Bovine parainfluenza virus type 3 (BPIV3) fusion and hemagglutinin-neuraminidase glycoproteins make an important contribution to the restricted replication of BPIV3 in primates - PubMed (original) (raw)

Bovine parainfluenza virus type 3 (BPIV3) fusion and hemagglutinin-neuraminidase glycoproteins make an important contribution to the restricted replication of BPIV3 in primates

A C Schmidt et al. J Virol. 2000 Oct.

Abstract

This study examines the contribution of the fusion (F) and hemagglutinin-neuraminidase (HN) glycoprotein genes of bovine parainfluenza virus type 3 (BPIV3) to its restricted replication in the respiratory tract of nonhuman primates. A chimeric recombinant human parainfluenza type 3 virus (HPIV3) containing BPIV3 F and HN glycoprotein genes in place of its own and the reciprocal recombinant consisting of BPIV3 bearing the HPIV3 F and HN genes (rBPIV3-F(H)HN(H)) were generated to assess the effect of glycoprotein substitution on replication of HPIV3 and BPIV3 in the upper and lower respiratory tract of rhesus monkeys. The chimeric viruses were readily recovered and replicated in simian LLC-MK2 cells to a level comparable to that of their parental viruses, suggesting that the heterologous glycoproteins were compatible with the PIV3 internal proteins. HPIV3 bearing the BPIV3 F and HN genes was restricted in replication in rhesus monkeys to a level similar to that of its BPIV3 parent virus, indicating that the glycoprotein genes of BPIV3 are major determinants of its host range restriction of replication in rhesus monkeys. rBPIV3-F(H)HN(H) replicated in rhesus monkeys to a level intermediate between that of HPIV3 and BPIV3. This observation indicates that the F and HN genes make a significant contribution to the overall attenuation of BPIV3 for rhesus monkeys. Furthermore, it shows that BPIV3 sequences outside the F and HN region also contribute to the attenuation phenotype in primates, a finding consistent with the previous demonstration that the nucleoprotein coding sequence of BPIV3 is a determinant of its attenuation for primates. Despite its restricted replication in the respiratory tract of rhesus monkeys, rBPIV3-F(H)HN(H) conferred a level of protection against challenge with HPIV3 that was indistinguishable from that induced by previous infection with wild-type HPIV3. The usefulness of rBPIV3-F(H)HN(H) as a vaccine candidate against HPIV3 and as a vector for other viral antigens is discussed.

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Figures

FIG. 1

FIG. 1

Genomes of the rHPIV3-FBHNB and rBPIV3-FHHNH chimeras and of the parent viruses, rHPIV3 JS and BPIV3 Ka, shown schematically (not to scale). The F and HN genes were exchanged as a single restriction fragment between rHPIV3 and rBPIV3, using _Sgr_AI and _Bsi_WI sites that had been introduced preceding the M and HN gene end sequences, respectively.

FIG. 2

FIG. 2

Confirmation of the identity of recombinant viruses by RT-PCR of viral RNA and _Eco_RI digestion. RT-PCR products of viral RNA were prepared with a primer pair that recognized conserved regions on either side of the F and HN genes in both BPIV3 and HPIV3. Digestion with _Eco_RI resulted in a unique pattern of restriction fragments for each of the four viruses. In the schematic diagrams on the left, horizontal lines symbolize the amplified viral sequences and vertical bars show the positions of _Eco_RI sites. The expected size (in nucleotides) of each restriction fragment is indicated above the line. Numbers below each line correspond to sequence positions in the antigenomic RNA of BPIV3 Ka, HPIV3 JS (GenBank accession no. AF178654 and Z11575), or the indicated chimeric derivative. On the right, a 1% agarose gel of the _Eco_RI digestion of PCR products confirms the identities of parental and chimeric viruses. The asterisks indicate gel bands that contain comigrating restriction fragments. Positions of molecular weight markers (MW) are indicated in nucleotides.

FIG. 3

FIG. 3

Multicycle replication of chimeric and parental viruses in simian LLC-MK2 cells. Multicycle replication (MOI of 0.01) of the three chimeras rHPIV3-FBHNB, rBPIV3-FHHNH, and rHPIV3-NB is compared with the replication of the BPIV3 Ka and rHPIV3 parents. Virus titers are shown as mean log10 TCID50 per milliliter ± standard error of triplicate samples. The lower limit of detection of this assay is 10 TCID50, as indicated by the dotted horizontal line.

FIG. 4

FIG. 4

Mean titers of chimeric and parental viruses in nasopharyngeal swabs of infected rhesus monkeys over the course of infection. Virus titers are shown as mean TCID50 per milliliter in LLC-MK2 cells ± standard error for groups of four or six monkeys infected with the same virus. Data are from the same experiment as shown in Table 1. (A) Mean titers of rHPIV3-FBHNB compared to rHPIV3 and BPIV3 Ka titers; (B) mean rBPIV3-FHHNH titers compared to those of BPIV3 Ka and rHPIV3, which are the same values in panel A but are presented separately to facilitate comparison. Day 5 titers are not shown because they were much lower than day 4 and day 6 titers, most likely due to technical problems during the sample collection.

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References

    1. Bailly J E, McAuliffe J M, Durbin A P, Elkins W R, Collins P L, Murphy B R. A recombinant human parainfluenza virus type 3 (PIV3) in which the nucleocapsid N protein has been replaced by that of bovine PIV3 is attenuated in primates. J Virol. 2000;74:3188–3195. - PMC - PubMed
    1. Bailly J E, McAuliffe J M, Skiadopoulos M H, Collins P L, Murphy B R. Sequence determination and molecular analysis of two strains of bovine parainfluenza virus type 3 that are attenuated in primates. Virus Genes. 2000;20:173–182. - PubMed
    1. Buchholz U J, Granzow H, Schuldt K, Whitehead S S, Murphy B R, Collins P L. Chimeric bovine respiratory syncytial virus with glycoprotein gene substitutions from human respiratory syncytial virus (HRSV): effects on host range and evaluation as a live-attenuated HRSV vaccine. J Virol. 2000;74:1187–1199. - PMC - PubMed
    1. Clements M L, Belshe R B, King J, Newman F, Westblom T U, Tierney E L, London W T, Murphy B R. Evaluation of bovine, cold-adapted human, and wild-type human parainfluenza type 3 viruses in adult volunteers and in chimpanzees. J Clin Microbiol. 1991;29:1175–1182. - PMC - PubMed
    1. Clements M L, Sears S D, Christina K, Murphy B R, Snyder M H. Comparison of the virologic and immunologic responses of volunteers to live avian-human influenza A H3N2 reassortant virus vaccines derived from two different avian influenza virus donors. J Clin Microbiol. 1989;27:219–222. - PMC - PubMed

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