Annales d'immunologie最新文献

We have tried to modify the lectin-immunotest first described by Guesdon and Avrameas by use of inactivated ConA for the preparation of the conjugates. The inactivation of ConA took place by removal of the bivalent cations Mn++ and Ca++ by means of dialysis against oxalic acid at pH 4.0. The inactivated ConA was coupled to an anti-rabbit IgG antibody using glutaraldehyde. Following the antigen-antibody reaction the lectin could be reactivated. The Con A was again able to react with horseradish peroxidase. The reactivation and the binding of the enzyme can be performed as a one-step procedure. In comparison to the conjugate of the same antibody and active ConA the conjugate of inactivated ConA showed the same sensitivity in the determination of the corresponding antigen but the concentration of the antibody in the working dilution had to be increased.

The helper activity of GAT (L-glutamic acid 60-L-alanine 30-L-tyrosine 10)-specific T cells from BALB/c mice has been characterized. The isotypic pattern of the PFC response (plaque-forming cell) obtained in vitro is different depending on the origin of the T helper cells. T helper cells from primed spleen induce a very predominant IgG1 PFC response while T helper cells from primed lymph node induce IgG1-IgG2a- and IgG2b-PFC responses. When primed spleen cells are added to primed lymph node cells in vitro, the IgG2a- and IgG2b-PFC responses are abolished. This results suggests the presence of isotype-specific suppressor cells in the spleen of immunized mice.

We have studied by ultrastructural histo-autoradiography, in a primary immunological response, the behavior of three types of guinea-pig histiocytic cells exposed to 125I-flagellin, lymph node histiocytes, alveolar macrophages and skin Langerhans cells, making use of the experimental model of Nossal et al. (1964). Whereas latero cave lymph node histiocytes exposed to 125I-flagellin by in vivo injection of the labelled antigen into the hind foot of the guinea-pig trap the flagellin as do 20% of alveolar macrophages incubated in vitro in a culture medium containing 125I-flagellin, skin Langerhans cells exposed in vivo (intradermal and hypodermal injection of the antigen) and in vitro, as was done for alveolar macrophages, are never labelled. These results suggest that, despite the fact that it belongs to the mononuclear phagocytic system, the Langerhans cell is not a common essentially phagocytic macrophage but represents a cell lineage involved in more complex immune reactions requiring the cooperation of sensitized lymphocytes.

A partial anti-tumour protection can be induced by transfer of peritoneal cells from mice pretreated with Corynebacterium parvum, in the two experimental tumours studied: a mammary carcinoma syngeneic to C3H mice and a lymphosarcoma syngeneic to XVII mice. This protection is abolished by heating the peritoneal cells at 70 degrees C for 30 min, by a 2,200-rad irradiation or by a non-lethal irradiation of the recipient mice. Transfer of normal peritoneal cells did not produce any anti-tumour protection in C3H mice but induced the same effect as stimulated cells in XVII mice. The difference in these results could be explained by the routes of injection of peritoneal and tumour cells: intraperitoneally in C3H mice and intravenously in XVII mice. It cannot be excluded that the protective effect induced by the injection of the stimulated peritoneal cells could be produced by the anti-tumour activity of the transferred C. parvum phagocytized by these cells.

Two BALB/c mice were immunized 4 times with a mixture of adw2 and ayw4 subtypes of HBs antigens. Their spleens were then hybridized with mouse myeloma cell line NS1. Using three different radioimmunoassays (RIA), 264 independent hybridomas were screened for anti-HBs activity. By at last one of these techniques, 95% of the colonies were positive. Selected colonies were cloned and supernatants studied by RIA and immunodiffusion techniques for specificity characterisation. Some clones recognised the common "a" subtype, and other were directed to more restricted specificities. Ascites fluid was active on RIA up to 10(7) dilution (up to 29.000 UI/ml). These monoclonal antibodies may be powerful reagent for the diagnosis and understanding of viral hepatitis B.

An attempt was made to detect antibodies against type I and/or II collagen in sera from patients with rheumatoid arthritis, systemic lupus erythematosus (SLE) and leprae. This study was performed with an immunoenzymatic technique: ELISA (enzyme-linked immunosorbent assay). The following steps were performed: bovine collagen type I or II was adsorbed on glass beads; possible free sites were saturated by incubating the beads with sheep serum; then, the antibodies specifically bound to collagen were detected by a peroxidase-labelled anti-immunoglobulin; the immune complexes at the surface of the beads were revealed by a substrate specific for peroxidase and of great stability: Trinder's reactive. Using conditions previously shown to be optimal, the prevalence of anti-collagen antibodies was as follows. In patients with lepromatous leprae the percentages of positive sera against collagen type I and II were 40% and 44%, respectively; in patients with tuberculoid leprae the percentages were lower: 10% and 30%, respectively. Ten per cent of the SLE patients had antibodies against collagen type I, half of the prevalence noted for anti-collagen type II antibodies (20%). Finally, 13.6% of the patients with rheumatoid arthritis had antibodies against collagen type I, a percentage very similar to that of the patients with anti-collagen type II antibodies (14.6%).

Several sugars can strongly inhibit the natural cytotoxicity (NK) exhibited by peripheral blood lymphocytes (PBL) against tumour cells. The inhibition of NK cytotoxicity mediated by mannose, glucose, galactose and saccharose was tested in human PBL. All these sugars significantly blocked (P less than 0.01 at 50 mM concentration) the NK cytotoxicity. The blocking is dose-dependent in a linear pattern. The sole preincubation of the attacking (NK) population is required for inhibition of cytotoxicity; no influence of the sugars were noticed on the K562 alone. In contrast, the NK cytotoxicity of PBL primed in mixed lymphocyte reaction and further cultured in presence of conditioned medium (CM), showed, aside of a clear increase (average 40% increase of specific 51Cr release, P less than 0.05) a full insensitivity to the tested sugars. Phytohaemagglutinin (PHA), interferon (IF) and an interleukin 2 (Il2) are contained in the CM. We showed that neither the PHA nor the IF preincubation of the cells can induce the insensitivity of NK for the sugars, thus our data strongly suggest that Il2 is implicated in the sugar insensitivity of the cytotoxicity of lymphocytes cultured with CM.

Antiidiotypic antibodies directed against the M-173 (IgG2a) mouse myeloma protein have been raised in syngeneic and allogeneic conditions. The antiidiotypic repertoires of several strains of mice have been compared by isoelectrofocusing, and a major idiotype has been identified by several antisera raised in allogeneic conditions in strains of mice which did not express the Igh-Ca allotype of the BALB background. Since this idiotype could be reformed in hybrid molecules containing the M-173 heavy chains and light chains which contained the J kappa 2 region, we propose that this determinant is dependent upon the J kappa 2, DH and JH regions, in addition, most probably, to a specific contribution of residues 45 and 54 of the heavy chains.

The authors describe a method for isolation of Toxoplasma gondii antigenic membrane components. Technical work is described into four points: 1) specific labelling of membrane proteins was realised by DD125ISA; 2) methacrylate microspheres were bound to unbroken toxoplasma membranes by the mean of an indirect technic with antibody molecules (double sandwich); these microspheres were used to modify the membrane density; 3) toxoplasmas bound to microspheres were broken by sonication; 4) microspheres fixed to membrane components were isolated by isopycnic ultracentrifugation on continuous sucrose gradient.

An anaemia and a decreased number of erythroid bone marrow cells were observed on the first weeks following the intravenous injection of 1 mg of BCG into normal mice. No such modification of erythropoiesis was detected in Nude mice. The graft of thymus from new born mice to Nude adults before BCG infection allowed to obtain a drop of packed-cell volume. In previously infected Nude mice, the injection of spleen cells providing from infected normal mice also modified the erythropoiesis. This effect was lost by pretreatment of injected spleen cells with anti-theta serum and complement. Thus, the conditions necessary to the development of the anaemia during BCG infection correlated with the conditions which allowed an immune response against infection.