Use of a flow-cell system to investigate virucidal dimethylmethylene blue phototreatment in two RBC additive solutions.
BACKGROUND
Limited photoinactivation kinetics, use of low-volume 30 percent Hct RBCs, and hemolysis have restricted the practicality of the use of dimethylmethylene blue (DMMB) and light for RBC decontamination. A flow-cell system was developed to rapidly treat larger volumes of oxygenated 45 percent Hct RBCs with high-intensity red light.


METHODS
CPD-whole blood was WBC reduced, RBCs were diluted in additive solutions (either Adsol or Erythrosol), and suspensions were subsequently oxygenated by gas overlay. Intracellular or extracellular VSV and DMMB were sequentially added. VSV-infected RBC suspensions (45% Hct) were passed through 1-mm-thick flow cells and illuminated. Samples were titered for VSV, stored for up to 42 days, and assayed for Hb, supernatant potassium, ATP, and MCV.


RESULTS
The use of oxygenated RBCs resulted in rapid and reproducible photoinactivaton of > or = 6.6 log extracellular and approximately 4.0 log intracellular VSV independent of additive solution. Phototreated Adsol RBCs exhibited more than 10 times greater hemolysis and 30 percent greater MCV during storage than identically treated Erythrosol RBCs. Phototreatment caused RBC potassium leakage from RBCs in both additive solutions. ATP levels were better preserved in Erythrosol than Adsol RBCs.


CONCLUSIONS
A rapid, reproducible, and robust method for photoinactivating model virus in RBC suspensions was developed. Despite improved hemolysis and ATP levels in Erythrosol-phototreated RBCs, storage properties were not maintained for 42 days.
Wagner
Stephen
S
Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross Biomedical Services, Rockville, Maryland 20855, USA. wagners@usa.redcross.org
Skripchenko
Andrey
A
Thompson-Montgomery
Dedeene
D
D000225:Adenine / Q000494:pharmacology*; D000255:Adenosine Triphosphate / Q000097:blood; D000818:Animals; D000998:Antiviral Agents / Q000097:blood / Q000494:pharmacology / Q000528:radiation effects*; D001793:Blood Preservation; D002522:Chlorocebus aethiops; D004912:Erythrocytes / Q000737:chemistry*; D005434:Flow Cytometry / Q000295:instrumentation*; D005947:Glucose / Q000494:pharmacology*; D006454:Hemoglobins / Q000737:chemistry; D006461:Hemolysis; D007552:Isotonic Solutions / Q000494:pharmacology*; D007700:Kinetics; D008353:Mannitol / Q000494:pharmacology*; D008751:Methylene Blue / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000528:radiation effects*; D010100:Oxygen / Q000494:pharmacology; D010108:Oxyhemoglobins / Q000737:chemistry; D010777:Photochemistry; D017319:Photosensitizing Agents / Q000097:blood / Q000494:pharmacology / Q000528:radiation effects*; D011188:Potassium / Q000097:blood; D015203:Reproducibility of Results; D012965:Sodium Chloride / Q000494:pharmacology*; D013535:Suspensions; D013997:Time Factors; D014709:Vero Cells / Q000821:virology; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000528:radiation effects; D038301:Virus Inactivation
C037839:ADSOL; D000998:Antiviral Agents; D006454:Hemoglobins; D007552:Isotonic Solutions; D010108:Oxyhemoglobins; D017319:Photosensitizing Agents; D013535:Suspensions; C435946:dimethylmethylene blue; C098027:erythrosol; D008353:Mannitol; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D005947:Glucose; D000225:Adenine; D011188:Potassium; D010100:Oxygen; D008751:Methylene Blue