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. 2016 Dec 20;12(12):e1006064. doi: 10.1371/journal.ppat.1006064

Fig 2. Primary Antigen-Specific CTL Responses Correlate with Adjuplex Dose and Tissue Inflammation.

Fig 2

C57BL/6 mice were vaccinated by intramuscular or subcutaneous injection of 100 μg ovalbumin in PBS alone or supplemented with 1, 5, 10 or 20% ADJ. 3–4 mice/group were sacrificed after 7 days. Injection sites were collected for routine histopathology, and spleens were collected for flow cytometry. Prior to FACS analysis, splenocytes were stimulated for 5 hours with SIINFEKL peptide in the presence of IL-2 and brefeldin-A. (A) Frequency of SIINFEKL-specific IFN-γ+ CD8 T cells among all splenocytes after IM vaccination. (B-F) Photomicrographs of histologic changes in the skeletal muscle at the injection site 7 days after IM injection at the indicated concentrations of Adjuplex. Images depict a progressive increase in the cellularity and extent of the inflammatory infiltrate at the injection site. (G) Frequency of SIINFEKL-specific IFN-γ+ CD8 T cells among all splenocytes after SQ vaccination. (H-L) Photomicrographs of histologic changes in the subcutaneous adipose tissue and skeletal muscle at the injection site 7 days after SQ injection at the indicated concentrations of Adjuplex. Images depict a progressive increase in the cellularity and extent of the inflammatory infiltrate at the injection site. Data represents two independent experiments.