Journal: NPJ Vaccines

Article Title: Plasmablast, memory B cell and T follicular helper cell responses after human papillomavirus vaccination: effect of dose number and age

doi: 10.1038/s41541-026-01408-w

Figure Lengend Snippet: HPV 16/18-specific IgG and IgM plasmablasts were enumerated from freshly isolated peripheral blood mononuclear cells at baseline (pre-vaccination), day 7 post-dose 1, and pre- and day 7 post-dose 2 or 3 of Gardasil 9 vaccination ( n = 20, 19, and 16 for the 4-8-, 9-14-, and 15-26-year-old groups, respectively). A Representative FluoroSpot readouts are shown for one participant in the 15–26-year-old group. Box and whisker plots show HPV 16-specific ( B ) IgG and ( C ) IgM secreting cells, with the lower, central, and upper lines denoting the minimum, median, and maximum values, respectively. Age groups are colour-coded, with each dot representing an individual. Statistical analyses evaluating the effects of dose number and age compared plasmablast numbers between baseline and post-dose 1, 2, or 3 timepoints. Paired one-way ANOVA with Dunnett’s adjustment and paired two-way ANOVA with Tukey’s adjustment were performed for pooled and age-stratified analyses, respectively. Statistically significant p-values ( p < 0.05) are highlighted in blue. Line graphs show the kinetics of HPV 16-specific ( D ) IgG and ( E ) IgM secreting cells across the four evaluated timepoints. yrs—years.

Article Snippet: Ninety-six-well plates were coated with HPV 16/18 VLPs (Merck Sharp and Dohme Corp., USA) at 4 °C overnight, washed and incubated with pre-stimulated PBMC for 20 h. During incubation, HPV 16/18-specific IgG antibodies secreted by B cells were captured by the HPV VLPs and the cells were removed by washing in PBS.

Techniques: Isolation, Whisker Assay

Journal: NPJ Vaccines

Article Title: Plasmablast, memory B cell and T follicular helper cell responses after human papillomavirus vaccination: effect of dose number and age

doi: 10.1038/s41541-026-01408-w

Figure Lengend Snippet: HPV 16/18-specific IgG Bmem were enumerated from freshly isolated peripheral blood mononuclear cells at baseline (pre-vaccination), day 14 post-dose 1, and pre- and day 14 post-dose 2 or 3 of Gardasil 9 vaccination ( n = 18, 19 and 17 for 4-8-, 9-14- and 15–26-year-olds, respectively). A Representative ELISpot readouts are shown for one participant in the 4–8-year-olds group. Box and whisker plots show ( B ) HPV 16- and ( C ) HPV 18-specific IgG Bmem responses, with the lower, central and upper lines denoting minimum, median and maximum values, respectively. Age groups are colour-coded with each dot representing an individual. Statistical tests to evaluate the effect of dose number and age compared Bmem numbers between baseline and post-dose 1, 2 or 3 timepoints. Paired One-way ANOVA with Dunnett’s adjustment and paired two-way ANOVA with Tukey adjustment were performed for pooled and age-stratified analyses, respectively. Statistically significant p values ( p < 0.05) are highlighted in blue. Line graphs show the kinetics of ( D ) HPV 16- and ( E ) HPV 18-specific IgG secreting cells across the four evaluated timepoints. yrs – years.

Article Snippet: Ninety-six-well plates were coated with HPV 16/18 VLPs (Merck Sharp and Dohme Corp., USA) at 4 °C overnight, washed and incubated with pre-stimulated PBMC for 20 h. During incubation, HPV 16/18-specific IgG antibodies secreted by B cells were captured by the HPV VLPs and the cells were removed by washing in PBS.

Techniques: Isolation, Enzyme-linked Immunospot, Whisker Assay

Journal: NPJ Vaccines

Article Title: Plasmablast, memory B cell and T follicular helper cell responses after human papillomavirus vaccination: effect of dose number and age

doi: 10.1038/s41541-026-01408-w

Figure Lengend Snippet: Frozen PBMCs were thawed and pre-stimulated in culture for 18 h with HPV 16 or HPV 18 VLPs, a mitogen (SEB) positive control, or cultured in medium only without stimulation (Medium). Stimulated cells were stained with the AIM antibody panel, and data were acquired on BD LSRFortessa III flow cytometer with gating performed in FlowJo. A Lymphocytes were first gated from all cells, followed by exclusion of dead cells and CD14+ monocytes. Total Tfh cells were then identified as FoxP3-CD4 + CD45RO + CXCR5+. B Activated cells were identified using AIM markers as OX40 + CD25 + , OX40 + PD-L1+ and PD-L1 + CD25+ across all stimulation conditions. Shown is a representative gating strategy from one subject in the 15–26-year-old group. VLPs - virus-like particles; SEB staphylococcal enterotoxin B; AIM activation-induced marker.

Article Snippet: Ninety-six-well plates were coated with HPV 16/18 VLPs (Merck Sharp and Dohme Corp., USA) at 4 °C overnight, washed and incubated with pre-stimulated PBMC for 20 h. During incubation, HPV 16/18-specific IgG antibodies secreted by B cells were captured by the HPV VLPs and the cells were removed by washing in PBS.

Techniques: Positive Control, Cell Culture, Staining, Flow Cytometry, Virus, Activation Assay, Marker

Journal: NPJ Vaccines

Article Title: Plasmablast, memory B cell and T follicular helper cell responses after human papillomavirus vaccination: effect of dose number and age

doi: 10.1038/s41541-026-01408-w

Figure Lengend Snippet: Activated HPV 16/18-specific Tfh cells were identified using an AIM flow cytometry assay with the phenotypes OX40 + CD25+, OX40 + PD-L1+ and PD-L1 + CD25+ within the total circulating Tfh cell pool. Box and whisker plots show activated HPV 16-specific Tfh cells based on A OX40 + CD25+, B OX40 + PD-L1+ and C PD-L1 + CD25+, and HPV 18-specific Tfh cells based on D OX40 + CD25+, E OX40 + PD-L1+ and F PD-L1 + CD25+. The lower, central, and upper lines indicate the minimum, median, and maximum values, respectively. Age groups are colour-coded, with each dot representing an individual. Statistical evaluation of the effects of dose number and age compared activated Tfh cell frequencies between baseline and post-dose 1, 2, or 3 timepoints. Unpaired one-way ANOVA with Dunnett’s adjustment and unpaired two-way ANOVA with Tukey adjustment were performed for pooled and age-stratified analyses, respectively. Statistically significant p values ( p < 0.05) are highlighted in blue. VLPs virus-like particles, SEB staphylococcal enterotoxin B, AIM activation-induced marker.

Article Snippet: Ninety-six-well plates were coated with HPV 16/18 VLPs (Merck Sharp and Dohme Corp., USA) at 4 °C overnight, washed and incubated with pre-stimulated PBMC for 20 h. During incubation, HPV 16/18-specific IgG antibodies secreted by B cells were captured by the HPV VLPs and the cells were removed by washing in PBS.

Techniques: Flow Cytometry, Whisker Assay, Virus, Activation Assay, Marker