Purification of porcine aminoterminal propeptide of type III procollagen from lymph and use for lymphatic clearance studies in pigs.
To investigate the lymphatic transport of the aminoterminal propeptide of type III procollagen (PIIINP) we established a thoracic duct-venous shunt in 6 pigs. Porcine PIIINP was purified, characterised, and compared with human PIIINP to ensure the suitability of the radioimmunoassay of human PIIINP for measurements in pigs. SDS-PAGE and radioimmunoinhibition assays show human and porcine PIIINP to be similar, thus indicating that the assay of human PIIINP is also reliable for determinations on pig serum and lymph. Intact PIIINP, as identified by gel filtration, accounted for 60% and 40% of the total PIIINP immunoreactivity in lymph and serum, respectively. The higher amount of total immunoreactivity and proportion of intact PIIINP in lymph compared with serum support the hypothesis that intact PIIINP is transported from peripheral tissue into the circulation by lymph. Two days after the shunt was established, the lymph was collected quantitatively hour-by-hour for 24 h. The flow was higher during the light periods than in the dark (p less than 0.01). The PIIINP concentration varied inversely with the flow, being higher in the dark hours (p less than 0.03). However, the total collected amount of PIIINP in lymph did not differ during the light and dark periods. Serum PIIINP remained unchanged over the 24 h. The lymphatic clearance of total PIIINP immunoreactive components was 6.2 ml serum/min and the lymphatic clearance of intact PIIINP was 9.1 ml serum/min, equal to 7 and 10 times the plasma volume/24 h, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
Jensen
L T
LT
Department of Rheumatology, University of Copenhagen, Hvidovre Hospital, Denmark.
Risteli
J
J
Nielsen
M D
MD
Henriksen
J H
JH
Olesen
H P
HP
Risteli
L
L
Lorenzen
I
I
D000818:Animals; D001692:Biological Transport; D005260:Female; D008196:Lymph / Q000737:chemistry*; D008208:Lymphatic System / Q000502:physiology*; D010446:Peptide Fragments / Q000302:isolation & purification / Q000493:pharmacokinetics; D011347:Procollagen / Q000302:isolation & purification / Q000493:pharmacokinetics; D013552:Swine / Q000378:metabolism*; D013897:Thoracic Duct
D010446:Peptide Fragments; D011347:Procollagen; C041876:procollagen Type III-N-terminal peptide