Degenerated human intervertebral discs contain autoantibodies against extracellular matrix proteins.

Degeneration of intervertebral discs (IVDs) is associated with back pain and elevated levels of inflammatory cells. It has been hypothesised that discogenic pain is a direct result of vascular and neural ingrowth along annulus fissures, which may expose the avascular nucleus pulposus (NP) to the systemic circulation and induce an autoimmune reaction. In this study, we confirmed our previous observation of antibodies in human degenerated and post-traumatic IVDs cultured in vitro. We hypothesised that the presence of antibodies was due to an autoimmune reaction against specific proteins of the disc. Furthermore we identified antigens which possibly trigger an autoimmune response in degenerative disc diseases. We demonstrated that degenerated and post-traumatic IVDs contain IgG antibodies against typical extracellular proteins of the disc, particularly proteins of the NP. We identified IgGs against collagen type II and aggrecan, confirming an autoimmune reaction against the normally immune privileged NP. We also found specific IgGs against collagens types I and V, but not against collagen type III. In conclusion, this study confirmed the association between disc degeneration and autoimmunity, and may open the avenue for future studies on developing prognostic, diagnostic and therapy-monitoring markers for degenerative disc diseases.

Pair-wise detection of eight common DNA polymorphisms of the human lipoprotein lipase gene by PCR-RFLP. Findings of the Olivetti Heart Study.

We developed a rapid procedure to analyse simultaneously two different DNA polymorphisms of the human LPL gene by PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The method involves PCR amplification of the gene fragments encompassing two polymorphic sites, direct digestion in the same PCR-tube of the amplification mixture with two restriction enzymes, and the analysis of the resulting DNA fragments by gel electrophoresis. In 422 participants of the 1994 follow-up examination of the Olivetti Heart Study, a total of eight common LPL gene polymorphisms have been analysed in pairs by this procedure: -93 T/G and D9N; V108V and T361T; N291S and PvuII; HindIII and S447X. Two of these polymorphisms (V108V and T361T) were analysed for the first time. This method is suitable for the routine analysis of clinical samples of varying DNA content and practically halves the times and costs of screening for these LPL polymorphisms.