Inactivation of WBCs in RBC suspensions by photoactive phenothiazine dyes: comparison of dimethylmethylene blue and MB

BACKGROUND: The transfusion of blood components containing WBCs can cause unwanted complications, which include virus transmission, transfusion‐associated GVHD, alloimmunization, febrile reactions, and immunomodulation. Phototreatment with 4 μM of dimethylmethylene blue (DMMB) and 13 J per cm2 of wh...

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Bibliographic Details
Published in:Transfusion (Philadelphia, Pa.) Pa.), 2000-08, Vol.40 (8), p.968-975
Main Authors: Skripchenko, A.A., Wagner, S.J.
Format: Article
Language:English
Subjects:
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Biological and medical sciences
Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis
Coloring Agents - pharmacology
DMMB = dimethylmethylene blue
DNA Fragmentation
Erythrocytes
FBS = fetal bovine serum
Flow Cytometry
Humans
LDA = limiting dilution analysis
Leukocytes - metabolism
Leukocytes, Mononuclear
Light
Medical sciences
Methylene Blue - analogs & derivatives
Methylene Blue - pharmacology
Phenothiazines - pharmacology
Photosensitizing Agents - pharmacology
PI = propidium iodide
Propidium - pharmacokinetics
TA-GVHD = transfusion-associated GVHD
TCGF = T-cell growth factor
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
Virus Activation - drug effects
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Summary:BACKGROUND: The transfusion of blood components containing WBCs can cause unwanted complications, which include virus transmission, transfusion‐associated GVHD, alloimmunization, febrile reactions, and immunomodulation. Phototreatment with 4 μM of dimethylmethylene blue (DMMB) and 13 J per cm2 of white light irradiation has previously been shown to be an effective way to inactivate different models of enveloped and nonenveloped viruses in RBC suspensions, with minimum damage to RBCs. The present study compares WBC photoinactivation in buffy coat after DMMB or MB phototreatment under virucidal conditions. STUDY DESIGN AND METHODS: Buffy coat diluted to 30‐percent Hct was treated with the dye and white light. Isolated WBCs were assayed for cell proliferation and viability by an assay using a tetrazolium compound, limiting dilution analysis, DNA fragmentation, and flow cytometry assays. RESULTS: DMMB and 2.5 J per cm2 of light phototreat‐ment can inactivate T cells to the limit of detection by limiting dilution analysis (>4.76 log reduction). No WBC proliferation activity was observed after DMMB and 3.8 J per cm2 of light. DNA degradation after DMMB phototreatment was light dependent. In addition, DMMB phototreatment induced apoptosis in WBCs. In contrast, MB phototreatment under virucidal conditions did not cause significant changes in the viability of WBCs. Neither DNA degradation nor signs of apoptosis were observed after MB phototreatment. CONCLUSION: DMMB phototreatment inactivates T‐lymphocytes, the cells that cause GVHD.
ISSN:0041-1132
1537-2995
DOI:10.1046/j.1537-2995.2000.40080968.x