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Biomedical subjects

G Mercier

Publications and source records attributed to G Mercier.

At least 73 records · Page 4Linked to original sources

Modulation of CD4 expression on lymphoma cells transplanted to mice fed (n - 3) polyunsaturated fatty acids.

Groups of adult AKR mice were fed well defined fats controlled diet regimens. These consisted of either saturated (beef tallow: 'BT') or (n - 3) polyunsaturated (fish oil: 'FO') fatty acids supplementation to basal mix mouse food. In other groups, the basal mix was given without any fat supplement ('NF'). Six weeks or more after the initiation of these diet regimens, mice received intraperitoneal injection of histocompatible RDM-4 lymphoma cells. Ascites RDM-4 tumors were harvested approximately two weeks later, and some of their physicochemical properties were studied. It was repeatedly found that: (1) the tumor grew considerably faster in the FO-fed donor than in the BT- or NF-fed donors; (2) cell membrane fluidity, content of C20(n - 3) and of C22(n - 3) fatty acids were significantly higher in the FO groups than in both BT and NF groups, while the content of C20(n - 6) and 22:4(n - 6) fatty acids was concomitantly decreased; (3) expression of the CD4 cell surface marker was always significantly diminished in the FO groups, whereas other markers such as CD8, H2K, Thy-1 and LFA-1 were not affected. Similar results were obtained, whether fats constituted from 1% to 16% by weight of the food intake. Use of a recently selected line of the RDM-4 lymphoma, exhibiting higher CD4 marker expression, resulted in similar observations. On the other hand, CD4 expression on cells from lymphoid organs of healthy adult AKR mice was not detectably modulated by the dietary fats.

Animals↗

Bacterial leaching of heavy metals from sewage sludge-bioreactors comparison.

More than 50% of municipal sewage sludges cannot be used on agricultural land because of their heavy metals content. Therefore, microbial leaching of heavy metal from municipal sludge was studied in a continuously stirred tank reactor without recycling (CSTR) or with sludge recycling (CSTRWR) at residence times of 1, 2, 3 and 4 days. The reactor CSTRWR is supposed to be more efficient for bacterial process due to the recycling of active bacteria from the settling tank to the reactor. The CSTRWR and the CSTR with 1 g litre(-1) FeSO(4).7H(2)O addition were equally efficient because of copper reprecipitation or recomplexation in the settling tank of the CSTRWR. In the CSTR, about 62% of copper and about 77% of zinc were dissolved in 3 days residence time compared to 50% of copper and 64% of zinc in the CSTRWR, if 3 g litre(-1) FeSO(4).7H(2)O was added. Thus with larger amount of substrate, the CSTR was more efficient than the CSTRWR. Residence time and pH were the main factors for zinc solubilization while for copper, the redox potential was also a major factor. The effect of FeSO(4).7H(2)O concentration on bacterial activity to solubilize heavy metals was also studied, increased concentration of FeSO(4).7H(2)O yielded better copper solubilization while it had no effect or a negative effect on zinc. This supports the hypothesis of a direct mechanism for zinc solubilization and of an indirect mechanism for copper solubilization.

Journal Article↗

Peripheral blood lymphocytes resistant to Epstein-Barr virus immortalization manifest high natural killer (NK) type activity against NK-resistant target cells.

Epstein-Barr virus (EBV) readily immortalizes human peripheral blood lymphocytes (PBL) in vitro. We found recently that PBL from two EBV-seropositive healthy adults were exceptionally resistant to immortalization by EBV. In contrast to PBL from other EBV-seropositive donors sensitive to immortalization by EBV (S-PBL), the "resistant" PBL (R-PBL) respond to EBV infection with an early interleukin-2 (IL-2) synthesis and high interferon gamma (IFN gamma) production. In order to determine whether these differences in cytokine responses between R-PBL and S-PBL could be associated with a detectable difference in lymphocyte cytotoxicity, we compared the natural killer (NK) activity of R-PBL and S-PBL effectors by using both NK-sensitive (i.e. K562) and NK-resistant (i.e. Raji) targets. We found that, while effectors from EBV-infected R-PBL and S-PBL cultures exhibited comparable NK activity against the K562 targets, they differed remarkably in their cytolytic activity against Raji cells. At days 3 and 5 of culture, effectors from EBV-infected R-PBL showed a significantly higher lytic activity against Raji targets, whereas S-PBL did not. Culture of EBV-infected R-PBL and S-PBL effectors in the presence of recombinant IL-2 (rIL-2) for 5 days resulted in increases of their lytic activity against Raji cells, whereas pretreatment of these effectors with recombinant IFN gamma (rIFN gamma) was found to increase only R-PBL cytotoxicity. These results suggest that the resistance of R-PBL to EBV immortalization could be associated with a lymphokine-mediated early cellular cytotoxic response of the NK/LAK (lymphokine-activated killer cell) type against EBV-infected cells.

Adult↗

Lymphokine release as measurement of anti-mouse hepatitis virus type 3 (MHV3) cellular reactions in various mouse lines exhibiting differential susceptibilities to MHV3-induced paralysis.

We found that susceptibility to Murine Hepatitis Virus, type 3 (MHV3)-induced paralysis is controlled by genes of the H-2 complex. In this article, we compared MHV3 antigen specific cellular reactions, in congenic mice harbouring different H-2 genes (or gene). In a first set of experiments, paralysis susceptible (B10.A x A/J)F1, partly susceptible (B10.AQR x A/J)F1 and resistant (B10.Q x A/J)F1 hybrids were infected with live MHV3. Three weeks or more post-infection (p. i.), the spleens and peritoneal exudate (PE) cells from the mice were put into culture. Killed MHV3 was added to cultures, and antigen specific lymphokine production and utilization were measured: IL-1 production by PE cells after 24 hr in culture, IL-2 production by splenocytes after 24 hr in culture, IL-2 utilization (as appraised by splenocyte proliferation) after 96 hr in culture. No clearcut difference, resulting from genetic disparity, could be observed in the antigen-specific responses. In a second set of experiments, mice were primed with ultra-violet radiation killed MHV3. In that case, increases of IL-1 production by PE cells, of IL-2 production by splenocytes and splenocyte proliferation were always observed, compared to PE cells and splenocytes from non-primed (control) donor mice. However, in latter case, addition of MHV3 antigen to cultures did not result in augmentation of antigen specific IL-2 production and utilization. Here again, no genetic effect was observed. We conclude from these results that MHV3 infection elicited strong lymphokine responses, but that antigen-specific IL-1 and IL-2 production did not correlate with the susceptibility to MHV3-induced paralysis.

Animals↗

alpha-IFN treatment does not induce Ki-ras expression in hairy-cell leukemia patients.

alpha-Interferon (IFN) is effective in the treatment of hairy-cell leukemia (HCL), but the treatment is sometimes over a long period. Biological changes such as the increase of tumorigenicity can occur rapidly in vivo as a result of beginning this treatment; an increase in c-Ki-ras oncogene expression has also been observed. In order to determine whether the findings observed in vitro would be duplicated in an in vivo system, we decided to analyze the Ki-ras RNA and protein levels in the lymphocytes of three HCL patients, compared with these levels in seven normal donors and one non-treated HCL patients. Ki-ras was not activated by IFN, at least not in lymphocytes. Therefore, the data suggest that the drug could be used for long-term therapy with relatively low risk to the patients.

Gene Expression Regulation↗

Differential interleukin-2 and interferon-gamma production by human lymphocyte cultures exceptionally resistant to Epstein-Barr virus immortalization.

Epstein-Barr virus (EBV) readily immortalizes human peripheral blood lymphocytes (PBL) in vitro. However, during the past several years, we found that PBL from two exceptional EBV-seropositive healthy adult individuals were refractory to immortalization by EBV. We report here a study aimed at learning about the immunobiological features which differentiate these EBV-resistant (R) PBL from others which are susceptible (S) to EBV immortalization. Results of this investigation indicate that: (a) Following EBV infection, R-PBL produced significantly higher amounts of interferon gamma (IFN-gamma) than S-PBL. There were however no differences in regard to interferon alpha production between these two types (R and S) of EBV-infected cultures. (b) R-PBL had a maximal interleukin-2 (IL-2) production by S-PBL occurred at least 48 hr later, i.e., at Day 7. (c) The percentage of non-B cells expressing the IL-2 receptor was also higher in EBV-infected R-PBL than S-PBL. (d) In contrast, expression of IL-2 receptors after EBV infection was higher on B cells from S-PBL than on B cells from R-PBL. Interestingly, no differences were noted in regard to IL-2 receptor expression between R-PBL and S-PBL treated with mitogens (i.e., phytohemagglutinin and pokeweed mitogen). (e) Finally, using anti-IL-2 and anti-IFN-gamma antibodies in EBV-infected R-PBL cultures, we were able to obtain EBV-induced immortalization of these cultures. Taken together, these results suggest that an early IL-2 synthesis and high IFN-gamma production by EBV-infected PBL play an important role against lymphocyte immortalization by EBV.

Antigen-Antibody Reactions↗

Tac expression induced by Epstein-Barr virus is restricted on non-transformable B lymphocytes.

During the course of a comparative study dealing with the immortalization of lymphocytes from a large number of normal healthy donors, we found that B cells of two of these individuals could not be immortalized by Epstein-Barr virus (EBV) under the standard conditions. The expression of the Tac antigen on the membrane of EBV-infected B cells from these two donors was compared with that of B cells from EBV immortalization-susceptible ones. The method used was two-colour immunofluorescence cytofluorometer analysis. We found that the Tac antigen expression was significantly and repeatedly reduced in the case of the two immortalization-resistant donors. This difference might be related to a genetic control of the resistance to EBV-immortalization.

B-Lymphocytes↗

Enhancement of antigen-specific interleukin 2 production by adding liposomes to rabies antigens for priming.

Antigen-specific IL-2 production was assessed, using splenocytes from rabies immune mice incubated for 24 h with rabies virus antigen. The antigenic material used for in vivo priming was either purified glycoprotein from rabies virus, or the inactivated virus. The time between priming, harvesting and restimulation of the splenocytes was 7 days. It was found that when antigenically inert liposomes were injected, together with antigenic material, to the prospective splenocyte donor mice, IL-2 production was enhanced. This augmentation was observed particularly when priming was performed with the inactivated rabies virus.

Animals↗

Dexamethasone stimulates expression of transposable type A intracisternal retroviruslike genes in mouse (Mus musculus) cells.

Dexamethasone treatment enhances expression of transposable intracisternal type A particles (IAP) at RNA and proteins levels in a murine retrovirus-transformed cell line (Ki-BALB). This effect was ascertained by electron microscopic numeration of IAP. By sequence comparison, we located glucocorticoid-responsive elements in IAP long terminal repeats. Their regulatory potential was tested on the promoter activity of an IAP long terminal repeat construct coupled with the chloramphenicol acetyltransferase gene. Our findings suggest that the IAP activation by dexamethasone occurs at the level of transcription.

Animals↗

Induction, by adriamycin and mitomycin C, of modifications in lipid composition, size distribution, membrane fluidity and permeability of cultured RDM4 lymphoma cells.

Adriamycin and mitomycin C were previously found to modulate the sensitivity of lymphoma cells to lysis by certain effectors of immunity and this modulation was dependent on drug concentration. In the present studies, RDM4 lymphoma cells were treated with different concentrations of the two drugs for 24 h in culture. These treatments resulted in changes in the lipid composition, membrane fluidity, cell size distribution, and permeability to 51CrO4, Trypan blue, Acridine orange and trimethylaminodiphenylhexatriene (TMA-DPH) of the cells. Changes in some of these parameters, as a function of drug concentration, resulted in dose-response curves which were bell-like shaped, hence paradoxical similarities between non-drug-treated cells and cells treated with higher drug concentrations were observed.

Animals↗

Protection from mouse hepatitis virus type 3-induced acute disease by an anti-nucleoprotein monoclonal antibody. Brief report.

Fusion of MHV-3-immune splenocytes from MHV-3-resistant A/J murine strain, with NS myeloma cells produced several hybridomas. Among eight hybridoma clones, the 1E7A4H1 clone secreted kappa IgG2a apparently directed against the nucleoprotein of the MHV-3 virion. The monoclonal antibody was able to neutralize the in vitro cytopathic effect of MHV-3 on cultured L2 cells, and was detected by indirect immunofluorescence on MHV-3-infected cultured YAC cells. In addition, it conferred a significant protection against MHV-3-induced acute disease, if injected intraperitoneally to C57BL/6 mice before inoculation with MHV-3.

Animals↗

The association of the rabies glycoprotein with liposome (immunosome) induces an in vitro specific release of interleukin 2.

BALB/c mice were primed by receiving a unique intraperitoneal injection of rabies virus antigens presented as complete inactivated virus (P.V. strain) or as purified glycoproteins either in the aggregated form or in physical combination with liposomes (i.e., in the form of "immunosomes"). The splenocytes of these mice were restimulated, 6-15 days after priming, in culture with rabies virus antigens, and antigen-specific IL-2 production was measured. It was found that rabies antigens presented as immunosomes were as active as the inactivated virus, whereas equivalent amounts of purified glycoproteins were inactive. The optimal amounts of rabies immunosomes used for priming was found to be 0.5 to 0.05 micrograms per mouse.

Animals↗

Drug-concentration dependent sensitivity to lysis by antibody and complement, of adriamycin- or mitomycin-treated murine lymphoma cells.

Murine lymphoma cells (RDM4) were pretreated in culture, with Adriamycin (ADM) and with Mitomycin C (Mit C), at different concentrations, for 20 h and carefully washed. In a first series of experiments, they were labelled with 51Cr and used as target to the lytic action of alloantiserum and complement (Ab + c), as appraised by the specific 51Cr release test. It was found that exposures of the prospective target cells to high drug concentrations (e.g. 32 micrograms/ml of ADM or 20 micrograms/ml of Mit C) enhanced their susceptibility to Ab + c killing, whereas exposures to low concentrations (i.e. 0.1 microgram/ml of ADM, or 1 microgram/ml of Mit C) protected them from Ab + c mediated lysis. In a second experimental series, similarly treated RDM4 were pulsed with 3H-TdR after their exposure to Ab + c. In this case, the cytotoxic effect was appraised as "inhibition" of isotope incorporation. Using this second criterion, the protective action of pretreatment with low drug concentrations was confirmed, and was particularly clearcut in the case of ADM. The protective effect of pretreatment with low concentration of ADM (0.1 microgram) was also confirmed in vivo, with the "Winn assay".

Animals↗

Modification, by a poly I:C injection, of organ distribution of intravenously injected murine lymphoma cells and of blood coagulability.

51Cr-or 111In-labelled murine lymphoma cells were injected IV in control and poly I:C-treated mice. The organ distribution (lung, spleen, liver) of radioactivity was measured 2 h after injection. The results showed that if cell injection was performed 1 day after poly I:C treatment, the modifications of organ distribution did not fit with the expectations from a reinforcement of the NK function in vivo. In NK-suppressed mice, poly I:C affected the distribution of radioactivity in spleen and liver in the same manner as in normal mice, suggesting that the action does not entirely depend on the NK system. Additionally to that, poly I:C injections affected coagulability of the plasma from treated mice, by prolonging the coagulation time. It is concluded that poly I:C exerts a complex action on circulation and fixation of lymphoma cells.

Animals↗

Enhancement of murine lymphoma cell lysability by CTL and by LAK cells, after treatments with mitomycin C and with adriamycin.

Murine lymphoma cells (RDM4) were treated, in culture, with adriamycin (ADM) and with mitomycin C (Mit C) at various concentrations for 20 h. They were then used as 51Cr-labelled target cells and tested for their sensitivity to the killing action both of specifically allosensitized cytotoxic T lymphocytes (CTL) and of lymphokine-activated killer (LAK) cells. They were also used as "cold target" inhibitors, in cytotoxicity assays involving CTL or LAK and 51Cr-labelled target cells. The results showed that treatments with different concentrations of Mit C enhanced the sensitivity of RDM4 cells to the killing actions of both CTL and LAK cells. ADM slightly enhanced sensitivity to the killing action by CTL, but to a much lesser extent than Mit C did. However, ADM proved to be almost as efficient as Mit C in its sensitizing action, in the case of lysis by LAK cells. Treatments with both drugs did not seem to modify the capacity of RDM4 cells to act as "cold target competitors" in cytotoxicity assays involving CTL killers, but did so in experiments involving the lytic action exerted by LAK cells.

Animals↗

Production of human and murine interleukin-2 by toxic shock syndrome toxin-1.

Toxic shock syndrome toxin-1 (TSST-1), isolated from Staphylococcus aureus strains associated with toxic shock syndrome (TSS), is known as a potent mitogen and interleukin-1 inducer. The potential of TSST-1 as an interleukin-2 (IL-2) inducer was tested on human peripheral blood lymphocytes (HPBL) and murine spleen lymphocytes (MSL). These cells were incubated with TSST-1 and the supernatants analysed for IL-2 production. Preincubation of IL-2-dependent indicator cells (IC) with a monoclonal antibody specific for murine IL-2 receptors inhibited their proliferation by supernatants of TSST-1-treated MSL, thus strongly suggesting that they contain IL-2. The concentrations of TSST-1 required for HPBL or MSL to produce IL-2 ranged between 10(-1) and 10(-4) micrograms/ml. The amount of IL-2 units/ml varied little from one experiment to another. In contrast, IL-2 production by PHA-stimulated HPBL or Con A-stimulated MSL showed great variability and dependence on mitogen concentration. T-cell depleted MSL exposed to TSST-1 produced less IL-2. Experiments with germ-free mice and TSST-1-primed mice demonstrated that IL-2 production is not related to TSST-1 antigenicity.

Adult↗