J. B. Dixon

The immune response to Echinococcus granulosus: Sequential histological observations of lymphoreticular and connective tissues during early murine infection

This paper describes the organ weight changes and the early (0 to 56 days) histological and ultrastructural changes in the lymphoid structures and injection sites after subcutaneous infection of Balb/c mice with Echinococcus granulosus protoscolices (psc). Local lymph nodes enlarged markedly, reaching maximal weight 5 to 14 days after infection (pi). Expansion of the thymus-dependent areas of draining lymph nodes was first recognized at 3 days pi and reached a maximum between 14 and 21 days pi. Lymphoproliferation within B-cell-dependent areas was less extensive and of shorter duration. Comparable, but diminished, changes were seen in contralateral lymph nodes and spleen. Lymphoid cell depletion was observed in T-dependent areas of lymphoid organs by 56 days pi. Local inflammatory reactions to psc, at the site of injection, were intense, involving neutrophils, eosinophils, macrophages and mast cells. Viable psc were present until at least 6 weeks pi. The central role of the T-lymphocyte in recognition of, and response, to E. granulosus psc is discussed.

Regulation of macrophage-mediated larvicidal activity in Echinococcus granulosus and Mesocestoides corti (Cestoda) infection in mice

Killing of metacestodes by normal or post-infection macrophages and the regulation of this activity by cytokines were studied in vitro. The protoscolecidal activity of normal macrophages against Echinococcus granulosus was inhibited by a product of naive T-enriched lymphocytes co-cultured with protoscoleces (PSC). By contrast, supernates from co-cultures of Mesocestoides corti tetrathyridia (MCT) and T-enriched or B-enriched normal lymphocytes increased killing of MCT by normal macrophages. Larvicidal activity (against both PSC and MCT) was enhanced by high concentrations of macrophage-activating factors produced by Con A-stimulated rat lymphocytes (Con A-LK), but was reduced by low concentrations of these factors. Activation by synergism between Con A-LK and recombinant interferon-gamma(r. IFN-gamma) was demonstrated in macrophage-mediated killing of MCT at high effector to target ratio. Cytokine-activation of normal or post-MCT infection macrophages was compared. Macrophages from both 8 and 20 week post-infection mice were refractory to lymphokines from lymphocyte-MCT cultures and displayed greatly reduced killing of MCT. Macrophage activation by Con A-LK and r.IFN-gamma was also impaired, implying a general defect in the ability of these post-infection macrophages to respond to macrophage activating signals. The data indicate that two different mechanisms may exist by which metacestodes regulate potentially larvicidal effector mechanisms. E. granulosus can elicit the production of lymphokines suppressive for PSC killing, whereas M. corti appears directly to induce a refractory state in effector macrophages.

Occurrence and biochemical characteristics of cestode lymphocyte mitogens

The cestodes Echinococcus granulosus, Taenia multiceps, T. pisiformis, T. hydatigena, Hymenolepis diminuta, Moniezia expansa and Anoplocephala perfoliata all produced substances that stimulated thymidine incorporation by whole blood lymphocyte cultures. This mitogenic activity was demonstrated in metacestode cyst fluids, live protoscoleces and scoleces, parasite culture supernatants, and extracts of adult parasites. T. multiceps metacestode cyst fluid mitogen adhered to, but would not pass through, cellulose dialysis tubing. This adherence was reduced or prevented by D-glucose and by proteins. The mitogen was weakly anionic. With Sephadex G75 gel filtration, its elution volume was greatly decreased when the elution buffer contained 0.5 M D-glucose. Mitogenic activity was protease resistant and could be separated by gel filtration from all the cyst fluid proteins.

Modification of cellular immunity by Taenia multiceps (Cestoda): accessory macrophages and CD4 + lymphocytes are affected by two different coenurus factors

Taenia multiceps coenurus fluid was analysed by fast protein liquid chromatography in order to separate the factors responsible for previously reported modification of immunological activity in macrophages and T-cells. One factor, F7, was found to be mitogenic for murine L3T4+ T-cells, to be macrophage dependent, to require macrophage compatibility at the I region of the H2 complex, to increase the sensitivity of T-cells to regulatory signals from macrophages and to increase the rate of generation of splenic rosette-forming cells (RFC) against sheep red cells. A second factor, F24, was found to alter macrophages so as to render them suppressive, rather than stimulatory, for parasite-activated and Con A-activated lymphocyte transformation, to depress the rate of generation of RFC and to antagonize the mitogenic effect of F7. The combined actions of these two factors are, therefore, sufficient to explain the known immunomodulatory effects of the metacestode.

The effects of tumour necrosis factor on host-parasite relations in murine Mesocestoides corti (Cestoda) infection.

The regulatory role of tumour necrosis factor (TNF) was investigated in murine infection with tetrathyridia of Mesocestoides corti. Recombinant TNF alpha reduced macrophage larvicidal activity in vitro. M. corti primed mice for TNF release in response to bacterial lipopolysaccharide (LPS) in vivo. TNF activity was amplified 100-fold at 14 days post-infection (p.i.), with a further rise at day 28 p.i. Maximal inflammatory reaction was observed histologically in the liver at the height of TNF activity. Hepatic necrosis was located within inflammatory foci, but not within the vicinity of the parasite itself, suggesting that TNF may contribute to the pathogenesis of infection. Peritoneal cells from infected mice, when stimulated with tetrathyridia in vitro, showed a 4-fold increase in TNF alpha activity at day 14 p.i. However, when peritoneal cells were stimulated with LPS in vitro, a marked increase in TNF alpha secretion was observed at 2 months post-infection followed by a slow decline. It is suggested that impaired macrophage effector function, previously attributed to endogenous endotoxin, which gains access to peritoneal macrophages through an inability of the liver to detoxify endotoxin, may be mediated through TNF alpha.

Differential regulation of murine Mesocestoides corti infection by bacterial lipopolysaccharide and interferon-γ

Many liver-invasive parasites cause extensive liver damage which may result in an impaired ability to catabolize endotoxin. The influence of endogenous endotoxin on the progress of liver-invasive parasitic diseases has been investigated in murine Mesocestoides corti infection. Invasion of liver tissue by tetrathyridia resulted in extensive parenchymal destruction with fibrosis. In association with this, undetoxified endotoxin, in potentially biologically active concentration, was found on peritoneal macrophages, 5 months post-M, corti infection. Host susceptibility was influenced by the Lps gene for responsiveness to lipopolysaccharide (LPS). The parasite burden of LPS-responsive (C3H/HeN) mice was significantly increased in the livers of these mice when compared to LPS-resistant (C3H/HeJ) mice. LPS reduced the ability of normal peritoneal macrophages to kill tetrathyridia, when co-cultured in vitro. LPS also abrogated the ability of recombinant interferon-gamma (r.IFN-gamma) to enhance macrophage larvicidal activity. These in vitro findings were confirmed in vivo. Daily intraperitoneal administration of LPS, at low concentration, caused a 4-fold increase in parasite burden in the liver, while r.IFN-gamma at optimal concentration reduced parasite burden by 57%. Post-infection macrophages have previously been shown to be refractory to cytokine-activation for larval killing. In this report, we conclude that (1) this refractoriness may be due to the presence of undetoxified endotoxin on post-infection macrophages and (2) endotoxin may reduce host resistance by abrogating effector macrophage response to IFN-gamma.