Radio-immunoassay for formyl methionyl leucyl phenylalanine. II. Demonstration of an enterohepatic circulation of immunoreactive bacterial chemotactic peptides in man.
Bacterial chemotactic peptides (F-met-oligopeptides) are secreted by several species of commensal enteric bacteria and can be assayed by bioassay techniques in human colonic luminal fluid. We have previously demonstrated intestinal absorption and enterohepatic circulation of radiolabelled F-met peptides introduced into rat colon, and an eightfold increase in absorption and biliary excretion in rats with experimental colitis. This paper describes the application of a radio-immunoassay to measurements of formyl oligopeptides in human faecal dialysates, colonic and systemic venous blood and bile. All samples were fractionated by reverse-phase high performance liquid chromatography (HPLC) prior to assay. Immunoreactivity was found in faecal dialysates (5-700 nmol/L F-met-leu-phe equivalents) and bile samples (3-150 nmol/L) from normal subjects. After HPLC fractionation, up to five distinct peaks of immunoreactivity were identified. One of these co-chromatographed with authentic F-met-leu-phe; the others probably represented either closely related peptides or peptides of different chain lengths originating from the same F-met-leu-phe precursor protein. Colonic venous blood from two patients with ulcerative colitis contained immunoreactive peptide (10-30 nmol/L) and substantial immunoreactivity was found in ileostomy fluid and bile from two patients with primary sclerosing cholangitis. These results suggest the presence of an enterohepatic circulation of bacterial F-met oligopeptides in man and provide a basis for studies of the role of such pro-inflammatory peptides in patients with inflammatory bowel disease and associated hepatobiliary disorders.
Roberts
E C
EC
Wellcome Medical Research Institute, University of Otago Medical School, Dunedin, New Zealand.
Hobson
C H
CH
Anderson
R P
RP
Chadwick
V S
VS
D000818:Animals; D001646:Bile / Q000737:chemistry; D002630:Chemotactic Factors / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D004764:Enterohepatic Circulation / Q000502:physiology*; D005243:Feces / Q000737:chemistry; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000097:blood*; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods*; D051381:Rats
D002630:Chemotactic Factors; D009240:N-Formylmethionine Leucyl-Phenylalanine