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Cellular and humoral immune responses to heat shock protein 65 are both involved in promoting fatty-streak formation in LDL-receptor deficient mice

Jacob George, MD^*, Arnon Afek, MD, Boris Gilburd, PhD, Yehuda Shoenfeld, MD and Dror Harats, MD

^* Department of Cardiology and the Cardiovascular Research Laboratory, Tel Aviv Medical Center, Tel Aviv, Israel
Institute of Pathology, Sheba Medical Center, Tel Hashomer, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel
The Research Unit of Autoimmune Diseases, Sheba Medical Center, Tel Hashomer, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel
Institute of Lipid and Atherosclerosis Research, Sheba Medical Center, Tel Hashomer, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel

View larger version (12K): [in a new window]   Figure 1 Characterization of heat shock protein 65 (HSP65) reactivity of lymphocytes and immunoglobulin G (IgG) employed for the transfer experiments. Reactivity of lymph-node cells (A) and splenocytes (Spl) (B) was determined by thymidine incorporation, whereas IgG binding was defined by enzyme-linked immunosorbent assay. BSA = bovine serum albumin.

View larger version (12K): [in a new window]   Figure 2 Fatty-streak formation in mice immunized with heat shock protein 65 (HSP65) or bovine serum albumin (BSA). Sections from mice immunized and boosted twice with HSP65 or BSA were stained with oil-red O. Lesion area was assessed by an unbiased observer using a grid.

View larger version (109K): [in a new window]   Figure 3 Immunohistochemical detection of heat shock protein (HSP)60/65. Immunoglobulin G (IgG) was purified from the sera of mice immunized with HSP65. Purified IgG was cross-reactive with HSP60 employing enzyme-linked immunosorbent assay. The IgG from HSP65 immunized mice was used to immunostain aortic sinus sections from low density lipoprotein receptor-deficient mice with the Histomouse kit. (A) Positive staining of the endothelial cells; (B) a similar section stained with control mouse IgG.