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Microvesicles are a source of contaminating cellular proteins found in purified HIV-1 preparations

  1. Author:
    Bess, J. W.
    Gorelick, R. J.
    Bosche, W. J.
    Henderson, L. E.
    Arthur, L. O.
    1. Year of Conference: 1997
  2. Conference Name: Conference on Retroviruses and Opportunistic Infections
    1. Pages: 139 (abstract no. 405)
  4. Type of Work: Meeting Abstract
  1. Abstract:

    Identification and quantitation of cellular proteins and other macromolecules associated with HIV-1 particles are complicated by the presence microvesicles that copurify with sucrose-gradient purified virions. These microvesicles bud from the surface of cells and copurify with retroviruses during sucrose gradient centrifugation because they exhibit a range of densities that includes the same density as retroviruses. HIV-1 infected and uninfected human T-cell lines were propagated and virus and microvesicles were purified from clarified cell culture supernatants by sucrose density gradient centrifugation or centrifugation through 20% sucrose pads. The concentrations of HIV-1 p24(CA), HLA-DR, and beta2-microglobulin (beta2-M) were determined and the ratios of HIV-1 p24(CA) to HLA-DR and beta2-M were found to vary with respect to the HIV-1 isolate, host cell, cell culture conditions and other factors. Electron microscopic analysis of microvesicles revealed amorphous particles ranging in size from 50 nm to 1000 nm in diameter. In addition to cellular proteins, including HLA-DR and beta2-M, microvesicles were found to contain a substantial amount of RNA and DNA. Analysis of fraction from sucrose gradient centrifugation revealed that gp120 copurified with HIV and was not detected in some cellular proteins, microvesicles from HIV-1-infected H9 cells did not appear to contain HIV-1 gp120(ENV).

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