Different effects of sandimmun and neoral on migration of lymph cells to allograft and lymphoid tissue.
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Animals↗
Biomedical subjects
W L Olszewski
Publications and source records attributed to W L Olszewski.
Rapid reconstitution of a lymphoid population of lethally irradiated rats following vascularized bone marrow transplantation is associated with engraftment of donor and stimulation of host hematopoietic cells.
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Animals↗
Concentrations of electrophoretic and size subclasses of apolipoprotein A-I-containing particles in human peripheral lymph.
When cultured cells are exposed to plasma, the initial acceptors of unesterified cholesterol are small lipid-poor apolipoprotein A-I (apoA-I)-containing high density lipoproteins (HDLs) with pre-beta electrophoretic mobility. These are converted by lecithin:cholesterol acyltransferase into larger spheroidal cholesteryl ester-rich HDLs with alpha mobility. To study the determinants of the concentration of small pre-beta HDLs in tissue fluids, we collected prenodal peripheral lymph from 34 fasted normal men. By crossed immunoelectrophoresis, the concentration of pre-beta HDLs in lymph averaged 20% of that in plasma. On multiple regression analysis, pre-beta apoA-I concentration in lymph was directly related to pre-beta apoA-I concentration in plasma and independently to alpha apoA-I concentration in lymph. Similar results were obtained when the same apoA-I-containing particles were quantified by size exclusion chromatography. Lymph pre-beta apoA-I concentration was low in a subject with familial lecithin:cholesterol acyltransferase deficiency, despite a normal plasma pre-beta apoA-I concentration, but was normal in a subject with familial lipoprotein lipase deficiency. These results suggest that the concentration of small pre-beta HDLs in human tissue fluids is determined only in part by the transfer of pre-beta HDLs across capillary endothelium from plasma. Local production, by remodeling of spheroidal alpha HDLs in tissue fluids, may be equally important. Lipolysis of triglyceride-rich lipoproteins by lipoprotein lipase appears to have little effect.
Adult↗
Haemostatic factors in human peripheral afferent lymph.
Peripheral afferent lymph was obtained by cannulation of a collecting vessel in 17 healthy men (mean age 26 years). Lymph/plasma ratios of all vitamin K-dependent factors were lower than expected from molecular weight. Factor VII, factor IX and tissue factor pathway inhibitor (TFPI) lymph/plasma activity ratios were higher than antigen ratios. Activated factor VII (FVIIa) and TFPI-Xa complex concentrations were higher in lymph than plasma, and the raised FVIIa did not appear to be due to cannulation. The fibrinogen lymph/plasma activity (Clauss) ratio averaged about 20% of the antigen ratio. The result of an ELISA for D-dimer was higher in lymph than plasma, often more than five-fold. This high level in lymph was not explored but may indicate proteolysis of fibrinogen and fibrin with release of D-like and D-dimer-like fragments in interstitial fluid.
Adult↗
Kinetics of bone marrow repopulation in lethally irradiated rats after transplantation of vascularized bone marrow in syngeneic hind limb.
Grafting of the recipient with bone marrow cell suspension provides only few stromal cells and no autologous environment for cooperation between hemopoietic and stromal cells. Vascularized bone marrow grafts provide the recipient with bone marrow hemopoietic and stromal cells in their natural spatial relationship. It is expected that such a graft will resume its function soon after transplantation and almost immediately repopulate bone marrow cavities of the irradiated recipient as well as supply the sites of cytopoiesis with the necessary growth factors. The aim of the study was to investigate the process of repopulation of bone marrow cavities and lymphoid organs of irradiated recipient by bone marrow cells from syngeneic hind limb transplant. Lewis rats received total body irradiation of 8 Gy followed by orthotopic transplantation of syngeneic limb or i.v. infusion of equivalent amount of bone marrow cells. In control experiments the hind limb was shielded with lead plate during total body irradiation. Ten days after irradiation and hind limb transplantation the yield of nucleated cells from tibia was reaching values of normal animals. In rats receiving i.v. bone marrow cell infusion it was 40% of control values and in rats repopulated from shielded own hind limb it was 60% of controls. In all groups a higher percentage of early and immediate normoblasts and a reduced pool of juvenile and segmented neutrophils was observed. Thirty days after irradiation and repopulation procedures all parameters were returning to normal levels in each group. The results indicate that bone marrow cell transplantation in hind limb graft is highly effective in lethally irradiated animals in reconstituting bone marrow.
Animals↗
Lipid and apolipoprotein concentrations in prenodal leg lymph of fasted humans. Associations with plasma concentrations in normal subjects, lipoprotein lipase deficiency, and LCAT deficiency.
The extent to which lipid and apolipoprotein (apo) concentrations in tissue fluids are determined by those in plasma in normal humans is not known, as all studies to date have been performed on small numbers of subjects, often with dyslipidemia or lymphedema. Therefore, we quantified lipids, apolipoproteins, high density lipoprotein (HDL) lipids, and non-HDL lipids in prenodal leg lymph from 37 fasted ambulant healthy men. Lymph contained almost no triglycerides, but had higher concentrations of free glycerol than plasma. Unesterified cholesterol (UC), cholesteryl ester (CE), phosphatidylcholine (PC), and sphingomyelin (SPM) concentrations in whole lymph were not significantly correlated with those in plasma. HDL lipids, but not non-HDL lipids, were directly related to those in plasma. Lymph HDLs were enriched in UC. However, as the HDL cholesterol/non-HDL cholesterol ratio in lymph exceeded that in plasma, whole lymph nevertheless had a lower UC/CE ratio than plasma. Lymph also had a significantly higher SPM/PC ratio. The lymph/plasma (L/P) ratios of apolipoproteins were as follows: A-IV > A-I and A-II > C-III and E > B. Comparison with the L/P ratios of seven nonlipoprotein proteins suggested that apoA-IV was predominantly lipid free. Concentrations of apolipoproteins A-II, A-IV, C-III, and E in lymph, but not of apolipoproteins A-I or B, were positively correlated with those in plasma. The L/P ratios of apolipoproteins B, C-III, and E in two subjects with lipoprotein lipase (LPL) deficiency, and of apolipoproteins A-I and A-IV in a subject with lecithin:cholesterol acyltransferase (LCAT) deficiency, were low relative to those in normal subjects. Thus, the concentrations of lipids, apolipoproteins, and lipoproteins in human tissue fluid are determined only in part by their concentrations in plasma. Other factors, including the actions of LPL and LCAT, are at least as important.
Adult↗
[Post-traumatic lymphatic and venous drainage changes in persistent edema of lower extremities].
Mechanical injury of soft tissues and bones of the lower extremity is followed by chronic edema at the site of trauma and distally to it. This complication affects almost every patient with a fracture of the lower limb. The question is whether posttraumatic edema is due to lymphatic obstruction, venous thrombosis or both, or a local cytokine and growth factor hyperactivity at the fracture site. The aim of study was to assess the venous and lymph outflow in patient with chronic postraumatic edema of the lower limbs. A group of 19 patients with chronic edema lasting for more than 3 months was evaluated. Limb circumference, tissue tone measurements, skin temperature and Doppler enhanced ultrasonography were all taken down for the 19 patients in the evaluated group. Limb circumference was measured at the following level: foot, ankle, calf and thigh. Results showed an increase of circumference in comparison with the healthy extremity at each evaluated level of: 1.20 +/- 1.65 cm, 1.63 +/- 1.41 cm, 1.40 +/- 1.72 cm and 0.30 +/- 1.90 cm. Local temperature increase compared to the healthy extremity was also noted (0.93 +/- 0.81 degree C and 0.37 +/- 0.21 degree C measured at ankle and calf level). Tissue tone measurements and tone index (a quotient of tone measurement values in the extremity with edema and in the healthy extremity) were also increased by 0.86 +/- 0.57, 0.85 +/- 0.34 and 0.86 +/- 0.28, when measured with 40 g, 110 g and 180 g weights respectively. In 17 cases (89.5%) lymphoscintigraphy demonstrated an increased lymphatic outflow compared to the contralateral extremity. A marked increase in the inguinal lymph nodes was also noted. In the remaining 2 cases (10.5%) extravasation of the contrast medium into the skin indicated lymph outflow disorders. Only in 5 cases (26.3%) ultrasonography indicated deep vein thrombosis. The obtained results indicate that the pathophysiology of chronic postraumatic edema is linked with an inflammatory and restorative reaction at the fracture site. Only in a limited number of cases deep vein thrombosis and damaged lymphatic vessels are responsible for postraumatic edema.
Chronic Disease↗
Continuous alloantigen elimination--a concept for allograft tolerancer.
The "enhancement" phenomenon evoked in rats by administration of donor lymphocytes and recipient-anti-donor-lymphocyte antibodies prior to organ transplantation brings about "continuous alloantigen elimination". This new concept is based on the experimental data from studies on the distribution of radiolabelled alloantigen and alloantibody in the recipient lymphoid organs and suggests that immediately after organ transplantation to the "enhanced" recipient, the circulating antibodies bind to the donor transplantation antigens both on endothelial cells and those shed into recipient blood circulation. The formed alloantigen-alloantibody complexes are trapped in the spleen and after opsonization and binding to the migrating splenocytes are transported to the liver for further processing. The process of continuous binding of alloantigen and elimination as alloantibody-alloantigen complexes in the lymphoid organs of the recipient prevents activation of the recipient effector cells and their migration to the graft. The organ grafts survive above 100 days. Although there is no evident rejection in the allograft itself a phenomenon of "rejection alert" is seen in the lymphoid tissue. The lymphocytes originating from the organ graft donor injected intravenously are rejected in the "enhanced" recipient spleen and nodes within 6 hours. The "physiological" site of elimination (rejection) of alloantigens are the lymphoid organs, as it happens with invading pathogens. The allotransplant is spared since its alloantigens are "blocked" by anti-donor-specific antibodies, the complexes are continuously detached and washed-off by the percolating blood and eliminated in the lymphoid tissues and liver.
Animals↗
Bacteriological studies of blood, tissue fluid, lymph and lymph nodes in patients with acute dermatolymphangioadenitis (DLA) in course of 'filarial' lymphedema.
Filarial lymphedema is complicated by frequent episodes of dermatolymphangioadenitis (DLA). Severe systemic symptoms during attacks of DLA resemble those of septicemia. The question we asked was whether bacterial isolates can be found in the peripheral blood of patients during the episodes of DLA. Out of 100 patients referred to us with 'filarial' lymphedema 14 displayed acute and five subacute symptoms of DLA. All were on admission blood microfilariae negative but had a positive test in the past. Blood bacterial isolates were found in nine cases, four acute (21%) and five subacute (26%). In 10 acute cases blood cultures were found negative. Six blood isolates belonged to Bacilli, four to Cocci and one was Sarcina. To identify the sites of origin of bacterial dissemination, swabs taken from the calf skin biopsy wounds and tissue fluid, lymph and lymph node specimens were cultured. Swabs from the calf skin biopsy wound contained isolates in nine (47%) cases. They were Bacilli in nine, Cocci in three, Acinetobacter and Erwinia in two cases. Tissue fluid was collected from 10 patients and contained Bacilli in four (40%) and Staphylococci in three (30%). Lymph was drained in four patients and contained isolates in all samples (100%). They were Staphylococcus epidermis, xylosus and aureus, Acinetobacter, Bacillus subtilis and Sarcina. Three lymph nodes were biopsied and contained Staphylococcus chromogenes, xylosus, Enterococcus and Bacillus cereus. In six cases the same phenotypically defined species of bacteria were found in blood and limb tissues or fluids. In the 'control' group of patients with lymphedema without acute or subacute changes all blood cultures were negative. Interestingly, swabs from biopsy wound of these patients contained isolates in 80%, tissue fluid in 68%, lymph in 70% and lymph nodes in 58% of cases. In healthy controls, tissue fluid did not contain bacteria, and lymph isolates were found only in 12% of cases. This study demonstrates that patients with acute episodes of DLA reveal bacteremia in a high percentage of cases. Diversity of blood and tissue bacterial isolates in these patients points to a breakdown of the skin immune barrier in lymphedema and subsequently indiscriminate bacterial colonization of deep tissues and spread to an blood circulation.
Adolescent↗
Fast lymphoid reconstitution after vascularized bone marrow transplantation in lethally irradiated rats.
BACKGROUND: Bone marrow (BM) transplantation for treatment of hematological and solid malignancies is routinely carried out in conjunction with radio- and chemotherapy. Many patients achieve complete remission of the malignant process; however, their lymphohematopoietic recovery remains in most cases incomplete. This is probably due to the functional changes in the recipient BM stromal cells subsequent to myeloablative therapy. Transplantation of BM hematopoietic cells in a spatial relationship with stromal cells would give an insight into the kinetics of hematological repopulation of the recipient. The aim of this study was to investigate the lymphopoietic reconstitution of irradiated rats after vascularized bone marrow transplantation (VBMT) in comparison with i.v. bone marrow cell (BMC) infusion. METHODS: Lewis rats were totally irradiated with 8Gy and repopulated with syngeneic BMC introduced i.v. or in orthotopic hind limb graft. Ten days after irradiation and BMC graft BM, peripheral blood (PB) and mesenteric lymph nodes (MLN) were collected. The yield and the phenotype of cells were analyzed. RESULTS: VBMT brings much higher cell repopulation of BM cavities of lethally irradiated rats than BMC infusion. Orthotopic hind limb graft promotes also rapid lymphocyte replenishment of PB and MLN of lethally irradiated syngeneic recipients. The population rate of BMC, PB lymphocytes, and MLN lymphocytes was higher after VBMT than BMC injection in suspension. The percentage of T and B lymphocytes in PB and MLN on day 10 after VBMT was comparable with control values. Reconstituted PB lymphocytes showed two subsets of CD4+ cells: "bright" and "dull." All CD4+ cells in PB lymphocytes of i.v. BMC infused recipients expressed low level of these molecules ("dull" subset). CONCLUSIONS: The results of our studies indicate that the presence of stromal cells in their close relationship with stem cells is essential for the fast lyphohematopoietic repopulation of irradiated recipients. The population of CD4+dull cells may represent immature cells. These cells were not found in MLN of VBMT rats. All MLN CD4+ cells represented the "bright" subset, what suggests that the process of cell maturation may occur in the lymphoid organs.
Adoptive Transfer↗
[Bacterial infections of skin and soft tissues in filariasis].
Adenolymphangitis is a common occurrence in filarial lymphedema. Damage to the lymphatics and lymph nodes by F. bancrofti is followed by obliteration of lymph vessels and lymph stasis. Obstruction of lymphatics prevents the bacteria penetrating skin to be evacuated with lymph stream to regional lymph nodes. Colonization of dermis, subcutis and lymphatics evokes clinical symptoms of adenolymphangitis. The question arises which strains of bacteria are responsible for the acute and chronic types of adenolymphangitis. The most probable strains responsible for this condition belong to the cocci and probably the bacillus strains.
Adult↗
The mechanism of immunological "enhancement" in allografts--distribution of alloantigen and alloantiserum in the recipient.
The active or passive "enhancement", by providing the recipient with donor cellular antigens and recipient antiserum against these antigens, has been known for long to prolong the survival time of organ allografts in rodents. Donor-specific blood transfusions seem to prolong the graft survival in animals and man evoking a similar protective mechanism. We investigated the mechanism of the effect of administering the recipient donor cells and antibodies against these cells on prolongation of heart graft survival. AUGust rat (RTIc) cells and WIStar rat (RTIu) antibodies against AUG antigens were labeled with 51Cr and I 125, respectively. They were administered i.v. into prospective WIS recipients of AUG hearts -11 and -10 days before grafting, respectively. The distribution and level of specific radioactivity were measured in the recipient lymphoid organs on days -9, -7 and -1 as well as on days +1 and +5 after transplantation. The highest levels of radioactivity of both donor cells and host antibodies against these cells were found on they -9 in recipient spleen but not liver, bone marrow and lymph nodes. After heart or skin transplantation, the radioactivity decreased in spleen to increase in liver and bone marrow. No specific accumulation of antibodies against donor antigens was detected in the grafts. These findings point to a role of antigen-antibody complexes located preferentially in the spleen on the downregulation of in vivo response to donor transplantation antigens. A concept of "continuous alloantigens elimination" is presented in which a systemic process of shed alloantigen inhibition and degradation in concert with blocking of organ surface antigens is presented.
Animals↗
DNA from rejecting allografts can be detected in recipient nonlymphoid tissues.
The main source of donor DNA in recipients of allograft are "passenger" cells. They are claimed to be responsible for the posttransplantation microchimerism and prolongation of allograft survival. We have noticed that beside of the cellular microchimerism, donor DNA can be found in the recipient tissues at the time of rejection of allograft. In this study we provide evidence for presence in the recipient of both, DNA in "passenger cells" and free DNA in tissues at terminal stage of rejection. Male BN (RTIn) rat heart or skin were transplanted to female LEW (RTII) rats followed by a vascularized bone marrow in hind-limb transplant. CsA was given in a dose of 17mg/kg b.w. for 30 days, then rats were followed until day 100 unless rejection occurred earlier. LEW blood, spleen, mesenteric node and bone marrow cells were stained with moAb OX27 specific for BN but not LEW. Genomic male DNA was isolated and amplified with SRY oligonucleotide. At day 30 and 100 cellular microchimerism was detected in blood, spleen, nodes and bone marrow cells. Donor DNA was detected in recipient skin, liver and heart extracts, beside of lymphoid organs, at the time of rejection of allograft but not when rats were maintained on CsA. Taken together, donor DNA can be detected in recipient tissues at the time of heart or skin rejection. It seems to be released from cells of rejecting grafts and not from "passenger" cells representing only a minor cellular mass compared with the graft.
Animals↗
Skin allografts--lymph veiled (dendritic) cells are responsible for initiation of rejection in canine skin/SCID mouse chimera model.
Skin allografts are acutely rejected despite of intensive immunosuppressive therapy. Resistance of skin dendritic cells to immunosuppressive drugs and irradiation may be responsible for this phenomenon. Skin allograft is a site of interaction between the dendritic cells and lymphocytes of the donor and host origin and "direct" and "indirect" pathway of antigen presentation. Increasing evidence supports the significant role for the "indirect" allorecognition in graft rejection. To investigate a critical role of skin dendritic cells in the "indirect" allorecognition and graft destruction we have used a canine skin to severe-combined-immunodeficient (SCID)-mice transplantation model. At the time the skin grafts were deprived of own dendritic (Langerhans) cells, SCID mice were reconstituted with allogeneic canine whole lymph leukocytes, lymph lymphocytes, lymph veiled (dendritic) cells or peripheral blood mononuclear cells, and an early phase of skin rejection was evaluated in histopathological studies. We demonstrated that circulating canine allogeneic veiled cells facilitate recruitment of T lymphocytes into skin graft and promote an extensive graft destruction, compared to the less expressed effect of allogeneic peripheral lymph lymphocytes or blood mononuclear cells. These drug and radiation-resistant dendritic cells may be responsible for initiation of the difficult to control rejection process.
Animals↗
Hepatocyte transplantation--in vitro cytotoxic reaction of autologous granulocytes and mononuclears to isolated hepatocytes.
UNLABELLED: Hepatocyte (HC) transplantation (tx) may be useful for bridging patients to whole organ transplantation and for providing metabolic support during liver failure and for replacing whole organ transplantation in certain liver metabolic diseases. In specific situations where the death rate of host hepatocytes is high, the transplanted cells can repopulate the native liver. Successful transplantation of hepatocytes is hampered by lack of proper cellular (stromal) and humoral (cytokines) environment at the site of implantation. We have found that another factor responsible for low in vivo survival rate of transplanted HC is their rapid destroying by host granulocytes and monocytes. AIM. In this study we investigated the in vitro process of destruction of HC by granulocytes and mononuclear cells, the phenotypes of effector mononuclears and the tempo of HC lysis. METHODS: In vitro cell-mediated cytotoxicity, HC-PMN and HC-PBM rosette formation rate and HC lysis, as well as phenotypes of HC-adhering cells were investigated. RESULTS: Granulocytes formed rosettes with HC almost immediately after the beginning of incubation and were found highly cytotoxic to HC. The cellular mechanism of lysis was not mediated by serum natural antibodies. Also the in vitro mixed HC-granulocyte 51Cr test showed high granulocyte cytolytic activity. Monoclonal antibodies to class I and II antigens, CD11/18 and 54 did not block the granulocyte cytotoxicity. Blood mononuclear cells also formed rosettes with HC and were cytotoxic to them, but the level of cytotoxicity was lower than of granulocytes. ED1+ monocytes revealed highest cytolytic activity toward HC. Hepatocyte contained only trace levels of endotoxin and no chemotactic activity of granulocytes and monocytes toward HC could be observed. CONCLUSIONS: A random physical contact of blood leukocytes seems necessary for adhesion to isolated HC. Taken together, granulocytes and monocytes recognize intercellular surface molecules on HC "exposed during isolation" from the hepatic trabeculae as "non-self" and lyse HC by direct contact.
Animals↗
Tolerance induction following allogeneic vascularized bone marrow transplantation--the possible role of microchimerism.
We have noticed that bone marrow transplanted in a vascularized limb graft, providing a continuous supply of donor bone marrow cells (BMC), may prolong the survival time of a skin graft from the same donor. The question arises whether the microchimerism raised plays a role in the prolonged survival of skin allografts. The aim of the study was to follow the development of microchimerism after allogeneic vascularized bone marrow transplantation (VBMTx) concomitantly with the rejection process of transplanted skin. Brown Norway (BN) rats served as donors and Lewis rats as recipients of VBMTx and free skin flap allografts. A hind limb was transplanted, followed by a full-thickness skin graft on the dorsum. Cellular microchimerism was investigated in recipients of VBMTx and skin grafts in blood, spleen, mesenteric lymph node, and bone marrow with the monoclonal antibody OX27 directed against MHC class I polymorphic RT1 on BN cells and quantitatively analyzed in a FACStar. In the VBMTx group, the free skin flap survived 70 days after weaning off cyclosporine A (CsA). An intravenous infusion of BMC in suspension equivalent to that grafted in the hind limb did not prolong skin graft survival after cessation of CsA therapy. Donor-derived cells could be detected in VBMTx recipients as long 70 days after weaning off CsA but not in recipients of i.v. suspension BMC grafting.
Animals↗
Hepatocyte transplantation--granulocytes and mononuclear cells recognize the surface of isolated autologous hepatocytes as non-self and destroy them.
Successful transplantation of hepatocytes (HC) is hampered by lack of a proper cellular (stromal) and humoral (cytokines) environment at the site of implantation. We have found that another factor responsible for the low survival rate of transplanted HC is their rapid destruction by host granulocytes. In this study we have investigated the mechanism of the rapid elimination of transplanted hepatocytes in a syngeneic and allogeneic recipient. Only 10% of the radioactivity of syngeneic (LEW-LEW) or allogeneic (BN-LEW) HC was recovered in recipient lymphoid tissues 6 h after i.v. infusion. Pretreatment of the recipient with Endoxan or sublethal irradiation brought about an increased accumulation of HC radioactivity in lungs and spleen, indicating the entrament of live cells in these organs. Only a few HC could be seen in the liver 6 h after intraportal infusion. Granulocytes were found to be cytotoxic to HC in vitro (39%). Monoclonal antibodies to class I and II antigens, CD 11/18 and 54, did not block the granulocyte cytotoxicity. Granulocyte and HC cluster formation could be seen on smears from a drop of mixed cultures, the rates of formation increasing with the time of incubation. Two out of 34 monoclonal antibodies to HC surface molecules partly blocked the granulocyte cytotoxicity. No evident differences in the elimination rate and in in vitro cytotoxicity were seen between the syngeneic and allogeneic HC.
Animals↗
A novel effect of cyclosporine A on lymphocyte migration to allograft and recipient lymphoid tissue.
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Animals↗