PubMed Health⌕ Search

Biomedical subjects

K Chadee

Publications and source records attributed to K Chadee.

At least 55 records · Page 3Linked to original sources
PubMed1992

Modulation of tumor necrosis factor production by macrophages in Entamoeba histolytica infection.

The macrophage-derived mediator tumor necrosis factor alpha (TNF) is a cytokine with pleiotropic effects. TNF exhibits potent immunologic and inflammatory properties in parasitic diseases. The present study examined the production of TNF by macrophages isolated from gerbils infected with Entamoeba histolytica and by naive macrophages in response to amoebae in vitro. Amoebic liver abscess-derived macrophages produced low constitutive basal levels of TNF; in response to lipopolysaccharide (LPS) stimulation, TNF production was enhanced by 14-, 11-, and 6-fold at 10, 20, and 30 days postinfection, respectively. Amoebic liver abscess-derived macrophages pretreated with either recombinant gamma interferon (IFN-gamma) or the cyclooxygenase inhibitor indomethacin augmented TNF production in response to soluble amoebic proteins and LPS. Kupffer cells and peritoneal and spleen macrophages from infected animals did not release TNF constitutively in vitro. However, TNF production in response to LPS stimulation was significantly higher at 10 and 20 days postinfection. Macrophages from infected and naive animals pretreated with recombinant IFN-gamma or indomethacin produced increased amounts of TNF in response to LPS but not in response to soluble amoebic protein stimulation. Pretreatment of naive macrophages with amoebic proteins inhibited LPS-induced TNF production by 69 to 79%; the effect of the amoebic proteins was partially reversed by indomethacin pretreatment. In contrast, IFN-gamma- and LPS-activated naive macrophages produced enhanced levels of TNF in response to live amoebae and soluble amoebic proteins. Our results demonstrate that TNF production by macrophages is altered during E. histolytica infection and in response to amoebae and suggest a role for IFN-gamma and prostaglandin E2 in regulating TNF production during the infection.

Animals↗
PubMed1992

Entamoeba histolytica modulates TNF-alpha, IL-1 alpha/beta and c-fos gene expression in macrophages.

E. histolytica infections induce a state of transient suppression of cell-mediated immunity. As macrophages are involved in host defense in amebiasis, we determined whether soluble amebic lysates (Eh) can modulate TNF-alpha, IL-1 alpha/beta and c-fos gene expression in naive bone marrow-derived macrophages (BM delta). By Northern analysis, the RNA production of these genes after 0, 0.5, 1 and 3 h exposure to Eh was determined and compared to lipopolysaccharide (LPS) stimulation. In response to Eh, TNF-alpha mRNA was increased two fold while IL-1 alpha/beta RNA levels were increased 6- and 19-fold, respectively. Pretreatment of BM delta with H7, a PKC inhibitor, abrogated Eh induced TNF-delta gene expression and reduced IL-1 alpha/beta gene expression 3.5- to 4-fold over control levels. We conclude that E. histolytica stimulates BM delta to induce TNF-alpha gene expression through a PKC-dependent pathway and IL-1 alpha/beta gene expression partially through PKC and another yet undetermined pathway(s).

Animals↗
PubMed1992

The fast release of mucin secretion from human colonic cells induced by Entamoeba histolytica is dependent on contact and protein kinase C activation.

The mucus producing colonic cell line, LS174T, was used as a model to study E. histolytica-induced mucin secretion. E. histolytica trophozoites in contact with the mucus layer overlying the LS174T cells and in response to PMA, a protein kinase C activator, and Ca2+ ionophore A23187 which elevates intracellular Ca2+ ([Ca]i), caused a time-dependent (0.25-2.00 h) release of mucin. PKC inhibitors, H7 and staurosporine inhibited E. histolytica (37 and 75%) and PMA (46 and 100%)-induced mucin secretion, whereas in response to Ca2+ ionophore mucin secretion was augmented (56 and 17%). Both PMA and E. histolytica-induced the translocation of the PKC enzyme from the cytoplasm to the membrane fraction with increased enzyme activity. These results suggest that even though mucin secretion can be induced by PKC and Ca(2+)-dependent pathways, E. histolytica evokes the fast release of mucins by a PKC-dependent mechanism.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1992

Entamoeba histolytica alters arachidonic acid metabolism in macrophages in vitro and in vivo.

Entamoeba histolytica infections are associated with a state of transient suppression of cell-mediated immunity. Macrophages, the most important cells in host defence and control of invasive amoebiasis, in infected animals have been found to be deficient in effector functions and accessory cell potential. However, little is known of the cellular mechanisms responsible for the down-regulation. This study investigated whether macrophage dysfunction in amoebiasis is associated with altered macrophage arachidonic acid (AA) metabolism. Resident peritoneal macrophages (PMO) from naive gerbils produced enhanced levels of prostaglandin E2 (PGE2) and leukotriene C4 (LTC4) in response to live E. histolytica trophozoites, to diffusible excretory/secretory products released from live amoebae in millicells and to freeze-thawed soluble amoebic proteins that were inhibitable by indomethacin (INDO) and nordihydroguaiaretic acid (NDGA), respectively. In contrast to PMO from naive gerbils, PMO from animals with amoebic liver abscesses at 10, 20 and 30 days post-infection (p.i.) released high basal levels of PGE2 and LTC4. In response to zymosan stimulation, PMO from infected animals produced two- and fourfold less PGE2 and LTC4, respectively, as compared to uninfected controls. AMO showed high constitutive basal release of PGE2 and LTC4. In response to amoebic and zymosan stimulation, AMO at 10 days p.i. produced significantly higher levels of PGE2 than AMO at 20 days p.i., while AMO at 30 days p.i. were refractory to stimulation to produce higher than basal levels of PGE2. Early (10 days) and late (20-30 days) AMO were refractory to amoebic and zymosan stimulation for enhanced LTC4 release. Pretreatment of AMO with AA substrate restored optimal PGE2 and LTC4 biosynthesis, but the cells were generally unresponsive to zymosan stimulation to produce augmented levels of LTC4. These results strongly suggest that intrinsic or secreted products or both from E. histolytica can induce profound alteration of eicosanoid formation in cyclooxygenase and lipoxygenase pathways in macrophages from naive and infected animals and that AA metabolism by AMO is sequentially modified during the course of the disease.

Animals↗
PubMed1991

Down-regulation of murine lymphocyte responsiveness to mitogens after treatment with antigens of Entamoeba histolytica.

Injection of mice with pathogenic Entamoeba histolytica (strain HM1-IMSS) antigens resulted in a decreased capacity of splenocytes to respond to mitogen-induced blastogenesis following a challenge with concanavalin A (Con A), phytohemagglutinin (PHA), and lipopolysaccharide (LPS), whereas no inhibition was observed in mice that had previously been injected with equivalent amounts of non-pathogenic E. histolytica-like Laredo antigens. Depletion of adherent cells in the splenocyte preparation indicated that these cells were not a major contributor to the observed immunosuppression. Quantification of splenic T-lymphocyte subsets demonstrated a significant decrease in Thy-1+ and Lyt-1+ cells, but Lyt-2+ cells were not affected. Splenocytes treated with pathogenic amoebic antigens in vitro affected the capacity of these cells to respond optimally to Con A- and LPS-induced blastogenesis but not to that induced by PHA. These findings demonstrate that amoebic antigens affect lymphocyte function and may be important co-factors in the immunoregulation and pathogenesis of amoebiasis.

Animals↗
PubMed1991

Mucin and nonmucin secretagogue activity of Entamoeba histolytica and cholera toxin in rat colon.

Depletion of colonic mucus occurs before invasion of the colonic mucosa by Entamoeba histolytica trophozoites. It is hypothesized that E. histolytica releases a mucus secretagogue; this was studied in a rat colonic loop model. In colonic loops exposed to live amebae, mucus secretion was quantitated by release of acid-precipitable [3H]glucosamine-labeled luminal glycoprotein and by specific immunoassay. Mucus secretion increased in dose-dependent fashion in response to greater than or equal to 1 X 10(5) trophozoites; cholera toxin (20 micrograms per loop), a known mucus secretagogue, elicited a similar response. Thin-section histological analysis of amebae and cholera toxin-exposed loops showed increased mucus release and streaming from mucosal goblet cells with cellular cavitation compared with control loops. Sepharose-4B chromatography of amebae and cholera toxin-stimulated glycoproteins demonstrated secretion of mucins and an 80%-90% increase in low-molecular-weight proteins. E. histolytica trophozoites and cholera toxin enhanced the secretion of preformed and newly synthesized mucin glycoproteins and stimulated colonic glycoprotein synthesis. The level of mucus secretion elicited by axenic E. histolytica strains correlated with their virulence in vivo and in vitro. The amebic secretagogue was released into the culture medium and was heat stable. Mucus secretagogue activity of E. histolytica may contribute to depletion or alteration of the protective mucus blanket, facilitating pathogenesis of invasive amebiasis.

Animals↗
PubMed1991

The interaction between intestinal mucus glycoproteins and enteric infections.

Adherence of pathogenic enteric organisms to specific receptors on mucosal surfaces is widely recognized as an important first step in the initiation of infectious diseases. The specific interactions whereby parasites and bacteria exploit mucus substrates for colonization, and the host uses them as a nonimmunological defense mechanism, is only now being unravelled. In this review, Sil-King Tse and Kris Chadee discuss various hypothetical models for interaction, including the role of the immune system in the regulation of mucus secretion.

Journal Article↗
PubMed1990

Monoclonal antibodies directed against the galactose-binding lectin of Entamoeba histolytica enhance adherence.

The Entamoeba histolytica galactose-binding lectin is a surface glycoprotein composed of 170- and 35-kDa subunits. Inhibition of this lectin with galactose or anti-170 kDa subunit polyclonal antibody blocks amebic adherence to target cells and colonic mucin glycoproteins. We describe the properties of 10 mAb with specificity for the 170-kDa subunit. Based on competitive binding studies, six nonoverlapping antigenic determinants on the lectin were identified. The effect of the mAb on adherence of amebic trophozoites to both Chinese hamster ovary (CHO) cells and human colonic mucins was measured. Antilectin antibodies directed against epitopes 1 and 2 enhanced adherence, with the number of amebae having at least three adherent CHO cells increasing with the addition of epitope 1 mAb from 26 +/- 9 to 88 +/- 2% and the binding of colonic mucins increasing from 34 +/- 1 to 164 +/- 3 pg/10(5) amebae. Antibody-enhanced adherence remained 90 to 100% galactose inhibitable, occurred at 4 degrees C and was not Fc mediated. Univalent Fab fragments of epitope 1 mAb augmented mucin binding by 238% and CHO cell adherence by 338%. The binding of purified lectin to CHO cells was increased from 1.1 +/- 0.1 to 2.4 +/- 0.3 ng/10(3) CHO cells by mAb directed to epitope 1, demonstrating that enhanced adherence was due to direct activation of the lectin. mAb to epitope 3 bound to the lectin only upon its solubilization from the membrane and had no effect on adherence. Adherence to CHO cells and mucins was inhibited from 50 to 75% by mAb to epitopes 4 and 5; epitope 6 mAb inhibited amebic adherence to CHO cells but not mucins. The pooled sera from 10 patients with amebic liver abscess blocked the binding to the 170-kDa subunit of mAb directed to all six epitopes. Striking individual variations in the effects of immune sera on adherence were observed. Although the sera of all 44 South African patients with amebic liver abscess had high titer anti-lectin antibodies, 16 patients' sera significantly (more than 3 SEM) enhanced adherence whereas 25 patients' sera significantly inhibited adherence. Antilectin antibodies exert profound functional effects on the interaction of E. histolytica with target cells and human colonic mucins. Exploration of the clinical consequences of adherence-enhancing and inhibitory antibody responses may give insight into the role of antilectin antibodies in immunity to invasive amebiasis.

Amebiasis↗
PubMed1990

Characterization of binding of Escherichia coli strains which are enteropathogens to small-bowel mucin.

Before an enteropathogen binds to the small bowel, it must interact with the small-bowel mucus (SBM) layer. To determine whether this interaction involves specific binding of diarrheagenic Escherichia coli, we used a quantitative assay with labeled, purified rabbit SBM. Binding of SBM from an adult rabbit was significantly greater to strain 162, an agglutinating E. coli strain, than it was to RDEC-1, a rabbit pathogen, and was significantly greater to strain 2348/PMAR, an enteropathogenic E. coli strain, than it was to strains 1392+ and 1392-, which are enterotoxigenic E. coli strains with and without colonizing fimbriae, respectively. Binding of strains RDEC-1, 2348/PMAR, and 162-4 was significantly greater to SBM than to bovine serum albumin. Binding of all strains increased in a linear fashion with increasing amounts of SBM and was reproducible (r = 0.85). Binding was significantly greater at pH 5.7 than at pH 7.4 or 8.0 for all five strains. Temperature did not alter the binding of any strain. Strains 162-4 and RDEC-1 bound significantly more to proximal SBM than to rabbit distal SBM, while strains 1392+ and 1392- bound significantly more to distal SBM. Oxidation of sugars from SBM significantly decreased the binding of all strains. Each pathogenic E. coli strain bound distinctively to SBM; the SBM sugars appeared to mediate this binding for all E. coli strains. Binding was also dependent on mucin characteristics, as binding varied by region of the gut (increased for proximal SBM for strains 162-4 and RDEC-1 and for distal SBM for strains 1392+ and 1392-). The developmental age of the gut significantly affected binding only of the rabbit pathogen RDEC-1.

Age Factors↗
PubMed1990

Binding of proteolytically-degraded human colonic mucin glycoproteins to the Gal/GalNAc adherence lectin of Entamoeba histolytica.

Rat and human colonic mucin glycoproteins bind to the Gal/GalNAc adherence lectin on the surface of Entamoeba histolytica in vitro, thus inhibiting the organism from adhering to and lysing the target cells. Human colonic mucin glycoproteins were isolated by Sepharose 4B gel filtration chromatography, they were proteolytically degraded with trypsin, pronase, and papain, and the glycoprotein fractions were reisolated by Sephacryl S-200 gel filtration chromatography. Binding of the mucin glycoprotein fractions to amoebae was quantitated by the inhibition of adherence of Chinese hamster ovary cells to the surface of the amoebae. Trypsin and papain digests caused 40 and 20% reductions, respectively, in the excluded fractions (void volume) that contained all the carbohydrates; pronase digests resulted in extensive degradation of the mucin glycoprotein with the carbohydrate fractions eluting over 40% of the gel bed volume. 3H-labelled mucin glycoprotein and sodium dodecylsulfate-polyacrylamide gel electrophoresis confirmed the presence of the high molecular weight carbohydrate-rich glycoproteins with no subunits in the excluded fractions and the absence of sugars in the included peptides. Only the high molecular weight carbohydrate-containing fractions bind amoebae and inhibit amoebic adherence to Chinese hamster ovary cells. The trypsin digested mucins in the excluded volume were more efficient than the native undigested mucins in binding amoebae. The carbohydrate-containing fractions of the pronase digests were the least effective in binding amoebae and inhibiting adherence of Chinese hamster ovary cells. This suggests that proteolytically-degraded colonic mucins that are glycosylated, as well as the undegraded native mucin glycoproteins of the gut, may play a protective role in binding to amoebae, thus preventing contact of amoebae with mucosal epithelial cells and potential invasion.

Acetylgalactosamine↗
PubMed1989

Human neutrophils activated by interferon-gamma and tumour necrosis factor-alpha kill Entamoeba histolytica trophozoites in vitro.

The interaction between cytokine-activated human neutrophils and Entamoeba histolytica trophozoites was studied as well as the mechanism(s) involved. Treatment of neutrophils with rIFN-gamma alone allowed them to kill 30% of E. histolytica trophozoites; however, rIFN-gamma and rTNF-alpha pretreatments in combination increased neutrophil killing to 70%. In the absence of direct contact between neutrophils and amebae, rIFN-gamma-treated neutrophils were shown to kill 70% of amebae, and rIFN-gamma- and rTNF-alpha-treated neutrophils killed 97% of amebae. Neutrophil enhancement of amebicidal activity following cytokine treatments was correlated with increased neutrophil resistance to amebic contact-dependent killing and was shown to be 73% H2O2 dependent.

Animals↗
PubMed1989

Macrophage killing of human papillomavirus type 16-transformed cells.

Papillomaviruses (HPV) are involved in proliferation of epithelial cells and are implicated in several human malignancies. We determined the capability of activated macrophages to exert cytostatic and/or cytolytic activities against HPV type 16 DNA-transformed cells. Both NIH-3T3 and A31-3T3 cells transformed with HPV DNA were susceptible to killing by activated macrophages; however, transformed and nontransformed cells were not susceptible to killing by soluble mediators. The HPV-transformed cells were as resistant as their nontransformed counterparts to recombinant TNF-alpha. These data suggest that the killing of the HPV-16-transformed 3T3 cells by macrophages occurred by a mechanism independent of TNF-alpha.

Animals↗
PubMed1989

Cytokine activation of murine macrophages for in vitro killing of Entamoeba histolytica trophozoites.

Macrophage-mediated effector mechanisms against the protozoan parasite Entamoeba histolytica were studied. Unstimulated macrophages were inefficient at killing E. histolytica trophozoites in vitro and were killed by the trophozoites. Conversely, immature cells of the mononuclear phagocyte lineage (promonocytes) were shown to display a strong spontaneous amebicidal activity. The acquisition of macrophage amebicidal activity following cytokine treatment was investigated. Gamma interferon, tumor necrosis factor alpha, and macrophage colony-stimulating factor 1, or combinations thereof, were shown to endow murine bone marrow-derived macrophages with significant amebicidal activity. Low doses of gamma interferon and tumor necrosis factor alpha and of gamma interferon and colony-stimulating factor 1 were shown to act synergistically in this phenomenon. This enhancement of amebicidal activity was shown to operate on bone marrow-derived macrophages, elicited peritoneal macrophages, and, to a much lesser extent, spleen macrophages. Although acquisition of amebicidal activity was associated with a strong respiratory burst, the addition of oxygen-free radical scavengers showed that the killing activity was approximately 45% H2O2 dependent. In addition, amebicidal activity by macrophages was shown to be contact dependent and was inhibited by 61% with the protease inhibitor tosyl lysyl chloromethyl ketone. Our results indicate that immunologic production of gamma interferon, tumor necrosis factor alpha, and colony-stimulating factor 1 could be important in the activation of macrophages for host defense against amebiasis and that promonocytes are strong effector cells against virulent amebae.

Amebicides↗
PubMed1989

Murine T-cell clones against Entamoeba histolytica: in vivo and in vitro characterization.

Eleven T-cell clones were raised from the spleens of BALB/c mice hyperimmunized against a crude soluble extract of Entamoeba histolytica trophozoites. Seven clones were of the Lyt-1+, and four of the Lyt-23+ phenotype. All clones proliferated in the presence of E. histolytica antigens but not to a purified protein derivative; five clones proliferated to a crude extract of the E. histolytica-like Laredo amoebae. Ten clones secreted T-cell growth factors in response to E. histolytica antigens. Two clones (Lyt-23+) mediated direct lymphocytotoxicity (73% and 86%) against amoebic trophozoites that was inhibited with rabbit anti-mouse TNF-alpha. Supernatants of five of the clones (all Lyt-1+) activated mouse peritoneal macrophages (Mphi) to kill E. histolytica trophozoites in vitro, seemingly independent of secreted reactive oxygen intermediates (O2- and H2O2) in the case of three clones supernatants. All of the clones that were activating Mphi to kill amoeba in vitro also mediated a local DTH reaction in mouse footpad. Our results demonstrate direct lymphocyte cytotoxicity via a cytolytic molecule antigenically related to TNF-alpha and lymphokines activating Mphi for amoebic killing by oxidative and non-oxidative mechanisms, the latter process mediated by a macrophage-activating factor (MAF) distinct from interferon-gamma (IFN-gamma).

Animals↗
PubMed1989

Clostridium difficile toxin A. Interactions with mucus and early sequential histopathologic effects in rabbit small intestine.

Clostridium difficile produces two toxins, A (enterotoxic) and B (cytopathic), that are implicated in the pathogenesis of pseudomembranous colitis. However, the relationship of the secretory effect and the early histopathologic events is still unclear. We examined the early histopathologic effect of purified C. difficile toxin A in rabbit ileal loops and correlated the mucosal damage with the secretory response. As early as 2 hours after inoculation, toxin A at 1.0 micrograms caused cytolysis of the basal portion of the apical epithelia cells. Electron microscopy showed the basal portions of the absorptive epithelial cells necrotic. No significant fluid accumulation occurred after 2 or 4 hours in ileal loops inoculated with either 0.1 or 1.0 micrograms toxin A. In contrast, with 3-10 micrograms toxin A there was significant hemorrhagic fluid accumulation after 2 hours that correlated with the severity of the histologic lesions. An early mucoid exudate was demonstrated not to be related to a mucus secretagogue effect of toxin A. Prior exposure of toxin A to rabbit intestinal mucus preparations did not alter its subsequent enterotoxic or cytopathic (Chinese hamster ovary cells) effects. These studies demonstrate that the early intestinal tissue damage by toxin A was dose and time dependent and involved disruption of the basal portions of the absorptive epithelial cells with marked separation of the cells along the mucosal surfaces that preceded the secretory response.

Animals↗
PubMed1988

Immunopathology of Entamoeba histolytica infections.

Entamoeba histolytica infects almost 10% of the world's population and results in about 100 000 deaths annually(1). Relatively little information is available concerning the immune response and the immunopathology elicited by this parasite, probably due in part to the lack of a truly appropriate animal model(2-4). However, there has been some progress - particularly concerning the interaction of this parasite with cells of the immune system(5,6). This review summarizes the salient features of the cellular immune response and immunopathology, largely from in vitro studies and studies using the gerbil model for invasive amoebiasis(7,8). Overall, the results suggest that invasive amoebtasis induces profound immune dysfunction both at the effector level of macrophages and on their accessory cell potential.

Journal Article↗
PubMed1988

Binding and internalization of rat colonic mucins by the galactose/N-acetyl-D-galactosamine adherence lectin of Entamoeba histolytica.

Purified rat colonic mucins inhibit Entamoeba histolytica in vitro adherence to and lysis of colonic epithelial cells by binding to the amoebic galactose/N-acetyl-D-galactosamine (Gal/GalNAc)-inhibitable adherence lectin. We found that 125I-labeled mucins demonstrated saturable Gal-specific binding to E. histolytica trophozoites (strain HM1:IMSS), with 2.8 x 10(3) binding sites per amoeba and a dissociation constant of 8.20 x 10(-11) M-1. Inhibition of parasite protein synthesis completely abrogated mucin binding; elevation of amoebic vesicle pH with ammonium chloride (10 mM) had no effect. Surface-bound 125I-labeled mucins were rapidly internalized and released from amoebae without evidence of proteolytic degradation. Three avirulent HM1 clones contained immunoreactive Gal/GalNAc lectin molecules with high-affinity binding of 125I-labeled mucins but exhibited markedly reduced rates of uptake and exocytosis of bound mucins. High-affinity binding by the Gal/GalNAc adherence lectin was followed by rapid internalization and eventual release of the colonic mucins. Additionally, defects in lectin surface expression and endocytosis were found in the avirulent clones.

Animals↗