PMC5632927_01
Bone marrow aspirate smear from patient #1, Wright-Giemsa, 1000x.
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fig1
Bone marrow evaluation showed 90% lymphoblasts, exhibiting slightly open chromatin and irregular nuclear contours (Figure 1).
Representative flow cytometric histograms of bone marrow aspirate from patient #1.
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fig2
CD3 (variable) and cytoplasmic CD3, CD5, CD7 (bright), CD34, CD45 (dim), and TCR gammadelta. The blasts were negative for CD1a, CD2, CD4, CD8, CD10, CD56, HLA-DR, TCR alphabeta, TdT, and MPO (Figure 2). Cytogenetic studies of the bone marrow showed that 90% of the metaphases had an abnormal karyotype: 47, XX, t(4;10)(q28;p12), cryp ins(11;10)(q23;p12p12), +17 (Figure 3(a)).
Cytogenetic and FISH results from patient #1. (a) Conventional karyotype with 47, XX, t(4;10)(q28;p12), +17. (b) Break-apart AF-10 probe on sequential G-banded to FISH metaphases shows that telomeric 3' AF10 moves to the 4q28, and the centromeric 5' AF10 signal inserts into the 11q23 region. (c) The MLL FISH is not rearranged. In (a), the arrows to 4p28 and 10p12 are for indication of t(4;10)(q28;p12); the arrow to chromosome 17 is for indication of +17.
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fig3
CD3 (variable) and cytoplasmic CD3, CD5, CD7 (bright), CD34, CD45 (dim), and TCR gammadelta. The blasts were negative for CD1a, CD2, CD4, CD8, CD10, CD56, HLA-DR, TCR alphabeta, TdT, and MPO (Figure 2). Cytogenetic studies of the bone marrow showed that 90% of the metaphases had an abnormal karyotype: 47, XX, t(4;10)(q28;p12), cryp ins(11;10)(q23;p12p12), +17 (Figure 3(a)). Of interest, the centromeric 5' AF10 signal is inserted into the 11q23 region (Figure 3(b)). By FISH, MLL was not rearranged (Figure 3(c)).
Real-time RT-PCR showing the MLL/MLLT10 fusion transcripts from patient #1. Delta Rn = normalized fluorescence reporter signal minus baseline; cycle number = cycle of PCR; purple is the positive control; green is the patient; red is no-template control; blue is the negative control (HL60 cell line).
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fig4
Molecular studies by real-time RT-PCR assay confirmed an MLL/MLLT10 (MLL/AF10) fusion transcript (Figure 4).
PMC5632927_02
Bone marrow aspirate smear from patient #2, Wright-Giemsa, 1000x.
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fig5
His bone marrow showed 89% blasts that were small to medium in size with a high nuclear to cytoplasmic ratio, fine chromatin, round to slightly irregular nuclei, and scant cytoplasm (Figure 5).
Representative flow cytometric histograms of bone marrow aspirate from patient #2.
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fig6
TCR gammadelta. They were negative for CD1a, CD2, CD4, CD8, CD10, CD34, TdT, MPO, and TCR alphabeta (Figure 6). Cytogenetic studies performed on the bone marrow showed normal a male karyotype without numerical or structural abnormalities. The patient received treatment per Total XVI (TOTXVI) protocol and achieved MRD negativity after induction on day 42.