Research Article

Comparison of Oral Tolerance to ApoB and HSP60 Peptides in Preventing Atherosclerosis Lesion Formation in Apob48/Ldlr Mice

Figure 2

Oral tolerance to peptides reduces the proliferative response of the splenocytes. Splenocytes were collected 3 days and at the end of the study after 10 weeks of high fat diet feed to induce atherosclerosis. (a) Splenocytes ( cells) were stimulated with either ApoB peptide (10 μg/mL), HSP60 peptide (10 μg/mL), or (concavalin A (10 μg/mL) in vitro for 72 h. Cell Proliferation was measured by Alamar blue reduction as quantified by measurement of fluorescence at 560 nm/590 nm. Splenocytes from each animal were plated in triplicates. ( animals/group). Values represent Mean ± SEM. (b) Splenic effector cells were generated from ApoB48/LDLr−/− mice immunized subcutaneously with the peptides. Treg cells were purified from spleen of ApoB- and HSP60-treated mice at the end of the study (after 10 weeks of high fat diet feed to induce atherosclerosis). Addition of purified Treg cells from oral tolerant mice is indicated at different ratios to effector cells. Proliferation of effector-cell alone is indicated by the black bar; proliferation index represents the percentage carboxyfluorescein succinimidyl ester reduction in culture stimulated with ApoB or HSP60 peptides (10 μg/mL) relative to unstimulated culture. , , .