Activation of granulocyte cytotoxic function by purified mouse colony-stimulating factors.

Highly purified mouse colony-stimulating factors (CSF) were tested for their effect on neutrophil cytotoxic function in a homologous antibody-dependent cell-mediated cytotoxicity (ADCC) assay in which TNP-coupled mouse thymoma cells coated with mouse anti-TNP antibodies were used as targets, and purified normal mouse bone marrow neutrophils or induced peritoneal neutrophils were used as effector cells. Biochemically pure granulocyte-macrophage (GM)- and granulocyte (G)-CSF enhanced the cytotoxic activity of neutrophils obtained from both sources, allowing them to kill target cells at low antibody concentrations. Furthermore, GM- and G-CSF showed an additive effect, suggesting either the presence of separate receptors for GM- and G-CSF or of separate subsets of neutrophils.

Attempts to modify lung granulomatous responses to Schistosoma japonicum eggs in low and high responder mouse strains.

A radioisotopic assay for acute granulomatous hypersensitivity (AGH) to lyophilized eggs of Schistosoma japonicum has been used to further examine responses to egg antigens in various inbred strains of mice. The ranking of responsiveness in mice from high (C57BL/6), intermediate (BALB/c) to low (CBA/H) was not influenced by high or low egg-sensitization regimens. However, the low responsiveness of responder mice sensitized with eggs by the intraperitoneal compared with the subcutaneous route of injection appears to be an egg dose-related phenomenon.

Enhancement of the intensity, persistence, and passive transfer of delayed-type hypersensitivity lesions by pertussigen in mice.

Pertussigen, a purified protein from Bordetella pertussis, was shown to increase delayed-type hypersensitivity (DTH) to protein antigens in mice. First, it caused an approximately twofold enhancement of the magnitude of 24-h DTH reactions. Second, the peak magnitude of DTH was delayed to 4-7 d after challenge, at which time it was five times more intense than in mice not receiving pertussigen.

Eosinophil activation by colony-stimulating factor in man: metabolic effects and analysis by flow cytometry.

Substantial increases in the killing capacity of human eosinophils after in vitro incubation with human placental conditioned medium (HPCM), a standard source of colony-stimulating factor (CSF), have recently been described. In this article, the interaction between HPCM and purified human eosinophils is analyzed by flow cytometry and by effects on iodination, superoxide production, and protein synthesis. HPCM increased the intensity of natural eosinophil autofluorescence (aFlu) (460 nm) after the absorption of ultraviolet light (360 nm) in a manner that was both time and dose dependent.

Delayed hypersensitivity induced by anti-T-cell-line antisera is enhanced by pertussigen and not restricted by histocompatibility genes.

Anti-T-cell-line antisera were raised by repeatedly injecting mice with syngeneic, antigen-specific, delayed hypersensitivity (DH) inducing cells grown as continuous T-cell lines in vitro. Many of these antisera could induce antigen-specific DH responses which, in some cases, were rather slight. For example, the DH reaction to azobenzenearsonate induced in A/J mice by an A/J antiserum produced against the syngeneic azobenzenearsonate-specific cell line AA3 was only a weak response.

Eosinophilia and acquisition of resistance to Nematospiroides dubius in mice sensitized with adult worms.

BALB/c mice develop resistance to challenge with N. dubius third stage infective larvae (L3) 2-3 weeks after the administration of N. dubius worms.

Activation of antibody-dependent cell-mediated cytotoxicity of human neutrophils and eosinophils by separate colony-stimulating factors.

Semi-purified human colony-stimulating factors (CSF) powerfully enhanced the antibody-dependent cell-mediated cytotoxicity (ADCC) by metrizamide gradient-purified human neutrophils and eosinophils. The stimulation was observed on three different tumor targets, was rapid (less than 1 hr) in onset, and CSF-stimulated cells needed direct contact with targets for killing. A subspecies of human CSF, CSF-alpha, with eosinophil and granulocyte-macrophage (GM) colony-stimulating activity enhanced both eosinophil and neutrophil killing.

Evidence for the control of eosinophilia by the major histocompatibility complex in mice.

The genetic control of eosinophilia has been studied in congenic strains of mice. Eosinophilia was induced with cyclophosphamide followed by keyhole limpet hemocyanin in complete Freund's adjuvant. After this treatment, BALB/c mice developed a high eosinophil response, whereas CBA, C57BL and A/J mice developed a low one.

Maturation in vivo of Schistosoma mansoni schistosomula after culture in vitro with granulocytes and antibody.

Seven experiments were carried out to test the relationship between the morphological assay for damage to schistosomula in vitro with toluidine blue and the loss of the ability of damaged organisms to mature in vivo. Schistosomula were prepared by penetration of rat skin and cultured for 12 to 38 h in the presence of various combinations of purified human eosinophils or neutrophils and heat-inactivated human antischistosomular serum. Samples were scored for microscopically detectable damage, and the remaining organisms were injected intravenously into normal mice.

Bullous pemphigoid blister fluid stimulates eosinophil colony formation and activates eosinophils.

An eosinophil stimulating material (ESM) is described in the blister fluids (BF) of six patients with bullous pemphigoid (BP). This ESM is similar in function to a subspecies of human colony stimulating factor (CSF), CSF-alpha, since (i) it stimulated the production from bone marrow cells of day 14 colonies (31 +/- 15, arithmetic mean +/- 1 s.e.

Enhancement of human blood eosinophil cytotoxicity by semi-purified eosinophil colony-stimulating factor(s).

Purified human blood eosinophils, when incubated in human placental conditioned medium (a source of colony-stimulating factors) [CSF]) demonstrate an enhanced ability to damage antibody- or complement-coated schistosomula. This enhancement represents a 4- to 10-fold increase of eosinophil schistosomicidal ability and a 10-fold lowering of the threshold for antibody or complement required in the killing reaction. The activity that enhances eosinophil cytotoxicity and the eosinophil colony-stimulating activity in the placental conditioned medium are eluted in the same fraction (CSF-alpha) after chromatography on Sephadex G-100 and phenyl-Sepharose columns, suggesting that these two activities might be associated with the same molecule.

Genetic control of eosinophilia in mice: gene(s) expressed in bone marrow-derived cells control high responsiveness.

A heterogeneity in the capacity of strains of mice to mount eosinophilia is described. BALB/c and C3H are eosinophil high responder strains (EO-HR) and CBA and A/J are eosinophil low responder strains (EO-LR), judged by the response of blood eosinophils to Ascaris suum, and the response of blood, bone marrow, and spleen eosinophils to keyhole limpet hemocyanin given 2 days after 150 mg/kg cyclophosphamide. Some of the gene(s) for high responsiveness appear to be dominant because (EO-HR X EO-LR)F1 mice were intermediate to high responders.

Cyclophosphamide pretreatment induces eosinophilia to nonparasite antigens.

Eosinophilia is usually regarded as a T cell-dependent hemopoietic response, although in most instances of immune responses by T cells there is no accompanying eosinophilia. BALB/c mice immunized with keyhole limpet hemocyanin (KLH) in complete Freund's adjuvant (CFA) were found not to develop eosinophilia, whereas pretreatment of such mice with 150 mg/kg cyclophosphamide (CY) induced marked eosinophilia in the blood (1000 to 3000/mm3) and bone marrow. CY as well as CY and KLH in CFA cause a leukocytosis and neutrophilia, but significant eosinophilia was seen only in mice receiving both CY and KLH-CFA.

An autoantibody reactive with nuclei of polymorphonuclear neutrophils: a cell differentiation marker.

An autoantibody that reacted with nuclei of polymorphonuclear neutrophils (PMN) was detected at titers of greater than 10 in sera of 25 of 50 patients with rheumatoid arthritis and 36 of 50 with autoimmune chronic active hepatitis but in none of 160 controls comprising 24 patients with alcoholic cirrhosis, 36 with multiple myeloma, and 100 healthy subjects. Through the use of enriched populations of hemopoietic cells, this antibody was shown to be cell-specific, reacting only with the nucleus of the mature neutrophil. It was unreactive with nuclei of progenitor cells in the myeloid series and with nuclei of eosinophils, monocytes, lymphocytes, and thymocytes.

Immune evasion by Schistosoma mansoni: loss of susceptibility to antibody or complement-dependent eosinophil attack by schistosomula cultured in medium free of macromolecules.

Schistosomula of Schistosoma mansoni, recovered either after penetration of cercariae through isolated rat skin or by mechanical transformation of cercariae, become fully resistant after 24-48 h of culture to damage by human blood eosinophils in the presence of human anti-schistosomular sera. Cultured schistosomula are also shown to lose their susceptibility to attack by human eosinophils in the presence of human complement. This resistance is related to the simultaneous reduction of the ability of human anti-schistosomular antibodies and human complement component C3 to bind to the surface of the cultured larvae.

Enhanced helminthotoxic capacity of eosinophils from patients with eosinophilia.

To determine whether eosinophils from patients with eosinophilia have an enhanced capacity to kill parasites, we compared purified eosinophils (mean purity, 89 per cent) from 30 patients with various degrees of eosinophilia and with or without infection with Schistosoma mansoni for the capacity to kill schistosomula, the larval stage of S. mansoni, in vitro. There was a significant correlation between peripheral eosinophil count and antibody-dependent, eosinophil-mediated death of parasites after 40 hours of culture (P < 0.

Mechanism of the interaction mediating killing of Schistosoma mansoni by human eosinophils.

Recent work on the role of eosinophils in immunity to Schistosoma mansoni is summarized. In vitro studies have shown that human eosinophils can kill antibody-covered schistosomula. The killing mediated by eosinophils is associated with adherence of these cells and subsequent degranulation and release of eosinophil major basic protein onto the larvae.

Functional studies on purified eosinophils and neutrophils from patients with Schistosoma mansoni infections.

Unpurified peripheral blood leucocytes or purified eosinophils and neutrophils from patients with schistosomiasis and from normal individuals were compared for their ability to interact with antibody coated schistosomula of Schistosoma mansoni. There was no difference in the ability of buffy coat cells or neutrophils from patients and from normal individuals to mediate antibody-dependent 51Cr release from labelled schistosomula. However, eosinophils from patients were significantly better than those from normal individuals in causing antibody-dependent 51Cr release.

Parasite immunity and the major histocompatibility complex.

Parasite infestations offer fertile ground for investigation of the relationship between immunity, disease and the major histocompatibility complex (MHC). However, due to the complexities of parasite life cycles and the success of parasites in evading the immune response, immune reactions against the parasite often do not parallel protective immunity, and immunity does not imply lack of disease. -- An additional level of complexity is introduced in some forms of parasite immunity by accessory effector cells, e.