Enhanced fatty streak formation in C57BL/6J mice by immunization with heatshock protein-65

Recent data suggest that the immune system is involved in atherogenesis. Th us, interest has been raised as to the possible antigens that could serve a s the initiators of the immune reaction. In the current work, we studied th e effects of immunization with recombinant heat shock protein-65 (HSP-65) a nd HSP-65-rich Mycobacterium tuberculosis (MT) on early atherogenesis in C5 7BL/6J mice fed either a normal chow diet or a high-cholesterol diet (HCD). A rapid, cellular immune response to HSP-65 was evident in mice immunized with HSP-65 or with MT but not in the animals immunized with phosphate-buff ered saline (PBS) alone. Early atherosclerosis was significantly enhanced i n HCD-fed mice immunized with HSP-65 (n=10; mean aortic lesion size, 45 417 +/-9258 mu m(2)) or MT (n=15; 66 350+/-6850 mu m(2)) compared with PBS-inje cted (n=10; 10 028+/-3599 mu m(2)) or nonimmunized (n=10; 9500+/-2120 mu m( 2)) mice. No fatty streak lesions were observed in mice fed a chow diet reg ardless of the immunization protocol applied. Immunohistochemical analysis of atherosclerotic lesions from the HSP-65- and MT-immunized mice revealed infiltration of CD4 lymphocytes compared with the relatively lymphocyte-poo r lesions in the PBS-treated or nonimmunized mice. Direct immunofluorescenc e analysis of lesions from HSP-65- and MT-immunized mice fed an HCD exhibit ed extensive deposits of immunoglobulins compared with the fatty streaks in the other study groups, consistent with the larger and more advanced lesio ns found in the former 2 groups. This model, which supports the involvement of HSP-65 in atherogenesis, furnishes a valuable tool to study the role of the immune system in atherogenesis.