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M L Rose

Publications and source records attributed to M L Rose.

At least 55 records · Page 3Linked to original sources
PubMed1998

A new enzyme-linked immunosorbent assay to measure anti-endothelial antibodies after cardiac transplantation demonstrates greater inhibition of antibody formation by tacrolimus compared with cyclosporine.

BACKGROUND: Chronic rejection or transplant-associated coronary artery disease (TxCAD) is the most serious complication after human cardiac transplantation. Previous studies, using Western blotting, have shown formation of antibodies against endothelial antigens of 56 and 58 kDa, which are associated with early TxCAD. These antigens were later identified as being vimentin and its breakdown products. The aims of the present study were to devise a robust assay for detection of anti-vimentin antibodies and to compare antibody formation in patients taking different immunosuppressive drugs. METHODS: 106 sequential serum samples from 19 patients taking tacrolimus and 68 sera from 12 patients taking cyclosporine were examined by enzyme-linked immunosorbent assay (ELISA) for anti-vimentin antibodies and Western blotting for reactivity against bands at 56/58 kDa. Serum samples were taken before transplantation and at 1, 3, 6, 9, and 12 months. RESULTS: The vimentin ELISA produced significantly higher numbers of positive episodes per patient (3.92+/-1.08) compared with use of Western blotting (2.54+/-0.52). Serum from patients taking tacrolimus contained significantly less antibodies measured by ELISA (15.8%) or Western blotting (6.5%) than sera from patients taking cyclosporine (46.8% for ELISA; P=0.001 and 21% by Western blotting, P=0.01). Intravascular ultrasound performed on six patients at 12 months showed a correlation between anti-vimentin antibody formation and detection of early coronary disease. CONCLUSIONS: The results demonstrate first, that differences in antibody profiles produced by different immunosuppressive drugs, and second, that detection of anti-vimentin antibodies may be a noninvasive method of detecting disease activity in transplanted vessels.

Antibodies↗
PubMed1998

Assessment of the contribution that direct allorecognition makes to the progression of chronic cardiac transplant rejection in humans.

BACKGROUND: Two populations of T cells contribute to allograft rejection. T cells with direct allospecificity are activated after recognition of intact MHC alloantigens displayed at the surface of donor passenger leukocytes carried within the graft. In contrast, T cells with indirect allospecificity recognize donor alloantigens as processed peptides associated with self (recipient)-MHC class II molecules. In small animal models of transplantation, direct pathway T cells dominate the acute rejection process and are rendered tolerant to the graft after the loss of donor passenger leukocytes. It has been argued that indirect pathway T cells contribute substantially to continual graft damage after passenger cell loss. The purpose of this study was to determine whether donor-specific tolerance could be detected in T cells with direct anti-donor allospecificity in human heart transplant recipients after prolonged graft residence. METHODS AND RESULTS: Alloreactive helper (HTLf) and cytotoxic (CTLf) T cells were enumerated by use of limiting dilution analysis. These assay systems were refined to make them specific for the direct pathway of allorecognition and more sensitive in the case of the HTLf assay. Recipient:anti-donor frequencies were generated in 10 long-term recipients of heart grafts with progressive chronic rejection and compared with those against equivalently HLA mismatched recipient:third-party controls. For HTLf, direct pathway donor-specific hyporesponsiveness was detected in 5 of the 10 recipients (HTLf<1:100,000). Of these 5 recipients, 4 also had low anti-donor CTLf (<1:100,000). In the 5th recipient, although the CTLf was >1:100,000, it was significantly lower than that estimated against the third-party control. CONCLUSIONS: Donor-specific hyporesponsiveness is demonstrated in 50% of recipients in both the HTLf and CTLf compartments of the direct alloresponse. Direct allorecognition therefore appears unlikely to be responsible for the progression of chronic rejection, implicating indirect allorecognition as the predominant immunological driving force. Furthermore, these data have potential implications for graft outcome, adjustment of immunosuppression, and recipient monitoring.

Antibody Specificity↗
PubMed1998

Endothelial cells as antigen-presenting cells: role in human transplant rejection.

The immunological properties of human endothelial cells suggest they perform a pivotal role in acute and chronic rejection following solid organ transplantation. In this review the basic features of acute and chronic rejection are described as are the cellular and molecular requirements for antigen presentation. Traditionally, antigen-presenting cells are considered to be bone marrow-derived cells. However, these conclusions have been derived from rodent models of allograft rejection where bone marrow-derived passenger leukocytes are the only source of donor major histocompatibility complex (MHC) class II in the grafted organ. In contrast, in humans, virtually all the microvascular and small vessel endothelial cells are 'constitutively' positive for MHC class II antigens. The phenotypic properties of human endothelial cells, their response to cytokines and their ability to stimulate resting T cells are described. Unlike bone marrow-derived antigen presenting cells (APCs), which utilise B7/CD28 interactions, human endothelial cells utilise lymphocyte function antigen 3 (LFA3)/CD2 pathways to stimulate T cells. They activate a CD45RO + B7-independent subpopulation of T cells. Their effect on allogeneic T cells is compared with other non-bone marrow-derived cells such as fibroblasts, epithelial cells and smooth muscle cells, which are unable to stimulate resting T cells. Evidence is presented suggesting that release of MHC and non-human leukocyte antigens (HLA) from endothelial cells stimulates an alloantibody and autoimmune response leading to chronic rejection.

Acute Disease↗
PubMed1998

Effect of HLA mismatching and antibody status on "homovital" aortic valve homograft performance.

BACKGROUND: Recipients of "homovital" aortic valve homografts are known to produce specific antibodies to human leukocyte antigen (HLA) determinants present on the cellular compartment of the valve tissue; however, the clinical significance of these antibodies is unknown. Data from 182 patients receiving homovital aortic valve homografts has been analyzed to determine the impact of HLA disparity and HLA antibody production on survival and function of the homograft. METHODS: Human leukocyte antigen mismatch data were available for 127 patients (mean follow-up, 6.02+/-0.26 years). Two patients were considered well matched for HLA A+B antigens (zero or one mismatch) compared with 125 poorly matched (two to four mismatches). Nine patients had a zero HLA-DR mismatch compared with 52 with one mismatch and 59 patients completely mismatched for DR antigens. RESULTS: There was no significant association between the degree of HLA mismatch for either class I or class II antigens whether the loci were considered alone or in combination (ie, A, B, DR, AB, or ABDR mismatching) with markers of long-term valve function including patient mortality, reoperation, valve degeneration, valve stenosis, presence of regurgitation, and postoperative New York Heart Association class. One hundred thirty-six of 167 (82%) were found to have produced antibodies after operation (mean time after operation, 6.42+/-0.58 years). In 61 cases both antibody specificity and donor HLA typing was available. In 92% of these, the antibodies were of the IgG subclass and were specific for the HLA class I molecules of the donor. The presence of HLA antibodies was associated with an increase in the frequency of mild valve stenosis (not significant) compared with those patients who did not develop HLA antibodies (antibody negative = 9.7%; panel reactive antibodies <50% = 29.1%; and panel reactive antibodies >50% = 22.2%; not significant). There was also an increased prevalence of valve degeneration associated with HLA antibodies. The actuarial freedom from valve degeneration for the 35 HLA antibody-negative patients was 100% at 1, 5, and 10 years compared with 100% at 1 year, 97% at 5 years, and 92% at 10 years for 55 patients with panel reactivity less than 50%, and 98% at 1 year, 94% at 5 years, and 88% at 10 years for the 77 patients who were highly sensitized (not significant). There was no correlation with other markers of long-term valve function. CONCLUSIONS: The influence of the immune response on valve function requires further studies involving large numbers of patients followed for a longer period of time. We believe prospective matching for HLA antigens is warranted to produce a well-matched cohort of patients for analysis and to reduce antibody sensitization, which would help to clarify this issue.

Actuarial Analysis↗
PubMed1998

The effects of monocytes on the transendothelial migration of T lymphocytes.

In vivo cell-mediated immune reactions are characterized by mixtures of monocytes and T cells. The purpose of this study was to investigate the role of monocytes on T-cell migration and induction of endothelial adhesion molecules. The in vitro model consisted of adding peripheral blood mononuclear cells (PBMC), T cells or mixtures of monocytes and T cells, to endothelial cells on a porous membrane and using flow cytometry to distinguish between the monocyte and lymphocyte components. PBMC and PBMC supernatants were highly potent at upregulating intercellular adhesion molecule-1 (ICAM-1) and inducing expression of vascular cell adhesion molecule-1 (VCAM-1) and E-selectin. Induction by supernatants was inhibited by antibodies to tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 (IL)-1 beta. Using monocyte-enriched populations, as few as one monocyte to 100 endothelial cells was sufficient to upregulate adhesion molecules. Fixed monocytes also induced adhesion molecules and expressed surface-bound cytokines. In contrast, highly purified unstimulated T cells were not found to induce adhesion molecules at 4, 6, 24 or 48 hr of coculture. Purified T cells showed low-level migration through resting (VCAM-1 negative) endothelium, which was approximately doubled by addition of small numbers of monocytes or TNF-alpha. In conclusion, monocytes, via cell surface or released cytokines play an essential role in allowing large-scale recruitment of T cells to inflammatory sites in vivo.

CD4-Positive T-Lymphocytes↗
PubMed1997

MHC class II expression on human heart microvascular endothelial cells: exquisite sensitivity to interferon-gamma and natural killer cells.

BACKGROUND: Immunocytochemical analysis of human organs in situ reveals differential expression of MHC class II antigens by microvascular endothelial cells (MVEC) and endothelial cells (EC) from large vessels. In view of the role of EC as initiators of allograft rejection, it is of interest to understand the regulation of MHC class II regulation by human MVEC. We have previously isolated, cultured, and characterized MVEC from the human heart, showing that although these cells were initially MHC class II positive, the antigens were lost after about 14 days in culture. These results suggest that basal expression in vivo is maintained by circulating factors. METHODS: Here we have compared the sensitivity of human heart MVEC, human umbilical vein endothelial cells (HUVEC), and adult large vessel EC (aorta, coronary artery, and pulmonary artery) to interferon (IFN)-gamma and natural killer (NK) cell-mediated induction of MHC class II antigens. MVEC and HUVEC were cultured with 1, 5, 10, 50, 100, and 500 U/ml of IFN-gamma for 4 days, the cells were washed, and flow cytometry was used to examine HLA-DR expression at days 1, 2, 4, 7, 10, 14, and 21. EC were also cultured with purified NK cells in the presence and absence of neutralizing antibodies to IFN-gamma, and MHC class II expression was analyzed. RESULTS: As little as 5 U/ml of IFN-gamma produced 98% positive cells in heart MVEC compared with 100-500 U/ml needed for the same effect in HUVEC or other large vessel EC (coronary, aorta, pulmonary). Class II expression was maintained longer by MVEC (for 17 days) compared with HUVEC (for 10 days). NK cells and supernatant from MVEC/NK cultures induced MHC class II antigens on MVEC and HUVEC in a dose-dependent fashion; the MVEC showed an enhanced sensitivity compared with the HUVEC. The NK effects were inhibited by neutralizing antibodies to IFN-gamma. The allostimulatory ability of MHC class II-positive EC was shown to be proportional to the amount of MHC class II on the cell surface. CONCLUSIONS: The results suggest that basal expression of MHC class II on human MVEC is maintained by circulating IFN-gamma and NK cells. This conclusion has implications for therapeutic interventions.

Adult↗
PubMed1997

Optimizing a limiting dilution culture system for quantifying the frequency of interleukin-2-producing alloreactive T helper lymphocytes.

BACKGROUND: The development of sensitive, specific, and reproducible techniques to quantify T cells with direct allospecificity has potential applications in the selection of bone marrow donors and in the monitoring of the antidonor alloresponse in patients after organ transplantation. Such data may provide an objective basis for altering existing immunosuppression, monitoring novel antirejection therapies, and predicting long-term graft outcome. We have previously published a correlation between donor antirecipient T helper frequencies (HTLf) and the severity of acute graft-versus-host disease after bone marrow transplantation. Using the same assay protocol, we have described the development of donor-specific hyporesponsiveness in a proportion of renal transplant recipients. However, several imperfections existed in the protocols used in these studies. Cellular interactions within the stimulator and the responder cell populations, and back stimulation of T cells within the stimulator cell population, could give rise to extraneous interleukin-2 and alter the validity or estimation of derived recipient antidonor HTLf. METHODS: Using peripheral blood mononuclear cells as the responding population and splenic mononuclear cells as the stimulating population, we have examined the possible effects of these cellular interactions on the results of limiting dilution analysis assays for HTLf measurement. RESULTS: These interactions have the ability to alter the validity or estimation of HTLf. We show that by depleting the responder population of HLA class II+ cells and depleting T cells from the stimulating population, these interactions are effectively abrogated. CONCLUSIONS: On the basis of the findings reported here, we describe an optimized HTLf assay which is sensitive, specific, and reproducible. This has obvious applications in the analysis of alloimmune responses in transplantation.

Bone Marrow Transplantation↗
PubMed1997

Point mutations in mitochondrial DNA of patients with dilated cardiomyopathy.

Mitochondrial (mt) DNA mutations are hypothesized to be involved in the pathogenesis of dilated cardiomyopathy, because the mtDNA encodes 13 polypeptides that are essential for oxidative phosphorylation, upon which the heart relies for energy. To test this hypothesis, we amplified the mitochondrial genome by long PCR and then used restriction analysis and direct sequencing to examine 58 unrelated patients with dilated cardiomyopathy and 49 controls for the detection of point mutations. The results demonstrated that point mutations were significantly more frequent in the mtDNA of patients than in that of controls (173 in 58 patients v 54 in 49 controls, 2=16.51, P<0.001). In addition to normal variants and mutations common to both patients and controls, 43 mutations were identified only in patients. All but four of these mutations were homoplasmic. Mutations involving the evolutionarily conserved residues of cytochrome c oxidase subunit I, NADH dehydrogenase 5, tRNAAla and tRNAArg were identified. As many as 13 point mutations were found in an 8-month-old patient. In conclusion, there exist significantly more point mutations in mtDNA of patients than in controls, suggesting that multiple mutations may exert a cumulative effect on heart function. Thus, by altering the function of respiratory enzyme subunits or tRNAs, mtDNA point mutations could be relevant for the pathogenesis of dilated cardiomyopathy.

Adult↗
PubMed1997

Anti-cell surface endothelial antibodies in sera from cardiac and kidney transplant recipients: association with chronic rejection.

The aetiologies of accelerated or transplant-associated coronary artery disease (TxCAD) following cardiac transplantation and chronic rejection following renal transplantation remain ill-defined. Previous studies have used Western blotting to demonstrate an association between the formation of anti-endothelial (anti-EC) antibodies and TxCAD after heart transplantation. However, Western blotting favours detection of cytosolic proteins. The objectives of this study were to determine whether flow cytometry, a method which detects antigens on the cell surface, could be used to detect anti-EC antibodies and also whether the observations would extend to renal transplant patients with chronic rejection. Flow cytometry was used to look for antibodies reactive with the surface antigens of macrovascular and microvascular endothelial cell lines in sera from 44 cardiac and 35 renal transplant recipients before and after transplantation. In addition, sera from normals (n = 20), patients with nontransplant CAD (n = 50) and patients with unrelated diseases (n = 40) were investigated. Of 23 cardiac recipients who had developed TxCAD at one or two years post-transplant, 61% had IgM and 13% had IgG anti-EC antibodies post-transplantation. In contrast, in 21 cardiac recipients who had not developed TxCAD 14% had IgM and 14% IgG anti-EC antibodies. There was little evidence for the presence of anti-EC antibodies in cardiac recipients before transplantation. Of 26 renal transplant recipients whose transplants failed due to chronic rejection, 42% had IgG and 19% IgM anti-EC antibodies post-transplantation. Of nine renal recipients whose grafts were either functioning normally or who had acutely rejected, none had IgG or IgM anti-EC antibodies either pre- or post-transplantation. The anti-EC antibodies were not found in normals and were rare (less than 4%) in the other disease groups; they do not appear to be autoantibodies. In conclusion, these results suggest the FACS assay could be an informative and rapid test to provide more information on chronic rejection following cardiac and renal transplantation.

Adolescent↗
PubMed1997

Dietary glycine prevents increases in hepatocyte proliferation caused by the peroxisome proliferator WY-14,643.

Peroxisome proliferators are a group of nongenotoxic carcinogens which include a number of hypolipidemic drugs, solvents, and industrial plasticizers. Although the mechanism by which they cause cancer remains unknown, one likely possibility is that they act as tumor promoters by increasing cell proliferation. Hepatic Kupffer cells represent a rich source of mitogenic cytokines (e.g., tumor necrosis factor alpha, TNF alpha) and are stimulated by peroxisome proliferators. Since glycine prevents activation of Kupffer cells, these experiments were designed to test the hypothesis that a diet containing glycine could block the mitogenic effect of the peroxisome proliferator [[4-chloro-6-(2,3-xylidino)pyrimidinyl]thio]acetic acid (WY-14,643). The effects of a glycine-enriched diet on WY-14,643-induced increases in cell proliferation after a single dose or after feeding WY-14,643 in the diet for 3 weeks were assessed. As expected, 24 h after a single dose of WY-14,643, rates of cell proliferation increased from basal values of 0.7 +/- 0.3% to 5.1 +/- 0.5%. Glycine largely prevented the increase caused by WY-14,643 with proliferation only reaching 1.9 +/- 0.4% (p < 0.05). Acyl CoA oxidase increased from 1.4 +/- 0.1 to 3.5 +/- 0.6 nmol of H2O2 min-1 (mg of protein)-1 (p < 0.05) indicating that peroxisome-specific enzyme activity was induced about 2-fold in livers of WY-14,643-treated rats after 24 h. Unlike cell proliferation, however, acyl CoA oxidase was not affected by dietary glycine, consistent with the hypothesis that cell proliferation and peroxisome proliferation occur via different mechanisms. After 3 weeks, dietary glycine reduced basal rates of cell proliferation by about 50% and completely prevented the sustained 5-fold increase in cell proliferation caused by feeding WY-14,643. Moreover, the 3-fold increase in TNF alpha mRNA caused by WY-14,643 was blocked completely by the glycine-enriched diet. Similarly, immunohistochemical staining for TNF alpha was increased 6-fold by WY-14,643, an increase which was prevented by dietary glycine. However, the 6-fold increase in acyl CoA oxidase activity was unaffected by glycine under similar conditions demonstrating that a diet containing 5% glycine prevents increased hepatocyte proliferation caused by a potent peroxisome proliferator without affecting induction of peroxisomes. These data demonstrate that a glycine-enriched diet prevents stimulated cell proliferation most likely by inhibiting TNF alpha production and raise the possibility that dietary glycine will be effective in preventing cancer caused by nongenotoxic carcinogens such as WY-14,643.

Animals↗
PubMed1997

Correlation of radionuclide and optical tracer assessments of fluid flow in artificial ventricles.

Characterization of flow properties of an artificial ventricle may aid development of designs to minimize thrombosis. Techniques for determining two such flow properties, viz. ventricular clearance rate and ejection fraction, are compared and validated here for polyurethane and silicone rubber ventricles operated in a mock circulatory loop at various stroke volumes and pulse rates. Ventricular clearance rats were measured both by clinical radionuclide tracer techniques and by an optical tracer method. Ejection fractions were measured by radionuclide imaging and validated by direct measurements of flow rate and ventricular volume. Results from the two methods for ventricular clearance are in close agreement. The optical tracer method is superior in spatial resolution, convenience and economy, but the radionuclide tracer method for ejection fraction gives excellent agreement with the absolute measurements.

Animals↗
PubMed1997

Kupffer cells are causally responsible for the mitogenic effect of peroxisome proliferators.

WY-14,643 [4-chloro-6-(2,3-xylidino)pyrimidinylthio-acetic acid] is a well-known non-genotoxic carcinogen and peroxisome proliferator that causes liver cancer in rodents by unknown mechanisms. Its ability to sustain elevated rates of hepatocyte DNA synthesis is most likely pivotal in the ultimate development of tumors. The source of this mitogenic stimulus following treatment of rats with WY-14,643 has been hypothesized to be Kupffer cells, the resident hepatic macrophages, since they are activated by peroxisome proliferators in vivo. Therefore, these studies were designed to determine if Kupffer cells are causally responsible for WY-14 643-induced increases in hepatocyte DNA synthesis in vivo. WY-14,643 (100 mg/kg) increased DNA synthesis 8-fold 24 h after treatment; however, inactivation of Kupffer cells with methyl palmitate, a nonhydrolyzable fatty acid ester and known Kupffer cell inhibitor, completely prevented the mitogenic effect of WY-14,643. On the other hand, the ability of WY-14,643 to induce peroxisomes was not affected by methyl palmitate. These data demonstrate that induction of peroxisomes is not dependent on factors from Kupffer cells and support the idea that stimulation of DNA synthesis and induction of peroxisomes occur via distinct mechanisms. Additionally, WY-14,643 increased liver mRNA transcripts of the hepatocyte mitogen tumor necrosis factor alpha (TNF alpha) more than twofold. This increase was also prevented by inactivating Kupffer cells with methyl palmitate. Therefore, it is concluded that Kupffer cells are causally responsible for WY-14,643-induced increases in hepatocyte DNA synthesis most likely by increasing production of TNF alpha, a hepatic mitogen.

Acyl-CoA Oxidase↗
PubMed1997

Role of endothelial cells in allograft rejection.

The immunological properties of endothelial cells suggest they perform a pivotal role in acute and chronic rejection following solid organ transplantation. Their constitutive expression of MHC class II molecules (which initiate allograft rejection by activating CD4 T cells) and accessory molecules allows them to present foreign antigen by both the direct and indirect route to the recipient's immune system. The costimulatory molecules used by endothelial cells appear to differ from those used by traditional antigen-presenting cells such as B cells and dendritic cells. Release of non-HLA antigens from damaged endothelial cells results in a chronic antibody response--possibly contributing to graft vasculopathy and chronic rejection. Further understanding of the factors that regulate MHC class II and accessory molecule expression on endothelial cells could lead to novel strategies of therapeutic intervention.

Acute Disease↗
PubMed1996

Nimodipine, a dihydropyridine-type calcium channel blocker, prevents alcoholic hepatitis caused by chronic intragastric ethanol exposure in the rat.

It has been shown recently that inactivation of Kupffer cells prevents free radical formation and early alcohol-induced liver injury, and that hypoxia subsequent to a hypermetabolic state caused by activated Kupffer cells is likely involved in the mechanism. Calcium is essential for the activation of Kupffer cells, which contain L-type voltage-dependent Ca2+ channels. Therefore, the purpose of this study was to determine whether a Ca2+ channel blocker, nimodipine, prevents early alcohol-induced liver injury in vivo and to evaluate its effect on intracellular calcium ([Ca2+]i) in Kupffer cells in vitro. Male Wistar rats were exposed to ethanol (10-12 g/kg/d) continuously for up to 4 weeks via intragastric feeding using an enteral model developed by Tsukamoto and French. In this model, ethanol causes steatosis, necrosis, and inflammation in only a few weeks. In the experimental group, nimodipine (10 mg/kg/d) was added to the diet and was shielded from direct light. Nimodipine had no effect on body weight over a 4-week treatment period, nor were mean ethanol concentrations or their cyclic pattern in urine affected. The mean urine ethanol values were 154 +/- 11 mg/dL in ethanol-fed and 144 +/- 38 mg/dL in ethanol + nimodipine-fed rats. After 4 weeks, serum aspartate transaminase (AST) levels were elevated in ethanol-treated rats to 183 +/- 78 U/L. In contrast, values only reached 101 +/- 9 U/L in rats given nimodipine + ethanol-values which were significantly lower. Steatosis and necrosis assessed histologically were also reduced significantly by nimodipine. Nimodipine (10 micrograms/kg) also blocked the swift increase in alcohol metabolism and elevated oxygen consumption in perfused livers from rats treated with alcohol in vivo. Further, in cultured Kupffer cells, nimodipine (1 mumol/L) largely prevented the elevation in [Ca2+]i caused by lipopolysaccharide (LPS) (LPS, 200 +/- 11 nmol/L; LPS + nimodipine, 94 +/- 31 nmol/L; P < .05). These results indicate that nimodipine prevents alcoholic hepatitis, possibly by inhibition of endotoxin-mediated Kupffer cell activation.

Animals↗
PubMed1996

Isolation, culture, and characterisation of MHC class II-positive microvascular endothelial cells from the human heart.

Immunological studies of human endothelial cells have been performed almost entirely on endothelial cells (EC) derived from foetal (umbilical vein) or adult (aorta) large vessels. Immunologically these EC have in common the fact that they are negative for major histocompatibility complex (MHC) class II antigens (HLA-DR), in contrast to microvascular cells within the heart which are constitutively positive for HLA-DR antigens. Here we describe a simple method for isolating microvascular EC from chunks of adult human ventricle using antibody (anti-HLA-DR) or Ulex lectin-coated magnetic beads. Cytospins of primary isolates reveal the majority of cells binding the beads are CD31+ve, CD36+ve, and HLA-DR+ve. After 14 weeks in culture, 19% of cells remained HLA-DR+ve but CD36 expression had disappeared. Cultures after 4-7 passages were analysed by flow cytometry and immunocytochemical staining of cytospins with mAbs against conventional endothelial antigens (EN4, PECAM-1, vWF, uptake of acetylated LDL) and nonendothelial antigens (CD45, cytokeratin, alpha-actin, desmin, MHC class I and class II). The observation that all the cells remained positive for CD31 and did stain strongly for cytokeratin confirmed their endothelial origin. There was some persistent, albeit low level, expression of HLA-DR which was upregulated by interferon-gamma, showing kinetics of induction similar to that found on large vessel EC. These microvascular cells should be very useful for those wishing to compare microvascular with macrovascular EC and for immunological studies of heart endothelial cells.

Adult↗
PubMed1996

Vascular adhesion molecules and immunogenicity in blood vessels used as coronary artery bypass grafts.

OBJECTIVE: The performance of coronary bypass grafts can be affected by a variety of circulating cell types. The initial event in any biological effect of such cells is adherence to the vascular endothelium prior to migration into the perivascular space. We aimed to investigate the expression of molecules that regulate cell adhesion in blood vessels employed as bypass conduits. METHODS: Segments of human saphenous vein, internal mammary artery, gastroepiploic artery and inferior epigastric artery were stained using specific monoclonal antibodies against the endothelial workers EN-4, Pal-E, von Willebrand factor small (vWF), and the cell adhesion molecules platelet-endothelium cell adhesion molecule (PECAM), intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), E-selectin, the leucocyte marker (CD45) and major histocompatibility complex (MHC) class I and II antigens, with visualisation by ABC immunoperoxidase method. RESULTS: All vessels had a strong expression of the endothelial specific antigens EN4, vWF, and PECAM as well as MHC class I. However, there was less expression of Pal-E, ICAM-1, E-Selectin and of the DR determinant of MHC class II. VCAM-1, DP and DQ determinants of MHC class II were expressed to a weaker extent. There were no marked differences in the expression of all the molecules examined between the four vessel types. CONCLUSION: Thus vessels used as bypass grafts are immunogenic and possess the potential to attract and interact with blood elements. Definition of the molecules responsible could offer opportunities for modulating the response to such interactions.

Adolescent↗