Accumulating monocytes in the vasculature of rat renal allografts: phenotype, cytokine, inducible no synthase, and tissue factor mRNA expression.
BACKGROUND
Necrotic patches and hemorrhagic lesions develop in the renal tissue between day 4 and day 5 after transplantation of fully allogeneic DA rat kidneys to LEW recipients. These lesions are at least in part due to destruction and obstruction of blood vessels. Damage of graft endothelial cells and blood coagulation are likely to be mediated by intravascular graft leukocytes. However, this cell population has not been thoroughly characterized before.


METHODS
We perfused untreated control kidneys, renal isografts, and allografts on day 4 after transplantation with phosphate-buffered saline/ethylenediaminetetraacetic acid to harvest leukocytes from both the blood stream as well as from the marginal intravascular pool. The mRNA expression of typical products of activated monocytes was analyzed in reverse-transcriptase polymerase chain reaction experiments. Graft monocytes were purified and their immunophenotype was investigated by flow cytometry.


RESULTS
Allograft rejection led to a 10-fold increase in the number of intravascular graft leukocytes compared to isografts. A mean number of about 100x10(6) leukocytes was harvested from a single allogeneic kidney, about 73% of these cells were monocytes and most of them displayed an activated phenotype. Compared to isografts, intravascular allograft leukocytes displayed an increased expression of tumor necrosis factor-alpha, inducible NO synthase and tissue factor.


CONCLUSIONS
Our study shows that large numbers of activated monocytes accumulate inside allograft vessels. As they express genes the products of which might damage the allograft by inducing cell death or thrombosis, we speculate that they directly participate in allograft destruction.
Grau
V
V
Institute of Anatomy and Cell Biology and Institute of Immunology, Philipps University Marburg, Germany.
Stehling
O
O
Garn
H
H
Steiniger
B
B
D000818:Animals; D016207:Cytokines / Q000235:genetics*; D015870:Gene Expression; D007668:Kidney / Q000098:blood supply; D016030:Kidney Transplantation / Q000502:physiology*; D007958:Leukocyte Count; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008297:Male; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011917:Rats, Inbred Lew; D011919:Rats, Inbred Strains; D012077:Renal Artery / Q000166:cytology; D012079:Renal Circulation / Q000502:physiology*; D012082:Renal Veins / Q000166:cytology; D013925:Thromboplastin / Q000235:genetics*; D013997:Time Factors
D016207:Cytokines; D012333:RNA, Messenger; D013925:Thromboplastin; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat