Korean J Lab Med.  2008 Apr;28(2):89-94. 10.3343/kjlm.2008.28.2.89.

Cytomorphology and Molecular Characterization of CLTC-ALK Rearrangement in 2 Cases of ALK-Positive Diffuse Large B-cell Lymphoma with Extensive Bone Marrow Involvement

• Choung HS
• Kim HJ
• Kim WS
• Kim K
• Kim SH

• KMID: 854834
• DOI: http://doi.org/10.3343/kjlm.2008.28.2.89

Abstract

Aanaplastic lymphoma kinase (ALK)-positive diffuse large B-cell lymphoma (DLBCL) is an unusual disease entity first reported in 1997 as DLBCL with expression of full-length ALK protein. The World Health Organization classification enlists the disease as a rare variant of DLBCL. Herein we describe two cases of ALK-positive DLBCL with cytomorphologic and molecular characteristics for the first time in Korea. The patients were 35-yr-old and 24-yr-old male patients. Immunohistochemical studies on the lymph nodes revealed large sized neoplastic cells with plasmablastic differentiation, which were negative for CD30 and positive for ALK with the characteristic granular staining in the cytoplasmic region. Extensive involvement of bone marrow was observed in both cases showing large, extremely atypical cells. Fluorescence in situ hybridization and molecular studies on the bone marrow aspirate specimens led to the detection of a clathrin (CLTC)/ALK rearrangement. Despite aggressive chemotherapy, the patients died 15 and 17 months after the diagnosis, indicating poor prognosis of the disease entity. This is the first report demonstrating the cytomorphologic findings of ALK-positive DLBCL cells on bone marrow aspirates.

Keyword

Diffuse large B-cell lymphoma; ALK-positive; Cytomorphology; CLTC-ALK; Fluorescence in situ hybridization

MeSH Terms

Adult
Bone Marrow/*pathology
Clathrin/*genetics
Fatal Outcome
*Gene Fusion
Humans
Immunohistochemistry
In Situ Hybridization, Fluorescence
Lymphoma, Large B-Cell, Diffuse/*genetics/metabolism/*pathology
Male
Protein-Tyrosine Kinases/*genetics
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, DNA

Figure

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  Fig. 1. (A) Bone marrow aspirate smear in Case 1 showing neoplastic cells with extremely large size and bizarre morphology (Wright-Giemsa stain, ×1,000). (B) Bone marrow biopsy section in Case 1 showed that the bone marrow was packed with the large neoplastic cells (H&E stain, ×1,000). (C) Bone marrow aspirate smear in Case 2 showing neoplastic cells with irregular morphology smaller than those in Case 1, but still large (Wright-Giemsa stain, ×1,000).

• 

  Fig. 2. Immunohistochemical stains on lymph node biopsy in Case 1. The large neoplastic cells showed positivity to CD79a (×200) (A), weak kappa restriction (×200) (B), and granular cytoplasmic positivity to ALK (×1,000) (C).

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  Fig. 3. Fluorescence in situ hybridization (FISH) study for the ALK gene rearrangement (Dual-color Break-apart Probe, Vysis). (A) Interphase FISH and (B) metaphase FISH in Case 1. Note that the unusually large sized neoplastic cells had two fusion, two red, and one green signal indicating polyploidy with ALK gene rearrangement, while relatively small-sized normal bone marrow hematopoietic cells had two fusion signals. (C) Interphase FISH in Case 2. Large neo-plastic cells had one fusion and one red signal indicating 5′ deletion of ALK gene.

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  Fig. 4. (A) RT-PCR revealed a chimeric CLTC/ALK fusion transcript at 270 bp. N, negative control; P, patient; M, 100 bp size marker. (B) Direct sequencing analysis of RT-PCR showed that the rearrangement occurred at intron 31 of CLTC and intron 20 of ALK.

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