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Biomedical subjects

H Bazin

Publications and source records attributed to H Bazin.

At least 181 records · Page 10Linked to original sources
PubMed1985

Rat monoclonal antibodies. IV. Easy method for in vitro production.

A simple technique for in vitro production of monoclonal antibodies is described. Two to 5 X 10(6) hybridoma cells are diluted in 300 ml of medium in a 2 litre 'roller bottle' and incubated for 2 weeks without subsequent adjustment of the cell concentration or renewal of the medium. The method is based on the observation that when hybridoma cell cultures are maintained in the same medium, although cell growth stops after a few days and the percentage of dead cells increases rapidly, the amount of monoclonal antibody present in the culture medium increases up to the 15th day, the yield reaching more than 10 mg per flask. The method can be used both for rat and mouse hybridoma cell culture.

Animals↗
PubMed1985

Rat monoclonal antibodies. III. A simple method for facilitation of hybridoma cell growth in vivo.

Hybridoma cells that did not grow when injected subcutaneously or intraperitoneally in histocompatible or Rnu/Rnu rats were injected intravenously into histocompatible recipients. Eight of the 9 cell lines injected in this way grew in several organs of the recipient 3-7 weeks later. Hybridoma cells proliferated mainly in the liver. When the liver homogenate of these animals was injected intraperitoneally into histocompatible recipients, hybridoma cells grew readily giving rise to ascites containing the expected monoclonal antibody for 6 of the 9 cell lines.

Animals↗
PubMed1985

Effect of aging on the hepatobiliary transport of dimeric immunoglobulin A in the male Fischer rat.

Recent studies have suggested that the hepatobiliary transport of serum dimeric immunoglobulin A constitutes an important secretory route for this ligand. However, the information that is currently available has been accumulated from studies in young animals or in patients with liver disease. Aging is known to result in (a) reduced hepatobiliary function(s), (b) an increased incidence of gastrointestinal infectious diseases, and (c) a marked decline in the immune response. We evaluated the effect of aging on the in vivo hepatic capacity to transport dimeric immunoglobulin A from blood to bile. Young adult rats (3-4 mo) secreted 125I-labeled dimeric immunoglobulin A into the bile at a rate sixfold greater than that measured in either mature (12 mo) or senescent (24-25 mo) animals. This age-related decline appears to be relatively independent of bile flow and bile acid secretion. Quantitative light and electron microscopic autoradiographic evidence suggests that aging may impair the rate at which this ligand is translocated across the hepatocytes to the bile canaliculi.

Aging↗
PubMed1985

Fc epsilon receptors on rat B-lymphocytes: specificity and binding kinetics.

The binding kinetics of radiolabelled rat IgE to Fc epsilon receptors (R) of rat B-cells have already been studied in IgE-stimulated animals. The receptors expressed after Nippostrongylus brasiliensis infection or 2 injections of 5 mg IgE/100 g body wt were class-specific: IgE binding was not hindered by rat IgM, IgD, IgA, IgG1, IgG2a, IgG2b and IgG2c in immunocompetitive-binding assays. The rat B-cell Fc epsilon R were not species-specific, since mouse IgE competes with rat IgE for binding to these receptors. The apparent number of Fc epsilon R on rat mesenteric lymph node B-cells varied with temp and was 1.1-2.4 X 10(5) at 4 degrees C and 5.9-7.7 X 10(5) at 37 degrees C. The experimental Ka was not influenced by temp and had an average value of 1.38 X 10(8) M-1. At 4 degrees C the IgE binding to B-cell Fc epsilon R had an association rate of 4.9 X 10(3) M-1 sec-1 and a dissociation rate of 4.65 X 10(-5) sec-1. After a very strong stimulation produced by injecting 5 mg IgE/100 g body wt every 24 hr for 5 days, the equilibrium binding curve became diphasic, indicating a probable heterogeneity of the B-cell Fc epsilon R.

Animals↗
PubMed1985

Possible involvement of terminal complement complex in active Heymann nephritis.

We investigated whether the appearance of various complement components in renal deposits of immune complexes correlated with the development of proteinuria in rats with active Heymann nephritis. Sequential kidney biopsy specimens and serum samples were obtained from Lewis rats immunized with Fx1A in complete Freund's adjuvant. Circulating antibodies against purified auto-antigen renal tubular epithelial glycoprotein, as measured by ELISA, were found in the circulation together with a diffuse granular deposition of IgG1, IgG2a, and IgG2b in the glomeruli within 2 weeks after immunization. Biopsy specimens taken 4 weeks after immunization showed diffuse deposition of C4 and C3, which indicated that activation of complement by the classical pathway had occurred. The detection of the C5b-9 complex of complement in glomerular deposits coincided with the development of abnormal proteinuria indicating that the glomerular damage in this autoimmune disease may be caused by complement-mediated lesions in the glomerular capillary walls.

Animals↗
PubMed1985

Antibody specificity and antigen characterization of rat monoclonal antibodies against Streptococcus mutans cell wall-associated protein antigens.

Monoclonal antibodies to Streptococcus mutans OMZ175 (serotype f) cell wall-associated antigens (wall-extracted antigens [WEA]) were derived from the fusion of Lou C plasmocytoma rat cells (IR 983 F) and spleen cells from Wistar R inbred rats immunized with WEA. Four cell lines producing monoclonal antibodies directed against a component of S. mutans WEA have been established. All four monoclonal antibodies reacted only with two antigens of WEA from S. mutans OMZ175 by Western blotting and immunoprecipitation techniques, enzyme-linked immunosorbent assay (ELISA), and competitive ELISA. Western blot analysis of WEA showed that the four monoclonal antibodies recognized two related cell wall-associated proteins with apparent molecular weights of 125,000 and 76,000. Immunoprecipitation of whole cells with the monoclonal antibodies confirmed the surface localization of the two antigens. The ELISA and competitive ELISA were used to analyze the distribution of the epitopes on seven S. mutans serotypes. All S. mutans serotypes were found to express the recognized epitopes; however, different reactivity patterns could be distinguished among the various strains tested, and the four monoclonal antibodies reacted only weakly with S. mutans serotypes d and g.

Animals↗
PubMed1985

Modulatory effects of Freund's adjuvant treatment on mast cell histamine release and homocytotropic antibody synthesis.

The present study examines the influence of Freund's complete adjuvant (FCA) injections on sensitized PVG rats with respect to serum levels of IgE and IgG2 alpha antibodies and total IgE (all assessed by radioimmunoassays) and the capacity of serosal mast cells to release histamine on challenge in vitro with 'immunological' secretagogues (specific antigen, anti-IgE, concanavalin A) or with compound 48/80. The rats were immunized with 10 micrograms ovalbumin (OA); alum, Bordetella pertussis vaccine, or silica gel were employed as adjuvants. Treatment with FCA was performed by single intraperitoneal injections 3, 2, or 1 week(s) before or 1 or 2 weeks after sensitization. Tests were conducted 3 weeks after sensitization. The results show that the effect of FCA treatment varied reproducibly with the adjuvant employed for sensitization and with the timing of the FCA administration. FCA treatment could either increase, fail to affect, or decrease total serum IgE and OA-IgG2 alpha antibody levels as well as serosal mast cell responsiveness, whereas OA-IgE antibody responses were decreased or not affected. Moreover, serum levels of OA-IgE and OA-IgG2 alpha antibodies and total IgE were affected by FCA treatment independently of each other. Finally, serosal mast cell responsiveness to a given secretagogue could be influenced by the FCA treatment apparently independently of that to other secretagogues. A salient finding was that effects of FCA treatment on mast cell responsiveness did not necessarily conform to effects on antibody synthesis. Collectively, these data support the opinion that the mechanisms of action of the IgE-promoting adjuvants employed differ and suggest that the expression of serosal mast cell responsiveness to each examined secretagogue can be regulated separately. They also suggest that the serosal mast cell sensitizing capacity of homocytotropic antibodies may not be adequately quantified by immunochemical methods employing reagents prepared against IgE and IgG2 alpha protein.

Alum Compounds↗
PubMed1985

Selective depression of thymus-independent anti-DNP antibody responses induced by adult but not neonatal splenectomy.

The effect of splenectomy was assessed on rats' capacity to respond to 2,4-dinitrophenol (DNP) conjugated to spider crab haemocyanin (MSH) or hydroxyethyl starch (HES). The response to DNP-HES was profoundly suppressed in all immunoglobulin classes and subclasses by adult splenectomy. This loss of responsiveness increased with time after removal of the spleen. By contrast no significant effect on the anti-DNP-MSH response was caused by splenectomy. Experiments in congenitally athymic rats confirmed the thymus dependency of anti-DNP antibody responses of all classes and subclasses induced by DNP-MSH. DNP-HES responses, however, were elevated in athymic animals. The effect of adult splenectomy in these animals was considerably less than that produced in euthymic rats. Neonatal splenectomy in euthymic rats only resulted in minor impairment of these rats' capacity to respond to DNP-HES as adults, indicating the ability of rats to compensate after neonatal but not adult splenectomy. These data are taken to show that the spleen plays an important role in responses against this thymus-independent type II antigen which is only partially replaceable by other compartments of the immune system.

2,4-Dinitrophenol↗
PubMed1985

Fc gamma receptors on rat eosinophils: isotype-dependent cell activation.

Fc receptors for rat IgG subclasses (IgG2a, IgG2c, and IgG1) were studied on rat eosinophils by rosette formation with erythrocytes coated with monoclonal immunoglobulin (Ig) or anti-Ig antisera in a reverse assay. Inhibition experiments revealed that IgG2a and IgG2c bind to the same receptor (IgG2a/IgG2c Fc receptor), distinct from the receptor for IgG1. In addition to the recent demonstration of the blocking effect of IgG2c antibodies in immunity to schistosomes, the present results show that the existence of this common receptor led to the specific inhibition by IgG2c of IgG2a-mediated eosinophil peroxidase release. Kinetic experiments on Schistosoma mansoni-infected rat eosinophils indicate that the IgG2a/IgG2c Fc receptors were occupied by cytophilic antibodies of the IgG2a isotype during the early phase of infection and by IgG2c thereafter. By rosette experiments it was possible to displace both in vivo and in vitro cytophilically bound IgG2a from its receptor. These results confirm, therefore, the major role played by antibodies in the modulation of eosinophil effector function during schistosomiasis. They underline, moreover, the possible isotypic regulation of cell activation.

Animals↗
PubMed1985

B-cell production and differentiation in adult rats.

The B-cell development in a group of rats was suppressed for the first 45 days of life by serial administration of rabbit anti-rat IgM and IgD antibody. Total or near total suppression of B lymphopoiesis was achieved. At 45 days, suppression was stopped by injection of IgM and IgD rat paraproteins. The sequence of B-cell and plasma cell development following suppression was assessed by immunohistological analysis of spleen lymph nodes and small intestinal lamina propria. The main findings are listed below. Complete reconstitution of B-cell numbers occurs within 8 days, at which stage germinal centres are also present. B lymphopoiesis in the red pulp of the spleen differs from that reported for bone marrow. Cells develop expressing surface sIgM and sIgM with IgA, but not sIgD. sIgD-positive cells first appear in splenic follicles 2 days after stopping suppression, but their appearance in lymph nodes is delayed until after 3 days. At this stage, sIgD-positive cells become apparent in the splenic red pulp. IgM plasma cells appear from day 4. IgA plasma cells in the gut appear in small numbers at day 6, and gradually increase to normal numbers by day 14. sIgG2c expression in the splenic marginal zone did not approach normal levels, even 2 weeks after suppression was stopped.

Animals↗
PubMed1985

Extracellular release of rat eosinophil peroxidase (EPO) I. Role of anaphylactic immunoglobulins.

The release of intracellular peroxidase (EPO) was investigated in order to evaluate rat eosinophil activation by various immunoglobulin (Ig) isotypes. After successive incubations with purified rat IgG1, IgG2a, IgG2b, IgG2c, IgE, or IgM and their respective anti-Ig antisera, eosinophils released significant amounts of EPO (up to 26% of the intracellular content) only in the case of Ig with anaphylactic activities (IgG2a and IgE). Other classes and subclasses were unable to induce EPO exocytosis. Selective depletion and reconstitution experiments suggested that mast cells were not required in this process. Similar levels of EPO could be released after interaction of eosinophils with antigen-antibody complexes (IgG2a monoclonal antibody and Schistosoma mansoni antigen) immobilized on nonphagocytosable surfaces. These results indicate that EPO exocytosis can be obtained after cell activation with specific antibodies, and that this mechanism is independent of phagocytosis. A kinetic study of eosinophils from S. mansoni-infected rats revealed that IgG2a and IgE cytophilic antibodies induced EPO release after incubation with either specific antisera or specific antigen, which suggests the in vivo relevance of such findings. The present work underlines the parallelism of interaction of anaphylactic-type Ig with eosinophils and with mast cells. Moreover, EPO release seems to represent an interesting marker of eosinophil activation, because close relationships were established between the present findings and previous work on the effector function of rat eosinophils.

Anaphylaxis↗
PubMed1985

Genetic factors in non-specific bronchial reactivity in rats.

Two bronchoconstrictory agents, serotonin (5 HT) and carbachol, were investigated in different inbred rat strains in order to delineate possible factors influencing the bronchial reactivity. The challenge was given intravenously and a single dose was given to an individual animal. Inbred strains of rats differed significantly from each other in their reactivity to 5 HT and to carbachol. IC rats were good reactors to both 5 HT and carbachol. RA rats were intermediate reactors to both agents. OM/N rats had a good reaction to 5 HT, but showed only a minor bronchoconstriction after carbachol. BN and LE rats were poor reactors to both agents. The strain reactivities to the 2 provocation agents were not related. Breeding studies, using a good reactor, IC, and a poor reactor, DA strain, showed that the bronchial reactivity to 5 HT was inherited with a pattern that fitted with the autosomal recessive way of inheritance, high reactivity being recessive.

Animals↗
PubMed1985

The rapid rejection of allogeneic lymphocytes by a non-adaptive, cell-mediated mechanism (NK activity).

The fate of allogeneic lymphocytes (AO or DA) transferred to non-immune PVG recipients was studied in the light of previous evidence (Heslop & McNeilage, 1983; Rolstad & Ford, 1983) that allogeneic lymphocytes can be rapidly destroyed in certain strain combinations of rats and mice by a mechanism that is distinct from either T-cell mediated immunity or an alloantibody response. AO lymphocytes injected into PVG recipients were discriminated from syngeneic lymphocytes within 15-30 min of i.v. injection, as testified by the excess release of 51Cr into the lymph plasma of the recipient. The following experiments were intended to distinguish between natural antibody and natural killer (NK) cells as the mechanism responsible for the allogeneic lymphocyte cytotoxicity (ALC) displayed by PVG rats. Nude rats treated from birth with anti-mu chain serum and shown to be lacking B and T lymphocytes, as well as being profoundly deficient in immunoglobulin, displayed more aggressive ALC than did control nude rats which, in turn, showed stronger ALC than did euthymic rats. Serum from PVG nude rats exerted no inhibitory or destructive effect on allogeneic lymphocytes in an antibody-dependent cellular cytotoxicity system, an assay of graft-versus-host activity, or when injected into 3-4-week-old PVG rats which had not yet developed ALC. Treatment of nude rats with anti-asialo GM 1 antiserum depressed ALC and NK activity in parallel, thus adding to a wide range of circumstances in which ALC and NK activity are closely correlated. In conclusion, ALC is implemented by a non-adaptive, cell-mediated mechanism independent of immunoglobulin, but the precise identity of the effector cell in the recipients' lymphatic tissues remains to be settled.

Animals↗