid cells andmacrophages

Plasma cells and RS

cells negative

Immuno-

globulin

B cells and plasma

cells

Paraffin c to lasmic

Frozen surface

L26

CD20

B cells occasional

macrophages

Plasma cells negative

Poor reactivity in

decalcified tissue

8/4/2019 20090114-1 Lymph Node Intro (1)

16/31


Tissue Sections

Antibody /

CD Anti en


CD79 B cells Plasma cells negative

LN1

CDw75

Germinal center B

cells

Better in B5

LN2CD74

B cells,interdigitating

dendritic cells,

macrophages

Better in B5

8/4/2019 20090114-1 Lymph Node Intro (1)

17/31


Tissue Sections

Antibody /

CD Anti en


MB1

CD45RA

B cells and some T

cells

Plasma cells negative

CD3 T cells May require

pretreatment

Leu-22CD43 T cells tissue CD5useful in small

lymphocytic

neoplasms

8/4/2019 20090114-1 Lymph Node Intro (1)

18/31


Tissue Sections

Antibody /

CD Anti en


UCHL1

CD45RO

T cells May see nonspecific

cytoplasmic staining

Leu-7

CD57

GC T cells and some

NK cells

Better in frozen tissue

TCR betachain

T cells Requires proteolyticdigestion; better in

frozen tissue

8/4/2019 20090114-1 Lymph Node Intro (1)

19/31


Tissue Sections

Antibody /

CD Anti en


LeuM1

CD15

Granulocytes,

macrophages

Carcinomas and

CMV infected cellsmay be reactive

BerH2

CD30

Activated B and T

cells, plasma cells

Less reactive in B5,

embryonal ca. and

few other non-hematolymphoid

neoplasms positive

8/4/2019 20090114-1 Lymph Node Intro (1)

20/31


Tissue Sections

Antibody /

CD Anti en


KP1

CD68

Macrophages

myeloid cells

Reactive in

melanoma, MFH,granular cell tumors

and renal cell ca.

Lysozyme Macrophages

myeloid cells

Reactive with many

non-hematolymphoidneoplasms

8/4/2019 20090114-1 Lymph Node Intro (1)

21/31


Tissue Sections

Antibody /

CD Anti en


S100 Langerhans cells,

interdigitating andsome follicular

dendritic cells

Reactive with many

non-hematolymphoidneoplasms

Mac-387 Macrophages

myeloid cells

Reactive with some

squamous cell ca.

8/4/2019 20090114-1 Lymph Node Intro (1)

22/31


Tissue Sections

Antibody /

CD Anti en


bcl-2 Non-germinal center

B cells, most T cellsand plasma cells

Best in B5;

differentiate benignfrom malignant

follicular lesions

Myeloper-

oxidase

Myeloid cells Most sensitive and

specific myeloid

marker

8/4/2019 20090114-1 Lymph Node Intro (1)

23/31

Antibodies Employed in Fresh or

Frozen Tissue

Antibody /

CD Anti en


CD45 Hematolymphoid

cells

Plasma cells and

ALCL may benegative

Immuno-

globulin

B cells Monoclonal Ig less

sensitive than

polyclonal Ig

Precursor Blymphomas and

leukemias may be

negative

8/4/2019 20090114-1 Lymph Node Intro (1)

24/31


Frozen Tissue

Antibody /

CD Anti en


CD19 B cells Earliest expressed B

cell antigen

CD20 B cells Most plasma cells

unreactive

CD79 B cells May be negative inplasma cells

8/4/2019 20090114-1 Lymph Node Intro (1)

25/31


Frozen Tissue

Antibody /

CD Anti en


CD10

CALLA

Precursor and GC

B cells

Follicular vs. other

low grade MLEwings and MFH

positive

CD21 Mantle and marginal

zone B cells

Most mantle and

marginal zone MLCD23 CLL / SLL reactive

Mantle cell ML

unreactive

Upregulated by EBV

infection

8/4/2019 20090114-1 Lymph Node Intro (1)

26/31


Frozen Tissue

Antibody /

CD Anti en


CD32 Mantle zone B cells Reactive with MCL

and some follicularML, myeloid

leukemias

CD35 Mantle and marginal

zone B cells

Similar to CD21

CD38 Lymphoid progenitor

cells, NK cells and

plasma cells

One of few markers

expressed by plasma

cell neoplasms

8/4/2019 20090114-1 Lymph Node Intro (1)

27/31


Frozen Tissue

Antibody /

CD Anti en


CD2, CD3,

CD5 andCD7

T cells Pan T cell antigens

CD7 earliestexpressed and most

commonly deleted

CD1a Cortical thymocytes Good for precursor T

cell neoplasms;thymomas rich in

CD1A cells

8/4/2019 20090114-1 Lymph Node Intro (1)

28/31


Frozen Tissue

Antibody /

CD Anti en


CD4 Helper/Inducer T

cells macrophagesand dendritic cells

May be expressed in

some plasma cellneoplasms

CD8 Cytotoxic/Suppressor

T cells and NK cells

Minority T subset;

may be increased in

early phase viralinfections or late HIV

infection

8/4/2019 20090114-1 Lymph Node Intro (1)

29/31


Frozen Tissue

Antibody /

CD Anti en


CD11c Myelomonocytic

cells

Hairy cell leukemia

and monocytoid Bcell ML

CD13 Myelomonocytic

cells

Most AML M1-M5

CD14 Monocytes andmacrophages

Many M4-M5 AML

CD33 Myeloid cells and

monocytes

Most myeloid

leukemias

8/4/2019 20090114-1 Lymph Node Intro (1)

30/31


Frozen Tissue

Antibody /

CD Anti en


CD16 NK cells and

granulocytes

CD56 NK cells and a few T

cells

Reactive with neural

and neuroendocrine

cells and neoplastic

plasma cellsCD57 Some NK cells and T

cells

8/4/2019 20090114-1 Lymph Node Intro (1)

31/31


Frozen Tissue

Antibody /

CD Anti en


CD25 Activated T cells, B

cells and monocytes

ATLL, HCL, ALCL

and some other T andB cell ML

CD34 Progenitor cells and

endothelial cells

Stem cell marker

Good for vascular

neoplasmsTdT Precursor cells in

marrow and cortical

thymocytes

Lymphoblastic

leukemia / l m homa

20090114-1 Lymph Node Intro (1)

Documents

Transcript of 20090114-1 Lymph Node Intro (1)