Summary
A concentrated live retrovirus is required for in vitro experiments. A cuprammonium-regenerated cellulose hollow fiber, termed BMM, originally developed for biohazardous viral removal, was used to concentrate two different retroviruses, an ecotropic murine leukemia virus (MuLV) and human immunodeficiency virus (HIV). The BMM was useful for concentrating live virus suspension 10- to 30-fold from 500–1000 ml of culture supernatant. The ecotropic MuLV concentrated by BMM was demonstrated to be viable and biologically intact by XC plaque-forming assay and reverse transcriptase assay. The concentrated MuLV reached a much higher titer in the spleen in mice than the original one. The virus concentration assessed by p24 antigen for HIV was clearly higher than that of the original culture supernatant of HIV-infected cell lines. Since BMM hollow fibers trapped viruses by the sieving mechanism but not by adsorption, the viral particles were recovered by washing and the total live virus recovery rate was high, about 50%. Furthermore 60 min sufficed to handle 1000 ml of supernatant in the case of a filtration area of 0.03 m2. These results show that the BMM provides us with a rapid, safe and efficient method for concentrating live retroviruses.
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Manabe S, Kamata Y, Iijima H, Kamide K (1987) Some morphological characteristics of porous polymeric membranes prepared by “micro-phase separation method”. Polymer J 19: 391–404
Hamamoto Y, Harada S, Kobayashi S, Yamaguchi K, Iijima H, Manabe S, Tsurumi T, Aizawa H, Yamamoto N (1989) A novel method for removal of human immunodeficiency virus:filtration with porous polymeric membranes. Vox Sang 56: 230–236
Manabe S, Tsurumi T, Ishikawa G, Satani M, Tamashiki T, Hamamoto Y, Yamaguchi K, Kobayashi S, Yamamoto N (1989) Mechanism of human immunodeficiency virus (HIV) removal by regenerated cellulose hollow fiber (BMM). Membrane 14: 77–83
Sekiguchi S, Ito K, Kobayashi M, Ikeda H, Tsurumi T, Ishikawa G, Manabe S, Satami M, Yamashiki T (1989) Possibility of hepatitis B virus (HBV) removal from human plasma using regenerated cellulose hollow fiber (BMM). Membrane 14: 253–261
Tsurumi T, Osawa N, Hirasaki H, Hirasaki T, Yamaguchi K, Manabe S, Yamashiki T (1990) Mechanism of removing monodisperse gold particles from a suspension using cuprammonium regenerated cellulose hollow fiber (BMM hollow fiber). Polymer J 22: 304–311
Tsurumi T, Osawa N, Hitaka T, Hirasaki T, Yamaguchi K, Manabe S, Yamashiki T (1990) Structure of cuprammonium regenerated cellulose hollow fiber (BMM hollow fiber) for virus removal. Polymer J 22: 751–758
Yamaguchi K, Hamamoto Y, Manabe S, Yamamoto N (1991) Microparticle removability of the regenerated cellulose hollow fiber (BMM): An electron microscopic evaluation. J Electron Microsc 40: 337–345
Pattengale PK, Taylor CR, Twomey P, Hill S, Jonasson J, Beardsley T, Haas M (1982) Immunopathology of B-cell lymphomas induced in C57BL/6 mice by dualtropic murine leukemia virus (MuLV). Am J Pathol 107: 362–377
Jolicoeur P (1991) Murine acquired immunodeficiency syndrome (MAIDS): and animal model to study the AIDS pathogenesis. FASEB J 5: 2398–2405
Morse HC III, Chattopadhyay SK, Makino M, Fredrickson TN, Hugin AW, Hartley JW (1992) Retrovirus-induced immunodeficiency in the mouse: MAIDS as a model for AIDS. AIDS 6: 607–620
Chattopadhyay SK, Morse HC III, Makino M, Ruscetti SK, Hartley JW (1989) Defective virus is associated with induction of murine retrovirus-induced immunodeficiency syndrome. Proc Natl Acad Sci USA 86: 3862–3966
Chattopadhyay SK, Sengupta DN, Fredrickson TN, Morse HC III, Hartley JW (1991) Characteristics and contributions of defective, ecotropic, and mink cell focus-inducing viruses involved in a retrovirus-induced immunodeficiency syndrome of mice. J Virol 65: 4232–4241
Johnson VA, Byington RE, Kaplan JC (1990) Reverse transcriptase (RT) activity assay. In: Aldovini A, Walker BD (eds) Techniques in HIV research. Stockton Press, New York, pp 98–102
Sasaki Y, Honda M, Makino M, Sasaki T (1993) Mycoplasmas stimulate replication of human immunodeficiency virus type 1 through selective activation of CD4+ T Imphocytes. AIDS Res Hum Retroviruses 9: 775–780
Rowe WP, Pugh WE, Hartley JW (1970) Plaque assay techniques for murine leukemia viruses. Virology 42: 1136–1139
Hartley JW, Rowe WP (1975) Clonal cell lines from a feral mouse embryo which lack host-range restriction for murine leukemia viruses. Virology 65: 128–134
Cloyd MW, Hartley JW, Rowe WP (1981) Genetic study of lymphoma induction by AKR mink cell focus-inducing virus in AKR × NFS crosses. J Exp Med 154: 450–458
Makino M, Morse HC III, Fredrickson TN, Hartley JW (1990) H-2-associated and background genes influence the development of a murine retrovirus-induced immuno-deficiency syndrome, MAIDS. J Immunol 144: 4347–4355
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Makino, M., Ishikawa, G., Yamaguchi, K. et al. Concentration of live retrovirus with a regenerated cellulose hollow fiber, BMM. Archives of Virology 139, 87–96 (1994). https://doi.org/10.1007/BF01309456
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DOI: https://doi.org/10.1007/BF01309456