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Biomedical subjects

D Rigal

Publications and source records attributed to D Rigal.

At least 55 records · Page 3Linked to original sources

Spontaneous programmed cell death (PCD) process of lymphocytes of FIV-infected cats: cellular targets and modulation.

Unstimulated lymphocytes from FIV-infected cats undergo spontaneous apoptosis in vitro as indicated by internucleosomal DNA fragmentation and hypodiploid DNA content of nuclei. Unlike what is reported in HIV-infected individuals, we observed that cell death of cat lymphocytes was inhibited by activation. Spontaneous apoptosis was reduced by the addition of cat serum and after activation by phorbol ester (PMA), superantigens (SEB, SEA), and to a lesser extent by mitogens such as concanavalin A and pokeweed mitogen. In contrast, apoptosis of lymphocytes from FIV-infected, but not from control cats was increased in the presence of calcium ionophore (ionomycin). Analysis of the phenotype of cells undergoing apoptosis revealed that cell death is not restricted to a cell subpopulation but involved all lymphocyte subsets. These data suggest that the mature lymphocytes of FIV-infected cats appear programmed to die by apoptosis unless rescued by specific agents, such as protein kinase C activators or mitogens.

Animals↗

Spontaneous programmed cell death (PCD) process of lymphocytes of FIV-infected cats: pharmacological modulation in vitro.

We previously reported that unstimulated lymphocytes in culture from FIV-infected cats undergo spontaneous apoptosis in vitro as indicated by internucleosomal DNA fragmentation and hypodiploid DNA content of nuclei. Unlike what is reported in HIV-infected individuals, we observed that cell death of cat lymphocytes was inhibited by activation. Spontaneous apoptosis was reduced by the addition of cat serum, interleukins [interleukin (IL)1, Il2, IL6 and interferon-gamma (IFN gamma)] and after activation by phorbol ester [phorbol myristyl acetate (PMA)], superantigens [staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin A (SEA)], and to a lesser extent by mitogens such as Concanavalin A and pokeweed mitogen, IN contrast, apoptosis of lymphocytes from FIV-infected, but not from control cats was increased in the presence of calcium ionophore (ionomycin). In this study, we studied the spontaneous programmed cell death (PCD)-inducing pathways, and the mechanisms of action of PMA, SEB and SEA. Spontaneous lymphocyte apoptosis of FIV-infected cats was inhibited by cycloheximide, ZnSO4 and N-acetyl-cystein. The preventive effect of SEB and SEA was inhibited by actinomycin, but not by inhibitors of kinases. Calyculin, an inhibitor of phosphatase, had no effect either on spontaneous apoptosis, or on the action of PMA, SEB and SEA. Ionomycin-induced apoptosis was found sensitive to PMA and cytokines. In FIV-infected cats, these data suggest that the mature lymphocytes appear programmed to die by apoptosis, unless rescued by specific agents, such as protein kinase C activators or growth factors, and that spontaneous PCD seems to be dependent of de nove protein synthesis (see effect of cycloheximide). The effects of PMA, SEB and SEA are probably mediated by de novo proteins which for PMA, undergo a phosphorylation involving serine-threonine and/or tyrosine groups. Our data suggest a clear difference between lymphocytes from FIV-infected cats and lymphocytes from HIV-infected humans, with regard to their metabolic regulations.

Animals↗

[Allergy and preservatives. Apropos of 3 cases of allergy to benzalkonium chloride].

BACKGROUND: Ophthalmologists are not always aware of the hypersensitising potential of benzalkonium chloride (preservative widely used in ophthalmology). OBSERVATIONS: Hypersensitivity to benzalkonium chloride was asserted by allergologic exploration in the three cases reported. Contact allergy, a frequent phenomenon, was diagnosed in two cases. The third case represents one of the few cases described in ophthalmology of immediate hypersensitivity associated to vital risk: chemosis and angioneurotic edema is reported in a woman who had been already presented an anaphylactic shock to a quaternary ammonium family anaesthetic. DISCUSSION: The ocular allergologic evaluation is specified. Risk to benzalkonium chloride sensitisation factors (ophthalmology, presence of benzalkonium chloride in numerous daily used products) are reviewed. The risk of severe immediate allergy is increased due to the relationship of benzalkonium chloride to other quaternary ammonium compounds. CONCLUSION: Best risk prevention remains allergen eviction.

Adult↗

[Bacterial contamination of eyedrops in clinical use].

PURPOSE: The aim of this study is to assess the importance of bacterial contamination of multidose eyedrops in a routine clinical setting. MATERIAL AND METHODS: 406 eyedrop vials were cultured about one week after clinical use: 204 collected from an Ophthalmic Department, and 202 from a Nursing Home. The microbiological analysis was performed on the tip and the residual eyedrop, counting the number of bacterial colonies. RESULTS: 66 (16.3%) from the 406 analyzed vials were contaminated, and 5.4% out of these were severely affected. There was no significant difference between the "Ophthalmic Department" and the "Nursing Home". Commensal germs were the most frequently encountered in both groups. 4 gram negative organisms were isolated from the "Nursing Home" group. CONCLUSION: These results are in agreement with the literature. Comparison between our two groups is difficult because the eyedrops and uses were different. However, we notice the presence of gram negative organisms in the "Nursing Home". These severe contaminations due to opportunistic pathogen organisms are rare (0.75%), probably underestimated, and represent a real infectious risk during instillation. The study of the contamination site shows that the eyedrop is more often contaminated than the tip. This can be in relation to germ desicsation and to an aspiration phenomenon of contaminated fluid at the tip level. At last, the role of preservatives is not sufficient to ensure the sterility of multidose eyedrops during their use, and this justifies safer (single dose or filtration system) eyedrop vials.

Adult↗

Ganciclovir ophthalmic gel (Virgan; 0.15%) in the treatment of herpes simplex keratitis.

PURPOSE: Ganciclovir is a broad-spectrum virustatic agent. Its efficacy and safety after ocular application have been demonstrated in studies of herpetic keratitis in rabbits. Two strengths of ganciclovir gel (0.05 and 0.15%) were compared with 3% acyclovir ointment in the treatment of superficial herpes simplex keratitis in humans. METHODS: Two multicenter randomized clinical trials were carried out in Africa (Trial 1) and Europe (Trial 2). Sixty-seven patients (Trial 1) and 37 patients (Trial 2) from herpetic ulceration were recruited. RESULTS: The results showed no statistically significant difference between the treatment groups, although the healing rates tended to be better in the group receiving 0.15% ganciclovir gel, with healing rates of 85% (Trial 1) and 83% (Trial 2) as compared with 72% (Trial 1) and 71% (Trial 2) in the group receiving acyclovir ointment. Local tolerance was found to be superior with the gel formulation of ganciclovir with fewer complaints of discomfort (stinging, burning) or blurred vision after application of the drug. Systemic absorption of the drug was low. No hematologic changes were detected. CONCLUSIONS: These findings support the efficacy of ganciclovir gel in the treatment of ulcerative herpes simplex keratitis and demonstrate its superior local tolerance when compared with acyclovir ointment.

Acyclovir↗

Human platelet antigen frequencies of platelet donors in the French population determined by polymerase chain reaction with sequence-specific primers.

To prevent human platelet alloimmunization, Blood Transfusion Centres have to develop a strategy close to the erythrocytes' one. The first step of this strategy is to perform the HPA typing of donors with an accurate method. We applied the PCR-SSP to type 800 platelet donors in the HPA-1 and HPA-5 systems and 350 in the HPA-2 and HPA-3 ones. This study reports the human platelet antigen frequencies of four platelet-specific alloantigen systems in the French population. The results are quite similar to those currently published for Caucasian population frequencies. Low prevalences are observed for the HPA-1b, (2%), HPA-2b (0.6%) and HPA-5b (2%) groups. Furthermore, this study confirms the need to type donors and recipients in the HPA-1 system at least, in case of post-transfusion pupura and platelet refractoriness to platelet transfusion therapy.

Antigens, Human Platelet↗

[Contribution of dual CD13/CD14 markers in combination with CD34 for the collection of peripheral hematopoietic stem cells].

We evaluated the reliability of a flow cytometry technique for counting mononuclear cells (MNCs) in cytapheresis products. Eighty freshly-prepared samples of peripheral stem cells were studied using a dual immunolabeling technique with antibodies to CD13/CD14, and were also labeled with anti-CD34. Results of this immunophenotype determination were compared to those of the conventional method for counting MNCs under the microscope. Dual CD13/CD14 labeling was found to be a simple and reliable method for counting MNCs in the presence of immature and stimulated cells. When used in combination with CD34 labeling, the dual immunolabeling method helped improve the evaluation of the quality of peripheral stem cell grafts.

Antigens, CD34↗

RU 41 740 (Biostim) and IL-4, or IL-13, have opposite effects on CD14, CD23, HLA-DR and HLA-DQ on monocytes.

RU 41 740 (Biostim) is a glycoprotein extract obtained from Klebsiella pneumoniae. Its immunostimulating properties on monocytes have been established in vivo and in vitro. To confirm its spectrum of action at molecular level we studied its role on the modulation of four molecules involved in antigen presentation (HLA-DR, HLA-DQ), uptake of endotoxin (CD14) and activation (CD23). These four molecules are known to be modulated by interleukins IL-4 and IL-13. We found that HLA-DR, HLA-DQ, CD14 and CD23 were differentially regulated by biostim and IL-4 or IL-13. Surprisingly, Biostim inhibited the IL-4 or IL-13-induced expression of CD23, HLA-DQ and HLA-DR, while it did not have any action on these molecules by itself. We therefore hypothesize that Biostim, through the action on its receptor, could interact with the IL-4 receptor and IL-13 receptor and/or inhibit the IL-4 and IL-13 receptor transducing signal.

Adjuvants, Immunologic↗

[Apheresis tolerance and acceptability in the child weighing 30 kg or less, with the exception of infants].

Over the ten past years, we performed 336 apheresis among 51 children who were 19 months to 15 years old (10 to 30 kg body weight). 3 types of apheresis were carried out. 14 red blood cell exchange, 293 plasma exchanges and 29 peripheral blood stem cell collections (CSP). 5 different types of continuous or discontinuous flow machines have been used. Technical adaptations depending on patient blood volume, hematocrit, type of machine used and apheresis performed permitted us to obtain a very good tolerance and acceptability. According us, the apheresis should be used each time this treatment is needed in the child, because of the very low frequency of side effects.

Adolescent↗

Expression of a NK cell-restricted epitope on decidual large granular lymphocytes.

Decidual large granular lymphocytes (DLGL) are the most abundant lymphoid cell type found in the first trimester maternal decidua. The function of DLGL remains controversial, although freshly isolated DLGL have been shown to exert a weak NK activity. We report here the phenotypic characterization of two DLGL subpopulations by immunofluorescence, using mAb against CD56, PEN5 as well as adhesion molecules potentially involved in cell-cell contact between DLGL and trophoblasts. DLGL are CD56bright and express the CD2, CD11a, CD18, CD38 and CD50 molecules, dimly the CD54 molecule, and poorly the CD102 and CD69 molecules. A strong expression of the polysialylated form of N-CAM (or CD56) was also observed on the surface of DLGL. Finally, 40% of CD56bright DLGL cells express the PEN5 epitope, which is selectively expressed on CD56dim cells NK in the peripheral blood. No other phenotypic difference was detected between the CD56brightPEN5+ and the CD56brightPEN5- DLGL populations. Our results show that DLGL are heterogeneous and suggest that the CD56brightPEN5+ DLGL subset belongs to the classical NK cell lineage.

Amino Sugars↗

Strong increase in the percentage of the CD8bright+ CD28- T-cells and delayed engraftment associated with cyclosporine-induced autologous GVHD.

Four children with acute lymphoblastic leukaemia had autologous bone marrow (BM) or peripheral stem cell (PSC) transplantation with low dose of cyclosporine (CsA, img/kg/d i.v. during the first 28 d) to induce an autologous GVHD (auto-GVHD). Two children did not have clinical auto-GVHD and they relapsed 3 and 4 months after treatment. The 2 other children had clinical signs of auto-GVHD (grade I and grade II); they both are in complete remission but after a first normal haematological recovery they had a prolonged period of aplasia until month 9 for 1 patient and still persistent at month 7 in the other case. We studied lymphocyte subsets reconstitution after transplantation in these patients. All patients had an important decrease in the CD4/CD8 ratio related both to a strong decrease in the CD4+ cells and a strong increase in the CD8+ cells. Most of the CD8+ cells were of the CD8bright+ CD28- phenotype. These CD8bright+ CD28- T-cells represented from 33% to 68% of the total lymphocytes. We discuss the role of these cells after autologous transplantation with CsA, and wonder if these cells could mediate cytotoxicity. In conclusion, among 4 children who received autologous BM or PBC transplantation with low dose of CsA, we observed a complete remission after an auto-GVHD and a prolonged period of aplasia in 2 patients and a relapse of leukaemia in 2 other patients. All these 4 patients had an increase in the CD8bright+ CD28- T lymphocytes.

Autoimmune Diseases↗

Allogenic BMT in children: differential lymphocyte subset reconstitution according to the occurrence of acute GVHD.

To acquire some biological markers associated with the occurrence of acute graft-versus-host disease (aGVHD) after allogeneic bone marrow transplant (BMT) in children, we have studied the lymphocyte subset reconstitution and the percentage of peripheral blood mononuclear cells bearing HLA-DR and HLA-DQ class II molecules. This study included 37 allogeneic BMT: either with (n = 17) or without (n = 20) aGVHD. Within 2 months after transplantation, we observed that patients with aGVHD had a unique mononuclear cell profile characterized by (i) a significant increase in the percentages of CD8bright+CD28- T cells (P = 0.05) and CD3+ T cells (P = 0.001), (ii) an important decrease in the percentage of CD56+ cells (P = 0.0001), and (iii) a decrease in the percentages of HLA-DQ+ and HLA-DR+ monocytes (P = 0.001) and HLA-DQ+ T lymphocytes (P = 0.0001), in comparison with patients without aGVHD. Moreover, statistical studies indicate that there was a positive correlation between CD8bright+CD28- and CD3+ T cells, whereas CD3+ T cells were negatively correlated to CD56+ cells. We did not find any statistical correlation between the percentages of HLA-DQ+ or HLA-DR+ cells and the percentages of these lymphocyte subsets. Therefore, in this study done in children, we suggest that patients with (i) less than 20% of DQ+ monocytes, (ii) less than 25% of CD56+ lymphocytes, and (iii) an enhanced percentage of CD8bright+CD28- T cells are strongly associated with aGVHD. Unfortunately, these biological markers of a GVHD may not precede the clinical manifestations of the disease.

Acute Disease↗

Lymphocyte subset abnormalities in German shepherd dog pyoderma (GSP).

Peripheral blood lymphocyte subpopulations were studied in 12 German shepherd dogs suffering from deep pyoderma (GSP). Twelve other healthy but matched dogs were used as controls. GSP was found to be associated with an imbalance in the CD4 and CD8 subsets (respectively 37.3 +/- 8.7% and 28.6 +/- 6.6%, as compared to 47.5 +/- 8.8% and 19.3 +/- 4.0% in the controls). The activation markers were not affected by GSP. Moreover, analysis of the B-cell populations showed a striking decrease in the level of CD21 cells (5.5 +/- 3.3% of CD21+ lymphocytes, compared to 12.2 +/- 6.0 in the controls). This study suggests that the immunological imbalance observed in GSP may be associated with defective helper cells, and provides further evidence that dogs suffering from GSP are not immunologically normal reactors.

Animals↗

Development of a simple, rapid and accurate in vitro whole blood technique for the detection and semi-quantification of FIV cellular viremia.

A new, simple, rapid and accurate culture technique is described for a semi-quantitative analysis of cellular viremia in FIV-infected cats. This assay can be carried out with small amounts of whole blood, and is based on the detection of FIV core gag antigen, which is released in culture supernatants. The amount of core antigen produced is measured with an enzyme-linked immunoassay using specific monoclonal antibodies. This whole blood technique (WB method) was compared with a culture method using isolated peripheral blood mononuclear cells (PBMC method). FIV could be detected in whole blood of all experimentally infected cats, but not from uninfected cats. This assay offers a number of advantages (small blood samples required, no leukocyte separation and lymphocyte purification procedures) and its reproducibility is very good. It provides a convenient in vitro cellular assay for viral semi-quantitation, well adapted for monitoring efficacy of prototype FIV vaccines or experimental antiviral drugs. Also, it could facilitate the study of the pathogenesis of FIV-related progressive immunodepression. Finally, it offers an alternative to serological techniques for diagnostic purposes in several circumstances: early viremia, maternal antibodies.

Animals↗

Autoantibodies to histone, DNA and nucleosome antigens in canine systemic lupus erythematosus.

Dogs can develop systemic lupus erythematosus syndromes that are clinically similar to those seen in humans. In contrast, previous observations suggest differences in their autoantibody reactivity patterns against histones and DNA which are components of the nucleosome in chromatin. The objective of this study was to assess comprehensively the levels of autoantibodies against histone, DNA and nucleosome antigens in a population of lupus dogs. The specificities of antibodies in lupus and control dog sera were determined using IgM- and IgG-specific reagents in an ELISA against a variety of chromatin antigens. When compared with control sera, IgG antibodies to individual histones H1, H2A, H3 and H4 were significantly higher in the lupus group. In contrast, we did not detect IgG antibodies specific for H2B, H2A-H2B, DNA, H2A-H2B-DNA or nucleosome in lupus dogs. There was no significant increase in any of the IgM specificities tested. Therefore, the reactivity pattern to nucleosome antigens in canine lupus is restricted to IgG antibodies against individual histones H1, H2A, H3 and H4. This stands in contrast with human and murine lupus, where autoantibodies are directed against a wide variety of nucleosomal determinants, suggesting that unique mechanisms lead to the expansion of anti-histone antibody clones in canine lupus. The high incidence of glomerulonephritis in dog lupus suggests that anti-DNA antibodies are not required for the development of this complication, whereas IgG anti-histone antibodies may be relevant to its pathogenesis.

Animals↗