Abstract 20: Apolipoprotein A-I Influences Regulatory T Cell Development and Proliferation in Homeostasis and Atherogenesis

Abstract

Apolipoprotein A-I (ApoAI) is the major protein component of HDL. HDL ApoAI is involved in the efflux of cholesterol from cells to maintain cellular cholesterol homeostasis. ApoAI also has anti-inflammatory properties. We previously showed that injecting mice fed an atherogenic diet with ApoAI decreased the number of activated CD4 lymphocytes. Since T regulatory lymphocytes (Treg) play a major role in inhibiting the immune response during atherosclerosis development, we wanted to determine if ApoAI influences Treg development, hypothesizing that ApoAI enhances Treg development. To test this hypothesis, we compared the numbers of Treg in ApoAI-/- mice to B6 mice, and found a 50% decrease in the numbers of Treg in the periaortic LNs (PaLN) of ApoAI-/- mice. BrdU labeling studies showed that ApoAI-/- Treg had a significant 30% reduction in proliferation, suggesting that in the absence of ApoAI and normal cholesterol homeostasis, Tregs have defective proliferation. Functionally, we discovered that ApoAI-/- Treg were significantly less suppressive than B6 Treg in reducing CD4 effector T cell proliferation, suggesting that ApoAI plays a role in both the development and function of Tregs. To determine if the addition of exogenous lipid-free ApoAI could rescue and promote Treg differentiation in ApoAI-/- mice, ApoAI-/- naïve T cells were incubated in vitro with TGFβ and exogenous ApoAI. Addition of ApoAI significantly increased development of naïve ApoA1-/- lymphocytes into Treg. To verify these results in vivo, we fed a novel Treg lineage tracker mouse (LT), Foxp3-YFP-Cre-Rosa26-RFP-ApoE-/- mice a western diet for 15 weeks and administered subcutaneous injections of ApoAI for the last 9 weeks of diet. These mice allow us to identify current functional Tregs and any exTregs that have lost active Treg function in vivo. We found that LT mice treated with ApoAI had a 37% decrease in exTregs and a concomitant 33% increase in current functional Tregs in the aorta. This was accompanied by decreased IFNγ and IL-17 production in PaLN, further confirming our in vitro findings that ApoAI promotes Treg development and function. In conclusion, we have identified a novel role for ApoAI by enhancing Treg development, emphasizing the immune properties of ApoAI for atheroprotection.