Production of active and passive anaphylactic shock in the WBB6F1 mouse,a mast cell-deficient strain

Summary The role of mast cells in active and passive anaphylactic shock was examined using the WBB6F₁ mouse, a genetically mast cell-deficient strain. Lethal anaphylactic shock occurred at high incidence rates in mice actively sensitized to bovine serum albumin (BSA). The reaction was specific to BSA since the shock could not be elicited by human or guinea pig serum albumin in these animals. Lethal shock could be prevented by CV-3988 but not by cyproheptadine, which suggests that the shock is mediated by PAF but not by histamine and serotonin. Similarly, lethal shock was provoked by homologous antigens in mice which had been passively sensitized with allogeneic anti-benzylpenicilloyl (BPO) IgG₁ monoclonal antibody or with allogeneic or xenogeneic anti-BSA antiserum, but not in those sensitized with allogeneic anti-BPO IgE monoclonal antibody. These findings suggest that mast cells are not necessarily required for anaphylactic shock in the mouse.     

Drug susceptibility of PCA in WBB6F1-W/Wv mice

Our previous study revealed that passive cutaneous anaphylaxis (PCA) can be produced in congenitally mast cell-deficient WBB6F1-W/W^v (abbreviated as W/W^v) mice on sensitization with undiluted or slightly diluted allogeneic and xenogeneic antisera but not on sensitization with allogeneic monoclonal immunoglobulin (Ig)E and IgG1 antibodies regardless of the antibody concentration [1]. In view of these findings, the present study was conducted to characterize PCA in this strain from its drug susceptibilities using mast cell-bearing WBB6F1-+/+ (abbreviated as +/+) and B6D2F1 mice as references. PCA in W/W^v mice mediated by a low dilution (1  4) of hyperimmune serum to bovine serum albumin of the B6D2F1 mouse origin was markedly suppressed by CV-6209, an antagonist of platelet-activating factor (PAF), but not by antihistamines such as cyproheptadine and oxatomide. In contrast, PCA in +/+ and B6D2F1 mice mediated by a high dilution (1  128) of the anti-serum (virtually by IgG1 antibody) was nearly completely suppressed by antihistamines but not by CV-6209. A remarkable difference between PCA in W/W^v and reference mice was also observed in the susceptibility to monoclonal anti mouse granulocyte (Gr-1) antibody PCA in W/W^v mice was potently suppressed by the 1- to 3-day pretreatment with this antibody but that in references was not at all. Putting these present results together with the previous finding that anti-granulocyte antibody greatly reduces circulatory Gr-1⁺ leukocytes, 1 to 3 days after the treatment [2], it is highly probable that PCA in W/W^v mice mediated by some antibody isotypes other than IgE and IgG1 is produced by PAF mainly released from Gr-1⁺ cells, while IgG1 antibody-mediated PCA in mast cell-bearing reference mice is evoked by histamine derived from mast cells. PCA homologous to that in W/W^v mice could also be produced in the reference mice on sensitization with undiluted or slightly diluted antiserum, when generalized blueing due to excess IgG1 antibody was removed by the oxatomide treatment be fore the antigen challenge. Received 10 December 1997; received after revision 2 February 1998; accepted 23 February 1998     

Carbohydrate interference of complement-dependent cell lysis

The antibody-mediated cytotoxicity of three autoreactive sera, an allogeneic hyperimmune serum and a xenogeneic hyperimmune serum was abrogated by the presence of either glucosamine, galactosamine, lactulose or lactose. This inhibition could be overcome in a dose-dependent fashion by increasing the amount of complement in the cytotoxicity assay, but not by increasing the amount of antibody. Furthermore, the inhibition was specific for these sugars in that isomers and N-acetylated derivatives were not inhibitory. The results suggest that these sugars directly blocked events of the complement cascade.     

Carbohydrate interference of complement-dependent cell lysis

Summary The antibody-mediated cytotoxicity of three autoreactive sera, an allogeneic hyperimmune serum and a xenogeneic hyperimmune serum was abrogated by the presence of either glucosamine, galactosamine, lactulose or lactose. This inhibition could be overcome in a dose-dependent fashion by increasing the amount of complement in the cytotoxicity assay, but not by increasing the amount of antibody. Furthermore, the inhibition was specific for these sugars in that isomers and N-acetylated derivatives were not inhibitory. The results suggest that these sugars directly blocked events of the complement cascade.     

Evidence for a selective cytotoxic effect of carrageenan on cells of the immune system in vivo and in vitro

Summary Carrageenan suppressed antibody production to sheep erythrocytes in the mouse only when injected together with or 24 h prior to antigen. Pretreatment of sensitized spleen cell cultures with carrageenan reduced the degree of stimulation obtained using antigen but not PHA.     

The abscence of intravascular clotting in rat anaphylaxis

Summary Intravascular fibrin formation could not be detected in various phase of IgE mediated anaphylactic shock of rats, either by using an isotope technique or testing the plasma samples by the ethanol gelation test.     

Die Unterdrückung des letalen anaphylaktischen Schocks bei der Maus

Summary The effect was investigated of Endoxan (cyclophosphamide) on anaphylactic shock in mice, induced with the aid of pertussis organisms. A complete inhibition by a dose of 4 mg/mouse can be proved. The formation of antibodies in the Endoxan-treated mice is depressed.     

Decrease of mast cells in regional lymph nodes in response to allogeneic antigens and syngeneic tumor antigens

Summary The absolute number of mast cells in regional lymph nodes decreases on the 5th day after stimulation by allogeneic lymphocytes and semisyngeneic leukaemic cells, despite the enlargement of stimulated lymph nodes. It is postulated that the reaction of lymphatic mast cells could be a sensitive test for tissue incompatibility, and probably also for the presence of tumor associated antigens.     

Roles for interleukin-2 (IL-2) and IL-4 in the generation of allocytotoxic T cells in the primary and secondary responses in vitro.

Roles for interleukin-2 (IL-2) and IL-4 in the generation of murine allocytotoxine T lymphocytes (allo-CTL) in the primary and secondary responses were studied in vitro. The generation of allo-CTL in the primary response was inhibited by anti-IL-2 monoclonal antibody (mAb), but was not inhibited by anti-IL-4 mAb. On the other hand, the generation of allo-CTL in the secondary response was partially inhibited by either anti-IL-2 or anti-IL-4 mAb, and it was almost completely inhibited by the combination of two mAbs. CD8+ cell-depleted splenocytes produced IL-2, but not IL-4, in response to alloantigens in the primary response, and these cells produced both IL-2 and IL-4 in the secondary response. Both exogenous IL-2 and IL-4 induced functionally active allo-CTL in the primary response from CD4+ cell-depleted splenocytes when these cells were stimulated with T cell-depleted allogeneic cells. These results suggest that the allo-CTL induction in the primary response is IL:-2-dependent and secondary allo-CTL induction is both IL-2 and IL-4-dependent, because unprimed CD4+ T cells produce IL-2, but not IL-4, whereas primed cells produce both IL-2 and IL-4 in response to alloantigens.     

Transfer of T-cell mediated immunity to Hymenolepis nana from mother mice to their neonates.

Administration of lymph node cells from Hymenolepis nana-infected mice into lactating mothers, or directly suckling neonates successfully transferred immunity to the neonates. The capacity of lymph node cells to transfer immunity was completely abrogated by pretreatment with anti-Thy-1.2 monoclonal antibody and complement.     

Transfer and induction of delayed hypersensitivity to methylated bovine serum albumin in the absence of adjuvant]

Delayed hypersensitivity to methylated bovine serum albumin may be induced in Mice without adjuvant, by injecting the antigen either with sensitized T lymphocytes, or by injecting just the antigen in Mice preptreated with Cyclophosphamid.     

Transfer of T-cell mediated immunity toHymenolepis nana from mother mice to their neonates

Administration of lymph node cells fromHymenolepis nana-infected mice into lactating mothers, or directly suckling neonates successfully transferred immunity to the neonates. The capacity of lymph node cells to transfer immunity was completely abrogated by pretreatment with anti-Thy-1.2 monoclonal antibody and complement.     

Roles for interleukin-2 (IL-2) and IL-4 in the generation of allocytotoxic T cells in the primary and secondary responses in vitro

Roles for interleukin-2(IL-2) and IL-4 in the generation of murine allocytotoxine T lymphocytes (allo-CTL) in the primary and secondary responses were studied in vitro. The generation of allo-CTL in the primary response was inhibited by anti-IL-2 monoclonal antibody (mAb), but was not inhibited by anti-IL-4 mAb. On the other hand, the generation of allo-CTL in the secondary response was partially inhibited by either anti-IL-2 or anti-IL-4 mAb, and it was almost completely inhibited by the combination of two mAbs. CD8⁺ cell-depleted splenocytes produced IL-2, but not IL-4, in response to alloantigens in the primary response, and these cells produced both IL-2 and IL-4 in the secondary response. Both exogenous IL-2 and IL-4 induced functionally active allo-CTL in the primary response from CD4⁺ cell-depleted splenocytes when these cells were stimulated with T cell-depleted allogeneic cells. These results suggest that the allo-CTL induction in the primary response is IL:-2-dependent and secondary allo-CTL induction is both IL-2 and IL-4-dependent, because unprimed CD4⁺ T cells produce IL-2, but not IL-4, whereas primed cells produce both IL-2 and IL-4 in response to alloantigens.     

Evidence for a role of IFN gamma in control of Listeria monocytogenes in T cell deficient mice.

The role of interferon (IFN) gamma in controlling chronic infections of Listeria monocytogenes (Listeria) was studied in athymic C57BL/6 nu/nu mice, and by treating thymectomized C57BL/6 +/+ mice with monoclonal rat CD4 and CD8-specific monoclonal antibodies (Mab). Mice treated with a combination of the two T cell subset antibodies were similar to athymic, nude mice in being able to control Listeria infection, keeping the titers below 3-5 log10 bacteria per organ, but they could not eliminate them completely. Treatment with antibodies to IFN gamma of nude or CD4+ + CD8+ - T cell-depleted mice suffering from chronic Listeria infection caused a marked increase of Listeria titers in liver and spleen. This result implies a role of IFN gamma in maintaining anti-Listeria resistance in mice lacking mature T cells.     

Potentiation of immunologic memory by Corynebacterium parvum and its mechanism]

Immune response to bovine serum albumin (BSA) at dose of 2,50 mg/kg which is rather a weak immunogen in Rabbits, when given intravenous was highly potentiated when the animals received a previous single intravenous infection of 2 mg/kg of C. parvum, followed by subsequent BSA anamnestic challenges for several months. Thus, the antibody amounts synthesized following the 1st anamnestic injection (3 weeks) were 0,260 mg/ml in the control versus, 0,800 mg/ml in the C. parvum pretreated groups; following the 2nd anamnestic challenge (12 weeks afterwards) 1 mg/ml in the control versus, 2,50 mg/ml in the treated groups following the 3rd anamnestic challenge (28 weeks afterwards) 1,3 mg/ml in the control versus 5 mg/ml in the C. parvum pretreated groups; following the 4th anamnestic challenge (52 weeks afterwards) 0,300 mg/ml in the control versus 0,800 mg/ml in the C. parvum treated groups. On the whole for the four first anamnestic challenges the differences at peak levels between the control and C. parvum treated groups were about to 4. Furthermore, the antibody molecules synthesized by the C. parvum treated animals were found to belong to IgG class. The results suggest that the immunological mechanisms mobilized are peculiar to C. parvum since they could not be reproduced either by BCG or by Freun'd adjuvant under similar conditions.     

Libération d'un composant du complément par des leucocytes humains in vitro

Summary Human blood leucocytes were shown to liberate active C1q in vitro. An agarose gel containing sensitized sheep red cells and a serum depleted of C1q has been used to detect C1q activity. A population of mononucleated cells which display such activity could be isolated by centrifugation on an albumin gradient.

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Ce texte est le résumé d'une communication faite à la Société Suisse de Biochimie le 17 mai 1969.     

Multiple organ-reactivity of monoclonal autoantibodies to mouse erythrocytes

Autoantibodies reacting with bromelain-treated autologous mouse red blood cells (Br-MRBC) are spontaneously produced by normal mice. In order to understand the biological significance of these autoantibodies, anti-Br-MRBC monoclonal autoantibodies have been prepared and studied for reactivity with a panel of frozen tissue sections from organs of normal mice by direct immunofluorescence. It has been found that the anti-Br-MRBC monoclonal autoantibodies are polyspecific, since they react with cells in multiple organs.     

Conjugates of adenine 9-α-D-arabinofuranoside monophosphate (ara-AMP) with lactosaminated homologous albumin are not immunogenic in the mouse

Summary Conjugates of adenine-9--D-arabinofuranoside (ara-A) or of ara-A monophosphate (ara-AMP) with asialofetuin or with heterologous lactosaminated serum albumin (L-SA) are strong antibody inducers. But ara-AMP conjugates prepared with homologous L-SA are not immunogenic, at least in mice.Acknowledgments. This work was supported by Consiglio Nazionale delle Ricerche, Progetto finalizzato, Controllo crescita neoplasica, grant 800154596. Ara-AMP was a gift from Dr M. L. Black, Warner-Lambert Co (Ann Arbor, MI). The excellent technical assistance of Mr Goffredo Nanetti is gratefully acknowledged.     

Multiple organ-reactivity of monoclonal autoantibodies to mouse erythrocytes

Summary Autoantibodies reacting with bromelain-treated autologous mouse red blood cells (Br-MRBC) are spontaneously produced by normal mice. In order to understand the biological significance of these autoantibodies, anti-Br-MRBC monoclonal autoantibodies have been prepared and studied for reactivity with a panel of frozen tissue sections from organs of normal mice by direct immunofluorescence. It has been found that the anti-Br-MRBC monoclonal autoantibodies are polyspecific, since they react with cells in multiple organs.16 October 1986     

Prevention of graft-versus-host-disease with preserved graft-versus-leukemia-effect by ex vivo and in vivo modulation of CD4+ T-cells

This is the first report showing that an epitope-specific ex vivo modulation of an allogeneic hematopoietic stem cell graft by the anti-human CD4 antibody MAX.16H5 IgG₁ simultaneously facilitates the anti-tumor capacity of the graft (Graft-versus-leukemia effect, GvL) and the long-term suppression of the deleterious side effect Graft-versus-host-disease (GvHD). To distinguish and consolidate GvL from GvHD, the anti-human CD4 antibody MAX16.H5 IgG₁ was tested in murine GvHD and tumor models. The survival rate was significantly increased in recipients receiving a MAX.16H5 IgG₁ short-term (2 h) pre-incubated graft even when tumor cells were co-transplanted or when recipient mice were treated by MAX.16H5 IgG₁ before transplantation. After engraftment, regulatory T-cells are generated only supporting the GvL effect. It was also possible to transfer the immune tolerance from GvHD-free recipient chimeras into third party recipient mice without the need of reapplication of MAX.16H5 IgG₁ anti-human CD4 antibodies. These findings are also benefical for patients with leukemia when no matched related or unrelated donor is available and provides a safer allogeneic HSCT, which is more effective against leukemia. It also facilitates allogeneic (stem) cell transplantations for other indications (e.g., autoimmune-disorders).