Analysis of immunosuppressive proteins in serum of liver-transplanted rats by using an anti-LSF-1 affinity column.
Liver suppressor factor one (LSF-1) is a 40-kDa immunosuppressive protein in the serum of rats 60 days after orthotopic liver transplantation (OLT) between the nonrejector combination of DA donors into PVG recipients. In the present study, the purification of proteins from rat OLT serum taken 60 days after transplantation was performed by affinity chromatography using the anti-LSF-1 polyclonal antibody (pAb). The assessment of column eluates using anti-LSF-1 and OLT serum was studied using rat heart and liver transplantation models. Rejection was not suppressed by the administration of OLT serum in heart or liver allografts. However, heart allografts treated with peak eluates (450 micrograms single shot im, dissolved in Intralipos) taken from the affinity OLT serum survived significantly longer than untreated rats (median = 36.5 days; n = 7 vs 6.5 days; n = 5, respectively, P = 0.011). The same treatment with anti-LSF-1 column eluates also prolonged liver allografts significantly (>200 days) than those in either the untreated group (median = 11 days; n = 7) or those which received only Intralipos (median = 10.5 days; n = 5, P = 0.019). Subsequent analysis of the N-terminal sequences of some of the proteins which were eluted from the affinity column revealed that the homology of a 30-kDa protein was identical to hemoglobin alpha-chain, a 59-kDa protein to granulocyte inhibitory factor, a 70-kDa and a 90-kDa to albumin and its precursor, respectively. Although the specific immunosuppressive component has not been isolated, our results suggested that the anti-LSF-1 column can extract immunosuppressive moiety of LSF-1 from OLT serum.
Kobayashi
S
S
First Department of Surgery, Juntendo University, Tokyo, Japan. kshig@med.juntendo.ac.jp
Goto
S
S
Lord
R
R
Shimizu
Y
Y
Swanson
C
C
Vari
F
F
Edwards-Smith
C
C
Chiba
S
S
Pan
T L
TL
Chen
C L
CL
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002846:Chromatography, Affinity; D016172:DNA Fingerprinting; D004591:Electrophoresis, Polyacrylamide Gel; D016027:Heart Transplantation / Q000502:physiology; D007166:Immunosuppressive Agents / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D016031:Liver Transplantation / Q000502:physiology*; D008297:Male; D011506:Proteins; D051381:Rats; D011919:Rats, Inbred Strains; D017386:Sequence Homology, Amino Acid
D000906:Antibodies; D007166:Immunosuppressive Agents; D011506:Proteins; C105479:liver suppressor factor 1