Aim To determine the effect of heat shock protein-65 (HSP65) on the anti-oxidant and anti-inflammatory properties of high density lipoprotein (HDL) in apolipoprotein E gene knocked-out (ApoE^－/－) mice.Methods Eight-week-old ApoE^－/－ mice were randomly divided into three groups: phosphate buffer as control group, 5 μg HSP65 group and 25 μg HSP65 group. The mice were boosted twice under the same protocol following three and six weeks. Sixteen weeks after the first immunization, serum lipids, paraoxonase-1 (PON-1) activity, myeloperoxidase (MPO) activity, high density lipoprotein inflammatory index (HII), the level of cytokines interleukin-10 (IL-10) and interferon-γ (IFN-γ) were assessed.Results With the increasing dose of HSP65, HDLC level, IL-10 expression and PON-1 activity reduced gradually, whereas HII, MPO activity, and the IFN-γ expression elevated. Compared with control group, markedly decreased HDLC, IL-10 level and PON-1 activity but a significant enhancement of the serum HII, MPO activity, and IFN-γ levels were resulted in 25 μg HSP65 group (all P<0.05).Conclusion HSP65 could induce inflammatory response and impair the anti-oxidant and anti-inflammatory properties of HDL, which was related to the dose of HSP65.