Journal: JCI Insight

Article Title: Circumventing cellular immunity by miR142-mediated regulation sufficiently supports rAAV-delivered OVA expression without activating humoral immunity

doi: 10.1172/jci.insight.99052

Figure Lengend Snippet: AAV1 injections were carried out on day 0 followed by sera collection every 2 weeks for a 12-week period. (A) qPCR detection of rAAV vector genome copies (GCs) in spleen, heart, liver, and injected skeletal muscle (n = 2). (B and C) ELISA quantification of circulating OVA expression (B) and anti-OVA IgG1 (C) of mice i.m. injected with rAAV1.OVA or rAAV1.OVA.miR142BS vectors (1 × 1011 GCs/mouse, n = 10). Sera were collected throughout a 12-week period. Box plots with whiskers represent mean ± SD and maximum and minimum values (n = 5). **P < 0.01, ***P < 0.001, ANOVA with Tukey’s post hoc test.

Article Snippet: After washing 4 times, peroxidase-conjugated rabbit anti-OVA polyclonal antibody (200-4333-0100, Rockland Immunochemicals) (1:5000 diluted) was added and incubated for 1 hour at room temperature.

Techniques: Plasmid Preparation, Injection, Enzyme-linked Immunosorbent Assay, Expressing

Journal: JCI Insight

Article Title: Circumventing cellular immunity by miR142-mediated regulation sufficiently supports rAAV-delivered OVA expression without activating humoral immunity

doi: 10.1172/jci.insight.99052

Figure Lengend Snippet: Timeline for the AAV-redosing experiment (A) and the description of the rAAV serotype and OVA transgene specific for each injection (B). C57BL/6 male mice, 6 weeks old, were injected i.m. with either rAAV1.OVA or rAAV1.OVA.142BS (1 × 1011 GCs/mouse). The same mice were then redosed with either rAAV8.OVA or rAAV8.OVA.142BS (1 × 1011 GCs/mouse) i.m. 4 weeks after the first injection (n = 5). (C and D) ELISA quantification of circulating OVA expression (C) and anti-OVA IgG (D) in vector-injected mice. Sera were collected throughout a 12-week period. Box plots with whiskers correspond to mean ± SD and maximum and minimum values. P values were determined by ANOVA with Tukey’s post hoc test for pairwise comparison. Asterisks (*) denote test of significance between injection groups 1 and 3, and hashes (#) denote test of significance between injection groups 2 and 4. P values for longitudinal comparisons were determined by repeated measures with 2-way ANOVA. Daggers (†) denote test of significance. *,#P < 0.05, **,##P < 0.01, ***,###,†††P < 0.001.

Article Snippet: After washing 4 times, peroxidase-conjugated rabbit anti-OVA polyclonal antibody (200-4333-0100, Rockland Immunochemicals) (1:5000 diluted) was added and incubated for 1 hour at room temperature.

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Expressing, Plasmid Preparation