Akzo Nobel N.v.

Irene LaRue-Kalisz

Mangalasseril G Sarngadharan

Vaniambadi S Kalyanaraman

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 (&gt;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.

Human T-cell line infected with HIV-2 which secretes functionally intact

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 (&gt;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) radioimmunoprecipitation 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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Frederick, MD, United States of America

Irene LaRue-Kalisz