Human T-cell line infected with HIV-2 which secretes functionall

Details Human T-cell line infected with HIV-2 which secretes functionall Human T-cell line infected with HIV-2 which secretes functionall

1996-07-03

1998-06-16

Achutamurthy, Ponnathapura

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving virus or bacteriophage

4352351, 435325, 530395, 4242081, C12Q 170, C12N 700, C07K 100, A61K 3921

Patent

active

057668442

ABSTRACT:
A novel cell line chronically infected with HIV-2.sub.NIHZ virus secretes the precursor of the viral envelope proteins in extracellular medium. The cells grow well in serum-free medium which enables the concentration of the conditioned medium of the cells to be greatly increased (>100 fold). Such a concentration step is otherwise impossible to achieve if the cells are grown under conventional conditions using 10% fetal calf serum. A specific affinity procedure is provided to purify the gp160 protein from the concentrated conditioned medium of cultures of 100 liter batches using a mouse monoclonal antibody to the HIV-2 gp41 protein, which is a part of the gp160 protein. This purified HIV-2 gp160 reacts with high specificity and sensitivity with the anti viral antibodies of HIV-2 infected humans by several criteria such as i) ELISA, ii) western blot analysis, and iii) radioimmuno-precipitation of metabolically labeled gp160 from the same cell line. The purified HIV-2 gp160 from this cell line is therefore a diagnostic tool for the detection of HIV-2 infection in humans. In view of the predicted oligomeric property of gp160 and its close conformational similarity to the virion envelope, the purified gp160 can also be useful for the development of vaccine strategies against HIV-2 in humans.

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Getchell et al., Continuous Production of a Cytopathic Human T-Lymphotropic Virus in a Permissive Neoplastic T-Cell Line, J. Clin. Microbiol., vol. 23, No. 4, pp. 737-742, see p. 740, col. 2, paragraph 1, Apr. 1986.
Robey et al., Prospect for prevention of human immunodeficiency virus infection: Purified 120-kDa envelope glycoprotein induces neutralizing antibody, Proc. Natl. Acad. Sci., vol. 83, pp. 7023-7027, see Abstract, Sep. 1986.
Rey et al., Characterization of Human Immunodeficiency Virus Type 2 Envelope Glycoproteins: Dimerization of the Glycoprotein Precursor during Processing, J. Virol. vol. 63, No. 2, pp. 647-658, see p. 647, column 2, second paragraph, Feb. 1989.

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