title: Genetic variants of chemokine receptor CCR7 in patients with systemic lupus erythematosus, Sjogren's syndrome and systemic sclerosis.
abstract: BACKGROUND
  The chemokine receptor CCR7 is a key organizer of the immune system. Gene targeting in mice revealed that Ccr7-deficient animals are severely impaired in the induction of central and peripheral tolerance. Due to these defects, Ccr7-deficient mice spontaneously develop multi-organ autoimmunity showing symptoms similar to those observed in humans suffering from connective tissue autoimmune diseases. However, it is unknown whether mutations of CCR7 are linked to autoimmunity in humans.
  
  
  RESULTS
  DNA samples were collected from 160 patients suffering from connective tissue autoimmune disease (Sjogren's syndrome, n = 40; systemic lupus erythematosus, SLE, n = 20 and systemic sclerosis, n = 100) and 40 health subjects (n = 40). All participants in this study were of German descent. Samples were screened for single nucleotide polymorphisms (SNP) by sequencing the coding region of the CCR7 gene as well asthe exon flaking intron sites and parts of the regions encoding for the 5'- and 3'-UTR. CCR7 variants were rare. We identified six different sequence variants, which occurred in heterozygosis. The identified SNP were observed at position -60 C/T (observed 1x), +6,476 A/G (7x), +6,555 C/T (15x), +6,560 C/T (6x), +10,440 A/G (3x) and +11,475 C/A (1x). Four of these variants (+6,476 A/G, +6,555 C/T, +6,560 C/T and +10,440 A/G) display allelic frequencies between 1% and 5 % and were present in both patients and control groups. The variants +6,476 A/G, +6,555 C/T, +6,560 C/T are located in the intron 2, while the +10,440 A/G variant corresponds to a silent mutation in exon 3. The variants -60 C/T and +11,475 C/A which are located at the 5'-UTR and 3-UTR respectively, display allelic frequencies below 1%. No correlation between these variants and the autoimmune diseases investigated could be observed. However, reporter gene expression assay demonstrated that the mutation at the -60 C/T position in homozygosis leads to reduced luciferase activity.
  
  
  CONCLUSIONS
  These results suggest that variants of CCR7 gene occur at an extremely low frequency in the German population and that neither Sjogren's syndrome, systemic lupus erythematosus, nor systemic sclerosis are associated with these variants. Nevertheless, the decreased luciferase activity observed in cells transfected with the promoter region bearing the -60 C/T mutation suggests that this CCR7 variant could potentially lead to increased susceptibility to autoimmunity.
authors.affiliation: Institute of Immunology, Hannover Medical School, Hannover, Germany. danielkahlmann@yahoo.de <danielkahlmann@yahoo.de>
authors.forename: Daniel
authors.initials: D
authors.lastname: Kahlmann
authors.forename: Ana Clara Marques
authors.initials: AC
authors.lastname: Davalos-Misslitz
authors.forename: Lars
authors.initials: L
authors.lastname: Ohl
authors.forename: Frauke
authors.initials: F
authors.lastname: Stanke
authors.forename: Torsten
authors.initials: T
authors.lastname: Witte
authors.forename: Reinhold
authors.initials: R
authors.lastname: Förster
mesh_terms: D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005787:Gene Frequency; D017930:Genes, Reporter / Q000235:genetics; D020022:Genetic Predisposition to Disease / Q000235:genetics*; D005820:Genetic Testing; D005858:Germany; D006801:Humans; D008156:Luciferases; D008180:Lupus Erythematosus, Systemic / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D054400:Receptors, CCR7 / Q000235:genetics*; D012595:Scleroderma, Systemic / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012859:Sjogren's Syndrome / Q000235:genetics*
chemical_list: C515370:CCR7 protein, human; D017931:DNA Primers; D054400:Receptors, CCR7; D008156:Luciferases