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D Rigal

Publications and source records attributed to D Rigal.

At least 73 records · Page 4Linked to original sources
PubMed1995

Abnormalities of lymphocyte subsets in canine systemic lupus erythematosus.

Canine systemic lupus erythematosus (SLE) is a disease clinically very similar to its human counterpart. But so far, no study has reported an accurate evaluation of the lymphocyte subsets in the canine disease. Here, we present a study in which lymphocyte subsets have been evaluated in the peripheral blood of 20 dogs suffering from spontaneous systemic lupus erythematosus (SLE) in active and inactive phases, before and during treatment with prednisone and levamisole. 22 healthy dogs have been used as a control population. We show that canine SLE in active phases is associated with a several lymphopenia (1050 +/- 520 10(6) cells/l versus 2130 +/- 1 020 10(6) cells/l in controls). A striking finding is the imbalance of the CD4 and CD8 subsets (respectively 56.7 +/- 10.7% and 10.9 +/- 3.8% of CD4+ and CD8+ lymphocytes versus 40.5 +/- 11.5% and 18 +/- 4.4% in controls) and a strong activation of T-cells in active phases (64.1 +/- 16.9% of 2B3+ cells versus 46.5 +/- 16.7%). Moreover, we observed a persistence of the T subset imbalance during spontaneous evolution. In contrast, the treatment induced in dogs showing a good response the correction of CD4/CD8 ratio and no clinical manifestations, whereas in low responders no such improvements were observed. Thus, this work suggests that the main immunological imbalance seen in SLE could be associated with defective suppressor cells and provides further evidence of similarity of human and dog SLE.

Animals↗
PubMed1995

[Granulated CD56 lymphocytes of decidual tissue].

The reproduction is a complex mechanism wich is subtly immunoregulated since the semiallogenic fetus is not rejected by the mother. At the feto-maternal tissue interface, where fundamental mechanisms of tolerance occur, a large and unusual lymphocytes population takes place. These cells recruited during the earlier stages of pregnancy are very granulated and express brightly the N-CAM molecule (or CD56) with numerous adhesion and activation molecules. Nevertheless, they seem fonctionnally inactivated. They phenotypically belong to the peripheral blood minor fraction of NK cells which are CD56++CD16-CD3-. Although their function in the establishment of the gestation is not clearly known, because of their phenotypical heterogeneity, they may have two distinct functions: cytotoxicity and suppression. Their cytokine pattern, preferentially of the Th1 type, inform us about their potential functions and their interactions with other cells populations in this tissue, allowing us to understand the main mechanism in the establishment of the pregnancy.

Abortion, Spontaneous↗
PubMed1994

Effects of intravenous immunoglobulins (IVIG) on peripheral blood B, NK, and T cell subpopulations in women with recurrent spontaneous abortions: specific effects on LFA-1 and CD56 molecules.

Polyspecific IgG given intravenously at high dose (IVIG) are increasingly used as an immunomodulating therapy in autoimmune diseases. However, very few studies have dealt with the action of IVIG on the expression of the leukocyte markers. During a clinical trial in which 13 young and healthy women received IVIG to prevent unexplained recurrent abortions we have evaluated by flow cytometry the action of IVIG on 17 clusters of leukocyte differentiation (CD). We found that the IVIG perfusions (0.5 g/kg) induced an increase in the number of polymorphonuclear and monocyte cells in the peripheral blood. This effect lasted 8 days. The IVIG treatment had no effect upon T cell populations stained with antibodies specific for CD2, CD3, CD4, CD8 and on CD4+CD45RA+, CD4+CD29+, CD8+CD28+, CD8+CD28- subpopulations. A weak decrease in the B cell number was observed. The most striking phenomenon was the decrease in the number of CD56+ cells, whereas CD16+ and CD57+ cells were unaltered. By the double-staining technique we showed that CD56+CD16+ cells became CD56-CD16+ cells. Moreover, IVIG decrease the expression level of the LFA-1 molecule on monocytes and lymphocytes. The other adhesion molecules studied remained steady (CD11b, CD49d, CD49e, CD29, CD28, and CD62L). This study has shown that IVIG have no effect on 15 of 17 CD used but downmodulate two adhesion molecules playing a key role in the immune system.

Abortion, Habitual↗
PubMed1994

Effects of PGE2 upon differentiation and programmed cell death of suspension cultured CD4-CD8- thymocytes.

Recently, several works have focused on the modulation of the immune response by arachidonic acid metabolites. Some of these metabolites, such as prostaglandins, have been shown to influence thymocyte "education" in vitro. However, the effect of one of them, prostaglandin E2 (PGE2), in the education of CD4-CD8- double negative immature thymocytes (DN cells) remained unclear. Using a flow cytometry analysis of DN cells cultured for 24 h in the presence of PGE2, we observed, compared with DN thymocytes cultured without PGE2, an increase in the CD4+CD8-CD3- immature thymocytes and in the CD4+CD8- and CD8+CD4- mature single positive thymocytes and a decrease in the DN and CD4highCD8high double positive thymocytes. Other differentiation thymocyte surface markers, such as CD3, CD5, TCR alpha beta, TCR delta gamma and HSAg, revealed an increasing number of thymocytes bearing these first four markers and a lower expression of the HSAg. Furthermore, we observed an accumulation of CD4lowCD8low thymocytes and an increasing proportion of hypodiploid nuclei. These two findings have been shown to be markers of the programmed cell death process. These findings suggest that PGE2 probably acts on thymocyte differentiation through at least two distinct pathways. On the one hand, PGE2 seems to promote differentiation of DN cells into CD4+CD8-CD3- immature cells and drive CD4+CD8+CD3+ thymocyte to a CD4+CD8- and CD8+CD4- mature phenotype. On the other hand, PGE2 is probably implicated directly or indirectly in the increase or the acceleration of the programmed cell death process of immature CD4+CD8+CD3+ thymocytes, which is linked to the positive and/or negative selection.

Animals↗
PubMed1994

A high percentage of HLA-DQ+ and HLA-DR+ mononuclear cells is associated with a low incidence of acute graft-versus-host disease after allogeneic bone marrow transplantation (BMT) in children.

In order to discover some biological markers of acute graft-versus-host disease (aGVHD), we have studied the percentage of peripheral monocytes and T lymphocytes bearing HLA-DR and HLA-DQ class II molecules. This study included 25 allogeneic BMT in children, either with (n = 10) or without (n = 15) aGVHD. Within 2 months after transplantation, a higher percentage of DQ+ and DR+ monocytes and of DQ+ T lymphocytes was observed in patients without aGVHD compared with patients with aGVHD. The most discriminating marker was the strong increase in the percentage of DQ+ monocytes in patients without aGVHD (P = 0.001). In a sequential study, we observed a low percentage of DQ+ and DR+ peripheral blood mononuclear cells (PBMC) as long as the clinical manifestations of aGVHD continued. We speculate if the modulation of DQ and DR molecules on PBMC after BMT is a consequence of the action of some lymphokines, and if it plays a role in the regulation of the acute GVH reaction. We conclude that MHC class II molecules on peripheral mononuclear cells may be reliable biological markers for the diagnosis of aGVHD.

Acute Disease↗
PubMed1994

Selective detection of human hepatitis B virus surface and core antigens in peripheral blood mononuclear cell subsets by flow cytometry.

The presence of hepatitis B surface protein (HBs) and hepatitis B core protein (HBc) was investigated, by flow cytometry, on the surface of peripheral mononuclear cells (PBMC) from cells of the following phenotype: CD3 (T lymphocytes), CD4 (T helper/ inducer), CD8 (T cytotoxic/suppressor), CD19 (B lymphocytes) and CD56 [natural killer (NK) cells] among eight patients suffering from chronic hepatitis B and five healthy HBV-negative subjects. This study demonstrated the presence of HBsAg and HBcAg on the lymphocyte surface for most of the patients. The mean percentage of labelled cells was 17% for HBsAg and 15% for HBcAg. Among the different lymphocyte subsets only B lymphocytes and the NK cells expressed HBsAg for 57% and 26% of cells, respectively. Similarly HBcAg was also detected among CD19 and CD56 cells only. Polymerase chain reaction (PCR) was used to search for the presence of hepatitis B virus (HBV) DNA and RNA in PBMC, using primers located in the S gene. HBV DNA was detected with variable intensity in the CD3, CD4, CD19 and CD56 subsets following their separation with a cell sorter. For HBV RNA the signal obtained after PCR and Southern blotting was higher for CD56 and CD19 cells than for CD3 cells and undetectable for CD4 cells. This study demonstrates that replication and transcription of the HBV can occur in CD19- and CD56-positive cells. Positive signals in CD3 cells may be due to contamination of this subpopulation by NK cells.

Antigens, CD↗
PubMed1994

Histone-reactive IgA antibodies in adult IgA nephropathy and other primary glomerulonephritis.

The levels of histone-reactive IgA antibodies in the sera of adult patients with IgA mesangial glomerulonephritis, membranous glomerulonephritis, membranoproliferative glomerulonephritis and idiopathic nephrotic syndrome (minimal change disease+segmental glomerulosclerosis+IgM nephropathy) were evaluated by an enzyme-linked immunosorbent assay. Increased levels of IgA antibodies to all five major histones (H1, H2A, H2B, H3, H4) were found in all four disease groups when compared to normal controls. These histone-reactive IgA antibodies were restricted to the IgA1 subclass and their levels did not correlate with the levels of total serum IgA, nor with serum creatinine, creatinine clearance, and 24-hour proteinuria. Increasing ionic strength resulted in only partial inhibition of the binding to histones and, in individual patients, levels of reactivity with individual histones were usually correlated. This study shows that elevated levels of IgA antibodies reactive with self antigens are present in primary glomerulonephritis and extends previous observations indicating that anomalies of the IgA system occur in various forms of primary glomerulonephritis and are not limited to IgA nephropathy.

Adult↗
PubMed1993

IgM and IgA rheumatoid factors in canine polyarthritis.

IgM and IgA rheumatoid factor (RF) were detected by ELISA using a purified dog IgG as antigen in normal controls (N = 84), dogs with unclassified polyarthritis (N = 95), dogs with rheumatoid arthritis (RA) (N = 22), dogs with systemic lupus erythematosus (SLE) (N = 35), dogs with leishmaniasis or heart worm disease (N = 20) and dogs with pyometra (N = 16). Frequency and titre of IgM and IgA RF are low and comparable (P < 0.05) in dogs with unclassified polyarthritis or RA: respectively 24.2% and 27.3% for IgM RF and 21.0% and 18.2% for IgA RF; the mean titre being respectively 0.781 +/- 0.581 and 0.649 +/- 0.365 for IgM RF, and 0.774 +/- 1.331 and 0.740 +/- 1.169 for IgA RF. The frequencies of IgM and IgA RF are a little higher in dogs with SLE (IgM RF: 37.1%, IgA RF: 25.7%) and higher in dogs with leishmaniasis or heart worm disease (45.0% and 30.0%), especially in dogs with pyometra (68.7% and 37.5%). So, although dogs can produce IgM and IgA RF, these auto-antibodies are uncommon in dogs with RA. Furthermore, when RF are present their titre is much lower than in human RA.

Animals↗
PubMed1993

Monitoring of early events of experimental woodchuck hepatitis infection: studies of peripheral blood mononuclear cells by cytofluorometry and PCR.

The peripheral blood mononuclear cells (PBMC) of woodchucks experimentally infected by woodchuck hepatitis virus (WHV) were examined simultaneously for the presence of membrane associated WHV antigens by cytofluorometry, and for WHV DNA and RNA sequences by the polymerase chain reaction (PCR). Four woodchucks were inoculated: two with a well-defined infectious inoculum and two with an inoculum obtained from an animal at the late incubation phase, which was positive for WHV DNA by PCR but still devoid of WHV markers. Infection was demonstrated in all four inoculated woodchucks by the appearance at different times of WHV DNA and WHV antigens in both leucocytes and serum. WHV DNA was first detected by PCR either in the serum (two cases) or in leucocytes (two cases). The mean percentage of cells positive for membrane associated WHsAg or WHcAg detected by cytofluorometry were 37% +/- 25 and 17% +/- 15 respectively. After 8 weeks, all inoculated animals were WHsAg positive in serum. These data suggest that PBMC are involved in the early events of hepadnavirus infection. They also show that sera which are positive by PCR for WHV DNA may transmit viral infection even while still seronegative for WHV markers and for WHV DNA by dot blot.

Animals↗
PubMed1993

[Demonstration of cell death process by apoptosis in cat lymphocytes infected by FIV (feline immunodeficiency virus)].

Feline immunodeficiency virus (FIV), the causative agent of feline AIDS, induces a disease syndrome in cats characterized by a decreased lymphocyte-proliferative response to mitogens at all stages of infection and selective depletion of CD4 lymphocyte subsets. In this work, we report that peripheral blood lymphocytes isolated from FIV-infected cats undergo a spontaneous death, in vitro, according to a programmed cell death (PCD) or apoptosis. This phenomenon has also been seen in peripheral blood lymphocytes from HIV-infected humans and SIV-infected macaques. Four different techniques were used to document PCD in FIV-infected cats. DNA gel electrophoresis has shown a DNA fragmentation pattern with DNA fragments displaying sizes corresponding to multiples of oligonucleosomes DNA length unit (180 bp). Transmission electron microscopy revealed condensation of both nuclear chromatin and cytoplasm. An increase in the percentage of fragmented DNA was demonstrated by Burton's technique. In addition, flow cytometric analysis detected a cell population with condensed chromatin. The spontaneous PCD in FIV-infected cats could not be inhibited by RNA synthesis inhibitors or protein synthesis inhibitors. Our results could have implications for understanding the pathogenesis of FIV-infection and establishing specific strategies against apoptosis in cats and humans.

Analysis of Variance↗
PubMed1992

Demonstration of woodchuck hepatitis virus infection of peripheral blood mononuclear cells by flow cytometry and polymerase chain reaction.

Peripheral blood mononuclear cells (PBMCs) from 10 woodchuck hepatitis virus (WHV)-infected woodchucks were examined for the presence of WHV surface (WHs) and core (WHc) antigens (WHsAg and WHcAg) by cytofluorometry using fluorescein isothiocyanate-conjugated anti-WHs and anti-HBc-purified immunoglobulins from woodchuck and human sera. The presence of viral DNA and RNA was detected in the serum and PBMCs from the same blood samples by polymerase chain reaction (PCR) with two primer sets located in the S and C genes of the WHV genome. Seven animals were found positive for both WHsAg and WHcAg on the surface of PBMCs: four WHV-chronic carriers, two WHsAg-positive animals with acute WHV infection, and one woodchuck which was bled during the incubation phase of WHV infection and which became WHsAg-positive only 1 month later. Sixteen to 71% of the studied leukocyte population expressed WHsAg with a low density of expression whereas 7 to 72% expressed WHcAg with a high density of expression. Only two cases were positive for WHsAg without WHcAg on PBMCs, one WHV chronic carrier and one anti-WHs-positive animal. All woodchucks positive for WHcAg and/or WHsAg by cytofluorometry were positive also for WHV DNA and RNA in PBMCs by PCR. The tenth animal was found negative for both viral antigens as well as for WHV DNA and RNA in PBMCs despite the presence of persistent viral DNA in the serum as detected by PCR. Five healthy woodchucks devoid of WHV serological markers served as negative controls. These results obtained with a novel approach further confirm, in the woodchuck model, that a significant proportion of PBMCs are probably permissive for WHV replication. The possible immunopathogenic implications of the phenomenon are discussed.

Animals↗
PubMed1992

Immunosuppressive property of a very high purity antihaemophilic preparation: a low molecular weight component inhibits an early step of PHA induced cell activation.

Immune deficiency has been reported in haemophiliac patients receiving antihaemophilic factor VIII preparations, but the mechanisms involved in the immunosuppression are not fully understood. By using the proliferative response of peripheral blood mononuclear cells to phytohaemagglutinin (PHA) as a test system, we investigated the inhibitory influence of a very high purity antihaemophilic factor (AHF) preparation on T cell proliferation and on T lymphocyte activation molecules. We observed that this preparation reduced significantly the PHA-induced mononuclear cell proliferation, independently of the monocyte concentration. The AHF preparation did not act through a cytotoxic mechanism or a steric hindrance of PHA. The AHF preparation had no effect on the immediate expression of T lymphocyte activation molecules such as CD54 (ICAM-1). In contrast, the very high purity AHF reduced the induced expression of two early T cell activation molecules: CD25 (interleukin-2 receptor) and CD71 (transferrin receptor). The very high purity AHF also had the capacity to inhibit the up-regulation of two late activation antigens, CD38 and CD11a/CD18, and to inhibit the induced expression of HLA-DR molecule, defined also as a late T cell activation molecule. The CD45R expression level, used as a control marker, was not changed after AHF exposure. The very high purity AHF therefore influenced an early step of cell proliferation. We have also shown that the immunoregulatory properties of the preparation were not restricted to the factor VIII itself, but resulted from the presence of dialysable and low molecular weight components in the preparation.

Antigens, CD↗
PubMed1992

Modification of human long-term bone marrow cultures: establishment of a functional stromal microenvironment devoid of myeloid progenitors.

Differences in the plastic adhesive properties of bone marrow (BM) cells were used to initiate modified stromal layers (MSL) from long-term cultures by removing non-adherent cells shortly (4 to 18 hours) after initial seeding. Following this early modification, adherent cells generated a confluent layer after 21 days of culture. Cellular characteristics of volume and spontaneous fluorescence determined by flow cytometry showed that the MSL included 82% fibroblastic stromal cells, 8% macrophages and 10% myelomonocytic cells. Furthermore, clonogenic assays revealed that the MSL were devoid of hematopoietic progenitor cells. MSL were found to sustain long-term myelopoiesis for at least 7 weeks from exogenously added hematopoietic progenitors isolated from bone marrow (CD34+ cells), thereby demonstrating their functionality. The present experimental model appears of interest for the study of interactions between defined populations of hematopoietic cells and cells of the adherent layer. Of importance, our present modifications of human long-term bone marrow culture are technically simple and do not involve manipulation of the stromal cells.

Antigens, CD↗
PubMed1992

Canine systemic lupus erythematosus. I: A study of 75 cases.

We studied 75 cases of canine systemic lupus erythematosus (SLE) presenting with at least four criteria of the American Rheumatism Association (ARA), including antinuclear antibodies (ANAb). This disease mainly affects male German shepherds of an average age of 5 years. The most common clinical signs are polyarthritis (91% of cases), and renal (65%) and cutaneo-mucous disorders (60%). Hemolytic anemia is rare (13%). ANAb are present, often at high levels (> 256 up to 10(6) by indirect immunofluorescence on mouse blood smears). The titers are correlated with the severity and the stage of the disease. As double-stranded DNA Ab are rare and as antihistone Ab are frequent, the former could be replaced by the latter in the ARA criteria applied to the SLE dog. Another category of ANAb, named anti-type 1, also seems useful in diagnosing canine SLE. As for therapy, long-term remissions (up to 9 years without treatment) were obtained in 55.6% of 27 SLE dogs treated by levamisole. At first, levamisole was associated with induction corticotherapy, then administered alone and finally discontinued. Side effects were uncommon and transient.

Animals↗
PubMed1992

Canine systemic lupus erythematosus. II: Antinuclear antibodies.

The frequency and the specificities of antinuclear antibodies (ANAb) were studied in dogs with systemic lupus erythematosus (SLE) and compared to those found in normal dogs and in dogs with various infectious diseases. Whole ANAb were detected by immunofluorescence. Anti-double-stranded DNA Ab were found in only 2% of SLE dogs, whereas anti-single-stranded DNA Ab were present in 21.4% of SLE dogs and in 26.8% of dogs with infectious disease. Antihistone Ab were frequently observed in SLE dogs (71%) and are essentially directed against trypsin-resistant epitopes of H3, H4 and H2A. The Western blots of nuclear extracts of HeLa cells were recognized mainly by type 1 Ab (30%, reacting with bands of 43, 36, 35, 34, 30 and 27 kDa) and by anti-Sm Ab (12%) associated with anti-RNP Ab. Anti-SSA and anti-SSB Ab were rare.

Animals↗
PubMed1992

Selective detection of human hepatitis B virus surface and core antigens in some peripheral blood mononuclear cell subsets by flow cytometry.

The presence of HBs and HBc antigens was investigated, by flow cytometry, on the surface of peripheral mononuclear cells (PBMC) from the following phenotype: CD3 (T lymphocytes), CD4 (T helper/inducer), CD8 (T cytotoxic/suppressor), CD19 (B lymphocytes) and CD56 (NK cells) among 8 patients suffering from chronic hepatitis B and 5 healthy HBV-negative subjects. This study demonstrated the presence of HBsAg and HBcAg on the lymphocyte surface for most of the patients. The mean percentage of labelled cells was 17% for HBsAg and 15% for HBcAg. Among the different lymphocyte subsets only B lymphocytes and the NK cells expressed HBsAg for 57% and 26% of cells, respectively. Similarly HBcAg was also detected among CD19 and CD56 cells only. PCR was used to search for the presence of HBV DNA and RNA in PBMC, using primers located in the S gene. HBV DNA was detected with variable intensity in the CD3, CD4, CD19 and CD56 subsets following their separation with a cell sorter. For HBV RNA the signal obtained after PCR and Southern blotting was higher for CD56 and CD19 cells than for CD3 cells and undetectable for CD4 cells. This study demonstrates that replication and transcription can occur in CD19 and CD56 cells. Positive signals in CD3 cells may possibly be due to contamination of this subpopulation by NK cells.

Antigens, CD↗
PubMed1992

[Study of the immune profile of pregnant women].

During pregnancy, the fetus is an hemiallograft perfectly tolerated by the mother. With the aim of elucidating the mechanism of this tolerance, we have looked to see whether lymphocyte subsets are modified by pregnancy. Using monoclonal antibodies and flow cytometry, we have studied the changes in the immune profile of normal pregnant women and compared them with non pregnant women. In pregnant women, a significant decrease in the percentage of CD3+ cells is observed in the first trimester (61.1 +/- 14.7% versus 73.8 +/- 6.8%; p less than 0.001). The same data are obtained for CD4+ cells (38.2 +/- 10.7% versus 44.0 +/- 7.0%; p less than 0.05) and CD8+ cells (22.8 +/- 5.6% versus 28.0 +/- 8.9%; p less than 0.05). On the other hand, B lymphocytes (CD19+), monocytes (Leu M3+) and natural killer (NK) cells (Leu7+) remain stable during pregnancy. CD11a+ cells decreased during the 1st and 2nd trimesters. Lastly, activated T lymphocytes (CD3+DR+, CD8+DR+) are not modified. Using absolute numbers, a significant decrease is shown only for CD3+ cells in the 2nd and 3rd trimesters and for CD11a+ cells in the 2nd trimester of pregnancy. The decrease of T lymphocyte subsets, NK and CD11a+ cells during pregnancy partially explains the tolerance for the fetus.

Antigens, CD↗