Progressive multifocal leukoencephalopathy (PML) development is associated with mutations in JC virus capsid protein VP1 that change its receptor specificity - PubMed (original) (raw)

. 2011 Jul 1;204(1):103-14.

doi: 10.1093/infdis/jir198.

Carl Reid, Manuela Testa, Margot Brickelmaier, Simona Bossolasco, Annamaria Pazzi, Arabella Bestetti, Paul Carmillo, Ewa Wilson, Michele McAuliffe, Christopher Tonkin, John P Carulli, Alexey Lugovskoy, Adriano Lazzarin, Shamil Sunyaev, Kenneth Simon, Paola Cinque

Affiliations

Progressive multifocal leukoencephalopathy (PML) development is associated with mutations in JC virus capsid protein VP1 that change its receptor specificity

Leonid Gorelik et al. J Infect Dis. 2011.

Abstract

Progressive multifocal leukoencephalopathy (PML), a fatal demyelinating disease caused by JC virus (JCV) infection of oligodendrocytes, may develop in patients with immune disorders following reactivation of chronic benign infection. Mutations of JCV capsid viral protein 1 (VP1), the capsid protein involved in binding to sialic acid cell receptors, might favor PML onset. Cerebrospinal fluid sequences from 37/40 PML patients contained one of several JCV VP1 amino acid mutations, which were also present in paired plasma but not urine sequences despite the same viral genetic background. VP1-derived virus-like particles (VLPs) carrying these mutations lost hemagglutination ability, showed different ganglioside specificity, and abolished binding to different peripheral cell types compared with wild-type VLPs. However, mutants still bound brain-derived cells, and binding was not affected by sialic acid removal by neuraminidase. JCV VP1 substitutions are acquired intrapatient and might favor JCV brain invasion through abrogation of sialic acid binding with peripheral cells, while maintaining sialic acid-independent binding with brain cells.

PubMed Disclaimer

Figures

Figure 1.

Figure 1.

Frequencies of PML-associated substitutions identified in JCV VP1 from the CSF of PML patients. The number of PML patients harboring a JCV with particular VP1 mutation or deletion in the CSF is displayed. Mutations were identified by comparing the sequences to either VP1 sequences from matched urine samples (when available) or to 253 sequences isolated from the urine of non-PML individuals as reported in GenBank and assembled elsewhere [9]. The most dominant mutation is plotted for each patient (ie, mutations present in a low number of clones are not displayed) (for a comprehensive list, see

Table 3). NOTE: PML indicates progressive multifocal leukoencephalopathy; JCV, JC virus; VP1, viral protein 1; CSF, cerebrospinal fluid.

Figure 2.

Figure 2.

JCV VLPs carrying PML-associated mutations lose the ability to hemagglutinate red blood cells (RBCs) and bind sialylated gangliosides. (A) Ability of mutant VLPs to hemagglutinate red blood cells from different blood groups. RBCs were incubated with serial 2-fold dilutions of various VLPs, starting from 100 μg/mL. Minimum hemagglutination (HA) concentration is the lowest concentration of VLP that still agglutinated RBCs. HA results were examined by visual inspection. All HA reactions were conducted in duplicates. Mean ± SD for the minimum HA concentration is calculated based on the results obtained from hemagglutination of 4 to 7 different blood group donor RBCs (A, B, O, and AB). VLPs displaying the minimum HA concentration of 100 μg/mL (dotted line) did not cause any hemagglutination at this concentration (ie, highest VLP concentration tested). (B) Specificity of JC VLPs for sialylated gangliosides. VLPs binding to an array of gangliosides were detected with a 2-step process involving the detection of VLPs bound to a ganglioside with VP1-specific murine antibodies and anti-murine IgG HRP–labeled antibodies followed by development with TMB substrate. Numbers represent percent increase in the optical density obtained with the specific VLP relative to that obtained without VLP (ie, background) and were calculated as follows: % specific VLP binding = 100%*(OD450(with VLP)–OD450(without VLP))/OD450(without VLP). Statistically significant (P < .05) interaction between a VLP and ganglioside in comparison to the background interaction between the VLP and MeOH-treated well is denoted by a red cell background. Results are depicted as a mean ± SD binding for each VLP to each of the gangliosides or to the control for several independent experiments conducted; N denotes number of independent measurements for each VLP and all gangliosides. Schematic structure of ganglioside is shown to reveal core binding structure bound by various VLPs. NOTE. JCV indicates JC virus; VLPs, virus-like particles; PML, progressive multifocal leukoencephalopathy; VP1, viral protein 1; TMB, tetramethylbenzidine.

Figure 3.

Figure 3.

JCV VLPs carrying PML-associated mutations bind differently to lymphoid vs CNS cells. (A) Glial cell line SVG-A, (B) primary human astrocytes, (C) Jurkat cells, (D) primary human kidney tubular epithelial cells (HRPTEC), or (E and F) peripheral blood mononuclear cells (PBMCs) were first incubated with different VLPs (as indicated on the x-axis) and then with anti-VP1 antibodies followed by staining with fluorescently labeled antibodies. Binding of VLPs to B (E) and T lymphocytes (F) was evaluated after costaining of PBMCs with anti-CD20 and anti-CD3 antibodies and gating on the corresponding population. The ratio of mean fluorescent intensity (MFI) of cells stained with anti-VP1 antibodies in the presence of VLPs relative to that of cells stained only with the detection antibodies in the absence of VLPs (background) is plotted for each VLP. Mean MFI ratio ± SD is calculated based on the results from several independent experiments (n = number of experiments). Dotted lines (MFI ratio = 2) represents a 2-fold increase in VLP binding over the background and deemed as being significant. NOTE. JCV indicates JC virus; VLPs, virus-like particles; PML, progressive multifocal leukoencephalopathy; CSF, cerebrospinal fluid.

Figure 4.

Figure 4.

Binding of JCV VLPs carrying PML-associated mutations to glial cells is neuraminidase treatment–insensitive. (A) SVG-A glial cells (top row) or Jurkat lymphoid cells (bottom row) were first either pretreated with neuraminidase (blue line) for 60 min at 37°C to remove all terminal α2–3–, α2–6–, and α2-8–linked sialic acid residues or mock-treated (red line) followed by incubation at 4°C with the indicated VLP and staining with the detection antibodies as described in the previous figure. One representative experiment out of 4 conducted is shown. Staining of the cells with Sambucus n igra agglutinin (SNA) lectin and Maackia amurensis lectin II (MAL II) proved neuraminidase treatment effectiveness in sialic acid removal (data not shown). (B) Sialic acid dependence as compared with VLP binding by various assays. The degree of binding was evaluated as a percent of wild-type VLP binding value in the assay (Figures 2 and 3) and were calculated as follows: 100%*(VLP VALUE – BASELINE VALUE)/(WT VALUE – BASELINE VALUE); hemagglutination assay values were LOG10 transformed first. Values were ranked accordingly: “–” < 10; 10 < “+” < 20; 20 < “++” < 40; 40 < “+++” < 60; 60 < “++++” < 80; and “+++++” > 80. NOTE. JCV indicates JC virus; VLPs, virus-like particles; PML, progressive multifocal leukoencephalopathy; HA, hemagglutination; SA, sialic acid binding; HRPTEC, human renal proximal tubular epithelial cells; astrocytes, primary human astrocytes. Dependence on sialic acid binding is denoted as NO, no dependence; or Part., partial dependence. Positive values are denoted by red cell backgrounds.

Comment in

References

    1. Major EO. Reemergence of PML in natalizumab-treated patients—new cases, same concerns. N Engl J Med. 2009;361:1041–3. - PubMed
    1. Cinque P, Koralnik IJ, Gerevini S, Miro JM, Price RW. Progressive multifocal leukoencephalopathy in HIV-1 infection. Lancet Infect Dis. 2009;9:625–36. - PMC - PubMed
    1. Egli A, Infanti L, Dumoulin A, et al. Prevalence of polyomavirus BK and JC infection and replication in 400 healthy blood donors. J Infect Dis. 2009;199:837–46. - PubMed
    1. Kean JM, Rao S, Wang M, Garcea RL. Seroepidemiology of human polyomaviruses. PLoS Pathog. 2009;5:e1000363. - PMC - PubMed
    1. Liu CK, Wei G, Atwood WJ. Infection of glial cells by the human polyomavirus JC is mediated by an N-linked glycoprotein containing terminal alpha(2-6)-linked sialic acids. J Virol. 1998;72:4643–9. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources