Preparation of virus-free pyridoxylated hemoglobin from the blood of HBV or HTLV-I healthy carriers.
One of our major aims was to find ways to utilize outdated or virus-contaminated blood. Pyridoxylated hemoglobin (PLP-Hb), a possible substitute for red cells as an artificial oxygen carrier was prepared from outdated human blood. By conjugation with polyethylene glycol (PEG), the biological half life was increased about 3 folds at 82% blood replacement in rats without significant side effects in vivo or in vitro. We next tried to prepare virus-free PEG-PLP-Hb from HBV or HTLV-I positive blood. A considerable amount of HBV (Dane particles) could be removed from HBV-positive red cells by washing and during the preparation of PEG-PLP-Hb. When the hemoglobin preparations containing Dane particles were filtered through a porous cellulose filter, BMM-30 (30 nm pore size, Asahi Chemical Industry Co., Ltd., Osaka, Japan), HBV-DNA in the filtered fractions became less than 0.33% of the initial amount. More than 96% of blood leukocytes could be removed with a leukocyte removal filter, Sepacell R-500 (Asahi Medical Co., Ltd., Tokyo, Japan). The leukocytes collected from filtrated fractions of HTLV-I positive blood did not survive beyond 3 days. Since transmission of HTLV-I occurs by cell to cell contact and is rare in cell-Free condition, it is unlikely that the PLP-Hb prepared from HTLV-I positive blood which is deprived of leukocytes transmits HTLV-I infection.
Sekiguchi
S
S
Research Division, Hokkaido Red Cross Blood Center, Sapporo, Japan.
Ito
K
K
Kobayashi
M
M
Ototake
N
N
Kosuda
M
M
Kwon
K W
KW
Ikeda
H
H
D000818:Animals; D001769:Blood / Q000382:microbiology*; D001781:Blood Component Removal / Q000379:methods*; D001802:Blood Substitutes; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D055598:Chemical Phenomena; D002621:Chemistry; D002627:Chemistry, Physical; D004279:DNA, Viral / Q000032:analysis; D004912:Erythrocytes / Q000166:cytology; D005078:Exchange Transfusion, Whole Blood; D005374:Filtration; D006678:HIV / Q000302:isolation & purification*; D006207:Half-Life; D006454:Hemoglobins / Q000302:isolation & purification* / Q000493:pharmacokinetics; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007962:Leukocytes / Q000166:cytology; D008297:Male; D011092:Polyethylene Glycols / Q000493:pharmacokinetics; D011732:Pyridoxal Phosphate / Q000031:analogs & derivatives; D051381:Rats
D001802:Blood Substitutes; D004279:DNA, Viral; D006454:Hemoglobins; D006514:Hepatitis B Surface Antigens; C030939:polyhemoglobin-pyridoxal-5-phosphate; C050907:pyridoxalated-hemoglobin-polyethylene glycol conjugate; D011092:Polyethylene Glycols; D011732:Pyridoxal Phosphate