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Nm (expressed as millioptical density units [mOD]). (A) Blocking of virus entry with purified gCt (OE), gDt (s), or gHt-gL (F); (B) blocking of virus entry with gDt alone (s), gHt-gL (F), or a mixture of 40 nM gD (concentration which gave 50 inhibition of entry) with various concentrations of gHt-gL (!).FIG. 5. Immunoblot (Western blot) analysis of serum samples from rabbits immunized with gHt-gL
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Ed to nitrocellulose, and reacted with R136 (lane 1), R137 (lane 2), R138 (lane 3), or R139 (lane 4).precipitated with LP11, separated by SDS-PAGE, and analyzed by Western blotting, probing for gH (Fig. 3A, lane 1) and gL (Fig. 3A, lane 2) on individual nitrocellulose strips. Both proteins were detected, showing that the complex was reactive with LP11. Similar results were obtained in assays using
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Ed to nitrocellulose, and reacted with R136 (lane 1), R137 (lane 2), R138 (lane 3), or R139 (lane 4).precipitated with LP11, separated by SDS-PAGE, and analyzed by Western blotting, probing for gH (Fig. 3A, lane 1) and gL (Fig. 3A, lane 2) on individual nitrocellulose strips. Both proteins were detected, showing that the complex was reactive with LP11. Similar results were obtained in assays using
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Ed to nitrocellulose, and reacted with R136 (lane 1), R137 (lane 2), R138 (lane 3), or R139 (lane 4).precipitated with LP11, separated by SDS-PAGE, and analyzed by Western blotting, probing for gH (Fig. 3A, lane 1) and gL (Fig. 3A, lane 2) on individual nitrocellulose strips. Both proteins were detected, showing that the complex was reactive with LP11. Similar results were obtained in assays using
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Eb Avg secondary scorec Group mortalityI IIMock (PBS) gD gHt-gL Mock (PBS) gD gHt-gL10.5 5.5 2.8 22.3 5.8 4.16.5 0 0 19.6 05/10 0/10 0/9 5/5 0/5 0/FIG. 6. Blocking of HSV entry by rabbit antibodies to gHt-gL. (A) HSV1(hrR3) was incubated for 90 min at 37 with various concentrations of rabbit anti-gHt-gL serum R136, R137, R138, or R139, and the serum-virus mixture was added to Vero cell monolayers
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Th HSV-1(hrR3) (22) and measuring -galactosidase activity at 5 h postinfection. The neutralization titer represents the dilution of antiserum needed to reduce -galactosidase activity to 50 of the maximum seen with no serum added. b Virus infection was measured by plaque formation by HSV-1(KOS) on Vero cells, using the black plaque assay. The neutralization titer represents the dilution of antiser
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Th HSV-1(hrR3) (22) and measuring -galactosidase activity at 5 h postinfection. The neutralization titer represents the dilution of antiserum needed to reduce -galactosidase activity to 50 of the maximum seen with no serum added. b Virus infection was measured by plaque formation by HSV-1(KOS) on Vero cells, using the black plaque assay. The neutralization titer represents the dilution of antiser
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Days 3 to 8. Score range was 0 to 10.molecular-weight forms (Fig. 5B). The composition of these bands remains to be determined. All four sera exhibited significant titers of complement-independent HSV-1 neutralizing activity (Table 1). In addition, these sera also neutralized HSV-2, albeit at a much reduced potency. These results indicated that the immunizing protein had biologic activity. In addi