Search PubMed⌕ Search

Biomedical subjects

G Bernard

Publications and source records attributed to G Bernard.

At least 19 recordsLinked to original sources
PubMed2002

Non-expression of HLA-A*2901102 N is caused by a nucleotide exchange in the mRNA splicing site at the beginning of intron 4.

We describe the identification of the novel human leukocyte antigen (HLA) blank allele A*2901102 N which was detected in an individual of mixed race. The serological HLA class I typing was A1; B7,44 whereas PCR-SSP indicated the presence of an additional A*29 allele. The pedigree analysis demonstrated that the new blank allele segregated with the haplotype A*29null B*07, inherited from the individual's Vietnamese father. A single G-->Tau transversion was detected at position 1 of intron 4, which is a highly conserved nucleotide position in vertebrate splice donor sites. Accordingly, it is very likely that this nucleotide exchange inhibits the splicing of intron 4, resulting in a premature stop codon further downstream. Despite this alteration, transcription into mRNA was demonstrated.

Alleles↗
PubMed2001

CD99 signals caspase-independent T cell death.

Death signaling by Fas and TNF receptors plays a major role in the control of activated mature T cells. However, the nature of the death receptors, which may be used by the immune system to control T cells that have not acquired susceptibility to Fas ligand or TNF, is not established. In this study, we demonstrate that engagement of distinct epitopes on CD99 rapidly induces T cell death by a novel caspase-independent pathway. A new mAb to these CD99 epitopes, Ad20, induces programmed cell death of transformed T cells as determined by morphological changes, phosphatidylserine exposure on the cell surface, and uptake of propidium iodide. In general, ligation of CD99 induced kinetically faster and more profound death responses as compared with the impact of anti-Fas and TNF-related apoptosis-inducing ligand (TRAIL). Ad20-induced programmed cell death was observed with seven of eight T cell lines examined, and notably, only two of these were distinctly responsive to anti-Fas and TRAIL. CD99-mediated death signaling proceeded independently of functional CD3, CD4, CD45, and p56(lck), revealed distinctions from CD47-mediated T cell death responses, and was not influenced by interference with CD47 signaling. In contrast to the effect on transformed T cell lines, Ad20-induced death responses were not observed with normal peripheral T cells. Thus, our data suggest that CD99 is linked to a novel death pathway that may have biologic relevance in control of early T cells.

12E7 Antigen↗
PubMed2001

Distinct hematopoietic support by two human stromal cell lines.

OBJECTIVE: The hematopoietic microenvironment is complex, and the role of myofibroblast in its function is crucial. In order to obtain a stable model reflecting this particular cell type, we have previously established human bone marrow cell lines from primary myofibroblastic Stro1(+) population (pStro1(+)). We placed HPV16 E6 and E7 expression under the control of different promoters. Here, we have characterized and studied the hematopoietic support for two cell lines corresponding to the promoters alpha-SM (alphaSM-56 line) and SV40 (SV40-56 line). MATERIALS AND METHODS: The expression profile was analyzed at the RNA level by gene array and at the protein level by Western blot, flow cytometry, and ELISA. Hematopoietic support determined using colony-forming unit (CFU) and stroma-adherent colony-forming cell (SA-CFC) assays. RESULTS: The phenotype of cell lines was not significantly modified compared with primary myofibroblastic cells. They secreted a broad spectrum of hematopoietic cytokines and nonspecific mediators. The two lines allowed the growth of hematopoietic precursors and had different support capabilities. CONCLUSIONS: We have extensively characterized two novel human bone marrow stromal cell lines. They retained a myofibroblastic phenotype and have substantial but different hematopoietic support capabilities. These lines provided a basis for determining stromal factors involved in stem-cell regulation.

Antigens, CD34↗
PubMed2001

Genes, isochores and bands in human chromosomes 21 and 22.

The recently available DNA sequences from chromosomes 21 and 22 enabled us to define the relationships of different band types with isochores and with gene concentration and to compare these relationships with previous results. We showed that chromosomal bands appear as Giemsa or Reverse bands depending not on their absolute GC level, but on the composition GC level relative to those of adjacent contiguous bands. We also demonstrated that the GC-richest, and gene-richest H3+ bands are characterized by a lower DNA compaction compared with the GC-poorest, gene-poorest L1+ bands. Moreover, our results indicate that the human genome contains about 30,000 genes.

Chromosome Banding↗
PubMed2000

CD99 engagement: an effective therapeutic strategy for Ewing tumors.

CD99 is a Mr 32,000 transmembrane molecule that shows a high level of expression on cells of the hemopoietic system as well as on Ewing tumor cells. Within the hematopoietic system, CD99 has been implicated in cell adhesion and cell death, participating in this way in the differentiation of T-cell precursors. In this study, we demonstrate that engagement of CD99 significantly inhibits the in vitro and in vivo growth ability of Ewing tumor cells by delivering an apoptotic stimulus and reducing the malignant potential of these cells. Moreover, we show that anti-CD99 monoclonal antibodies may be advantageously used in association with conventional anticancer agents. These results provide a novel entry site for therapeutic intervention, which may have application in the care of patients with Ewing tumor, and warrant additional studies to clarify the molecular mechanisms activated by CD99 engagement.

12E7 Antigen↗
PubMed2000

Caspase-independent commitment phase to apoptosis in activated blood T lymphocytes: reversibility at low apoptotic insult.

Little is known about the mechanisms of programmed death triggered in T lymphocytes by stimuli that can bypass caspase activation. Anti-CD2 monoclonal antibody and staurosporine are such apoptosis inducers because they operate in the presence of broad-spectrum caspase inhibitors BOC-D.fmk and Z-VAD.fmk. A system was devised, based on the isolation according to density of activated blood T cells progressively engaged in the apoptotic process. This allowed definition of a sequence of caspase-dependent and caspase-independent apoptogenic events that are triggered by anti-CD2 and staurosporine. Thus, a commitment phase to apoptosis was defined that is entirely caspase independent and that is characterized by cell volume loss, partial chromatin condensation, and release into the cytosol and the nucleus of mitochondrial "apoptosis-inducing factor " (AIF). Committed cells were viable, displayed a high mitochondrial inner transmembrane potential (triangle upPsim), and lacked large-scale and oligonucleosomal DNA fragmentation. Mitochondrial release of AIF was selective because cytochrome c was retained in mitochondria of the very same cells. Mitochondrial release of cytochrome c occurred later, at the onset of the execution phase of apoptosis, concurrently with triangle upPsim collapse, poly (ADP-ribose) polymerase cleavage, and DNA fragmentation. The apoptogenic events of this commitment phase are reversible if the strength of the stimulus is low and of short duration.

Apoptosis↗
PubMed2000

CD99 (E2) up-regulates alpha4beta1-dependent T cell adhesion to inflamed vascular endothelium under flow conditions.

CD99/E2 is an integral transmembrane protein which forms, together with Xga, a distinct family whose genes are located in the pseudoautosomal region. The number of T cells that firmly bound to vascular endothelial cells under physiological shear stress increased 2-14-fold upon CD99 stimulation, and bound cells became much more resistant to detachment forces and spread. T cell arrest occurred within 1 min and was dependent on the alpha4beta1-VCAM-1 pathway. In contrast, the alphaLbeta2-ICAM-1 pathway remained unactivated. This was observed with T cell lines and with activated peripheral blood lymphocytes, and was limited within the resting peripheral CD4+ T cells to the memory subset, while virgin cells were unaffected. This discloses a stepwise regulation of the T cell extravasation cascade.

12E7 Antigen↗
PubMed2000

Fertility after hysteroscopic myomectomy: effect of intramural myomas associated.

OBJECTIVE: The aims of this retrospective study were to evaluate the subsequent fertility and outcome of pregnancies after hysteroscopic myomectomy according to (a) the characteristics of submucous myomas and (b) the association with intramural myomas. MATERIALS AND METHODS: From July 1994 to June 1997, 119 patients had hysteroscopic myomectomy including 31 infertile women. Among these 31 patients, the mean number of removed myomas by hysteroscopy was 1.4 (range 1-4) and the mean diameter of fibroid was 20 mm (range 10 to 50). RESULTS: Eleven out of 31 women (35.5%) became pregnant. Thirteen pregnancies were observed including nine term deliveries, three miscarriages and one premature labor at 24 weeks of amenorrhea. A difference in delivery rate was found between patients with one submucous myoma resected and those with two or more (p=0.02). No difference in pregnancy and in delivery rates was observed according to size and location of submucous myomas. In contrast, in patients without intramural myomas, the delivery rate (p<0.03) was significantly greater and the delay of conception (p=0.05) was significantly shorter than those found in patients with intramural myomas. CONCLUSION: Our study suggest that fertility after hysteroscopic myomectomy depend on (a) the number of submucous myomas resected and (b) the association with intramural fibroids.

Abortion, Spontaneous↗
PubMed2000

Analysis of the microenvironment necessary for engraftment: role of the vascular smooth muscle-like stromal cells.

This is a review of recent data concerning the phenotype of human and murine stroma, as grown in long-term cultures. Using data on cytoskeletal and extracellular matrix protein expression, a sequential model of differentiation from mesenchymal (stem) cells to vascular-smooth muscle-like stromal cells is proposed. This model would apply, at least in the mouse, to stromal cells generated from different sites of hematopoiesis (bone marrow, fetal liver, spleen, and yolk sac). The in vivo counterparts of vascular-smooth muscle-like stromal cells in the different sites of definitive hematopoiesis are discussed.

Animals↗
PubMed2000

[Integrins and cell cycle control by the environment].

Integrins insure cell adhesion to extra-cellular matrix components; they are thus involved in tissue architecture. They also can insure intercellular adhesions by binding to surface molecules from the immunoglobulin superfamily. Integrins binding to their ligands induce cytoskeleton reorganisation and, consequently, they gather into focal adhesion contacts. This greatly strenghthens mechanical forces. Nevertheless, integrins can also participate in cell locomotion and, moreover, tranduce within cells signals that can extensively influence cell metabolism, cell cycle and apoptosis. Doing so, they can interact with signals from other cellular receptors, such as soluble growth factors. They are therefore key molecules to integrate intrinsic and extrinsic events of the cellular behavior. They profoundly influence oncogenesis and the metastatic process.

Animals↗
PubMed1999

Vascular smooth muscle differentiation of murine stroma: a sequential model.

Previous studies by our group showed that stromal cells from human long-term marrow cultures were mesenchymal cells following a vascular smooth muscle pathway. The present study using 58 immortalized stromal lines from different hematopoietic sites was conducted to verify whether this hypothesis also held true for murine stroma. Principal components analysis performed using cytoskeletal and extracellular matrix proteins allowed the segregation of five factors explaining more than 70% of the variance. Factor I, including osteopontin and vimentin, and factor II, laminins and fibronectins, were representative of the mesenchyme. The remaining three factors were representative of vascular smooth muscle: factor III, including alphaSM actin, SM alpha actinin, SM22alpha, EDa+ fibronectin, and thrombospondin-1; factor IV, metavinculin and h-caldesmon; and factor V, smooth muscle myosin SM1 and desmin. All lines expressed factors I and II; 53 lines expressed factor III, 35 lines expressed factor IV; and 11 lines expressed factor V. A second principal components analysis including membrane antigens indicated the cosegregration of vascular cell adhesion molecule-1 with osteopontin and that of Ly6A/E with vimentin, whereas CD34 and Thy-1 appeared to be independent factors. The heterogeneity of vascular smooth muscle markers expression suggests that harmonious maintenance of hematopoiesis depends on the cooperation between different stromal cell clones.

Animals↗
PubMed1999

Chronic pulmonary paracoccidioidomycosis (South American blastomycosis): high-resolution CT findings in 41 patients.

OBJECTIVE: To assess the pulmonary parenchymal findings on high-resolution CT in 41 patients with the chronic form of paracoccidioidomycosis (South American blastomycosis). SUBJECTS AND METHODS: The study included 41 consecutive patients in whom chronic paracoccidioidomycosis had been proven. All patients underwent high-resolution CT (1-mm collimation, high-spatial-frequency reconstruction algorithm) at 12 equally spaced intervals through the chest. The images were analyzed by two radiologists, and each final decision was reached by consensus. RESULTS: Thirty-eight (93%) of the 41 patients had CT scans with abnormal findings. The findings included interlobular septal thickening in 36 patients (88%), 1-25 mm diameter nodules in 34 (83%), peribronchovascular interstitial thickening in 32 (78%), centrilobular opacities in 26 (63%), intralobular lines in 24 (59%), ground-glass opacities in 14 (34%), cavitation in seven (17%), air-space consolidation in five (12%), traction bronchiectasis in 34 (83%), and paracicatricial emphysema in 28 (68%). In approximately 90% of patients, the abnormalities were bilateral and symmetrical and involved all lung zones. CONCLUSION: High-resolution CT findings of paracoccidioidomycosis consist predominantly of interstitial abnormalities and nodules associated with traction bronchiectasis and paracicatricial emphysema in a bilaterally symmetrical distribution.

Adult↗
PubMed1999

[Mechanical and metabolic complications of hysteroscopic surgery: report of a retrospective study of 352 procedures].

Our objective was to determine, in a retrospective study of 352 operative hysteroscopies: (a) the rates and the types of complications and (b) the risk factors of peroperative perforations. The most important complications represented 1.7% including two haemorrhage, one symptomatic metabolic abnormalities and three uterine perforations with bowel injuries (0.8%). Furthermore, minor complications were observed in 9.3% including non symptomatic metabolic abnormalities (5.5%) and uterine perforations without visceral injury. Among mechanical complications, the majority were uterine perforations (4%). No relation was found between menopausal status of the patients and the occurrence of uterine perforation. In contrast, the perforation rate was statistically greater in patients treated for synechia than those found for myoma (p < 0.0001). Furthermore, the perforation rate was statistically higher for resection of myomas as compared with endometrial resection (p < 0.0001) or polyp resection (p < 0.0008). Moreover, in our experience, the perforation rate depended on hysteroscopic experience of surgical operators.

Adult↗
PubMed1998

[Laparoscopy-assisted hysterectomy and laparoscopic preparation. Apropos of a series of 177 cases].

Our objective was to determine the limits of laparoscopic-assisted vaginal hysterectomy (LAVH) and the value of a preoperative scoring system to determine the operative approach to hysterectomy. Between January 1991 and December 1996, 152 out of 177 patients had LAVH and 25 had laparoconversion. The mean operating time was 163 min. The overall postoperative complication rate was 8.4%. The hospital stay was 4.8 days for LAVH versus 6.2 days for laparoconversion (p < 0.01). For each patient, a preoperative scoring system was established according to uterine size, previous laparotomy, uterine mobility, pelvic adhesions and endometriosis stage. The laparoconversion rate increased according to the score, as it was 7.8% for a score < or = 7 and 80% for a score > 7. LAVH offers a technique to convert some abdominal hysterectomies into vaginal hysterectomies. The use of the preoperative scoring system may help to determine patients who may benefit from the laparoscopic route and those with a high risk of laparoconversion.

Adult↗
PubMed1997

Apoptosis of immature thymocytes mediated by E2/CD99.

E2/CD99 is a 32-kDa transmembrane molecule that does not belong to any known family of proteins. It appears to regulate adhesion properties of T cells as previously reported, in particular, the induction of homotypic adhesion in CD4+ CD8+ thymocytes. Apoptosis induced via E2/CD99 displays characteristic morphologic features, but includes early mitochondrial alterations and phosphatidylserine exposure at the outer leaflet of the plasma membrane. It is not followed by detectable DNA fragmentation, and its time course is much longer than apoptosis induced via the Fas/CD95 pathway. It requires 18 h for completion. E2/CD99-induced apoptosis does not require any RNA or protein synthesis and still occurs following blockage of the Fas pathway. It is, however, dependent on CPP32 and IL-1beta-converting enzyme-type cysteine proteases, as shown by blockade with their respective specific inhibitors. This effect is restricted to double-positive thymocytes carrying an intermediate density of CD3 and including all CD69+ cells. Thus, E2/CD99 apears to mediate a distinctive apoptotic signal at a critical stage of thymocyte differentiation, i.e., when positive selection is known to occur.

12E7 Antigen↗
PubMed1997

Integrin-associated protein (CD47) is a comitogenic molecule on CD3-activated human T cells.

IAP is a glycoprotein functionally and physically associated with some integrins, i.e., the leukocyte response integrin and the beta3 integrin chain on placenta, platelets, and polymorphonuclear cells. IAP may act as a transducer element in activation mediated via these integrins. Since IAP is present at high density on peripheral T lymphocytes we have investigated its involvement in T cell activation. We tested three mAbs against IAP, namely B6H12, BRIC126, and 2D3, which recognize two distinct epitopes. IAP cross-linking with B6H12 or BRIC126, but not 2D3, transduces costimulatory signals within highly purified CD3-activated T lymphocytes, i.e., enhancement of proliferation, CD25 expression, and IL-2 secretion, while no effect was observed upon CD2 stimulation. However, we could not observe any functional association between IAP and integrins on peripheral T cells. In an attempt to explore further the activation signal delivered by IAP, we show here that IAP cross-linking with the comitogenic B6H12 mAb induces the phosphorylation on tyrosine of several proteins, one of which is identified as p56(lck) protein tyrosine kinase. Moreover, we observed that IAP is associated with p56(lck) on PMA-activated, but not on resting, T cells. These data suggest that on T cells, IAP may be involved directly via a specific ligand in cell-matrix or cell-cell interactions. Such interactions could trigger protein tyrosine phosphorylation pathways, which play an important role in both maturation and activation of T cells.

Antibodies, Monoclonal↗