Article Title: CTLA-4 Blockade, during HIV Virus-Like Particles Immunization, Alters HIV-Specific B-Cell Responses
Figure Lengend Snippet: CTLA-4 blockade amplifies serum HIV Gag- and Env-specific antibody responses. Post-prime serum was collected 13 days after the first VLP immunization; Post-boost 1 serum was collected 11–13 days after the second VLP immunization. Post-prime and Post-boost 1 time points analyzed serum from 10 mice per immunization group from Cohorts 1 and cohorts 2. The Post-boost 2 time point analyzed serum from the 15 mice (5 per group) in Cohort 2 and was collected 7 days after the third VLP immunization. ( A ) Total Env-specific IgG and ( B ) total Gag-specific IgG were analyzed through ELISA. Quantitative ELISAs with standard curves for IgG1, IgG2b, IgG2c, and IgG3 were performed on all serum samples from Cohorts 1 and 2 to detect HIV Env- and Gag-specific IgG subtypes. Shown at each of the following time points: ( C ) post-prime, ( D ) Env post-prime Gag, ( E ) post-boost 1 Env, ( F ) post-boost 1 Gag, ( G ) post-boost 2 Env, and ( H ) post-boost 2 Gag IgG subtypes. The results above are the cumulative results of two independent experiments. P-values were determined by one-way ANOVA and Tukey post-hoc analysis for multiple comparisons. A line on top of two groups indicates statistical differences between these two groups. **** p
Article Snippet: Quantitative IgG Subtype ELISAMicro-vinyl plates (96-well, Corning Costar, Corning, NY, USA) were coated with a standard curve of purified IgG1 (BD Pharminogen #554121, San Diego, CA, USA), IgG2b (eBioscience eBMG2b #14732-82, San Diego, CA, UAS), IgG2c (GeneTex #GTX35043, Irvine, CA, USA), or IgG3 (eBioscience #14-4742-82, San Diego, CA, USA) protein while remaining wells were coated with 0.5 μg/mL BaL gp120 recombinant protein (NIH AIDS reagent #4961, Germantown, MD, USA) in a sodium bicarbonate buffer and incubated overnight.
Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay