Table 2.
Assay and serum pool | Neutralization titer for virus |
||||
---|---|---|---|---|---|
HPV16 | HPV31 | HPV33 | HPV45 | HPV58 | |
L2-based assay | |||||
PBS, pool 1 | <50 | <50 | <50 | <50 | <50 |
PBS, pool 2 | <50 | <50 | <50 | <50 | <50 |
HPV16 VLP, pool 1 | 172,080 | <50 | <50 | <50 | <50 |
HPV16 VLP, pool 2 | 306,550 | <50 | <50 | <50 | <50 |
L2 11-88×5, pool 1 | 7,500 | 50 | 4,315 | 2,780 | 5,370 |
L2 11-88×5, pool 2 | 13,760 | 765 | 4,130 | 2,640 | 4,025 |
L2 11-88×5, pool 3 | 4,095 | <50 | <50 | 365 | 700 |
L2 11-88×5, pool 4 | 10,730 | 7,865 | 27,145 | 7,805 | 13,820 |
L2 11-88×5, pool 5 | 20,900 | 2,600 | 1,605 | 5,890 | 3,395 |
L2 11-88×5, pool 6 | 27,630 | 480 | 210 | 5,005 | 10,940 |
L2 11-88×5, pool 7 | 3,420 | 190 | 22,385 | 1,375 | 1,140 |
L1-based assay | |||||
PBS, pool 1 | <50 | <50 | <50 | <50 | <50 |
PBS, pool 2 | <50 | <50 | <50 | <50 | <50 |
HPV16 VLP, pool 1 | 7,600 | <50 | <50 | <50 | <50 |
HPV16 VLP, pool 2 | 60,605 | <50 | <50 | <50 | <50 |
L2 11-88×5, pool 1 | <50 | <50 | <50 | <50 | <50 |
L2 11-88×5, pool 2 | <50 | <50 | <50 | <50 | <50 |
L2 11-88×5, pool 3 | <50 | <50 | <50 | <50 | <50 |
L2 11-88×5, pool 4 | <50 | <50 | 50 | 50 | <50 |
L2 11-88×5, pool 5 | <50 | <50 | <50 | 50 | <50 |
L2 11-88×5, pool 6 | <50 | <50 | 50 | <50 | <50 |
L2 11-88×5, pool 7 | <50 | <50 | 150 | 50 | <50 |
P value, 1-7 (L2-based vs L1-based assay) | 0.001 | 0.011 | 0.012 | 0.002 | 0.001 |
Cross-neutralization titers obtained with “L1-based” and “L2-based” assays. Sera were obtained from mice that had been immunized with L2 (11-88×5) and shown to be immune to in vivo challenge with HPV16, -31, -45, or -58. Sera from 5 animals that underwent identical treatment were pooled and assessed for in vitro neutralization with the newly described L2-based neutralization assay or the “L1-based” assay. As specificity controls, pooled sera from HPV16 VLP-immunized animals were also assayed. The assay was performed in duplicate, and the average titer is shown. Triplicate experiments were not possible due to limited sample volume. The bottom row indicates the P value of the difference between the two groups of 7 pools for each virus type. Titers that were found to be <50 were assigned a value of 25 for statistical analysis. The P value is based on the Mann-Whitney test with Gaussian approximation.