Chromatography of Venezuelan Equine Encephalomyelitis Virus Strains on Calcium Phosphate

Pedersen, Carl E.; Slocum, Donald R.; Levitt, Neil H.

doi:10.1128/am.24.1.91-95.1972

Cited by 6 publications

(3 citation statements)

References 12 publications

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“…Since elution profiles are strongly influenced by changes in pH or gradient dimensions, it is difficult to compare our results with those reported by other investigators who have used similar, but not identical, techniques for alphaviruses. Pedersen et al (12) reported that the elution peaks for VEE strains eluted from calcium phosphate columns at pH 7.4 ranged from 0.25 to 0.32 M phosphate. Fleming (7) reported that SF virus strains were eluted from hydroxylapatite columns by 0.6 to 0.7 M phosphate at pH 8.0 to 8.2, similar to our results (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The applications of column chromatography methods, using calcium phosphate or its derivatives, to the purification and characterization of numerous viruses have been reported (4,5,7,(11)(12)(13). This paper describes procedures using the hydroxylapatite form of calcium phosphate to effect satisfactory separations of selected alphavirus strains including Western equine encephalomyelitis virus (WEE), Eastern equine encephalomyelitis virus (EEE), and Semliki Forest virus (SF) strains.…”

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confidence: 99%

“…Different conditions of pH and phosphate molarity were required for different alphaviruses. Previously published methods (12) for chromatographic separations of Venezuelan equine encephalomyelitis virus (VEE) on the brushite form of calcium phosphate were not useful for separating EEE or SF strains. In the present study hydroxylapatite was substituted for brushite since it was reported to afford better recoveries of some viruses, especially under alkaline conditions (5).…”

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Chromatographic Separations of Alphavirus Strains by Hydroxylapatite

Jahrling

Beall

1977

J Clin Microbiol

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Hydroxylapatite column chromatography methods were developed to characterize selected alphavirus populations. Different conditions of pH and phosphate molarity were required to obtain satisfactory elution profiles and separations for Western equine encephalomyelitis virus strains, compared with Eastern equine encephalomyelitis virus and Semliki Forest virus strains. Raising the pH of the buffers effected earlier elutions of all viruses. Selection of phosphate gradients with more gentle slopes and adjustment to the proper pH effected better separations of virus subpopulations. Elution profiles were not affected by 0.85% NaCl, 10% fetal calf serum, or 1% bovine serum albumin, which are common constituents of virus-stabilizing diluents. Passage of Western equine encephalomyelitis and Semliki Forest viruses in BHK-21, Vero, or duck embryo cell cultures or in suckling mouse brains did not usually affect elution profiles, unless passage also resulted in a shift in the plaque size marker. Essentially all infectious virus applied to the column was recoverable in appropriate fractions. This permitted accurate determinations of heterogeneity within alphavirus populations. For Western equine encephalomyelitis large-plaque (LP) and small-plaque (SP) virus populations, it was possible to detect ratios of 1 LP in a population of 10 6 SP, and 1 SP in 10 3 LP by using linear phosphate gradients. When stepwise elution procedures were used, it was possible to detect ratios of 1 SP in a population of 10 5 LP. Hydroxylapatite column chromatography therefore appears to be a useful tool for characterizing alphaviruses and for isolating minority subpopulations of viruses of biological or epidemiological importance from apparently homogeneous virus stocks.

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Chromatographic Separations of Alphavirus Strains by Hydroxylapatite

Jahrling

Beall

1977

J Clin Microbiol

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Selective Clearance of a Benign Clone of Venezuelan Equine Encephalitis Virus from Hamster Plasma by Hepatic Reticuloendothelial Cells

Jahrling

Gorelkin

1975

Journal of Infectious Diseases

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A benign small-plaque clone of Venezuelan equine encephalitis virus was efficiently removed from the blood of inoculated hamsters by adsorption to cells of the hepatic reticuloendothelial system. More than 99% of infectious small-plaque virus, intrinsically labeled with 32P, was cleared from the blood within 30 min of inoculation; 47.6% of the 32P-labeled small-plaque virus inoculum was concentrated in the liver. In contrast, only 0.8% of a virulent large-plaque clone of the virus was cleared from the blood. The affinity of small-plaque virus for liver tissue was confirmed by electron microscopy, since inoculated small-plaque virions were readily visualized in phagocytic vacuoles of hepatic endothelial and Kupffer cells, where they appeared to be undergoing degradation. In contrast, large-plaque virus was not visualized in the liver. A critical determinant of virulence for viruses that do not replicate in hepatic reticuloendothelial cells may be the efficiency with which they are removed from the blood by adsorption to such cells.

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Effects of Antigen-Antibody Complexes on the Primary Immune Response in Rhesus Monkeys

et al. 1974

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A model for the enhancement of the primary humoral immune response of rhesus monkeys to marginal or weakly antigenic vaccines is presented. Our procedure used the complexing of formalin-inactivated Venezuelan equine encephalomyelitis (VEE) virus vaccine with specific, homologous, immune gamma globulin (IgG) at equivalence. Equivalence was determined by combining the concentrated, isotopically labeled ( 3 H) virus with various concentrations of specific Sephadex-fractionated IgG. Enhancement of VEE virus antibody response in monkeys was obtained from preparations containing a marginal concentration of antigen that was complexed at equivalence with homologous IgG as compared with antigen alone. Protection in Swiss mice closely paralleled the antibody response pattern observed in monkeys, since complexes at equivalence provided 30 to 50% greater protection against challenge than antigen alone.

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Chromatography of Venezuelan Equine Encephalomyelitis Virus Strains on Calcium Phosphate

Cited by 6 publications

References 12 publications

Chromatographic Separations of Alphavirus Strains by Hydroxylapatite

Chromatographic Separations of Alphavirus Strains by Hydroxylapatite

Selective Clearance of a Benign Clone of Venezuelan Equine Encephalitis Virus from Hamster Plasma by Hepatic Reticuloendothelial Cells

Effects of Antigen-Antibody Complexes on the Primary Immune Response in Rhesus Monkeys

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