Neutralization of poliovirus by antibody-mediated polymerization

Brioen, Paul; Dekegel, D.; Boeyé, Albert

doi:10.1016/0042-6822(83)90159-9

Cited by 48 publications

(43 citation statements)

References 10 publications

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“…After incubation, the mixtures were centrifuged in order to pellet virus aggregates, but not individual virions or small complexes. At physiological ionic strength (Table 1, lines 1 to 3), more than 90~o of the virus radioactivity was pelleted as expected from previous results (Brioen et al, 1983;Thomas et al, 1985). When the ionic strength was reduced from p. = 0.13 to ~t = 0.002 (lines 4 to 6), the aggregates represented only about 50~o of the input when Ca 2+ or Mg 2÷ was present, and even less (25 ~) in the presence of EDTA.…”

Section: Discussionsupporting

confidence: 51%

“…Some monoclonal antibodies form stable complexes with single virions, whose infectivity is consequently reduced (Icenogle et al, 1983); our 35-1f4 antibody (Brioen et al, 1982) causes the virions to aggregate (Brioen et al, 1983), thereby lowering the infectivity of the virus preparation without irreversible damage to the virions (Thomas et al, 1985). In both mechanisms, the antibody remains bound to the (clusters of) neutralized virions.…”

Section: Discussionmentioning

confidence: 99%

“…1). It must be remembered, however, that the 35-1f4 antibody also fails to form stable complexes with individual, native virions (Brioen et al, 1983), so that the lack of binding to the 80S particles does not prove that these particles have lost the epitope recognized by antibody 35-1f4. The antigenicity of the 80S particles recovered from a sucrose density gradient was studied using Protein A-aided immunoprecipitation.…”

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confidence: 99%

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Hit-and-Run Neutralization of Poliovirus

Brioen

Rombaut

Boeyé

1985

Journal of General Virology

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SUMMARYAt physiological ionic strength, monoclonal antibody 35-1f4 has previously been shown to neutralize poliovirus type 1 by antibody-mediated polymerization. At low ionic strength, this antibody neutralized the virus by a hit-and-run mechanism: the virions were converted to non-infectious, empty capsids devoid of antibodies. These empty capsids resembled those formed by thermal denaturation of native polio virions in their sedimentation coefficient (80S), antigenicity (H) and isoelectric pH (6.3).Neutralizing antibodies appear to interact with poliovirus in different ways. Some monoclonal antibodies form stable complexes with single virions, whose infectivity is consequently reduced (Icenogle et al., 1983); our 35-1f4 antibody (Brioen et al., 1982) causes the virions to aggregate (Brioen et al., 1983), thereby lowering the infectivity of the virus preparation without irreversible damage to the virions (Thomas et al., 1985). In both mechanisms, the antibody remains bound to the (clusters of) neutralized virions. In the present paper, we show the existence of yet another mechanism of the hit-and-run type : the virion is disrupted to a noninfective, empty capsid and the antibody is released.The rate of neutralization of T-even phages has been shown to decrease with increasing ionic strength (Jerne & Skovsted, 1953;Cann & Clark, 1956). We are not aware of similar findings with animal viruses. As we will show in the present paper, the ionic strength influences not only the degree of poliovirus neutralization, but also the nature of the antibody-virion interaction.Nine solutions were prepared with three levels of ionic strength, the highest of which corresponded to physiological saline (Table 1). At each level, either Mg -'÷, Ca 2÷ or EDTA was present at a concentration of 0.1 mM. Radiolabelled poliovirus type 1 was reacted with monoclonal antibody 35-1f4 in each of these media. After incubation, the mixtures were centrifuged in order to pellet virus aggregates, but not individual virions or small complexes. At physiological ionic strength (Table 1, lines 1 to 3), more than 90~o of the virus radioactivity was pelleted as expected from previous results (Brioen et al., 1983;Thomas et al., 1985). When the ionic strength was reduced from p. = 0.13 to ~t = 0.002 (lines 4 to 6), the aggregates represented only about 50~o of the input when Ca 2+ or Mg 2÷ was present, and even less (25 ~) in the presence of EDTA. When the ionic strength was further reduced by omission of the basal salts mixture (lines 7 to 9), the sedimentable material was down to 19 to 27~ of the input radioactivity. The results clearly show that the ionic strength is a major factor determining the extent of virus aggregation. The additional influence of EDTA was only evident at I1 = 0-002.Neutralization experiments were carried out in PBS (150 mM-NaCI, 13 mM-KC1, 0.3 mMCaC12, 0.5 mM-MgC12, 10raM-sodium phosphate pH 7.3) or in 0.1 mM-EDTA pH 7.3. The residual infectivity was measured in both media, and the results are given in Table 2. Even though the vi...

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Section: Discussionsupporting

confidence: 51%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Hit-and-Run Neutralization of Poliovirus

Brioen

Rombaut

Boeyé

1985

Journal of General Virology

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“…It is known that the binding of PV with the neutralizing monoclonal antibody 35-1f4 can neutralize the virus by polymerization. The two paratopes of the 35-1f4 monoclonal antibody are binding on two different virion particles and not on two epitopes on the same virion [30]. In this way, large aggregates are formed which are too large to be injected into the capillary, which has a diameter of 50 mm or/and are sedimenting to the bottom of the tube whereby they cannot be injected anymore into the capillary.…”

Section: Resultsmentioning

confidence: 99%

Identification of poliovirions and subviral particles by capillary electrophoresis

et al. 2010

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Poliovirions, purified from infected cell extracts with anion-exchange chromatography, can be analyzed and identified by CE in untreated fused silica capillaries using UV detection. Other subviral particles can be eluted as well from the same infected cell extract using a higher salt concentration buffer on the ion-exchange chromatography. Virions can be identified because of their conversion into empty capsids upon heating at 561C. As a result of heating, the viral genome is released from the capsid. Here, we show that during this incubation some intermediate particles were found. The latter were identified by enzymatic peak shift analysis. The high salt concentration eluate subviral particles were analyzed with preincubation affinity CE together with their sensitivity for RNase and proteinase K treatment. Electropherograms of the higher salt concentration eluate display a mixture of at least four different subviral particles. One particle proved to have an [N1, H] antigenicity and was resistant to RNase and proteinase K digestion. The remaining particles were all sensitive to proteinase K treatment. This CE method proved to be valuable in the detection, identification and analysis of poliovirions and poliovirus particles offering an alternative powerful, cheap, fast and easy analysis method.

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“…Since each aggregate gives rise to only one p.f.u, reduced infectivity results. It was recently demonstrated (Icenogle et al, 1983;Brioen et al, 1983) that reductions in poliovirus survival, caused by neutralizing monoclonal antibodies, were directly proportional to the number of virus particles entering the aggregates.…”

confidence: 99%

A Hypothesis Accounting for the Effect of the Host Cell on Neutralization-resistant Virus

Kjellén

1985

Journal of General Virology

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SUMMARYEvidence is presented showing thata monkey anti-enterovirus 71 immune serum contains several antibody populations which differ in their mode of function. One population reduces infectivity, although inefficiently, by interactions at exposed antigenic sites and can be detected by measuring residual virus infectivity after mixtures of virus and antibody have been all0wed_.to interact. Another antibody population, which is unaffected by the immunosorbent Staphylococcus aureus (Cowan I strain), appears to attach to its antigenic site(s) only after interactions between enterovirus 71 and host cells have already begun. In view of the transience of (presumed) conformational changes in the invading viruses, demonstration of this type of antibody activity requires a particular host cell system. This second type of antibody neutralization could be detected on RD cells but not on green monkey kidney cells.In comparative plating experiments with partly neutralized vesicular stomatitis virus and echovirus 4, a marked difference in response was found between the two cell types used (Kjell6n & Schlesinger, 1959; Kjell6n &von Zeipel, 1984). The obvious dependence of plaque assay results upon the cells used may help to shed light on controversial questions regarding the nature of the interactions between virus and antibody that result in neutralization.Enterovirus 71 was used in the present study. The strain 52343, notorious for its large 'nonneutralizable' fraction on Vero and green monkey kidney (GMK) cells (von Zeipel, 1979) and the monkey hyperimmune serum, C-898, were kindly supplied by Dr G. von Zeipel (Stockholm, Sweden). Monolayers of cells were prepared and plaque assays performed as described previously (Kjell6n &von Zeipel, 1984). Simultaneous titrations of stocks of enterovirus 71 yielded a similar number of plaques (p.f.u./ml) on monolayers of either GMK cells (GMK-AH1 ; G/inalp, 1965) or rhabdomyosarcoma cells (RD;McAllister et al., 1969). Nine triplicate titrations of a stock of enterovirus 71 yielded on average 5.65 × 105 p.f.u./ml on RD cells and 5.89 x 105 on GMK-AH1 cells with a range of 0.55 to 1.50 in the ratio of these titres.To ascertain the neutralizing capacity of the heat-inactivated immune serum (IS), a suspension of about l0 s p.f.u, of enterovirus 71 was mixed with IS dilutions in phosphatebuffered saline (PBS) of 10 -2, 10 -3 and 10 -4 in a total volume of 2 ml; mixtures were then incubated for 2 h to complete virus-antibody combinations and seeded onto monolayers of GMK or RD cells. The hyperimmune serum neutralized virus infectivity, with a nonneutralizable fraction in all mixtures of about 10 -2 and 10 -1 of control virus for RD and GMK cells respectively. All mixtures showed surviving non-neutralizable fractions of virus which were independent of antibody concentration over the range studied, indicating excess of antibody at all dilutions (Dulbecco et al., 1956). The differences in surviving fractions of infectivity (SFI) between the two cell strains were statistically highly significan...

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Neutralization of poliovirus by antibody-mediated polymerization

Cited by 48 publications

References 10 publications

Hit-and-Run Neutralization of Poliovirus

Hit-and-Run Neutralization of Poliovirus

Identification of poliovirions and subviral particles by capillary electrophoresis

A Hypothesis Accounting for the Effect of the Host Cell on Neutralization-resistant Virus

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