Figure 1.
Adhesion molecule activation observed in malaria-altered uRBCs. (A) Laminin-α5 adherence was assessed by passing RBCs incubated in culture medium (control) and P. falciparum in vitro cultures (MC) over laminin-α5–coated Ibidi μ-slides. Lu/BCAM-mediated adhesion was tested by Lu/BCAM blocking antibody (MC+Lu/BCAM). Mean ± standard error of the mean (SEM); n = 4-7 (left). Cells were stained with the DNA dye Hoechst to distinguish iRBCs from uRBCs. Laminin-α5 adhesion frequency of MC-derived uRBCs was normalized to control RBCs (right). (B) RBCs from age-matched controls (n = 5) and patients with malaria (n = 6) were stained with Hoechst for iRBC detection and tested for laminin-α5 adherence. Mean ± standard deviation (SD); Mann-Whitney test. (C) RBCs exposed to malaria-conditioned medium (CM) overnight were assessed for Lu/BCAM-mediated laminin-α5 adherence. Mean ± SEM; n = 4-7. (D) RBCs incubated with heat-treated (1 hour, 56°C) (HI CM) were passed over laminin-α5 coated μ-slides, to determine the heat stability of the laminin adhesion–inducing factor in CM. Mean ± SEM, n = 4. (E) Quantification of HA interaction was performed by passing RBCs derived from control and MC conditions on HA-coated μ-slides. CD44-mediated interaction was assessed by CD44 blocking antibody. The frequency of HA interaction of uRBCs was normalized to control RBCs. iRBCs were stained with Hoechst. Mean ± SEM, n = 3-4. (F) Frequency of HA interaction of uRBCs in patients with malaria was compared with those from age-matched controls. Samples were stained with Hoechst for iRBC-uRBC distinction. Mean ± SD; n = 4; Mann-Whitney test. (G) CD44 mediated HA interaction of CM-exposed RBCs was assessed by passing CM RBCs and CD44-blocked CM RBCs over HA-coated μ-slides. HA-rolling of treated cells were normalized to control RBCs. Mean ± SEM; n = 4-7. (H) Heat instability of the factor that mediates HA interaction was tested under flow conditions with RBCs incubated with HI CM. Mean ± SEM, n = 5. (I) Microscopy images of laminin-α5 adherent control RBCs (i), malaria culture RBCs (ii), and CM-exposed RBCs (iii). Hoechst staining (blue) indicates iRBCs. Images were taken under a ×40 oil-immersion objective. Bars represent 20 μm. Wilcoxon matched-pairs signed-rank test, unless stated otherwise. The error bar denotes SEM or SD.

Adhesion molecule activation observed in malaria-altered uRBCs. (A) Laminin-α5 adherence was assessed by passing RBCs incubated in culture medium (control) and P. falciparum in vitro cultures (MC) over laminin-α5–coated Ibidi μ-slides. Lu/BCAM-mediated adhesion was tested by Lu/BCAM blocking antibody (MC+Lu/BCAM). Mean ± standard error of the mean (SEM); n = 4-7 (left). Cells were stained with the DNA dye Hoechst to distinguish iRBCs from uRBCs. Laminin-α5 adhesion frequency of MC-derived uRBCs was normalized to control RBCs (right). (B) RBCs from age-matched controls (n = 5) and patients with malaria (n = 6) were stained with Hoechst for iRBC detection and tested for laminin-α5 adherence. Mean ± standard deviation (SD); Mann-Whitney test. (C) RBCs exposed to malaria-conditioned medium (CM) overnight were assessed for Lu/BCAM-mediated laminin-α5 adherence. Mean ± SEM; n = 4-7. (D) RBCs incubated with heat-treated (1 hour, 56°C) (HI CM) were passed over laminin-α5 coated μ-slides, to determine the heat stability of the laminin adhesion–inducing factor in CM. Mean ± SEM, n = 4. (E) Quantification of HA interaction was performed by passing RBCs derived from control and MC conditions on HA-coated μ-slides. CD44-mediated interaction was assessed by CD44 blocking antibody. The frequency of HA interaction of uRBCs was normalized to control RBCs. iRBCs were stained with Hoechst. Mean ± SEM, n = 3-4. (F) Frequency of HA interaction of uRBCs in patients with malaria was compared with those from age-matched controls. Samples were stained with Hoechst for iRBC-uRBC distinction. Mean ± SD; n = 4; Mann-Whitney test. (G) CD44 mediated HA interaction of CM-exposed RBCs was assessed by passing CM RBCs and CD44-blocked CM RBCs over HA-coated μ-slides. HA-rolling of treated cells were normalized to control RBCs. Mean ± SEM; n = 4-7. (H) Heat instability of the factor that mediates HA interaction was tested under flow conditions with RBCs incubated with HI CM. Mean ± SEM, n = 5. (I) Microscopy images of laminin-α5 adherent control RBCs (i), malaria culture RBCs (ii), and CM-exposed RBCs (iii). Hoechst staining (blue) indicates iRBCs. Images were taken under a ×40 oil-immersion objective. Bars represent 20 μm. Wilcoxon matched-pairs signed-rank test, unless stated otherwise. The error bar denotes SEM or SD.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal