PMC9846588_01
Lymph nodes histological and immunohistologic features. (A) The lymph
node architecture was effaced due to infiltrate of medium to large sized
atypical lymphatic cells with interfollicular growth pattern and cells
having blastoid morphology (HE staining, x40 magnification). ((B-E), x40
magnification) Immunohistochemical staining of the interfollicular
lymphatic infiltrate for CD20 (image (B)) and PAX-5 (image (C)) was
negative while CD38 (image D) and CD79a (image E) staining were
positive. Also, tumour cells were diffusely and strongly MUM1 positive
(image (F), x10 magnification). ((G-I), x20 magnification) The staining
for CD138 (image (G)), HHV-8 (image (H)) and EBER in situ hybridisation
(image (I)) were negative. The CD30 stained only blastoid cells and in
less than 5% of tumour cell population (image (J), x40 magnification).
There was high Ki-67 proliferation index, approximately 65% (image (K),
x10 magnification) and between tumour cells there was lot of histiocytes
(image (L), x10 magnification).
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TdT-, CD45+, Bcl-6-, CD10- and HHV-8-. The Epstein-Barr encoding region (EBER) in situ hybridisation was negative (Figure 1). CD68 staining showed lot of histiocytes and Ki-67 proliferation index was high, approximately 65% (Figure 1).
GeneScaning analysis of multiplex IgH and IgKappa PCR products. This figure
shows the B cell clonality analysis which is PCR based high resolution
fragment analysis of immunoglobulin heavy chain (IgH) and Ig kappa (Kappa)
light chain gene rearrangement using multiplex primers from BIOMED2
protocol. The top 2 electropherograms show polyclonal population
(characteristic Gaussian size distribution) of lymphocytes with multiple
PCR product using IgH FR2 (image (A)) and IgH FR3 (image (B)) primers.
However, the immunoglobulin light chain kappa gene clonality analysis
showed typical results for clonal population of lymphocytes. Arrows
indicate clonality peaks of specific size for IgKappa tube A (149 bp in
size) (image (C)) and IgKappa tube B (235 and 287 bp in size) (image (D)).
All the gene rearrangement analyses were performed in triplicates. The
orange, regularly spaced peaks represent the size standards.. Abbreviation: nt, nucleotides.
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The clonality analysis of IgH gene in FR2 and FR3 regions showed polyclonal distribution of PCR products (gaussian distribution of multiple peaks representing multiple PCR products) while the IgKappa gene clonality analysis showed only one prominent peak (monoclonal result consisting of one type of PCR product) (Figure 2).
GeneScaning analysis of multiplex TCR PCR products. Figure shows PCR
based T cell clonality analysis using multiplex primers for T-cell
receptor (TCR) gene rearrangement. In TCRbeta tube C (image (A)) high
resolution fragment analysis showed one prominent clonal PCR product of
307 bp in size (red arrow). For TCRgamma tube A (image (B)) high resolution
fragment analysis showed the monoclonal (biallelic) rearrangement
(154 bp and 156 bp in size-red arrows). All of the gene rearrangement
analyses were performed in triplicates. The orange, regularly spaced
peaks represent the size standards.. Abbreviation: nt, nucleotides.
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However, the T cell receptor analysis of TCR gamma region showed bi-clonal and TCR beta region monoclonal distribution of PCR products (Figure 3).