Search PubMed⌕ Search

Biomedical subjects

Marion Haubitz

Publications and source records attributed to Marion Haubitz.

At least 19 recordsLinked to original sources
PubMed2007

Engulfment of apoptotic cells by microvascular endothelial cells induces proinflammatory responses.

Circulating endothelial cells (CECs) have been detected in a variety of vascular disorders, but their interactions with healthy endothelium remain unknown. The aim of this study was to evaluate the response of human endothelial cells (ECs) to apoptotic or necrotic ECs in an in vitro model and to delineate pathogenetic pathways. Here we show that incubation of the human microvascular endothelial cell line (HMEC-1) with apoptotic ECs resulted in increased expression of chemokines and enhanced binding of leukocytes to HMEC-1 cells, whereas exposure of HMEC-1 cells to necrotic ECs caused no changes in leukocyte-binding affinity. Both apoptotic and necrotic cells were bound and engulfed by HMEC-1 cells and primary human umbilical vein endothelial cells (HUVECs). We therefore suggest that exposures to apoptotic and necrotic ECs induce different patterns of chemokine synthesis and leukocyte adhesion in healthy ECs. These data indicate that CECs are not only markers of vascular damage but may induce proinflammatory signals in the endothelium.

Apoptosis↗
PubMed2006

Circulating endothelial cells: a novel marker of endothelial damage.

Circulating endothelial cells (CECs) were first described over 30 years ago in smears of peripheral blood. Since then, more sophisticated techniques for CEC isolation have become available. In particular, immunomagnetic isolation and fluorescence-activated cell sorting (FACS) have been employed with success. We provide a short historical perspective and a comprehensive review on the subject. We review isolation and enumeration of CECs with an emphasis on CD146-driven immunomagnetic isolation and FACS. We describe, in great detail, advantages and pitfalls of both approaches and compare their specificity. Moreover, we provide a comprehensive list of clinical studies in this field and describe the possible clinical use of CECs. We also describe the phenotype of these cells and list typical surface markers. In addition, we review the phenotype of CECs and discuss mechanisms of detachment. We speculate about potential interactions between CECs and other cell subsets. We also describe other serum markers of endothelial damage and compare CECs with these markers. Finally, we highlight differences between circulating endothelial cells and endothelial progenitor cells. In summary, CECs must now be regarded as a sensitive and specific marker of endothelial damage. We emphasize that use of CECs in a clinical setting is on the horizon and pathogenetic clues may also be obtained.

Blood Cells↗
PubMed2006

Circulating endothelial cells as a prognostic marker in thrombotic microangiopathy.

BACKGROUND: Circulating endothelial cells (CECs) are a reliable marker of disease activity in a variety of vascular disorders. Damage to microvascular endothelial cells is a hallmark of thrombotic microangiopathy (TMA). The aim of this study is to identify and count CECs during the course of TMA and evaluate whether cell numbers may serve as a prognostic marker in patients undergoing plasma exchange. METHODS: Fifteen patients (8 women, 7 men) aged 31 to 66 years with TMA of different causes were studied before and after 4 sessions of plasma exchange. CECs were isolated by using anti-CD146-driven immunomagnetic isolation and counted after staining with Ulex Europaeus lectin-1. RESULTS: Numbers of CECs were markedly elevated in all patients before treatment (64 to 672 CEC/mL; mean, 320 +/- 205 CEC/mL) compared with healthy controls (0 to 16 CEC/mL; mean, 6.4 +/- 4.2 CEC/mL; P < 0.001). Patients with a favorable outcome had significantly greater initial CEC levels (mean, 426 +/- 175 CEC/mL; P < 0.001), and cell numbers decreased significantly after 4 treatments of plasma exchange (mean, 101 +/- 53 CEC/mL; P = 0.001). Patients with disease unresponsive to plasma exchange presented with lower initial CEC levels (mean, 108 +/- 36 CEC/mL), and numbers failed to decrease after plasma exchange (mean, 114 +/- 57 CEC/mL; P = 0.827). CONCLUSION: Markedly elevated numbers of CECs reflect severe and widespread endothelial damage in patients with TMA. Cell numbers at presentation and their degree of decrease after 4 sessions of plasma exchange could provide important prognostic clues.

Adult↗
PubMed2006

Direct evidence of endothelial injury during cardiopulmonary bypass by demonstration of circulating endothelial cells.

Endothelial activation is considered a key process in the development of a whole body inflammatory response secondary to cardiopulmonary bypass (CPB). Increased levels of a multitude of soluble mediators have been described as being released during and after cardiac surgery. Circulating endothelial cells have recently been established as a novel marker of endothelial damage in a variety of vascular disorders. Blood samples from 20 patients undergoing elective coronary artery bypass surgery were obtained preoperatively and 1, 6, 12, 24, and 48 h after termination of CPB. Control samples were obtained from ten healthy volunteers. Circulating endothelial cells (CEC) were isolated with immunomagnetic anti-CD146-coated Dynabeads, and counted in a Nageotte chamber. Low numbers of CEC were observed in healthy control volunteers (12 +/- 6 cells/mL; median: 9 cells/mL). CEC numbers were already significantly elevated in all patients before CPB, and there was a further significant increase after weaning from CPB (maximum increase at 6 h after CPB: 73 +/- 30 cells/mL; range: 30-153 cells/mL, p < 0.001). The number of CEC provides further and direct evidence that CPB is associated with a pronounced endothelial injury and/or damage. CEC appear to be most useful markers for vascular endothelial activation because they are specific, stable, and circulating components of injured vessel wall.

Aged↗
PubMed2005

Urine protein patterns can serve as diagnostic tools in patients with IgA nephropathy.

BACKGROUND: IgA nephropathy (IgAN) is the most common chronic glomerular disease in adults. End-stage renal disease (ESRD) develops in about 30% of the patients. Early intervention and consequent therapy may prevent or at least delay the development of ESRD in these patients. Up to now, the diagnosis could only be achieved with a renal biopsy. METHODS: The urine of 45 patients with IgAN was collected and screened for protein/polypeptide patterns with a novel high throughput method, capillary electrophoreses on-line coupled to a mass spectrometer (CE-MS). CE-MS allows the fast and accurate evaluation of up to 2000 polypeptides in one urine sample. The results in IgAN were compared to findings in 13 patients with membranous nephropathy (MN) and 57 healthy individuals. RESULTS: In the patients with IgAN, even when urinary protein excretion was within the normal range of regular tests, the polypeptide pattern in urine differed significantly from that of healthy controls and patients with MN, indicating a specific "IgAN" pattern of polypeptide excretion. Classification regarding discrimination of IgAN from healthy controls and from MN had a sensitivity of 100% and 77%, respectively. Specificity was 90% and 100%, respectively. Compared to patterns established earlier in patients with minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), or diabetic nephropathy (DN), sensitivity and specificity were 100%. Treatment of the patients was associated with changes of the pattern, possibly indicating a therapeutic effect. CONCLUSION: Proteomic analysis with CE-MS coupling permits fast and accurate identification and differentiation of polypeptide patterns in the urine of patients with IgAN, allowing differentiation from healthy controls and, probably, other renal diseases.

Adult↗
PubMed2005

Detection of acute tubulointerstitial rejection by proteomic analysis of urinary samples in renal transplant recipients.

This study investigates proteomic analysis of urinary samples as a non-invasive method to detect acute rejection of renal allografts. Capillary electrophoresis coupled to mass spectrometry (CE-MS) was used to analyze urinary samples in 19 patients with different grades of subclinical or clinical acute rejection (BANFF Ia to IIb), 10 patients with urinary tract infection and 29 patients without evidence of rejection or infection. A distinct urinary polypeptide pattern identified 16 out of 17 cases of acute tubolointerstitial rejection, but was absent in two cases of vascular rejection. Urinary tract infection resulted in a different polypeptide pattern that allowed to differentiate between infection and acute rejection in all cases. Potentially confounding variables such as acute tubular lesions, tubular atrophy, tubulointerstitial fibrosis, calcineurin inhibitor toxicity, proteinuria, hematuria, allograft function and different immunosuppressive regimens did not interfere with test results. Blinded analysis of samples with and without rejection showed correct diagnosis by CE-MS in the majority of cases. Detection of acute rejection by CE-MS offers a promising non-invasive tool for the surveillance of renal allograft recipients. Further investigation is needed to establish polypeptide patterns in vascular rejection and to explore whether changes in the urinary proteome occur before the onset of histologically discernible rejection.

Adult↗
PubMed2005

Circulating endothelial cells. Biomarker of vascular disease.

Recent research has recognised new populations of non-hematopoietic cells in the blood. One of these, circulating endothelial cells (CECs), often defined by the expression of membrane glycoprotein CD146, are rarely found in the blood in health, but raised numbers are present in a wide variety of human conditions, including inflammatory, immune, infectious, neoplastic and cardiovascular disease, and seem likely to be evidence of profound vascular insult. An additional population are endothelial progenitor cells, defined by the co-expression of endothelial and immaturity cell surface molecules and also by the ability to form colonies in vitro. Although increased numbers of CECs correlate with other markers of vascular disease, questions remain regarding the precise definition, cell biology and origin of CECs. For example, they may be damaged, necrotic or apopototic, or alive, and could possess procoagulant and/or proinflammatory properties. However, since these cells seem to be representative of in situ endothelium, their phenotype may provide useful information. Indeed, whatever their phenotype, there is growing evidence that CECs may well be a novel biomarker, the measurement of which will have utility in various clinical settings related to vascular injury. Despite this promise, progress is impeded by the diversity of methodologies used to detect these cells. Accordingly, results are sometimes inconclusive and even conflicting. Nevertheless, increased CECs predict adverse cardiovascular events in acute coronary syndromes, suggesting they may move from being simply a research index to having a role in the clinic. The objective of the present communication is to condense existing data on CECs, briefly compare them with progenitor cells, and summarise possible mechanism(s) by which they may contribute to vascular pathology.

Animals↗
PubMed2004

Serologic evidence of Chlamydia pneumoniae infection as a long-term predictor of cardiovascular death in renal transplant recipients.

BACKGROUND: Cardiovascular disease is the main cause of death with a functioning graft in renal transplant recipients. Elevated levels of C-reactive protein (CRP) and evidence of chronic Chlamydia pneumoniae infection have been linked to cardiovascular disease and survival in patients with normal renal function and patients with end-stage renal disease on dialysis. So far, no such data have been available in renal transplant recipients. METHODS: CRP, immunoglobulin (Ig)G and IgA antibodies to C. pneumoniae, and classic risk factors were compiled in 143 patients who underwent renal transplantation between January 1989 and April 1991. Samples were collected at transplantation, 1 year later, and at study end. Cardiovascular disease, death, and graft loss were documented during follow-up. RESULTS: A total of 44 patients died during a mean follow-up of 10 years. Cardiac events were responsible for 37% of deaths. Age, gender, number of antihypertensive drugs, and seropositivity for IgG and IgA antibodies to C. pneumoniae, but not CRP levels, were significantly associated with cardiac death. C. pneumoniae serology and CRP levels, however, did not influence graft survival. Age, presence of diabetes, calcium phosphorus ion product, number of antihypertensive drugs, serum creatinine at 1 year, and presence of chronic rejection were all negatively correlated with graft survival. CONCLUSIONS: Serologic evidence of chronic C. pneumoniae infection is associated with mortality as the result of cardiovascular disease in renal transplant recipients. CRP serum levels do not predict cardiac death in renal transplant recipients, in contrast with patients with normal renal function and patients on dialysis.

Adult↗
PubMed2004

Circulating endothelial cells as a marker of endothelial damage in allogeneic hematopoietic stem cell transplantation.

Damage to endothelial cells is the common feature of vascular disorders associated with hematopoietic stem cell transplantation (HSCT). Elevated numbers of circulating endothelial cells reflect the extent of endothelial damage in a variety of disorders but their use in HSCT has not been investigated so far. We studied 39 patients undergoing allogeneic HSCT with different conditioning regimens and 22 healthy controls. Circulating endothelial cells were enumerated with immunomagnetic isolation during the course of HSCT. After conditioning, cell numbers were significantly elevated (median 44 cells/mL) compared with baseline (median 16 cells/mL) and controls (median 8 cells/mL). Patients who received radiation had an earlier peak when compared with patients who received chemotherapy. Patients who received reduced-intensity conditioning had significantly lower cell numbers (median 24 cells/mL) than those who received standard conditioning. These observations provide a novel marker to investigate microvascular endothelial damage and the effects of different conditioning regimens in patients undergoing HSCT.

Adult↗
PubMed2004

No association of G-463A myeloperoxidase gene polymorphism with MPO-ANCA-associated vasculitis.

BACKGROUND: The activation of neutrophils and monocytes by ANCA, resulting in the release of reactive oxygen species and proteases like myeloperoxidase (MPO), is essential to the pathogenesis of ANCA-associated vasculitis. As the A allele of the G-463A MPO gene polymorphism is associated with diminished activity of MPO, it is conceivable that the presence of this allele protects against MPO-ANCA-associated vasculitis. METHODS: Allelic frequencies of the G-463A polymorphism were studied in 119 ANCA-associated vasculitis patients, 48 with MPO-ANCA and 71 with proteinase 3 (PR3)-ANCA. RESULTS: Allelic frequencies of MPO G-463A promoter polymorphism did not differ between MPO-ANCA- and PR3-ANCA-associated vasculitis patients. Moreover, allelic distribution was similar to that of the normal population. CONCLUSIONS: The data suggest that G-463A polymorphism does not seem to contribute to either MPO-ANCA- or PR3-ANCA-associated vasculitis formation.

Adult↗
PubMed2004

Proteomic patterns established with capillary electrophoresis and mass spectrometry for diagnostic purposes.

BACKGROUND: Proteomics applied in large scale may provide a useful diagnostic tool. METHODS: We developed an online combination of capillary electrophoresis with mass spectrometry, allowing fast and sensitive evaluation of polypeptides found in body fluids. Utilizing this technology, polypeptide patterns from urine are established within 45 minutes. About 900 to 2500 polypeptides as well as their concentrations are detected in individual urine samples without the need for specific reagents such as antibodies. To test this method for clinical application, we examined spot urine samples from 57 healthy individuals, 16 patients with minimal change disease (MCD), 18 patients with membranous glomerulonephritis (MGN), and 10 patients with focal segmental glomerulosclerosis (FSGS). RESULTS: One-hundred seventy-three polypeptides were present in more than 90% of the urine samples obtained from healthy individuals, while 690 polypeptides were present with more than 50% probability. These data permitted the establishment of a "normal" polypeptide pattern in healthy individuals. Polypeptides found in the urine of patients differed significantly from the normal controls. These differences allowed the distinction of specific protein spectra in patients with different primary renal diseases. Abnormal pattern of proteins were found even in urine from patients in clinical remission. CONCLUSION: The data indicate that capillary electrophoresis with mass spectrometry coupling provides a promising tool that permits fast and accurate identification and differentiation of protein patterns in body fluids of healthy and diseased individuals, thus enabling diagnosis based on these patterns.

Adolescent↗
PubMed2004

Circulating endothelial cells and vasculitis.

Systemic vasculitides are a heterogeneous group of disorders with inflammation of blood vessels as their common pathogenetic hallmark. They often pose difficulties with regard to diagnosis and monitoring of disease activity, both at the initial presentation and during follow-up. Novel markers of disease activity are therefore eagerly awaited. Circulating endothelial cells have recently emerged as one such marker and we have demonstrated their clinical use in ANCA-associated small-vessel vasculitis. Not only entire cells but also endothelial microparticles can be detected in vasculitis although their use is not established to date. Repair of endothelial damage is believed to occur via endothelial progenitor cells and their precise role in vasculitis is also unclear at present. Circulating endothelial cells may complement, rather than replace, conventional markers of disease activity. The ultimate aim of our studies may thus be a panel of various laboratory markers for systemic vasculitis.

Biomarkers↗
PubMed2003

Determination of peptides and proteins in human urine with capillary electrophoresis-mass spectrometry, a suitable tool for the establishment of new diagnostic markers.

The on-line coupling of capillary electrophoresis (CE) with electrospray-time-of-flight mass spectrometry (MS) has been used to obtain patterns of peptides and proteins present in the urine of healthy human individuals. This led to the establishment of a "normal urine polypeptide pattern", consisting of 247 polypeptides, each of which was found in more than 50% of healthy individuals. Applying CE-MS to the analysis of urine of patients with kidney disease revealed differences in polypeptide pattern. Twenty-seven polypeptides were exclusively found in samples of patients. Another 13, present in controls, were missing. These data indicate that CE-MS can be applied as powerful tool in clinical diagnostics.

Electrophoresis, Capillary↗