Research ArticleInfectious Disease

JC polyomavirus mutants escape antibody-mediated neutralization

See allHide authors and affiliations

Science Translational Medicine  23 Sep 2015:
Vol. 7, Issue 306, pp. 306ra151
DOI: 10.1126/scitranslmed.aab1720
  • Fig. 1. JCV neutralization serology (healthy adults).

    Serum samples from 96 individual adult subjects (rows) were serially diluted and initially tested for neutralization of a wild-type JCV-2A pseudovirus. Sixty serum samples that detectably neutralized JCV-2A were tested against additional JCV pseudoviruses. Sera that failed to detectably neutralize JCV-2A were excluded from further analysis and are not shown in the figure. The inverse log10 of the calculated EC50 is indicated with a color code. EC50 values ≤2 (blue cells) are considered neutralization-negative. For the precise VP1 sequence identity of JCV pseudovirus, see table S1.

  • Fig. 2. PML patient neutralization serology.

    Pre- and post-PML plasma samples from six patients were tested for neutralization of cognate (or near-cognate) wild-type (WT) and PML-mutant pseudoviruses. Specifically, patient 5029 was tested against viruses 5029w (cognate WT, blue) and 5029m (cognate mutant, red); 5031: 5031w/5031mb; 5040: 2A/5040m; 5053: 5053w/5053m; 5058: 2A/5147m; 5147: 5147w/5147m (see table S1). Testing of PML patient sera against additional pseudoviruses is shown in fig. S4. Patients whose disease progressed (left column) are indicated with (P), and patients who survived (right column) are indicated with (S). Dominant PML-associated mutations observed in each patient’s CSF are indicated. Y axes indicate neutralizing EC50. Error bars represent SEM for data from three independent experimental replicates, two of which were performed with blinding. Arrows below the x axes indicate the date of onset of PML symptoms. Date format is month/day/year.

  • Fig. 3. Preclinical evaluation of a candidate JCV VLP vaccine.

    Mice were given an intramuscular injection of VLPs based on the JCV genotype indicated in each row label. Four weeks later, sera from the mice were tested for neutralization of various pseudoviruses indicated in column labels. Numerical values represent EC50 neutralizing titers. Error represents SD for independent neutralization assays of sera from five replicate mice. Preimmune sera were nonneutralizing at a 1:100 dilution.

  • Fig. 4. JCV-neutralizing titers after vaccination of PML patient 5228.

    Patient 5228 was administered JCV VLPs subcutaneously at three time points (downward purple triangles). Recombinant IL-7 was also administered for two cycles each consisting of a subcutaneous weekly dose of 10 μg/kg (upward green triangles). The patient’s neutralizing titer against her cognate PML-mutant JCV (red squares) or inferred WT JCV (blue circles) was monitored over time. JCV load in the patient’s plasma (gray dots) was also monitored over time. See the Supplementary Materials for additional patient details.

  • Fig. 5. Evolution of PML lesions by magnetic resonance imaging in patient 5228.

    The patient was initially diagnosed with PML on 24 May 2012. The date of each scan is indicated along the top of the figure. Axial fluid-attenuated inversion recovery (FLAIR, row a), diffusion-weighted imaging (DWI, row b), and FLAIR-DWI merged images (row c) show the evolution of PML lesions. Row a: The first image on the left (July 2012) shows a high signal intensity lesion of the right parieto-occipital white matter extending contralaterally through the corpus callosum; another focus is present in the left subcortical temporal region. Subsequent examinations showed evolution of the signal alteration with progressive rapid extension of the lesions to the entire white matter bilaterally. In September 2012, the white matter was completely occupied by lesional and atrophic processes and enlargements of the ventricles and cortical sulci began to appear. The last examinations (January and March 2013) show progression of atrophy. Row b: Axial DWI images show the evolution of the hyperintense signal alterations corresponding to the front of progression of PML lesions (red arrows). The signal alteration is substantially reduced starting from November 2012 and is no longer visible at the March 2013 examination, implying lesion stabilization. Row c: Merged FLAIR and DWI sequences show, in yellow (black arrows), the initial fronts of advance of PML lesions and their regression over time.

Supplementary Materials

  • www.sciencetranslationalmedicine.org/cgi/content/full/7/306/306ra151/DC1

    Text

    Fig. S1. Transducibility of various cell lines.

    Fig. S2. An example of luminometry results for a pilot JCV neutralization assay using ART cells.

    Fig. S3. Neutralization assay validation.

    Fig. S4. PML patient neutralization serology (an expansion of Fig. 2).

    Fig. S5. Serological analysis of mice after a booster dose of JCV VLPs.

    Fig. S6. JCV-neutralizing titers after vaccination of PML patient 5228 (an alternative version of Fig. 4).

    Table S1. Characteristics of JCV pseudovirus stocks.

    Table S2. Patient characteristics.

    Table S3. PML patient neutralization serology (source data).

    Table S4. Neutralization serology of patient 5228 (source data).

    Table S5. Viremia of patient 5228 (source data).

    References (4048)

  • Supplementary Material for:

    JC polyomavirus mutants escape antibody-mediated neutralization

    Upasana Ray, Paola Cinque, Simonetta Gerevini, Valeria Longo, Adriano Lazzarin, Sven Schippling, Roland Martin, Christopher B. Buck,* Diana V. Pastrana*

    *Corresponding author. E-mail: buckc{at}mail.nih.gov (C.B.B.); pastrand{at}mail.nih.gov (D.V.P.)

    Published 23 September 2015, Sci. Transl. Med. 7, 306ra151 (2015)
    DOI: 10.1126/scitranslmed.aab1720

    This PDF file includes:

    • Text
    • Fig. S1. Transducibility of various cell lines.
    • Fig. S2. An example of luminometry results for a pilot JCV neutralization assay using ART cells.
    • Fig. S3. Neutralization assay validation.
    • Fig. S4. PML patient neutralization serology (an expansion of Fig. 2).
    • Fig. S5. Serological analysis of mice after a booster dose of JCV VLPs.
    • Fig. S6. JCV-neutralizing titers after vaccination of PML patient 5228 (an alternative version of Fig. 4).
    • Table S1. Characteristics of JCV pseudovirus stocks.
    • Table S2. Patient characteristics.
    • Table S3. PML patient neutralization serology (source data).
    • Table S4. Neutralization serology of patient 5228 (source data).
    • Table S5. Viremia of patient 5228 (source data).
    • References (4048)

    [Download PDF]

Navigate This Article