title: Skin fibroblasts from individuals with Chediak-Higashi Syndrome (CHS) exhibit hyposensitive immunogenic response.
abstract: BACKGROUND
  Chediak-Higashi Syndrome (CHS) is a rare autosomal recessive disease characterized by immunodeficiency, oculocutaneous albinism, neurological dysfunction, and early death. Individuals with CHS present with increased susceptibility to infections of the skin, upper-respiratory tract, gastrointestinal tract, and oral tissues. Classical CHS is caused by mutations in the gene encoding lysosomal trafficking regulator (LYST). Although defects in cytotoxic T cell lytic secretory granule secretion and neutrophil phagocytosis are suggested to contribute to the immunodeficiency in CHS, the underlying molecular mechanisms are unknown. We hypothesized that skin fibroblasts from CHS subjects exhibit impaired immune response due to defective trafficking of inflammatory factors.
  
  
  RESULTS
  Primary skin fibroblasts from CHS subjects or healthy controls were assessed for genes encoding inflammatory response factors using PCR array. At baseline, we found CD14, IL1R1 and TLR-1 were down-regulated significantly (≥2 fold change) and the genes encoding TLR-3, IL-1β and IL-6 were up-regulated in CHS cells compared to control cells. When challenged with E. coli lipopolysaccharide (LPS), CHS cells were less responsive than control cells, with only 8 genes significantly up-regulated (3-68 fold change) compared to baseline values, whereas 28 genes in control cells were significantly up-regulated at a much higher magnitude (3-4,629 fold change). In addition, 50% of the genes significantly up-regulated in LPS-treated control cells were significantly lower in LPS-treated CHS cells. IL-6, a fibroblast-derived proinflammatory cytokine essential for fighting infections was significantly lower in culture media of CHS cells with or without LPS. Furthermore, Western blot and immunofluorescent staining revealed that TLR-2 and TLR-4 were diminished on cell membranes of CHS cells and dissociated from Rab11a.
  
  
  CONCLUSIONS
  For the first time, results from our study indicate defective trafficking of TLR-2 and TLR-4 contributes to the hyposensitive response of CHS skin fibroblasts to immunogenic challenge, providing a potential therapeutic target for clinical intervention in CHS.
authors.affiliation: NIH/NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD, USA. le.wang@nih.gov.
authors.forename: Le
authors.initials: L
authors.lastname: Wang
authors.affiliation: NIH/NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD, USA. kamilark@yahoo.com.br.|University of Campinas - Piracicaba Dental School, Piracicaba, Sao Paulo, Brazil. kamilark@yahoo.com.br.
authors.forename: Kamila Rosamilia
authors.initials: KR
authors.lastname: Kantovitz
authors.affiliation: NIH/NHGRI - National Human Genome Research Institute, Bethesda, MD, USA. andrew.cullinane@nih.gov.
authors.forename: Andrew Robert
authors.initials: AR
authors.lastname: Cullinane
authors.affiliation: NIH/NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD, USA. fhnociti@gmail.com.|University of Campinas - Piracicaba Dental School, Piracicaba, Sao Paulo, Brazil. fhnociti@gmail.com.
authors.forename: Francisco Humberto
authors.initials: FH
authors.lastname: Nociti
authors.affiliation: NIH/NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD, USA. brian.foster@nih.gov.
authors.forename: Brian Lee
authors.initials: BL
authors.lastname: Foster
authors.affiliation: NIH/NHGRI - National Human Genome Research Institute, Bethesda, MD, USA. joseph.roney@nih.gov.
authors.forename: Joseph Concepcion
authors.initials: JC
authors.lastname: Roney
authors.affiliation: NIH/NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD, USA. anne.tran2@nih.gov.
authors.forename: Anne Bich
authors.initials: AB
authors.lastname: Tran
authors.affiliation: NIH/NHGRI - National Human Genome Research Institute, Bethesda, MD, USA. wintrone@mail.nih.gov.
authors.forename: Wendy Jewell
authors.initials: WJ
authors.lastname: Introne
authors.affiliation: NIH/NIAMS - National Institute of Arthritis and Musculoskeletal and Skin Diseases, Bethesda, MD, USA. martha.somerman@nih.gov.
authors.forename: Martha Joan
authors.initials: MJ
authors.lastname: Somerman
mesh_terms: D002478:Cells, Cultured; D002609:Chediak-Higashi Syndrome / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology*; D005260:Female; D005347:Fibroblasts / Q000276:immunology* / Q000473:pathology; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D055261:Immunogenetic Phenomena / Q000235:genetics / Q000276:immunology*; D007153:Immunologic Deficiency Syndromes / Q000175:diagnosis / Q000235:genetics / Q000276:immunology; D008297:Male; D012867:Skin / Q000276:immunology* / Q000473:pathology