title: Opioid peptide-expressing leukocytes: identification, recruitment, and simultaneously increasing inhibition of inflammatory pain.
abstract: BACKGROUND
  Inflammatory pain can be effectively controlled by an interaction of opioid receptors on peripheral sensory nerve terminals with opioid peptides released from immune cells upon stressful stimulation. To define the source of opioid peptide production, we sought to identify and quantify populations of opioid-containing cells during the course of Freund's complete adjuvant-induced hind paw inflammation in the rat. In parallel, we examined the development of stress-induced local analgesia in the paw.
  
  
  METHODS
  At 2, 6, and 96 h after Freund's complete adjuvant inoculation, cells were characterized by flow cytometry using a monoclonal pan-opioid antibody (3E7) and antibodies against cell surface antigens and by immunohistochemistry using a polyclonal antibody to beta-endorphin. After magnetic cell sorting, the beta-endorphin content was quantified by radioimmunoassay. Pain responses before and after cold water swim stress were evaluated by paw pressure thresholds.
  
  
  RESULTS
  In early inflammation, 66% of opioid peptide-producing (3E7+) leukocytes were HIS48+ granulocytes. In contrast, at later stages (96 h), the majority of 3E7+ immune cells were ED1+ monocytes or macrophages (73%). During the 4 days after Freund's complete adjuvant inoculation, the number of 3E7+ cells increased 5.6-fold (P < 0.001, Kruskal-Wallis test) and the beta-endorphin content in the paw multiplied 3.9-fold (P < 0.05, Kruskal-Wallis test). In parallel, cold water swim stress-induced analgesia increased by 160% (P < 0.01, analysis of variance).
  
  
  CONCLUSIONS
  The degree of endogenous pain inhibition is proportional to the number of opioid peptide-producing cells, and distinct leukocyte lineages contribute to this function at different stages of inflammation. These mechanisms may be important for understanding pain in immunosuppressed states such as cancer, diabetes, or AIDS and for the design of novel therapeutic strategies in inflammatory diseases.
authors.affiliation: Klinik für Anaesthesiologie und operative Intensivmedizin, Universitätsklinikum Benjamin Franklin, Freie Universität Berlin, Germany. rittner@medizin.fu-berlin.de
authors.forename: H L
authors.initials: HL
authors.lastname: Rittner
authors.forename: A
authors.initials: A
authors.lastname: Brack
authors.forename: H
authors.initials: H
authors.lastname: Machelska
authors.forename: S A
authors.initials: SA
authors.lastname: Mousa
authors.forename: M
authors.initials: M
authors.lastname: Bauer
authors.forename: M
authors.initials: M
authors.lastname: Schäfer
authors.forename: C
authors.initials: C
authors.lastname: Stein
mesh_terms: D000698:Analgesia; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D005456:Fluorescent Dyes; D006412:Hematopoietic Stem Cells / Q000276:immunology; D007150:Immunohistochemistry; D018189:Immunomagnetic Separation; D007249:Inflammation / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology; D017493:Leukocyte Common Antigens / Q000302:isolation & purification; D007962:Leukocytes / Q000378:metabolism*; D008214:Lymphocytes / Q000276:immunology; D008297:Male; D018847:Opioid Peptides / Q000096:biosynthesis*; D010146:Pain / Q000503:physiopathology*; D010147:Pain Measurement; D011863:Radioimmunoassay; D051381:Rats; D017208:Rats, Wistar; D001615:beta-Endorphin / Q000097:blood
chemical_list: D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D018847:Opioid Peptides; D001615:beta-Endorphin; D017493:Leukocyte Common Antigens