concanavalin-a and Eosinophilia

Acute, lethal graft-versus-host disease (GvHD) develops in B6D2F1 hybrid recipients of wild-type, C57BL/6, parental strain grafts; however, when interferon-gamma (IFN-gamma) gene knockout (gko) donors are used, the disease is prolonged and associated with a higher level of engraftment, particularly of T cells. Lesions containing large, mixed cellular infiltrates develop in the skin, liver, pancreas, salivary gland, lung and kidney. In our current study, we wished to determine whether GvHD features a preponderance of T helper 2 (Th2) cytokines in the absence of donor-derived IFN-gamma, and whether autoantibody production, commonly associated with chronic GvHD, also occurs. Because mitogen responsiveness is consistently suppressed in mice with acute GvHD, we wished to measure this response in recipients of IFN-gamma gko grafts. Our findings indicate that spleen cells from the latter produce interleukin (IL)-4, IL-5 and IL-13 in culture, but respond poorly to concanavalin A (Con A) and lipopolysaccharide (LPS). Their sera contain anti-nuclear antibodies (ANA), some of which are specific for double-stranded (ds)DNA and are predominantly immunoglobulin (Ig)M and IgG1. We also noted the presence of numerous eosinophils in the infiltrates developing within the target organs. In some respects, this syndrome bears resemblance to both systemic lupus erythematosus (SLE) and chronic GvHD. However, histological evidence of glomerulonephritis is lacking and proteinuria fails to develop in recipients of IFN-gamma gko grafts, suggesting that IFN-gamma may be necessary for the development of lupus nephritis. On a broader scope, our findings underscore the importance of IFN-gamma in the pathogenetic mechanism of GvHD, and demonstrate that the absence of this cytokine promotes the development of chronic GvHD and autoimmunity.

Topics: Animals; Antibodies, Antinuclear; Concanavalin A; Cytokines; Eosinophilia; Female; Graft vs Host Disease; Immunoglobulin G; Immunoglobulin M; Interferon-gamma; Interleukin-13; Interleukin-4; Interleukin-5; Lipopolysaccharides; Mice; Mice, Inbred C57BL; Mice, Knockout; T-Lymphocytes, Helper-Inducer

This study characterized the consequences of zinc-sufficient (Zn+, 60 mg zinc/kg diet, ad libitum), zinc-deficient (Zn-075 mg zinc/kg diet, ad libitum) and energy-restricted (ER, 60 mg zinc/kg diet which was restricted to match food intake of Zn- mice) diets on the in vivo and in vitro immune response of BALB/c mice during both primary and challenge infection with Heligmosomoides polygyrus. In Zn+ mice, both primary and challenge infection with H. polygyrus induced not only a strong Th2 response (IgE, IgG1, eosinophilia, IL-4, IL-5, IL-10), but also elements of a TH1 response (IgG3, IFN-gamma). Zinc deficiency significantly depressed Th2-dependent antibody production during both primary and challenge infection, and reduced mitogen and antigen-induced T cell proliferation during the challenge infection. Th2 cytokine production was reduced by zinc deficiency (IL-4), energy restriction (IL-5) and by zinc deficiency possibly in combination with energy restriction (IL-10) during the primary infection whereas TH1 cytokine production (IFN-gamma) was depressed during the challenge infection by zinc deficiency, possibly together with energy restriction. Both zinc deficiency and energy restriction reduced eosinophilia with the more profound effect being exerted by zinc deficiency. Thus, both zinc deficiency and its concurrent energy restriction modify immune responses in the mice during primary and challenge infection with H. polygyrus.

Topics: Animal Nutritional Physiological Phenomena; Animals; Antibodies, Helminth; Antigens, Helminth; Cell Division; Concanavalin A; Eosinophilia; Female; Immunoglobulin E; Immunoglobulin G; Interferon-gamma; Interleukin-10; Interleukin-4; Interleukin-5; Lipopolysaccharides; Mice; Mice, Inbred BALB C; Nematospiroides dubius; Strongylida Infections; Th1 Cells; Th2 Cells; Zinc

Subcutaneous angioblastic lymphoid hyperplasia with eosinophilia (SALH) was reviewed with respect to eosinophil chemotaxis. Lymphoid cells separated from the granuloma spontaneously released at least two different eosinophil chemotactic factors (ECF): low-molecular-weight and high-molecular-weight ECF according to the profile on gel filtration (LMW-ECF, about 500; HMW-ECF, 45,000 to 70,000). The cells, however, failed to produce chemotactic activity for macrophages and neutrophils. By analysis with monoclonal antibodies against lymphocyte subpopulations, the granuloma T cells, probably OKT4-positive cells, were shown to be responsible for spontaneous production of these two ECF. Furthermore, the blood mononuclear leukocytes were separated from the patients with SALH. An ECF closely resembling HMW-ECF was also spontaneously produced by the blood OKT4-positive T lymphocytes, whereas no LMW-ECF was released. Mononuclear leukocytes from healthy donors, however, could produce an ECF resembling HMW-ECF and chemotactic activities for macrophages and neutrophils by stimulation with concanavalin A (Con A). Protein synthesis appeared to be essential for spontaneous ECF and for Con A-induced ECF production. These results suggest that the granuloma OKT4-positive T lymphocytes of the patients with SALH are in activated condition to release LMW- and HMW-ECF, whereas the blood OKT4-positive T lymphocytes are in activated condition to release only HMW-ECF. Such spontaneous and prolonged production of HMW-ECF by the cells can be one of the diagnostic means of SALH.

Topics: Angiolymphoid Hyperplasia with Eosinophilia; Antigens, Differentiation, T-Lymphocyte; Antigens, Surface; Chemotactic Factors; Chemotactic Factors, Eosinophil; Concanavalin A; Eosinophilia; Granuloma; Humans; Molecular Weight; Puromycin; T-Lymphocytes; Time Factors

Delayed tissue eosinophilia in DNP-ovalbumin-induced allergic inflammatory skin lesions of guinea pigs was markedly enhanced by previous treatment with alum hydroxy gel (Alum) or Bordetella pertussis vaccine. This enhancement seemed due to increased production of a lymphocyte-derived eosinophil chemotactic factor (ECF) at the skin site. Treatment of animals with Alum potentiated antigen-induced in vitro ECF production by lymphoid cells from spleen and mesenteric lymph node of sensitized animals. The co-culture supernatants of lymphoid cells from Alum-treated animals also potentiated concanavalin A (Con A)-induced in vitro ECF production. The potentiating effect of Alum on ECF production seemed to be ascribed to the release of soluble factors from macrophages of the Alum-treated animals. The macrophage-derived soluble factor ECF-potentiating factor (ECF-PF) selectively potentiated ECF production but not macrophage chemotactic lymphokine production by Con A-stimulated lymphoid cells from normal animals. ECF-PF activity was associated with two separate m.w. fractions: one was 50,000 to 70,000 and the other was 10,000 to 20,000. The present study provides one of the explanations for enhanced ECF production by adjuvants, such as Alum and Bordetella pertussis vaccine.

Topics: Adjuvants, Immunologic; Aluminum Hydroxide; Animals; Bacterial Vaccines; Bordetella pertussis; Cell-Free System; Chemotactic Factors; Chemotactic Factors, Eosinophil; Concanavalin A; Eosinophilia; Guinea Pigs; Immunosorbent Techniques; Lymphocytes; Lymphokines; Macrophages; Male; Molecular Weight; Passive Cutaneous Anaphylaxis

Mechanisms for degranulation in human eosinophils were evaluated. Release of eosinophil cationic protein (ECP), a unique eosinophil granule constituent, was measured upon exposure of purified eosinophils to a large surface consisting of Sephadex beads coated with serum, which leads to complement activation. Extracellular release of approximately 15% of the cellular ECP occurred both with eosinophils from patients with eosinophilia and normal people. Almost all eosinophils isolated from patients with eosinophilia and normal people adhered to serum-treated Sephadex. The data suggest that interaction through C3 receptors is a prerequisite for ECP release from eosinophils when exposed to serum-treated Sephadex. Both cytochalasin B, cytochalasin D and hydrocortisone reduced the release of ECP. Neither the cytochalasins nor hydrocortisone inhibited the adherence of eosinophils to the Sephadex beads. Thus the inhibitory effect of these agents on ECP release is a direct effect on the degranulation process. ECF-A, histamine and colchicine did not affect the release mechanism. No direct relationship was found between degranulation and oxidative burst inasmuch as some soluble mediators induced a high respiratory burst without a concomitant ECP release. Our data suggest that mechanisms for degranulation are not fully identical in eosinophils and neutrophils.

Topics: Antigen-Antibody Complex; Blood Proteins; Cell Adhesion; Concanavalin A; Cytochalasin B; Cytochalasin D; Cytochalasins; Cytoplasmic Granules; Eosinophil Granule Proteins; Eosinophilia; Eosinophils; Humans; Hydrocortisone; Lactoferrin; Neutrophils; Oxygen Consumption; Peroxidase; Ribonucleases

An intradermal injection with 20 micrograms concanavalin A (Con A) induce a marked tissue eosinophilia peaking at 24 h after the injection in guinea-pigs. Two different eosinophil chemotactic factors (ECFs) were isolated from the Con A-induced skin reaction sites. A non-dialysable ECF with mol. wt of about 70,000 closely resembled delayed ECF-a, which had been isolated from active cutaneous anaphylactic skin lesions and confirmed to be a product of T lymphocytes by antigenic stimulation, by virtue of the antigenicity, the chromatographic profiles on Sephadex G-100 and on DEAE-Sephadex, affinity to Con A-Sepharose, and other physicochemical properties. The activity of the factor paralleled the intensity of tissue eosinophilia, suggesting that the factor may function for the tissue eosinophilia as a natural mediator. Although another ECF with a low mol. wt (dialysable) was isolated from the same skin lesions, the dialysable factor may not contribute to the tissue eosinophilia because the activity of the factor paralleled the intensity of tissue basophilia but not that of eosinophilia.

Topics: Animals; Chemotactic Factors; Chemotactic Factors, Eosinophil; Chemotaxis, Leukocyte; Chromatography, Gel; Chromatography, Ion Exchange; Concanavalin A; Dermatitis; Eosinophilia; Eosinophils; Guinea Pigs; Immunosorbent Techniques; Lymphokines; Molecular Weight; Skin

Previous studies have shown that eosinophils from eosinophilic individuals differ functionally from those of normal individuals. In order to test whether agents that might induce eosinophilia could also affect eosinophil function, we have compared the capacity of culture supernatants from mononuclear cells of eosinophilic or normal individuals to enhance eosinophil activity, as reflected by an increased killing of schistosomula of Schistosoma mansoni in vitro. An enhancing activity was detected, which increased both the antibody-dependent, and to some extent the antibody-independent killing of schistosomula by eosinophils, in the absence of complement. Under similar conditions, the supernatants failed to stimulate an otherwise undetectable neutrophil-mediated killing. The activity could be removed from the assay by washing, without reversing previous eosinophil stimulation, and was not directly toxic to the schistosomula. Preliminary characterization of the activity indicated that it was relatively heat-stable at 100 degrees C for 30 min, and had an estimated molecular weight of 35,000-45,000 as judged by G-200 Sephadex fractionation. The activity was produced by a nonlymphocytic, nonspecific esterase-containing adherent mononuclear cell in the absence of either Con A or antigenic stimulation. Significant enhancing activity was detectable after 1 h of culture and continued for at least 25 h. Protein synthesis was required for its production or release. Although the activity was detectable in supernatants from both eosinophilic and normal individuals, the supernatants that demonstrated highest activity and that could be titrated out furthest were generally derived from eosinophilic individuals, suggesting that there might be some association between eosinophilia and enhanced eosinophil function.

Topics: Animals; Cells, Cultured; Concanavalin A; Cytotoxicity, Immunologic; Emetine; Eosinophilia; Eosinophils; Humans; Lymphocytes; Monocytes; Schistosoma mansoni; Schistosomiasis

Studies were done on blood eosinophils from six patients with a transient eosinophilia, to see whether blood eosinophils were structurally or functionally different from blood eosinophils in eleven normal individuals. It was found that many of the patients' eosinophils were vacuolated, and some contained less specific granules than normal. These eosinophils also possessed Fc receptors for rabbit IgG. When the eosinophil counts returned to normal these abnormalities were no longer found. The nature of these alterations are discussed in relation to the properties of eosinophils in tissues and other types of phagocytic cells responding to stimulae. Suggestions are made about the mechanisms by which they could have come about. It was concluded that blood eosinophils in patients with an eosinophilia may be functionally mature or altered in response to unknown stimulae while they are in the blood.

Topics: Complement C3; Concanavalin A; Cytoplasmic Granules; Eosinophilia; Eosinophils; Erythrocytes; Female; Humans; Immunoglobulin G; Immunoglobulin M; Leukocyte Count; Male; Neutrophils; Vacuoles

The peripheral blood lymphocytes of a patient with massive hyperimmunoglobulinaemia E were used for in vitro studies. The serum IgE ranged from 140,000-210,000 u/ml. Peripheral blood lymphocytes had approximately 7% of cells staining for surface IgE. When these cells were cultured in vitro, IgE was produced as measured by the double antibody radioimmunoassay technique. The total IgE produced ranged from 140 to 484 units per 24 hr in different cultures. IgE production was greatest in the first 24 hr of culture and declined progressively thereafter. Some cultures still had measurable IgE at 48 hr. If the lymphocytes staining for surface IgE were the cells producing the IgE, it was estimated that between 1-7 and 2-8 molecules per cell per second were produced. No definite effect of concanavalin A, pokeweed mitogen or phytohaemagglutinin on in vitro IgE production could be demonstrated under the conditions of these experiments.

Topics: Cell Fractionation; Cell Membrane; Cell Survival; Cells, Cultured; Concanavalin A; Eosinophilia; Humans; Hypergammaglobulinemia; Immunoglobulin E; Lectins; Leukocyte Count; Lymphocytes; Receptors, Antigen, B-Cell; Time Factors