Evaluation of the Galalpha1-3Gal epitope as a host modification factor eliciting natural humoral immunity to enveloped viruses

Abstract

Human sera contain high levels of natural antibody (Ab) to Galalpha1-3Gal, a terminal glycosidic structure expressed on the surface of cells of mammals other than Old World primates. Incorporation of this determinant onto retroviral membranes by passage of viruses in cells encoding alpha-1-3-galactosyltransferase (GT) renders retroviruses sensitive to lysis by natural Ab and complement in normal human serum (NHS). Plasma membrane-budding viruses representing four additional virus groups were examined for their sensitivities to serum inactivation after passage through human cell lines that lack a functional GT or human cells expressing recombinant porcine GT. The inactivation of lymphocytic choriomeningitis virus (LCMV) by NHS directly correlated with host modification of the virus via expression of Galalpha1-3Gal and was blocked by incorporation of soluble Galalpha1-3Gal disaccharide into the inactivation assay. GT-deficient mice immunized to make high levels of Ab to Galalpha1-3Gal (anti-Gal Ab) were tested for resistance to LCMV passaged in GT-expressing cells. Resistance was not observed, but in vitro analyses of the mouse immune sera revealed that the antiviral activity of the sera was insufficient to eliminate LCMV infectivity on its natural targets of infection, macrophages, which express receptors for Ab and complement. Newcastle disease virus and vesicular stomatitis virus (VSV) were inactivated by NHS regardless of cell passage history, whereas Sindbis virus (SV) passaged in human cells resisted inactivation. Both VSV and SV passaged in Galalpha1-3Gal-expressing human cells incorporated this sugar moiety onto their major envelope glycoproteins. SV passaged in mouse cells expressing Galalpha1-3Gal was moderately sensitive to inactivation by NHS. These results indicate that enveloped viruses expressing Galalpha1-3Gal differ in their sensitivities to NHS and that a potent complement source, such as that in NHS, is required for efficient inactivation of sensitive viruses in vitro and in vivo.

FIG. 1

Expression of Galα1-3Gal on cultured cells. Cells used in these experiments were stained with purified anti-Gal Ab and analyzed by flow cytometry as described in Materials and Methods. Peaks depict Galα1-3Gal expression on the surfaces of L-929, N115, HeLa, BHK-21, SK-N-MC (SK) cells or SK-N-MC cells transduced with recombinant GT (SK-GT), and cells from GT-deficient (GT-KO) or GT-sufficient (GT-WT) mice.

FIG. 2

Expression of Galα1-3Gal on virion proteins. Purified SV (A and B) and VSV (C and D) passaged in SK-GT (lanes 1) or SK (lanes 2) cells were analyzed by Western blotting for expression of viral envelope proteins by binding of anti-viral Ab (B and D) or for expression of Galα1-3Gal by binding of IB4 lectin (A and C) as described in Materials and Methods. Sizes are indicated in kilodaltons.