Partitioning of TSE infectivity during ethanol fractionation of human plasma

Gregori, Luisa; Maring, J.-A.; MacAuley, Claudia; Dunston, B; Rentsch, Marianne Berger; Kempf, Christoph; Rohwer, Robert G.

doi:10.1016/j.biologicals.2003.08.004

Cited by 51 publications

(38 citation statements)

References 33 publications

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“…Such findings are consistent with earlier reports of low levels of prions in blood (21)(22)(23) and the replication of prions in the lymphoreticular system (24)(25)(26)(27)(28)(29). The distribution of prions in blood has been difficult to resolve, because the levels of infectivity are so low (21,(30)(31)(32). Whether some prions are nonspecifically bound to circulating lymphocytes, perhaps by PrP C , or whether they reproduce at low levels in a distinct set of lymphoid cells is unclear (33)(34)(35)(36).…”

Section: Prions In Bloodsupporting

confidence: 89%

Human prions and plasma lipoproteins

Safar

Wille

Geschwind

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Prions are composed solely of an alternatively folded isoform of the prion protein (PrP), designated PrP Sc . The polyoxometalate phosphotungstic acid has been used to separate PrP Sc from its precursor PrP C by selective precipitation; notably, native PrP Sc has not been solubilized by using nondenaturing detergents. Because of the similarities between PrP Sc and lipoproteins with respect to hydrophobicity and formation of phosphotungstic acid complexes, we asked whether these molecules are bound to each other in blood. Here we report that prions from the brains of patients with sporadic Creutzfeldt-Jakob disease (CJD) bind to very low-density (VLDL) and low-density (LDL) lipoproteins but not to high-density lipoproteins (HDL) or other plasma components, as demonstrated both by affinity assay and electron microscopy. Immunoassays demonstrated that apolipoprotein B (apoB), which is the major protein component of VLDL and LDL, bound PrP Sc through a highly cooperative process. Approximately 50% of the PrP Sc bound to LDL particles was released after exposure to 4 M guanidine hydrochloride at 80°C for 20 min. The apparent binding constants of native human (Hu) PrP Sc or denatured recombinant HuPrP(90 -231) for apoB and LDL ranged from 28 to 212 pM. Whether detection of PrP Sc in VLDL and LDL particles can be adapted into an antemortem diagnostic test for prions in the blood of humans, livestock, and free-ranging cervids remains to be determined. prion protein ͉ apolipoprotein B ͉ blood ͉ Creutzfeldt-Jakob disease P rions are composed solely of an alternatively folded isoform of the prion protein, designated PrP Sc . Despite numerous efforts to solubilize native PrP Sc , no procedure has been developed except for incorporation of the protein into liposomes (1, 2). The hydrophobic properties of PrP Sc not only have impeded biological investigations but also have prevented structural studies at atomic resolution.During the development of an immunoassay for PrP Sc that does not depend upon limited proteolysis to hydrolyze the precursor protein designated PrP C , we found that the Na salt of phosphotungstic acid (PTA) selectively complexes with PrP Sc (3). PTA is a water-soluble salt featuring the nearly spherical trianion [PW 12 O 14 ] 3Ϫ that belongs to a broad class of polynuclear transition metal-oxo complexes known as polyoxometalates (POMs; see ref. 4). Recently, two of us (J.G.S. and S.B.P.) investigated the mechanism of selective complex formation between Keggin-type POMs and PrP Sc using a series of POM analogues (5). The ability of POMs to complex with PrP Sc as well as lipoproteins (6) raised the possibility that these two hydrophobic proteins might copurify.We began by examining the distribution of human (Hu) PrP Sc in human plasma fractions spiked with sporadic CreutzfeldtJakob disease (sCJD) prions from human brain. CJD prions were found with very low-density lipoproteins (VLDL) and low-density lipoproteins (LDL) but not with high-density lipoproteins (HDL) or any other plasma component. In addition to b...

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Section: Prions In Bloodsupporting

confidence: 89%

Human prions and plasma lipoproteins

Safar

Wille

Geschwind

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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“…The hamster brain contains approximately 2.5 mg of total DNA/g, and brains taken from scrapie-infected hamsters at late-stage disease contain Ն10 10 ID 50 /g (6,36,26). The concentration of infectivity relative to hamster DNA is therefore 4 ϫ 10cutes [1]), then the number of ribosomal gene copies per 40 ng of DNA would increase proportionately.…”

Section: Discussionmentioning

confidence: 99%

“…The hamster brain contains approximately 2.5 mg of total DNA/g, and brains taken from scrapie-infected hamsters at late-stage disease contain Ն10 10 infectious doses which cause disease in 50% of animals inoculated (ID 50 )/g (6,36,26). Thus, a 40-ng sample of genomic DNA from hamster brain, which is the sample size used in these PCR experiments, can be calculated to contain 1.6 ϫ 10 5 ID 50 , or 1.1 ϫ 10 5 infectious doses (ID), since 1 ID 50 corresponds to 1.44 ID.…”

Section: Methodsmentioning

confidence: 99%

Absence of Spiroplasma or Other Bacterial 16S rRNA Genes in Brain Tissue of Hamsters with Scrapie

et al. 2006

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Spiroplasma spp. have been proposed to be the etiological agents of the transmissible spongiform encephalopathies (TSEs). In a blind study, a panel of 20 DNA samples was prepared from the brains of uninfected hamsters or hamsters infected with the 263K strain of scrapie. The brains of the infected hamsters contained >10 10 infectious doses/g. The coded panel was searched for bacterial 16S rRNA gene sequences, using primers selective for spiroplasma sequences, primers selective for mollicutes in general, and universal bacterial primers. After 35 PCR cycles, no samples were positive for spiroplasma or any other bacterial DNA, while control Spiroplasma mirum genomic DNA, spiked at 1% of the concentration required to account for the scrapie infectivity present, was readily detected. After 70 PCR cycles, nearly all samples yielded amplified products which were homologous to various bacterial 16S rRNA gene sequences, including those of frequent environmental contaminants. These sequences were seen in uninfected as well as infected samples. Because the concentration of scrapie infectivity was at a known high level, it is very unlikely that a bacterial infection at the same concentration could have escaped detection. We conclude that the infectious agent responsible for TSE disease cannot be a spiroplasma or any other eubacterial species.The identity of the causal agents of the transmissible spongiform encephalopathies (TSEs)-neurodegenerative diseases which include scrapie in sheep, Creutzfeldt-Jakob disease (CJD) in humans, bovine spongiform encephalopathy in cattle, and chronic wasting disease in deer and elk-has been a major source of controversy since these diseases were shown to be transmissible (41,43,17,38). Early experiments showing that TSE infectivity passed through bacterial filters (54) and was resistant to disinfection by common bacteriocides (27,10,41,42,9) seemingly eliminated bacteria as the agent. However, the discovery in the 1970s of spiroplasmas, very small, thermostable, wall-less, helical-fibrillar bacteria that pass through 0.2-m bacterial filters and show remarkable resistance to many common biocides including heat (47), provided a possible bacterial candidate for the infectious agent of TSEs. There have also been reports of spiral structures resembling spiroplasmas in brain tissue from CJD patients (2, 23, 39, 33), although other investigators have suggested these structures might be artifactual (24, 28). Moreover, no spiroplasmas could be cultured from CJD brain tissue (32). Intracranial inoculation of Spiroplasma mirum into new-born hamsters (31), suckling rats (5), or suckling mice (16) produces central nervous system disease, though not a progressive spongiform encephalopathy (16).More recently, Bastian et al. (3,4) have reported the presence of spiroplasma-specific 16S rRNA genes in brain tissue taken at autopsy/necropsy from TSE-infected humans and animals. PCR amplification of 16S rRNA genes, or "ribotyping," is a powerful method for detecting and identifying microbial agents in environm...

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“…fractionation, depth filtration and nanofiltration, were shown to be capable of eliminating TSE agents with an overall TSE reduction capacity of >12 LRF. Brain homogenate from scrapie-infected hamsters was used in these studies as a model for agents causing human TSE [10].…”

Section: Virus Elimination and Tse Clearance During Manufacture Of IVmentioning

confidence: 99%

“…It has previously been shown that IgG preparations with a high dimer content were poorly tolerated [5][6][7]. Nanofiltration was introduced as an additional physical virus removal step, which also effectively contributes to the removal of the transmissible spongiform encephalopathy (TSE) agents in experimental models [8][9][10]. Furthermore, IVIG-F10 contains no sucrose or other carbohydrates and its immunoglobulin A (IgA) content is greatly reduced.…”

Section: Introductionmentioning

confidence: 99%

Clinical Properties of a Novel Liquid Intravenous Immunoglobulin: Studies in Patients with Immune Thrombocytopenic Purpura and Primary Immunodeficiencies

Borte¹,

Davies²,

Touraine³

et al. 2004

Transfus Med Hemother

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Background: We have developed a novel liquid intravenous immunoglobulin (IVIG-F10). It consists of a nanofiltered 12% immunoglobulin G (IgG) solution, stabilized with nicotinamide, L-proline and L-isoleucine. The efficacy, tolerability, safety, and pharmacokinetics of this product were assessed in patients with chronic immune thrombocytopenic purpura (ITP) and primary humoral immunodeficiencies (PID). Patients and Methods: 33 chronic ITP patients with platelet counts of < 20 × 10⁹/l were treated with IVIG-F10 or Sandoglobulin at doses of 0.4 g/kg body weight on 5 consecutive days. The primary efficacy endpoint was an increase in platelet counts to ≧50 × 10⁹/l. Secondary endpoints were time to and duration of platelet response and regression of bleeding. 34 PID patients with X-linked agammaglobulinemia, common variable immunodeficiency or IgG subclass deficiency were treated for 6 months with IVIG-F10 or Sandoglobulin at doses of 0.3–0.8 g/kg, infused at 3- or 4-week intervals. The primary efficacy endpoint was the number of days absent from school/work. Secondary endpoints were feeling of well-being, days of hospitalization, and use of antibiotics. Results: In ITP patients, the primary endpoint was met by 12/16 patients on IVIG-F10 and by 12/17 patients on Sandoglobulin (p = 1.000). Results of the secondary endpoints were comparable in the two study groups. In the PID study, 10/17 patients on IVIG-F10 and 9/17 patients on Sandoglobulin missed days at school/work, with monthly mean absences of 0.7 and 0.6 days (p = 0.746), respectively. There were no significant differences in the outcome of the secondary endpoints. Pharmacokinetics showed constant peak and trough serum IgG levels in PID patients. The median half-life (t_1/2) of IgG was 33 days in the IVIG-F10 group and 41 days in the Sandoglobulin group. For anti-HBsAg, median t_1/2 values were shorter, i.e. 17 and 19 days for IVIG-F10 and Sandoglobulin, respectively. In the ITP study, adverse events related to study drug were suspected in 9 and 14 patients treated with IVIG-F10 and Sandoglobulin, respectively; in the PID study adverse events occurred in 8 and 9 patients, respectively. Viral safety was ascertained in both studies by serology supplemented with nucleic acid amplification testing. Serum levels of the stabilizers transiently increased after infusion of IVIG-F10, but were back to baseline at the following day. Conclusions: The clinical studies in patients with chronic ITP and PID showed that the efficacy, tolerability, safety, and pharmacokinetics of IVIG-F10 were comparable to the properties of Sandoglobulin.

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Partitioning of TSE infectivity during ethanol fractionation of human plasma

Cited by 51 publications

References 33 publications

Human prions and plasma lipoproteins

Human prions and plasma lipoproteins

Absence of Spiroplasma or Other Bacterial 16S rRNA Genes in Brain Tissue of Hamsters with Scrapie

Clinical Properties of a Novel Liquid Intravenous Immunoglobulin: Studies in Patients with Immune Thrombocytopenic Purpura and Primary Immunodeficiencies

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