Journal: Science Advances
Article Title: Lymph node–targeted DNA engages TBK1/IFN-I–driven innate immunity to induce potent T cell responses and durable memory in mice and NHPs
doi: 10.1126/sciadv.aec1416
Figure Lengend Snippet: C57BL/6J mice ( n = 5) were immunized as described in . Immunizations contained 5 μg of OVA protein and either 5 nmol AMP-DNA, 1 nmol AMP-CpG-7909, or the indicated comparator adjuvants (100 μg of alum; 10 μg of MPLA; 1:2 emulsions of IFA, AddaVax, and AddaS03). Cells were assayed 7 days post–booster dose. ( A ) Peripheral blood CD8 + T cells were assayed ex vivo for OVA-reactive TCRs using a SIINFEKL-specific tetramer. ( B and C ) Flow cytometry analysis of cytokine production by CD8 + (B) and CD4 + T cells (C) in peripheral blood. Shown are percentages of cytokine + cells among CD8 + or CD4 + T cells and representative flow cytometry dot plots of TNFα and IFNγ positive CD8 + T cells. ( D ) ELISpot analysis of splenocytes restimulated with OVA OLPs overnight. Shown is the frequency of IFNγ SFCs per 10 6 splenocytes. ( E and F ) Flow cytometry analysis of cytokine production by CD8 + (E) and CD4 + T cells (F) in perfused lung tissue. Representation same as (B) and (C). ( G to I ) Serum anti-OVA pan-IgG (G), IgG1 (H), and IgG2c (I) titers were determined against OVA protein. ( J ) Ratio of IgG2c:IgG1 antibody response from (H) and (I). Mock vaccines contained antigen only. Values depicted are means (SD). In panels where multiple groups are statistically compared to each other, both AMP-dT 50 and AMP-dA:dT 50 are significantly different to all other indicated groups as shown. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001, one-way ANOVA followed by Tukey’s post hoc analysis.
Article Snippet: Comparator adjuvants included Alhydrogel adjuvant 2% (alum, 100 μg, InvivoGen, catalog no. vac-alu), AS03-like squalene-based adjuvant (AddaS03, 1:2 dilution, InvivoGen, catalog no. vac-as03), MPLA (10 μg, InvivoGen, catalog no. vac-mpla), MF59-like adjuvant (AddaVax, 1:2 dilution, InvivoGen, catalog no. vac-adx), and IFA (1:2 dilution, InvivoGen, catalog no. vac-ifa).
Techniques: Ex Vivo, Flow Cytometry, Enzyme-linked Immunospot, Vaccines
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