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  • Brief Communication
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Polio vaccine samples not linked to AIDS

Nature volume 410, pages 1045–1046 (2001)Cite this article

A search through the archives clears early vaccines of starting the AIDS pandemic.

Abstract

It has been suggested that chimpanzee kidney cultures may have been used in the preparation of oral polio vaccine stocks used in Africa during the late 1950s, and so could have introduced the primate precursor of the immunodeficiency virus HIV-1 into humans1,2. Here we analyse frozen samples of the suspect vaccine by using the polymerase chain reaction (PCR) to amplify any HIV-1-related nucleic acids or chimpanzee mitochondrial DNA that might be present, but we have failed to detect either. Our findings do not support the hypothesis that HIV-1 was introduced by oral vaccination against poliovirus.

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Figure 1: Neighbour-joining tree of 12S mtDNA sequences (see ftp://ftp.pasteur.fr/pub/retromol/wistar01).

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Author information

Authors and Affiliations

  1. Unité de Rétrovirologie Moléculaire, Institut Pasteur, 28 rue du Dr Roux, Paris, 75724 cedex 15, France

    Philippe Blancou, Jean-Pierre Vartanian, Nicole Chenciner & Simon Wain-Hobson

  2. Department of Infectious Diseases, Roche Molecular Systems, 1145 Atlantic Avenue, Alameda, 94501, California, USA

    Cindy Christopherson & Shirley Kwok

  3. Department of Microbiology, New York University School of Medicine, 550 First Avenue, New York, 10016, New York, USA

    Claudio Basilico

Authors
  1. Philippe Blancou
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  2. Jean-Pierre Vartanian
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  3. Cindy Christopherson
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  4. Nicole Chenciner
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  5. Claudio Basilico
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  6. Shirley Kwok
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  7. Simon Wain-Hobson
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Corresponding author

Correspondence to Simon Wain-Hobson.

Additional information

Supplementary information

The online version of this article (doi:10.1038/35074171) contains supplementary material, which is available to authorized users.

Supplementary information

[This information is also available at: ftp://ftp.pasteur.fr/pub/retromol/wistar01/]

Methods

For the Alameda laboratory, two regions within the gag and pol genes of HIV-1/SIV were targeted for amplification. For the gag region, 3 upstream and 2 downstream primers were used to amplify a 155 bp fragment. The primers are all minor variants of the SK145-SKCC1B primers1 used in the AMPLICOR HIV-1 MONITOR version 1.5 Test. Two probes, SK102 and GAG102_O, were used to detect HIV-1 and SIVcpz (primarily US and ANT; and to a lesser extent GAB1), respectively. For the pol region, two upstream primers (one specifically for SIVcpzANT) and one downstream primer were used to amplify a 274 bp fragment. A single probe was used to detect both HIV-1 and SIVcpz.

Two hundred microliters of each sample were processed using the AMPLICOR HIV-1 MONITOR extraction protocol2. The equivalent of ~45 µl of the original sample, was analyzed in duplicate with both primer systems. One of the replicates for gag was amplified in the presence of 100 copies of IC to monitor inhibition; none was noted Following amplification, the gag products were analyzed on microwell plates coded with SK102, GAG102_O and, where appropriate, with the IC probe. The pol products were analyzed on plates coated with CC37.

Upstream gag primers were: WH43 AGTGGGGGGACATCAAGCAGCCATGCAAA, WH45 AGTGGGGGGACACCAGGCAGCTATGCAAA and SK145W AGTAGGGGGACATCAAGGAGCCATGCA. Downstream gag primers were: WH50 GGTACTAGTAGTTCCTGCTATGTCACTTCC and SKCC1W TGCTGGTTGTTCCTGCTATATCACTTCC. gag probes were SK102 GAGACCATCAATGAGGAAGCTGCAGAATGGGAT, GAG102_O GAAGTAATCAATGAGGAAGCAGCAGATTGGGAT. Upstream pol primers were: CC36B TAAAATTAGCAGGAAGATGGCCAGTAA and CC36Bant TAAAATTAGCCAGCAGATGGCCAGTAA. Downstream pol primers were: CC26B CCCCCAATCCCCCCTTTTCTTTTAAAATTGTG while the pol probe was CC37 TTTGGAATTCCCTACAATCCACAAAGTCAAGGAGT.

For the Paris laboratory total nucleic acids were extracted using Master Pure extraction kit (Epicentre) from half of the samples in order to keep some material left (total volumes varied from 300 µl to 1 ml). Briefly, 2 µg of tRNA (Sigma) were added as carrier and each sample was incubated 15 min at 56°C in the presence of proteinase K (100 µg/ml) then the acid nucleic lysates were purified. cDNA synthesis was performed on half of the extracts using UC1, UC12, cpzLTR3' and 3'ARN12S primers. To avoid HIV/SIV/polio contamination, the complete extraction and PCR amplifications procedures were performed in a HIV/SIV/polio free building.

Hot start PCR was applied in all reactions. The buffer conditions were as follows: 2.5 mM MgCl2, 50 mM KCl, 10 mM Tris-HCl (pH 8.3), 200 mM of each dNTP, 100 mM of each primer, 2.5 units of Taq DNA polymerase (Perkin Elmer) in a final volume of 100 ml. Annealing, extension and denaturation conditions were: an initial denaturation at 95°C for 5 minutes, then 35 cycles at 95°C (30 sec.), 55°C (30 sec.), 72°C (30 sec.) followed by a final step at 72°C for 10 minutes. Poliovirus VP1 (UG1 and UC1) and 3D (outer UG16 and UC12, inner UG7 and UC8) primers have been described3. Primers for SIVcpz were: cpzLTR5’ CTCAATAAAGCTTGCCTTGAGTG and cpzLTR3’ CCTGTTCGGGCGCCACTGCTAGAGAT. For mitochondrial 12S rDNA, 5'ARN12S CTATATACCGCCATCTTCAGCRAAC and 3’ARN12S TCTTRRTTTACTRCTAAATCCWCCTT as well as 12Sa and 12So4. PCR products were purified, cloned and sequenced.

References

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NUMT sequences

Chat pool 13

  • Cl 2

    GCTTAGCCCTAAACTGTAATAGTTACATTAACAAAACTATTCACCAGAATACTACAAGCAATAGCTTAAAACTCAAAGGACTTGGTGGTGCTTTATGTCCCTC

  • Cl 6

    GCTTAGCTGTAAACTTAAATAATTTAACAAACAAAATTATTCACCAGAGTATTACGAGCAAGAGCTTAAAACTCAAAGGACATGGCCATGCTTTATACTCCTC

  • Cl 8

    GCTTAGCCTTAAACCTCAATAGTTAAAACAACAAAACTACTCTCCAGAATACTACAAGCAACAGCTTAAAACTCAAAGGACTTGGCGGTGCTTCACATCCCTC

  • Cl 10

    GCTTAGCCCTAAACTCTAGTAGTTACATAAACAAAACAATTCCTCAGAATACTACAAGCAACGGCTTAAAACTCAAAGGACTTGGCGGTGCTTTGTATCCCTC

  • Cl 11

    GCTCAGCTCTAAACTCTAATAGTTATATTAACAAAACCACTCGCCAGAGTACTACAAGAAACAGCTGAAAACTCAAAGGACTTGGTGGTGCTTTATATCCCCC

  • Cl 21

    ACTTAGCCCTAAACTCTAATAGTTACATTAACAAAACCACTCACCAGAGTACTACGAGCAACAGCTTAAAACTCAAAGGATTTGGTGGTGCTTTATATCCGTC

  • Cl 23

    GCTTAGCCCTAAACCTCAATAGTTAAAACAACAAAACTACTCACCAGAATACTACAAGCAACAGCTTAAAACTCAACGGACTTGGCGGTGCTTCACATCCCCC

  • Cl 24

    GCTTAGCCCTAAACTGTAATAGTTACATTAACAAAACCATTAACCAGAATACTACAAGCAATAGCTTACAACTCAAAGGACTTGGTGGTGCTTTATGTCCCTC

  • Cl 27

    GCTTAGCCCTAAAACTCAATAGTTAAATAACAAAACTACTCGCCAGAATACTACAAGCAACAGCTTAAAACTCAAAGGACTTGACGGTGCTTTACTTCCCTC

  • Cl 28

    GCTTGGCCTTAAACCTCAGTAGTTAAATAACAAAACTACTTGCCAGAATACTACAAGCTACAGGTTAAAACTCAAAGGACTTGACGGTGCTTTACATCCCCC

  • Cl 30

    GCTTAGTCCTAAACTTCATTAGTTAAATTAACAAAACTACTCACCAGAATACTACAAGCAATAGCTTAAAACTCAAAGGACCTGGCGGTGCTTTATATCCCCC

  • Cl 32

    GCTTAGCCCTAAACTCTAGTAGTTACATAAACAAAACAATTCCTCAGAATACTACAAGCAACAGCTCAAAACTCAAAGGACTTGGCGGTGCTTTGTATCCCTC

  • Cl 33

    GCTTAGCCCTAAACCTCAATAGTTAGAACAACAAAACTACTCGCCAGAATACTACAAGCAACAGCTTGAAACTCAAAGGACTTGGCAGTGCTTCATATCCCTC

  • Cl 38

    GCTTAGTTCTAAACCCAAATAGTTCAACCAACAAAACTCTTCATCAGAGTACTATAAGCAACAGCTTAAGACTCAAAAGACTTGGTGGTGCTTTATATCCCTC

  • Cl 44

    GCTTGGCCCTAAACCTCAGTAATTAGATAACAAAACTACTCGCCAGAATACTACAAGCAACAGCTTGAAACTCAAAGGACTTGACGGTGCTTTACATCCCCC

  • Cl 46

    GCTTGGCCCTAAACTTCAACAATTAAATTAACAAAACTGCTCGCCAGAACACTACAAGCACTAGCTTAAAACTCAGAGGACTTGGCGGTGCTTCACATCCCTC

  • Cl 51

    GCTCAGCTCTAAACTCTAATAGTTAGATTAACAAAACCACACGCCAGAGTACTACAAGAAACAGCTGAAAACTCAAAGGACTTGGTGGTGCTTTATATCCCCC

  • Cl 64

    GCTTAGCCCTAAACCTCAATAATTAAAATAACAAAACTACTCGCCAGAATACTACAAGCAACAGCTTGAAACTCAAAGGACTTGGCGGTGTTTCACATCCCTC

Chat pool 16-A15

  • Cl 25

    GCTTGGCCCTAAACCTCAGTAATTAGATAACAAAACTACTCGCCGAATACTACAGGCAACAGTTGAAACTCAAAGTACTTGACGGTGCTTTACATCCCCC

Wch24 57C-40

  • Cl 03

    GCTTAGTTCTAAACCCAAATAGTTCAACCAACAAAACTCTCCATCAGAGTACTATAAGCAACAGCTTAAGACTCAAAAGACTTGGTGGTGCTTTATATCCCC

Chat type I Wy4B

  • Cl 148

    GCTTGGCCCTAAACCTCAGTAATTAGATAACAAAACTACTCGCCAGAATACTACAAGCAACAGCTTGAAACTCAGAGGACTTGACGGTGCTTTACATCCCC

  • Cl 140

    GCTTAGCCCTAAAACTCAATAGTTAAATAACAAAACTACTCGCCAGAATACTACAAGCAACAGCTTAAAACTCAAAGGACTTGACGGTGCTTTACATCCCC

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Blancou, P., Vartanian, JP., Christopherson, C. et al. Polio vaccine samples not linked to AIDS. Nature 410, 1045–1046 (2001). https://doi.org/10.1038/35074171

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