Efficient cell infection by Moloney murine leukemia virus-derived particles requires minimal amounts of envelope glycoprotein.

Retrovirus entry into cells is mediated by specific interactions between the retrovirally encoded Env envelope glycoprotein and a host cell surface receptor. Though a number of peptide motifs responsible for the structure as well as for the binding and fusion activities of Env have been identified, only a few quantitative data concerning the infection process are available. Using an inducible expression system, we have expressed various amounts of ecotropic and amphotropic Env at the surfaces of Moloney murine leukemia virus-derived vectors and assayed for the infectivity of viral particles. Contrary to the current view that numerous noncooperative Env-viral receptor interactions are required for cell infection, we report here that very small amounts of Env are sufficient for optimal infection. However, increasing Env density clearly accelerates the rate at which infectious attachment to cells occurs. Moreover, our data also show that a surprisingly small number of Env molecules are sufficient to drive infection, albeit at a reduced efficiency, and that, under conditions of low expression, Env molecules act cooperatively. These observations have important consequences for our understanding of natural retroviral infection as well as for the design of cell-targeted infection techniques involving retroviral vectors.

Screening for oxygen sensitivity in U.S. Navy combat swimmers.

The United States Naval Special Warfare Community uses oxygen tolerance testing to screen Navy combat swimmer candidates for sensitivity to the toxic effects of hyperbaric oxygen (HBO2). Between 1976 and 1997, 6,250 oxygen-tolerance tests were performed at the Naval Special Warfare Center and Naval Special Warfare Group One. Our review of these data found only six episodes of O2 toxicity for an incidence of 0.096%, an incidence considerably lower than the rate of 1.9% reported in an earlier review using data from the Naval Safety Center. Additionally, we reviewed data from the Naval Safety Center from 1986 to 1997 and found only one episode of O2 toxicity among 157,930 LAR V dives. Many factors other than individual sensitivity to HBO2 may contribute to the occurrence of O2 toxicity episodes during combat swimmer operations. The authors conclude that O2 tolerance testing of U. S. Navy SEAL candidates is not a useful screening test and recommend discontinuation of this test.

Antiviral activity of an intracellularly expressed single-chain antibody fragment directed against the murine leukemia virus capsid protein.

We have addressed the possibility that intracellularly expressed miniantibodies directed against the viral capsid protein can be used as antiretroviral agents in gene transfer experiments. R187 is a rat monoclonal antibody that has been reported to recognize the MuLV p30gag capsid polypeptide. We report here that it also binds to the Pr65gag precursor polyprotein. R187 has been cloned and expressed in the form of a single-chain variable fragment (scFv) that shows the same binding specificity as the parental antibody. When expressed intracellularly, the R187 scFv favors the production of viral particles showing reduced infectivity. It, however, exerts no detectable protective effect against infection. This was observed both when using replication-incompetent MuLV-derived vector and replication-competent wild-type MuLV. Although the intimate mechanism of the inhibition is not clear, this work raises the possibility that gene engineering of anti-capsid protein scFvs may offer an additional lead for gene therapy of severe retrovirus-linked diseases.