Limits...
In vitro and in vivo characterization of ultraviolet light C-irradiated human platelets in a 2 event mouse model of transfusion.

Zhi L, Chi X, Vostal JG - PLoS ONE (2013)

Bottom Line: UVC-based technology differs from UVA or UVB-based technologies in that it uses a specific wavelength at 254 nm without the addition of any photosensitizers.Previously, it was reported that UVC irradiation induces platelet aggregation and activation.Unlike UVB-platelets, UVC-platelets did not lead to lung injury or induce MIP-2 release.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Hematology, Division of Hematology, OBRR, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.

ABSTRACT
UV-based pathogen reduction technologies have been developed in recent years to inactivate pathogens and contaminating leukocytes in platelet transfusion products in order to prevent transfusion-transmitted infections and alloimmunization. UVC-based technology differs from UVA or UVB-based technologies in that it uses a specific wavelength at 254 nm without the addition of any photosensitizers. Previously, it was reported that UVC irradiation induces platelet aggregation and activation. To understand if UVC-induced changes of platelet quality correlate with potential adverse events when these platelets are transfused into animals, we used a 2-event SCID mouse model in which the predisposing event was LPS treatment and the second event was infusion of UVC-irradiated platelets. We analyzed lung platelet accumulation, protein content in bronchoalveolar lavage fluid as an indication of lung injury, and macrophage inflammatory protein-2 (MIP-2) release in mice received UVC-irradiated or untreated control platelets. Our results showed UVC-irradiated platelets accumulated in lungs of the mice in a dose-dependent manner. High-doses of UVC-irradiated platelets were sequestered in the lungs to a similar level as we previously reported for UVB-irradiated platelets. Unlike UVB-platelets, UVC-platelets did not lead to lung injury or induce MIP-2 release. This could potentially be explained by our observation that although UVC treatment activated platelet surface αIIbβ3, it failed to activate platelet cells. It also suggests lung platelet accumulation and subsequent lung damage are due to different and separate mechanisms which require further investigation.

Show MeSH

Related in: MedlinePlus

UVC irradiation of HPs activated αIIbβ3 integrin without affecting platelet surface P-selectin expression.Untreated and UVC irradiated HPs at varying UVC doses were labeled with PAC1-FITC or with a combination of CD41a-FITC and CD62P-PE and the binding was detected with a FACSCalibur cytometer. Shown here is flow cytometric quantification of the percentage of platelet cells positive for PAC-1 binding (A) or CD41a and CD62P staining (B). Mean ± SE, n=3-5.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3815158&req=5

pone-0079869-g002: UVC irradiation of HPs activated αIIbβ3 integrin without affecting platelet surface P-selectin expression.Untreated and UVC irradiated HPs at varying UVC doses were labeled with PAC1-FITC or with a combination of CD41a-FITC and CD62P-PE and the binding was detected with a FACSCalibur cytometer. Shown here is flow cytometric quantification of the percentage of platelet cells positive for PAC-1 binding (A) or CD41a and CD62P staining (B). Mean ± SE, n=3-5.

Mentions: Verhaar R et al. showed that increasing doses of UVC irradiation increased binding of PAC-1 monoclonal antibody to platelets [24]. This antibody selectively recognizes the high-affinity active conformation of platelet integrin αIIbβ3 [25-27]. We further characterized the effect of UVC irradiation on platelet activation by analyzing platelet PAC-1 binding and surface P-selectin expression. We found that, similar to UVB irradiation [21], UVC exposure of platelets did significantly increase PAC-1 binding (Figure 2A). At a dose of 0.2 J/cm2 UVC light, about 70% of platelets demonstrated PAC-1 binding (69.3 ± 2.2 % positive cells), whereas binding in untreated control platelets was barely detected (8.1 ± 1.9 % positive cells). When the UVC dose was increased by 6-fold to 1.2 J/cm2, no further increase in PAC-1 binding to UVC-platelets was observed under our experimental conditions. In contrast to the marked effect of UVC on activation of platelet αIIbβ3, we did not observe an effect of UVC on platelet surface P-selectin expression, which remained at about 30% of the cells positive after the platelets were exposed to 0.2 or 1.2 J/cm2 UVC irradiation (Figure 2B).


In vitro and in vivo characterization of ultraviolet light C-irradiated human platelets in a 2 event mouse model of transfusion.

Zhi L, Chi X, Vostal JG - PLoS ONE (2013)

UVC irradiation of HPs activated αIIbβ3 integrin without affecting platelet surface P-selectin expression.Untreated and UVC irradiated HPs at varying UVC doses were labeled with PAC1-FITC or with a combination of CD41a-FITC and CD62P-PE and the binding was detected with a FACSCalibur cytometer. Shown here is flow cytometric quantification of the percentage of platelet cells positive for PAC-1 binding (A) or CD41a and CD62P staining (B). Mean ± SE, n=3-5.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3815158&req=5

pone-0079869-g002: UVC irradiation of HPs activated αIIbβ3 integrin without affecting platelet surface P-selectin expression.Untreated and UVC irradiated HPs at varying UVC doses were labeled with PAC1-FITC or with a combination of CD41a-FITC and CD62P-PE and the binding was detected with a FACSCalibur cytometer. Shown here is flow cytometric quantification of the percentage of platelet cells positive for PAC-1 binding (A) or CD41a and CD62P staining (B). Mean ± SE, n=3-5.
Mentions: Verhaar R et al. showed that increasing doses of UVC irradiation increased binding of PAC-1 monoclonal antibody to platelets [24]. This antibody selectively recognizes the high-affinity active conformation of platelet integrin αIIbβ3 [25-27]. We further characterized the effect of UVC irradiation on platelet activation by analyzing platelet PAC-1 binding and surface P-selectin expression. We found that, similar to UVB irradiation [21], UVC exposure of platelets did significantly increase PAC-1 binding (Figure 2A). At a dose of 0.2 J/cm2 UVC light, about 70% of platelets demonstrated PAC-1 binding (69.3 ± 2.2 % positive cells), whereas binding in untreated control platelets was barely detected (8.1 ± 1.9 % positive cells). When the UVC dose was increased by 6-fold to 1.2 J/cm2, no further increase in PAC-1 binding to UVC-platelets was observed under our experimental conditions. In contrast to the marked effect of UVC on activation of platelet αIIbβ3, we did not observe an effect of UVC on platelet surface P-selectin expression, which remained at about 30% of the cells positive after the platelets were exposed to 0.2 or 1.2 J/cm2 UVC irradiation (Figure 2B).

Bottom Line: UVC-based technology differs from UVA or UVB-based technologies in that it uses a specific wavelength at 254 nm without the addition of any photosensitizers.Previously, it was reported that UVC irradiation induces platelet aggregation and activation.Unlike UVB-platelets, UVC-platelets did not lead to lung injury or induce MIP-2 release.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cellular Hematology, Division of Hematology, OBRR, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, United States of America.

ABSTRACT
UV-based pathogen reduction technologies have been developed in recent years to inactivate pathogens and contaminating leukocytes in platelet transfusion products in order to prevent transfusion-transmitted infections and alloimmunization. UVC-based technology differs from UVA or UVB-based technologies in that it uses a specific wavelength at 254 nm without the addition of any photosensitizers. Previously, it was reported that UVC irradiation induces platelet aggregation and activation. To understand if UVC-induced changes of platelet quality correlate with potential adverse events when these platelets are transfused into animals, we used a 2-event SCID mouse model in which the predisposing event was LPS treatment and the second event was infusion of UVC-irradiated platelets. We analyzed lung platelet accumulation, protein content in bronchoalveolar lavage fluid as an indication of lung injury, and macrophage inflammatory protein-2 (MIP-2) release in mice received UVC-irradiated or untreated control platelets. Our results showed UVC-irradiated platelets accumulated in lungs of the mice in a dose-dependent manner. High-doses of UVC-irradiated platelets were sequestered in the lungs to a similar level as we previously reported for UVB-irradiated platelets. Unlike UVB-platelets, UVC-platelets did not lead to lung injury or induce MIP-2 release. This could potentially be explained by our observation that although UVC treatment activated platelet surface αIIbβ3, it failed to activate platelet cells. It also suggests lung platelet accumulation and subsequent lung damage are due to different and separate mechanisms which require further investigation.

Show MeSH
Related in: MedlinePlus