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Vaccines and Antiviral Agents

Recombinant Respiratory Syncytial Virus That Does Not Express the NS1 or M2-2 Protein Is Highly Attenuated and Immunogenic in Chimpanzees

Michael N. Teng, Stephen S. Whitehead, Alison Bermingham, Marisa St. Claire, William R. Elkins, Brian R. Murphy, Peter L. Collins
Michael N. Teng
Respiratory Viruses Section and
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Stephen S. Whitehead
Respiratory Viruses Section and
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Alison Bermingham
Respiratory Viruses Section and
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Marisa St. Claire
Bioqual, Inc., Rockville, Maryland, 20850
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William R. Elkins
Experimental Primate Virology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, 20892, and
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Brian R. Murphy
Respiratory Viruses Section and
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Peter L. Collins
Respiratory Viruses Section and
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DOI: 10.1128/JVI.74.19.9317-9321.2000
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  • Table 1.

    rA2ΔNS1 and rA2ΔM2-2 are highly attenuated in both the upper and lower respiratory tracts of chimpanzees but are highly immunogenic

    Virus used to infect chimpanzeesaNo. of animalsDoseb (per site, log10PFU)Mean peak virus titerc(log10 PFU/ml) ± SE (Duncan grouping)Mean peak rhinorrhea scored (range, 0–4)Mean neutralizing antibody titer in serume (reciprocal log2)
    Nasopharyngeal swabTracheal lavageDay 0Day 28
    Wild-type RSV A2f 24.05.0 ± 0.35 (A)5.5 ± 0.40 (A)3.0<3.311.2
    rA2g 24.04.9 ± 0.15 (A)5.4 ± 0.05 (A)2.5<3.310.5
    rA2ΔSHg 34.04.6 ± 0.10 (A)3.8 ± 0.31 (B)1.0<3.310.2
    rA2ΔNS2g 44.03.8 ± 0.41 (B)1.4 ± 0.29 (C)1.03.410.6
    rA2cp248/404g 45.02.5 ± 0.25 (C)1.4 ± 0.37 (C)0.83.410.6
    rA2ΔNS145.01.6 ± 0.12 (D)1.2 ± 0.43 (C)2.0<3.39.8
    rA2ΔM2-245.01.5 ± 0.09 (D)<0.71.8<3.39.1
    • ↵a All recombinant-derived viruses contain the sites and HEK mutations (see the text), except for rA2ΔM2-2.

    • ↵b Chimpanzees were inoculated by the intranasal and intratracheal routes with the indicated amount of virus in a 1-ml inoculum per site.

    • ↵c Nasopharyngeal swab samples were collected daily for 10 days, and tracheal lavage samples were collected on days 2, 5, 6, 8, and 10. Mean peak titers were calculated and assigned to statistically similar groups by Duncan's multiple-range test (α = 0.05). Means in each column with different letters are significantly different.

    • ↵d The amount of rhinorrhea was estimated daily and assigned a score (0 to 4) that indicated extent and severity. Scores indicate severe (4), moderate (3), mild (2), trace (1), or no (0) rhinorrhea. Shown are the mean peak scores.

    • ↵e Serum RSV-neutralizing antibody titers were determined by a complement-enhanced 60% plaque reduction assay using wild-type RSV A2 and HEp-2 cell monolayer cultures incubated at 37°C. RSV-seronegative chimpanzee serum used as a negative control had a neutralizing antibody titer of <3.3 log2 reciprocal. Adult human serum used as a positive control had a neutralizing antibody titer of 11.4 log2 reciprocal.

    • ↵f Historic controls from the study of Crowe et al. (10).

    • ↵g Data from the study of Whitehead et al. (29).

  • Table 2.

    Infection of chimpanzees with rA2ΔNS1 or rA2ΔM2-2 induced significant protection against subsequent challenge with wild-type RSV A2 in the upper and lower respiratory tracts

    Immunizing virusInoculum dosea (log10PFU/ml)No. of animalsReplication of RSV challenge virus at the indicated sitebMean peak rhinorrhea score
    NasopharynxTrachea
    Mean no. of days of shedding ± SEMean peak titerc ± SEMean no. of days of shedding ± SEMean peak titer ± SE
    rA2ΔNS15.042.8 ± 0.751.7 ± 0.461.0 ± 0.411.8 ± 0.731.0
    rA2ΔM2-25.043.5 ± 0.872.3 ± 0.711.0 ± 0.711.7 ± 0.631.0
    rA2ΔNS2d 4.04ND1.9 ± 0.30ND2.2 ± 0.771.0
    cpts248/404e 4.723.5 ± 0.502.3 ± 0.250<0.71.0
    Nonee 28.5 ± 0.505.0 ± 0.356.0 ± 1.04.8 ± 0.303.0
    • ↵a Each virus was initially administered at the indicated dose in a 1.0-ml inoculum given intranasally and intratracheally.

    • ↵b On day 56, chimpanzees were challenged with wild-type RSV A2 administered at a dose of 104 PFU/ml in a 1.0-ml inoculum given intranasally and intratracheally. Nasopharyngeal swab samples were collected daily for 12 days, and tracheal lavage samples were collected on days 2, 5, 6, 8, and 12. ND, not determined.

    • ↵c Mean peak titers (log10 PFU/ml) were calculated by using the peak virus titer achieved in each animal.

    • ↵d Data from the study of Whitehead et al. (29).

    • ↵e Historic control animals from the study of Crowe et al. (10) were used.

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Recombinant Respiratory Syncytial Virus That Does Not Express the NS1 or M2-2 Protein Is Highly Attenuated and Immunogenic in Chimpanzees
Michael N. Teng, Stephen S. Whitehead, Alison Bermingham, Marisa St. Claire, William R. Elkins, Brian R. Murphy, Peter L. Collins
Journal of Virology Oct 2000, 74 (19) 9317-9321; DOI: 10.1128/JVI.74.19.9317-9321.2000

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Recombinant Respiratory Syncytial Virus That Does Not Express the NS1 or M2-2 Protein Is Highly Attenuated and Immunogenic in Chimpanzees
Michael N. Teng, Stephen S. Whitehead, Alison Bermingham, Marisa St. Claire, William R. Elkins, Brian R. Murphy, Peter L. Collins
Journal of Virology Oct 2000, 74 (19) 9317-9321; DOI: 10.1128/JVI.74.19.9317-9321.2000
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KEYWORDS

Antigens, Viral
HN Protein
Respiratory Syncytial Viruses
viral nonstructural proteins
Viral Proteins

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