Figure 1.
Nbeal2 deficiency significantly alters parasite sequestration, lung and brain pathology, and survival in PbA-infected mice. (A) Time course of platelets in peripheral blood of C57BL/6J mice after PbA infection compared with naive control mice (n = 7-10 mice/group) as determined by flow cytometry. (B) Platelets in whole blood of naive Nbeal2−/− mice normalized to naive C57BL/6J (B6) control mice as determined by flow cytometry (n = 20-24 mice/group). (C) Frequency of platelet-iRBC conjugates (left, n = 10-20 mice/group) in whole blood of C57BL/6J and Nbeal2−/− mice after PbA-GFP infection and time course of peripheral parasitemia as determined by staining and counting of thin blood smears (right, n = 10 mice/group). (D) Representative flow cytometry plots and quantification of the frequency of P-selectin+, CD63+, and CD41/CD61+ platelets as well as total platelets normalized to control mice in whole blood isolated from C57BL/6J (B6) and Nbeal2−/− mice at day 5 after infection with PbA (n = 10-26 mice/group). (E) Quantification of PF4 in the plasma of C57BL/6J and Nbeal2−/− mice at day 6 after infection with PbA (n = 10-14 mice/group) or naive (N) (n = 9-10 mice/group) as determined by ELISA. (F-H) Representative images and bioluminescence quantification of sequestered PbA schizonts expressing luciferase under the AMA-1 promoter in spleens (F), lungs (G), and brains (H) of PbA-infected C57BL/6J (B6) and Nbeal2−/− mice (n = 8 mice/group). Values are normalized to naive control mice from each respective group (n = 4 mice/group). (I-J) Lung permeability (I) and brain permeability (J) in C57BL/6J (B6) and Nbeal2−/− mice injected IV with 200 μL of 1% Evans blue dye at day 6 after infection with PbA (n = 9-13/group). Representative images and quantification of dye extracted from whole organs are shown. Optical density values are normalized to naive control mice from each respective group (n = 4 mice/group). (K) Survival curves of PbA-infected Nbeal2−/− (blue line) and C57BL/6J (B6, red line) mice (n = 15-16 mice/group). The gray shaded region represents the typical timeframe of death from ECM. Graphs in panels A-E and I-J represent the mean ± standard error of the mean. Boxes in panels F-H represent the median ± the 25th and 75th percentiles with minimum/maximum whiskers. Statistical analyses were performed using the Mann-Whitney U test (A-J) and log-rank Mantel-Cox test (K). Only statistically significant (P < .05) values are shown. For graphs in panels A and C, *P < .05, **P < .005, ***P < .0005, and ****P < .0001. Figures represent combined data from 2 (A; C, right; D, middle and right graphs; and E-J) or ≥3 (B; C, left; D, left; and K) independent experiments. P.I., postinfection; SSC, side scatter.

Nbeal2 deficiency significantly alters parasite sequestration, lung and brain pathology, and survival in PbA-infected mice. (A) Time course of platelets in peripheral blood of C57BL/6J mice after PbA infection compared with naive control mice (n = 7-10 mice/group) as determined by flow cytometry. (B) Platelets in whole blood of naive Nbeal2−/− mice normalized to naive C57BL/6J (B6) control mice as determined by flow cytometry (n = 20-24 mice/group). (C) Frequency of platelet-iRBC conjugates (left, n = 10-20 mice/group) in whole blood of C57BL/6J and Nbeal2−/− mice after PbA-GFP infection and time course of peripheral parasitemia as determined by staining and counting of thin blood smears (right, n = 10 mice/group). (D) Representative flow cytometry plots and quantification of the frequency of P-selectin+, CD63+, and CD41/CD61+ platelets as well as total platelets normalized to control mice in whole blood isolated from C57BL/6J (B6) and Nbeal2−/− mice at day 5 after infection with PbA (n = 10-26 mice/group). (E) Quantification of PF4 in the plasma of C57BL/6J and Nbeal2−/− mice at day 6 after infection with PbA (n = 10-14 mice/group) or naive (N) (n = 9-10 mice/group) as determined by ELISA. (F-H) Representative images and bioluminescence quantification of sequestered PbA schizonts expressing luciferase under the AMA-1 promoter in spleens (F), lungs (G), and brains (H) of PbA-infected C57BL/6J (B6) and Nbeal2−/− mice (n = 8 mice/group). Values are normalized to naive control mice from each respective group (n = 4 mice/group). (I-J) Lung permeability (I) and brain permeability (J) in C57BL/6J (B6) and Nbeal2−/− mice injected IV with 200 μL of 1% Evans blue dye at day 6 after infection with PbA (n = 9-13/group). Representative images and quantification of dye extracted from whole organs are shown. Optical density values are normalized to naive control mice from each respective group (n = 4 mice/group). (K) Survival curves of PbA-infected Nbeal2−/− (blue line) and C57BL/6J (B6, red line) mice (n = 15-16 mice/group). The gray shaded region represents the typical timeframe of death from ECM. Graphs in panels A-E and I-J represent the mean ± standard error of the mean. Boxes in panels F-H represent the median ± the 25th and 75th percentiles with minimum/maximum whiskers. Statistical analyses were performed using the Mann-Whitney U test (A-J) and log-rank Mantel-Cox test (K). Only statistically significant (P < .05) values are shown. For graphs in panels A and C, *P < .05, **P < .005, ***P < .0005, and ****P < .0001. Figures represent combined data from 2 (A; C, right; D, middle and right graphs; and E-J) or ≥3 (B; C, left; D, left; and K) independent experiments. P.I., postinfection; SSC, side scatter.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal