Dr. Anurag Mehta

Director Lab services

RGCI & RC

Carl Sternberg

Dorothy Reed

Sir Samuel Wilks

Thomas HodgkinCD15

Long history > 150 years

Enigmas

EBV

Origin of RS cell

Mélange of

Antigens

Milieu

Clustering

&

familial

Overlapping

features with

Several Lymphoma

types

Despite long history and not being infrequent

Difficult D/D

References: 1. American Cancer Society. Cancer Facts & Figures 2013. Atlanta, GA: American Cancer Society; 2013. 2. Types of non-Hodgkin lymphoma.

American Cancer Society Web site. http://www.cancer.org/cancer/non-hodgkinlymphoma/detailedguide/non-hodgkin-lymphoma-types-of-non-hodgkin-lymphoma.

Updated November 14, 2013. Accessed December 19, 2013. 3. Küppers R. Nat Rev Cancer. 2009;9(1):15-27. 4. International T-Cell Lymphoma Project.

J Clin Oncol. 2008;26(25):4124-4130. 5. Swerdlow SH et al, eds. WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th ed.

Lyon, France: IARC; 2008. Note: B-cell and T-cell sub-classifications are illustrated on slides 10 and 11, respectively.

Nodular lymphocyte-

predominant

Hodgkin lymphoma (NLPHL)

(5%)

Non-Hodgkin

lymphoma (NHL)

Hodgkin

lymphoma (HL)

Classical Hodgkin

lymphoma (cHL)

(95%)

Lymphoma

1. The Hodgkin lymphoma is separated into

2 types for several reasons.

2. One amongst them is the morphology of

neoplastic cells

• RS cells, Hodgkin Cells & variant

→ cHL

• LP Cells → NLPHL

3. Immunophenotypic attributes

4. And now also on the basis of cell

of origin

The classical Hodgkin Lymphoma is further

divided into subtypes on the basis of

1. Quantity of RS cells

2. Host response: Nature of the inflammatory

components

3. Fibrosis

• Banded fibrosis

• Birefringent, fibrillar

• Non-birefringent, amorphous

Hodgkin Lymphoma: Classifications

WHO 2008

Nodular lymphocyte predominant HL

Classical HL

1. Nodular sclerosis

2. Mixed cellularity

3. Lymphocyte depleted

4. Lymphocyte-rich

Although classification is mainly

based on histopathology, recent

studies have demonstrated that

these disease entities are

biologically different, with different

genomic alterations, gene-

expression patterns, cytokine

milieu and clinical behavior.

1. Devilard E, Bertucci F, Trempat P, et.al. Gene expression profiling defines molecular subtypes

of classical Hodgkin’s disease. Oncogene 2002, 21:3095-3102.

2. Levy A, Armon Y, Gopas J, Ariad S, et.al. : Is classical Hodgkin’s disease indeed a single entity?

Leuk Lymphoma 2002, 43:1813-1818.

LP CELL

What is the cell of origin of RS Cells & LP cell?

Difficult to identify till recently

• Neoplastic population is sparse

• Strange phenotypes

Rescued from apoptosis

and transforming events

Granzyme B /perforin

CD30

Extensive

reprogramming. USP

loss of the B cell

identity, so that these

“failed” B cells escape

the apoptosis

LMP1

How is a GCB cell with crippling mutation rescued/ why doesn’t it die

an apoptotic death?

1.LMP1 mimics an active CD40 receptor, a central costimulatory molecule for B cells

2.LMP2a carries a cytoplasmic motif that resembles the signaling module of the BCR

LP Cells

1. Express B-lymphocyte markers, including GCB Ag like BCL-6 and

activation-induced cytidine deaminase.

2. LP cells grow in a follicular pattern in assn. with FDCs & THFC

3. LP cells express functional BCR

4. A gene expression study of isolated LP cells indicate that these cells

resemble an intermediate developmental stage between germinal

centre and memory B cells.

Marafioti, T. et al. Origin of nodular lymphocyte-predominant Hodgkin's disease from a clonal

expansion of highly mutated germinal-center B cells. N. Engl. J. Med. 337, 453- 458 (1997).

Brune, V. et al. Origin and pathogenesis of nodular lymphocyte-predominant Hodgkin

lymphoma as revealed by global gene expression analysis. J. Exp. Med. 205, 2251- 2268

(2008).

RS and Hodgkin cells of cHL

1. B-cells of the germinal center origin with crippling mutations –

somehow rescued – loses several B cell phenotypes including

immunoglobulin receptors

2. Create a unique histology by to and fro exchange of cytokine-ligand

interactions

3. 40% of cases associated with EBV in Western countries; higher in

developing countries ~ 60%

Marafioti T, et al. Hodgkin and Reed-Sternberg cells represent an expansion of a single

clone originating from a germinal center B-cell with functional immunoglobulin gene

rearrangements but defective immunoglobulin Transcription. Blood. 2000;95(4):1443–

1450.

Familial Predisposition

1. Genetic predisposition observed in 1% of Hodgkin lymphoma.

2. 3- to 7-fold increased risk of developing the disease in siblings.

3. KLHDC8B loss of function mutations / SNP with reduced function

4. germline frame shift mutation of the NPAT gene was found in a

family with four members affected by NLPHL

5. Suspected germline alterations responsible for familial HL are

PVT1, REL, GATA3

and a germline variation leading to the deletion of serine 724 was found in several cases

suggesting an elevated risk for the disease (odds ratio = 4.11; P = .018). NPAT is thus far

the first gene implicated in nodular lymphocyte predominant Hodgkin lymphoma

predisposition.

Saarinen S1, Aavikko M et.al., Exome sequencing reveals germline NPAT mutation as

a candidate risk factor for Hodgkin lymphoma. Blood. 2011 Jul 21;118(3):493-8. doi:

10.1182/blood-2011-03-341560. Epub 2011 May 11.

Proc Natl Acad Sci U. S A. 2009 Sep 1; 106(35): 14920–14925.

Published online 2009 Aug 12.

Genetics

Mutations in a gene encoding a midbody kelch protein in familial and sporadic

classical Hodgkin lymphoma lead to binucleated cells

Stephen J. Salipante,a Matthew E. Mealiffe,b,1 Jeremy Wechsler,c

ABSTRACT

Classical Hodgkin lymphoma (cHL) is a malignancy of B-cell origin in which the neoplastic cells, known as

“Reed-Sternberg” (RS) cells, are characteristically binucleated. Here we describe a family where

multiple individuals developing cHL have inherited a reciprocal translocation between

chromosomes 2 and 3. The translocation disrupts KLHDC8B, an uncharacterized gene from a

region (3p21.31) previously implicated in lymphoma and related malignancies, resulting in its loss of

expression. We tested KLHDC8B as a candidate gene for cHL and found that a 5′-UTR polymorphism

responsible for decreasing its translational expression is associated with cHL in probands from other

families with cHL and segregates with disease in those pedigrees. In one of three informative sporadic

cases of cHL, we detected loss of heterozygosity (LOH) for KLHDC8B in RS cells, but not reactive T

lymphocytes, purified from a malignant lymph node.

HL: Definition

Characterized by presence of a tiny population of

neoplastic cells in a defined milieu

1.The Neoplastic cell- RS cell, Hodgkins cell,

variants and LP cells

2.The Milieu

Mature & small lymphocytes

Can have abundance of Histiocytes

Occasionally paucicellular for

lymphocytes

Some plasma cells??

Eosinophils

Delicate fibrosis between cells

The concept of context/ milieu is actually fluid. Not

something carved in stone. Most of the

lymphocytes are mature (95%). Exceptions occur.

Reactive and atypical lymphocytes sometimes make

up the milieu

Sohani AR, Jaffe ES, Harris NL et.al

Nodular lymphocyte-predominant Hodgkin

lymphoma with atypical T cells: a morphologic

variant mimicking peripheral T-cell lymphom

Am J Surg Pathol. 2011 Nov;35(11):1666-78.

10.1097/PAS.0b013e31822832de.

TypeLympho

cyte

Histio

cyteEo

Plasma

cell

Banded

collagen

Fibrous

tissueRS cell

NLPHL 5+ + to 3+ 0 0 0 0LP cells

±

LRCHL 4+ + to 3+ 0 to + 0 to + 0 0 +

NS + to 4+ + to 2+ + + + to 4+ + +, +++

MC 2+ to 3+ + to 3+ 2+ + 0 2+ 2+ to 3+

LD 0 to + 0 to + + to 2+ + 0 + to 4+ 2+ to 5+

Cellular Composition & types of HL

Nodular Lymphocyte Predominant Hodgkin Lymphoma (NLPHL)

1. 5-6% 0f all HL

2. Nodules of B cells + Spheroidal collection of FDCs + LP cells in center

3. Evolve from a nodular → nodular and diffuse→ diffuse.

4. All ages, including children. 4th decade

5. M:F = 2.5:1

6. Mostly Stage I

7. Cervical, axillary, inguinal Lymph nodes most often involved.

8. Rare involvement of mediastinum. Extranodal disease in NLPHL is

rarely observed, with the exception of splenic involvement, which can

be seen in 10%–15% of cases. Liver involvement occurs in 10% of

patients, and bone marrow or lung infiltration is found in 5%.

9. 10-15% in higher stages

Nodules in

NLPHL

Sclerosis can occur in older

nodes

NLPHL

Plasma cells in

NLPHL

Rosettes around the L P

cells

CD20 nodules

CD20 nodulesCD20 nodules &

LP Cells

LCA

CD20

CD20

PAX5

PAX5

CD3

CD57

CD4

CD57

EMA

CD23

BOB-1

CD30 expression

in NLPHL

OCT2

Nodular Lymphocyte Predominance Hodgkin Lymphoma:

Immunophenotype of LP/L&H Cells

1. CD45 + (95%)

2. CD30 - (10% weak +)

3. CD15 - (10% +)

4. CD20, PAX-5 + (95%)

5. Bcl-6, bcl-2 + (95%)

6. EMA +/- (70%)

7. CD3, CD43, CD10 - (0%)

8. Consistent lack of EBV markers

9. CD4 and CD57 co expressing T cells ring around the LP cells