PMC4706875_01
Gross photomicrograph depicting pupil-optic nerve (PO) section of enucleated eye which reveals diffuse thickening of the iris and part of the choroid layer.
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fig1
On gross examination, a cross section of the eye showed diffuse circumferential thickening of the choroid (Figure 1).
Blasts cells infiltrating the iris (H&E, magnification x20).
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fig2
Microscopic examination revealed dense infiltration of the choroid and iris and optic nerve margin by a monotonous mononuclear cell infiltrate morphologically resembling blasts (Figures 2 and 3).
Sheets of monotonous mononuclear blast cells (H&E, magnification x400).
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fig3
Microscopic examination revealed dense infiltration of the choroid and iris and optic nerve margin by a monotonous mononuclear cell infiltrate morphologically resembling blasts (Figures 2 and 3).
Flow cytometric immunophenotypic analysis of vitreous fluid: blast gate (P4 gate in the top line) represents the dim CD45+ cells with low side light scatter (SSC). Blast gate (P3 gate in the bottom line) represents the same population of blasts analyzed for cytoplasmic markers in a separate tube. The distinct cluster of blasts (red color population) exhibit the following immunophenotypic characteristics: positive for CD34, CD117, CD7, CD11b, CD13 (dim), and CD33; negative for CD19, CD10, CD20, CD3, CD5, CD15, CD16, CD61, CD235a, HLA-DR, CD14, cytoplasmic CD3, cytoplasmic CD79a, cytoplasmic MPO, and cytoplasmic TdT.
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fig4
By flow cytometry (Figure 4), the blasts were positive for CD34 and demonstrated myeloid immunophenotype expressing CD13, CD33, and CD11c.
CD34 immunohistochemistry: sheets of monotonous CD34 positive blast cells (magnification x400).
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fig5
By immunohistochemical staining (Figures 5-7), the blasts were positive for CD34 (Figure 5) with focal expression of variable intensity for CD117 (Figure 6) and strong and diffuse expression of TdT (Figure 7) but were negative for B- or T-lineage lymphoid antigen markers (CD20, CD3, and CD79a) (not shown).
CD117 immunohistochemistry: focal and variable intensity of CD117 expression is seen in the blasts (magnification x400).
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fig6
By immunohistochemical staining (Figures 5-7), the blasts were positive for CD34 (Figure 5) with focal expression of variable intensity for CD117 (Figure 6) and strong and diffuse expression of TdT (Figure 7) but were negative for B- or T-lineage lymphoid antigen markers (CD20, CD3, and CD79a) (not shown).
TdT immunohistochemistry: sheets of blasts showing nuclear expression of TdT (magnification x400).
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fig7
By immunohistochemical staining (Figures 5-7), the blasts were positive for CD34 (Figure 5) with focal expression of variable intensity for CD117 (Figure 6) and strong and diffuse expression of TdT (Figure 7) but were negative for B- or T-lineage lymphoid antigen markers (CD20, CD3, and CD79a) (not shown).
Fluorescence in situ hybridization assay on vitreous humor smear shows two BCR-ABL1 fusion signals, one BCR probe signal, and two ABL1 probe signals; thus a t(9;22) BCR-ABL1 gene rearrangement with gains of 9 and 22 [orange = 9q34 ABL1 probe; green = 22q11.2 BCR probe].
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fig8
FISH cytogenetic analysis on vitreous fluid (Figure 8) demonstrated presence of BCR/ABL1 [t(9;22)] gene rearrangement identical to the abnormalities detected in the original bone marrow (Figure 9) in which the diagnosis of CML was rendered.
Fluorescent in situ hybridization on original diagnostic bone marrow aspirate shows one BCR-ABL1 fusion signal, two BCR probe signals, and three ABL1 probe signals in 90.5% of interphase cells.
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fig9
CML. Conventional karyotyping and fluorescence in situ hybridization (FISH) on the bone marrow aspirate (Figure 9) showed evidence of t(9;22) BCR-ABL1 rearrangement and real time quantitative PCR was positive for M-BCR/ABL t(9;22), p210 fusion transcript. FISH cytogenetic analysis on vitreous fluid (Figure 8) demonstrated presence of BCR/ABL1 [t(9;22)] gene rearrangement identical to the abnormalities detected in the original bone marrow (Figure 9) in which the diagnosis of CML was rendered.