Figure 2.
Alteration to BM megakaryocyte form and function during experimental VL. (A) BM MKs were counted manually on hematoxylin and eosin–stained frozen sections in 10 random fields of view for uninfected (n = 4) and infected (n = 4) mice. TEM (FEI Tecnai 12G2) was performed on femur sections of uninfected and day-28–infected mice. Images were analyzed by creating outlines of demarcation membranes with Fiji ImageJ software (supplemental Figure 3). (B) Perimeters were then calculated on 25 uninfected and 46 infected mouse BM MKs. Data represent the mean ± standard error of the mean (SEM) across individual mice. The unpaired Student t test was applied to the means of the perimeter of membranes; ****P < .0001, in uninfected vs infected mice. (C) Representative TEM images of BM MKs in uninfected (i-ii) and infected (iii-iv) mice, showing the presence of multilobulated nucleus (N) and DMS (d) within the cytoplasm. Bar represents 5 μm. Original magnification ×2000. (D) Anti-CD41 mAb-induced ITP in infected and uninfected mice. A single intraperitoneal injection (0.2 µg/g body weight) of anti-CD41 mAb (clone, MWReg30) was administered, followed by collection of blood for platelet counts at the specified time points. Platelet count (E) and rate of decline and recovery of platelet count (F) in uninfected and infected mice after anti-CD41 mAb-induced thrombocytopenia. Data were pooled from 3 independent experiments with uninfected (n = 3), anti-CD41–treated uninfected (n = 9), infected (n = 5), and infected+anti-CD41–treated (n = 10) mice. (G) Representative TEM images of BM MK from infected anti-CD41–treated (i-ii) and uninfected anti-CD41–treated (iii-iv) mice, showing DMS and platelet areas. Bar represents 5 μm. Original magnification ×2000. (H) The perimeter of the demarcation membranes of anti-CD41–treated uninfected (n = 20) and anti-CD41–treated infected (n = 30) mice. The dotted line corresponds with the mean of the perimeter of the DMS in nontreated infected mice. Data are expressed as means ± SEM and analyzed with the unpaired Student t test. *P < .05.

Alteration to BM megakaryocyte form and function during experimental VL. (A) BM MKs were counted manually on hematoxylin and eosin–stained frozen sections in 10 random fields of view for uninfected (n = 4) and infected (n = 4) mice. TEM (FEI Tecnai 12G2) was performed on femur sections of uninfected and day-28–infected mice. Images were analyzed by creating outlines of demarcation membranes with Fiji ImageJ software (supplemental Figure 3). (B) Perimeters were then calculated on 25 uninfected and 46 infected mouse BM MKs. Data represent the mean ± standard error of the mean (SEM) across individual mice. The unpaired Student t test was applied to the means of the perimeter of membranes; ****P < .0001, in uninfected vs infected mice. (C) Representative TEM images of BM MKs in uninfected (i-ii) and infected (iii-iv) mice, showing the presence of multilobulated nucleus (N) and DMS (d) within the cytoplasm. Bar represents 5 μm. Original magnification ×2000. (D) Anti-CD41 mAb-induced ITP in infected and uninfected mice. A single intraperitoneal injection (0.2 µg/g body weight) of anti-CD41 mAb (clone, MWReg30) was administered, followed by collection of blood for platelet counts at the specified time points. Platelet count (E) and rate of decline and recovery of platelet count (F) in uninfected and infected mice after anti-CD41 mAb-induced thrombocytopenia. Data were pooled from 3 independent experiments with uninfected (n = 3), anti-CD41–treated uninfected (n = 9), infected (n = 5), and infected+anti-CD41–treated (n = 10) mice. (G) Representative TEM images of BM MK from infected anti-CD41–treated (i-ii) and uninfected anti-CD41–treated (iii-iv) mice, showing DMS and platelet areas. Bar represents 5 μm. Original magnification ×2000. (H) The perimeter of the demarcation membranes of anti-CD41–treated uninfected (n = 20) and anti-CD41–treated infected (n = 30) mice. The dotted line corresponds with the mean of the perimeter of the DMS in nontreated infected mice. Data are expressed as means ± SEM and analyzed with the unpaired Student t test. *P < .05.

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