SummarySince a dysregulated synthesis of tumor necrosis factor a (TNF-a) may be involved in the pathogenesis of autoimmune diseases, it was ofinterest to precisely locate the recently reported Ncol restriction fragment length polymorphism (RFLP) of the TNF-a region . However, by mapping of 56.8 kb of overlapping cosmid clones and direct sequencing, we could localize the polymorphic Ncol restriction site within the first intron of the TNF-0 gene and not in the TNF-a gene. To study whether regulatory mechanisms are affected by this polymorphism, we analyzed the TNF-a/TNF-0 production of phytohemagglutinin-stimuated peripheral blood mononuclear cells ofindividuals homozygous for the TNF-0 Ncol RFLP by ELISA and concomitant Northern blot analysis. On days 2-4 after stimulation with mitogen, the TNFB*1 allele corresponding to a 5.3-kb Ncol fragment presented with a significantly higher TNF-0 response. A mRNA analysis demonstrated that higher protein levels of TNF-0 correlate also with increased amounts ofTNF-0 transcripts . No allelic association was found in respect to TNF-a production . To further investigate a possible allelic influence on transcription, we determined the DNA sequence of 2 kb ofthe 5' portion of our cloned TNFB*2 allele and compared it with the available TNF-0 sequences. By computer-aided recognition motif search ofDNA binding factors, we report putative binding sites conserved between mouse and man in the 5' flanking region as well as in intron 1 ofthe TNF-0 gene, found also in other cytokine promoter sequences. In addition, by polymerase chain reaction amplification and sequencing of 740 by of the 5' part of TNF-0 of individuals typed homozygously for the Ncol RFLP, we could show that amino acid position 26 is conserved as asparagine in the TNFB*1 and as threonine in the TNFB* 2 sequence. A previously reported, EcoRI RFLP in the 3' untranslated region of TNF-0 does not segregate with either of the two alleles. Thus, four TNFB alleles can de defined at the DNA level .

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Our findings demonstrate that autoantibodies in CP target epitopes on both extra- and intracellular domains of BP180 and highlight the importance of testing for both IgG and IgA reactivity in these patients' sera.

II-2

Background Lupus erythematosus tumidus (LET) is a rare disease which was first described in 1909 but has not always been considered as a separate entity of cutaneous lupus erythematosus (CLE) in the international literature. Objectives To compare characteristic features of different subtypes of CLE and to analyse whether LET can be distinguished as a separate entity in the classification system of the disease. Methods The study involved 44 patients with CLE, including 24 patients with LET, 12 with discoid lupus erythematosus (DLE) and eight with subacute CLE (SCLE), from two centres in Germany. A core set questionnaire and an SPSS database were designed to enable a consistent statistical analysis. Results Location of skin lesions did not differ significantly between the CLE subtypes; however, the activity score was significantly lower in LET than in DLE (P < 0.01), and the damage score was significantly lower in LET than in SCLE (P < 0.01) and DLE (P < 0.01). Photosensitivity and antinuclear antibodies were confirmed to be different in LET compared with SCLE and DLE but without statistical significance. Moreover, histological analysis of skin biopsy specimens showed that abundant mucin deposition is significantly more present in LET compared with SCLE (P < 0.01) and DLE (P < 0.01) while prominent interface dermatitis and alteration of hair follicles were absent in LET. Conclusions Several significant differences were found between LET and other subtypes of CLE with regard to clinical, histological and laboratory parameters. These data strongly indicate that LET should be defined as a separate entity in the classification of CLE.