Tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, is believed to play an important role in multiple sclerosis (MS) pathogenesis. A bi-allelic polymorphism in the TNF-alpha promoter region (TNF alpha-308), has been reported to influence levels of TNF-alpha production. In the present study, we investigated the TNF alpha-308 polymorphism in 93 patients with MS, 17 patients with optic neuritis (ON) and 95 healthy individuals using an allele-specific PCR technique. Allelic genotype was compared with TNF-alpha mRNA expression levels and HLA class II phenotypes. No significant difference regarding the TNF alpha-308 polymorphism was observed between MS patients and controls. Specifically, the less common allele, TNF2, which is associated with higher expression levels of TNF-alpha, was somewhat less frequent among MS patients. In fact, analysis of 19 patients homozygous for the MS associated HLA-DR-DQ haplotype HLA-Dw2 showed that this haplotype does not carry the TNF2 allele. In addition, in 47 patients, the TNF-alpha alleles did not correlate with expression levels measured as numbers of TNF-alpha expressing cells. Thus, we found no evidence for an important role of TNF alpha-308 polymorphism for genetic susceptibility to MS.