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. 2001 Aug;75(16):7384-91.
doi: 10.1128/JVI.75.16.7384-7391.2001.

Host range of small-ruminant lentivirus cytopathic variants determined with a selectable caprine arthritis- encephalitis virus pseudotype system

I Hötzel  1 , W P Cheevers
Affiliations

Host range of small-ruminant lentivirus cytopathic variants determined with a selectable caprine arthritis- encephalitis virus pseudotype system

I Hötzel et al. J Virol. 2001 Aug.

Abstract

The small-ruminant lentiviruses ovine maedi-visna virus (MVV) and caprine arthritis-encephalitis virus (CAEV) cause encephalitis, progressive pneumonia, arthritis, and mastitis in sheep and goats. Icelandic MVV strains, which are lytic in tissue culture, have a wide species distribution of functional receptors, which includes human cells. In contrast, functional receptors for the nonlytic CAEV CO are absent from human cells. To determine if the wide species distribution of functional receptors is a common property of MVV strains or related to cytopathic phenotype, we tested the infectivity of viruses pseudotyped with the envelope glycoproteins of MVV K1514, CAEV CO, and lytic and nonlytic North American MVV strains to cells of different species. Replication-defective CAEV proviral constructs lacking the env, tat, and vif genes and carrying the neomycin phosphotransferase gene in the vif-tat region were developed for the infectivity assays. Cotransfection of human 293T cells with these proviral constructs and plasmids expressing CAEV, MVV, or vesicular stomatitis virus envelope glycoproteins produced infectious pseudotyped virus which induced resistance of infected cells to G418. Using these pseudotypes, we confirmed the wide species distribution of Icelandic MVV receptors and the narrow host range of CAEV. However, functional receptors for the two North American MVV strains tested, unlike the Icelandic MVV and similar to CAEV, were limited to cells of ruminant species, regardless of cytopathic phenotype. The results indicate a differential receptor recognition by MVV strains which is unrelated to cytopathic phenotype.

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Figures

FIG. 1
FIG. 1
Alignment of predicted amino acid sequence of MVV 85/34 (GenBank accession number U64439) and S93 (AF338226) Env precursors. Only the residues of S93 Env not conserved with 85/34 are shown. Dots represent amino acid identity with MVV 85/34 Env, and dash represents a gap introduced into the 85/34 sequence for optimal alignment. The amino acid residue positions are indicated on the right of the alignment. The boundaries of the leader peptide, SU, and TM subunits of the Env precursor are indicated above the alignment. The putative cleavage site between the leader peptide and SU is in the position homologous to the chemically defined amino terminus of CAEV 63 SU (15).
FIG. 2
FIG. 2
Structure of pCAEVneo10 and pCAEVneo11 proviral constructs compared to wild-type CAEV provirus. Positions of viral and neo genes are shown with boxes. Bars labeled Δ1, Δ2, and Δ3 indicate the deletions in the vif-tat and env genes and the 3′ U3 region, respectively, present in the pCAEVneo constructs. Deletion Δ2 also introduced a frameshift (fs) mutation in env. Bent dotted lines indicate the intron separating rev exons 2 and 3. The human CMV and SV40 early promoters are shown as arrows. R/U5, R and U5 regions of the 5′ LTR.
FIG. 3
FIG. 3
Semiquantitative RT-PCR assay of double-spliced rev transcripts in GSM cells infected with CAEVneo10 or CAEVneo11. (A) The CAEVneo10 and CAEVneo11 integrated proviral structure is shown to indicate the relative positions of primers P1, P2, and P3. Thick lines indicate the rev and predicted neo/rev exons. Bent dotted lines indicate introns. The expected sizes of amplicons obtained by RT-PCR for each transcript are shown on the right. (B) Ethidium bromide-stained agarose gel with RT-PCR products. The size of the amplified fragments is indicated on the left. Primers for PCR are indicated above the gel. The RNA used for RT-PCR is indicated above each lane. GSM, uninfected GSM cell RNA; neo10, CAEVneo10-infected GSM cell RNA; neo11, CAEVneo11-infected GSM RNA.
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