Page 166 - Haematologica March 2020

M. Xu et al.
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Figure 3. Tg(hsp70:p210BCR/ABL1) adult fish display abnormal myeloid cell expansion resembled chronic myeloid leukemia (CML)-like phenotypes. (A) May-Grunwald- Giemsa staining of peripheral blood (PB) cells (top) and kidney marrow (KM) blood cells (bottom) were obtained from wild-type (WT) (a, e) (n=55) and Tg(hsp70:p210BCR/ABL1) (b-d, f-h) (n=101) adult zebrafish. Blue arrows indicate accumulated promyelocytes in CML-chronic phase (CP)-like fish. Yellow arrows indicate neutrophils in CML-CP-like fish. Red arrows indicate myeloid precursors in CML accelerated phase (AP)-like fish. Black arrows indicate blasts in both CML-AP-like fish and CML-BP-like fish. The percentages indicate the ratio that number of fish with leukemia-like phenotype in total number of fish. Original magnification x400. (B) Statistical analysis. Blood cell counts of PB and KM were calculated manually based on their morphology. ANOVA; mean±Standard Error of Mean; *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001. (C) Large numbers of eosinophils (b, yellow arrow), lymphocytes (c, blue arrow), and platelets (d, green arrow) accumulated in CML-like Tg(hsp70:p210BCR/ABL1) adult fish PB compared with WT controls (a, n=55). The percentages indicate the ratio that number of fish with abnormal cells accumulated in total number of CML-like Tg(hsp70:p210BCR/ABL1) adult fish. Red arrows indicate neutrophils. Original magnification x400. Stains were May-Grunwald- Giemsa. (D) Histology of the spleen from CML-like Tg(hsp70:p210BCR/ABL1) adult fish showing disruption of the spleen architecture and massive invasion by hematopoi- etic cells compared with WT controls. Original magnifications x200 for each panel. Inset (red rectangles): granulocytic myeloid cells at higher magnification x1000. Stains were Hematoxylin & Eosin. (E) The morbidity rate of CML was higher in 1 Y-HS Tg(hsp70:p210BCR/ABL1) adult fish (n=35) compared with 1 Y-No HS Tg(hsp70:p210BCR/ABL1) adult fish (n=18), 97.14% and 44.44%, respectively. 1Y-No HS: 1-year-old Tg(hsp70:p210BCR/ABL1) adult fish without heat shock treatment. 1Y- HS: 1-year-old Tg(hsp70:p210BCR/ABL1) adult fish with heat shock treatment. The morbidity rates of Tg(hsp70:p210BCR/ABL1) transgenic fish in the HS group developing into CML-CP 1 phase was 17.14%, while that in the No-HS treatment group was 33.33%. The morbidity rate of Tg(hsp70:p210BCR/ABL1) transgenic fish in the heat shock treatment group developing into CML-CP 2 phase and CML-AP phase were 42.86% and 37.14%, compared with 0.00% and 11.11%, respectively, in the No-HS Tg(hsp70:p210BCR/ABL1) adult fish.
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haematologica | 2020; 105(3)