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P M Kaye

Publications and source records attributed to P M Kaye.

At least 19 recordsLinked to original sources
PubMed1992

Altered course of visceral leishmaniasis in mice expressing transgenic I-E molecules.

Previous studies had shown that the outcome of infection with Leishmania donovani was exquisitely sensitive to the influence of the major histocompatibility complex. In this study, we have examined the course of infection in non-obese diabetic (NOD) and NOD-E-3 mice, the latter expressing an I-E molecule as a result of transgenic introduction of the wild-type Ed alpha gene. Introduction of this transgene significantly altered the course of infection allowing for enhanced parasite multiplication in the viscera from day 14 to day 28. This was associated with both a delayed and reduced tissue granulomatous response in NOD-E-3 mice. In vitro, spleen cells from these mice produced equivalent levels of interferon (IFN)-gamma during the early phase of infection but this originated from populations having a different balance of T cells subsets. In NOD mice CD8+ T cells contribute substantially to the total levels of IFN-gamma produced, but in transgenic mice the contribution from this subset is significantly decreased. This is reflected in a reduction in the proportion of Leishmania-specific CD8+ T cells, which could only partially be accounted for by deletion of V beta 5- and V beta 3-expressing CD8+ T cells in NOD-E-3 mice. This study highlights the impact of the introduction of a class II gene product on disease outcome and unexpectedly on the functional potential of CD8+ T cells.

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PubMed1992

Leishmania donovani infection in scid mice: lack of tissue response and in vivo macrophage activation correlates with failure to trigger natural killer cell-derived gamma interferon production in vitro.

Infection of immunocompetent mice with Leishmania donovani is characterized by the development of a tissue granulomatous response, in vivo macrophage activation, and a predominantly Th1-type CD4+ T-cell response. To determine whether a recently described T-cell-independent pathway of gamma interferon (IFN-gamma) production involving the collaboration of macrophages and natural killer (NK) cells contributed to this pattern of events, we have investigated the responses of scid mice to L. donovani infection. The multiplication of parasites in the livers of scid mice progressed at a rate equivalent to that seen in BALB/c mice over the first 14 days of infection, but by day 28 scid mice had a fivefold-higher parasite burden. This infection was not, however, accompanied by any demonstrable histological response in the liver or by elevated major histocompatibility complex class II expression on splenic macrophages. In vitro, L. donovani was unable to trigger IFN-gamma production from scid spleen cell cultures under conditions which allowed efficient triggering by bacterial stimuli. Although L. donovani also failed to stimulate the release of tumor necrosis factor, an important macrophage-derived cofactor for IFN-gamma secretion by NK cells, exogenous recombinant tumor necrosis factor alpha could not restore the IFN-gamma response. Even with the potent synergistic effect of exogenous interleukin-2, L. donovani was unable to stimulate this pathway to the same extent as Listeria monocytogenes. Indeed, L. donovani inhibited the response to L. monocytogenes in a dose-dependent fashion. Experiments involving the transfer of supernatants and the use of neutralizing monoclonal antibodies have failed to find evidence that interleukin-10 is involved in this inhibition. These data suggest that NK cell-derived IFN-gamma is unlikely to participate in the early regulation of visceral leishmaniasis in the mouse.

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PubMed1992

Recombinant interleukin-1 alpha augments granuloma formation and cytokine production but not parasite clearance in mice infected with Leishmania donovani.

In vivo administration of various doses of recombinant interleukin-1 alpha to B10.D2/n mice chronically infected with Leishmania donovani resulted in enhanced formation of granulomas and in vitro production of gamma interferon. By direct microscopical enumeration, reduction in gross parasite burden in the viscera was not observed, however. These data highlight an important discordance between granuloma formation per se and parasite elimination and suggest that interleukin-1 deficiency alone cannot account for the chronicity of this disease.

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PubMed1991

Differential production of Th1- and Th2-derived cytokines does not determine the genetically controlled or vaccine-induced rate of cure in murine visceral leishmaniasis.

Recent studies with models of cutaneous leishmaniasis have provoked much interest in the role of CD4+ T cell subsets in determining the outcome of infectious disease. In Leishmania major infections, cure vs progressive disease correlates with the expansion of Th1-like or Th2-like CD4+ populations, respectively. We have investigated whether similar responses are associated with the differential patterns of infection seen in models of visceral leishmaniasis, caused by L. donovani. Splenic lymphocytes from infected Lsh congenic C57BL/10 (Lshs;H-2b) and B10.L-Lshr (Lshr;H-2b) mice and MHC congenic non-curing B10.D2/n (Lshs;H-2d) mice were examined for the production of cytokines representative of these CD4+ populations (IL-2, IL-3, IL-4, IL-5, and IFN-gamma). In all three strains examined, there was no evidence for the production of Th2-restricted cytokines. In addition, levels of serum IgE were depressed during the early phase of infection, indicative of in vivo IFN-gamma production. In the non-curing B10.D2/n strain, late phase of infection was associated with the decreased ability to produce cytokines in response to Ag and not with the production of IL-4 or IL-5 in response to Ag or mitogen. Serum IgE levels were also not raised above levels seen in uninfected controls. C57BL/10 mice were vaccinated with SDS-PAGE fractionated amastigote Ag bound to nitrocellulose and cytokine levels determined at various times after infection. The protocol used for vaccination was able to induce significant modulation of the course of infection in this strain and it was clear that IFN-gamma production in vitro provided an excellent correlate of rate of cure. Occasional individuals produced low levels of IL-5 in culture in response to parasite Ag, but this did not correlate with disease progression. Together, these data suggest that over-expansion of Th2-type cells and production of their specific cytokines (IL-4 and IL-5) is not a contributing factor to the variable long term course of L. donovani infection in these strains of mice.

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PubMed1991

Presentation of Leishmania donovani promastigotes occurs via a brefeldin A-sensitive pathway.

For the presentation of Leishmania promastigotes to polyclonal CD4+ T cells, a processing period within activated macrophages of 3-4 h is required. Presentation can be inhibited by both chloroquine and brefeldin A (BFA), the latter implicating a requirement for newly synthesized MHC class II molecules. This inhibition is both reversible and specific, in that BFA did not inhibit mixed lymphocyte reaction stimulation by these infected macrophages. Immunogold labeling demonstrated that class II was associated with the parasite-containing phagolysosome. The level of class II was not significantly altered in BFA-treated cells in the time period studied, suggesting that antigen may exist the phagolysosome and interact with class II in another cellular compartment.

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PubMed1991

Antigen presentation by dendritic cells provides optimal stimulation for the production of interleukin (IL) 2, IL 4 and interferon-gamma by allogeneic T cells.

Previous studies have shown that dendritic cells are the most potent inducers of T cell proliferation in vitro and that this is reflected in the release of interleukin (IL) 2 into culture supernatants during dendritic cell-T cell interaction. However, the role of the dendritic cells, and, indeed, of the antigen-presenting step, has not yet been explored with respect to other T cell-derived cytokines, in either a qualitative or relative fashion. In this study, therefore, we have examined the comparative role of different antigen-presenting cells (APC) as inducers of T cell cytokine release in allogeneic responses. We have confirmed that dendritic cells are the most effective inducers for IL2 and have shown that this is true not only in primary alloresponses, but also in alloresponder T cells maintained for extended periods and then rechallenged. Dendritic cells were also the most potent inducers of IL3 and interferon-gamma (IFN-gamma) in primary cultures. No IL4 was demonstrable irrespective of the type of presenting cells used, and both tissue macrophages and dendritic cells can induce synthesis of IL6. Likewise, in secondary alloresponses both dendritic cells and to a lesser extent tissue macrophages induce release of IL3, no IL4 is detectable, and activated macrophages and B cells raise IFN-gamma levels in the supernatants albeit to a lower concentration than that seen when dendritic cells are used as stimulators. The results were similar in the tertiary alloresponse except that (a) IL4 was now detectable in the supernatants but only where dendritic cells had been used as APC, and (b) both resting and activated macrophages induced IL2 and IFN-gamma. By the eighth cycle of allostimulation there is negligible IL2. Dendritic cells, tissue macrophages and activated B cells constitute a hierarchy of APC for IL3, IFN-gamma and IL4. These findings therefore demonstrate the role of dendritic cells as potent in vitro inducers of IL3, IL4 and IFN-gamma synthesis as well as of IL2.

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PubMed1991

Antigen processing and presentation: modelling with Leishmania.

Most foreign antigens are "processed" by APC into a form in which they can bind MHC molecules and be recognised by the TCR. Functional and immuno-gold labelling studies have been used to determine the putative sites of MHC-Ag interaction following uptake of Leishmania donovani into a phagolysosome. During infection with this parasite in vivo, upregulation of class II occurs. Based on studies using scid mice, NK cells appear to contribute little to this process. During chronic infection, however, APC function is diminished in spite of high class II levels. We propose that a lack of coordinate costimulator expression may lead to subsequent loss of T cell function late in infection, by a mechanism involving the induction of T cell unresponsiveness.

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PubMed1990

A modified colorimetric assay of macrophage activation for intracellular cytotoxicity against Leishmania parasites.

An in vitro method is described which colorimetrically assesses the activation of macrophages for intracellular cytotoxicity against the obligate intracellular parasite Leishmania donovani. The assay system uses a highly purified macrophage population derived from 10-day murine bone marrow cultures. These were infected in vitro as a suspension culture with viable L. donovani amastigotes and then exposed to activating agents. After 48 h the intracellular parasites were released by SDS lysis of the macrophages. Surviving Leishmania organisms were quantitated by their conversion of the chromophore MTT. The sensitivity of this method was comparable with the established method of [3H]dThd incorporation. This assay system has been used to show that there is a dual signal requirement (recombinant interferon-gamma and bacterial endotoxin (LPS] for effective activation of macrophages for leishmanicidal activity.

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PubMed1988

Acquisition of cell-mediated immunity to Leishmania. II. LSH gene regulation of accessory cell function.

The macrophage natural resistance gene. Lsh, regulates the ability of a selective population of tissue macrophages to control intracellular multiplication of Leishmania donovani by a T-cell independent mechanism. We show here, using mice congenic for Lsh, that this gene also contributes to the acquisition of T-cell-mediated immunity. Whereas both resistant and susceptible mice generate equivalent primary T-cell responses to infection, resistant mice show a rapid increase in accessory cell activity, allowing for greater subsequent T-cell expansion. This change in accessory cell function correlates with increased class II antigen expression relative to susceptible mice, both in vivo during early infection and in vitro in response to induction by interferon-gamma (IFN-gamma). Differences in vitro were independent of, but differentially affected by, amastigote infection.

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PubMed1987

Acquisition of cell-mediated immunity to Leishmania. I. Primary T-cell activation detected by IL-2 receptor expression.

In order to analyse the early stages of the T-cell response to Leishmania, bioassays for detecting low levels of IL-2 receptor expression both in bulk culture and under limiting dilution conditions have been used. Infection of C57BL/10 mice with Leishmania donovani amastigotes leads to the appearance of antigen-specific T lymphocytes bearing high-affinity IL-2 receptors 24-72 hr later. Phenotypic analysis by complement-mediated cytotoxicity indicates that these activated T cells comprise both L3T4+, Lyt2- and L3T4-, Lyt2+ populations. The data also suggest the existence of activated cells bearing both these markers. By both assay techniques, the appearance of receptor-positive populations appears transitory, with few such cells detectable at 7 days post-infection. The implications of these data for further studies of murine leishmaniasis are discussed.

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PubMed1987

Inflammatory cells in murine visceral leishmaniasis express a dendritic cell marker.

Immunohistological studies of the murine liver following Leishmania donovani infection have been performed. We describe here the identification of cells expressing a recently defined dendritic cell marker, as detected by monoclonal antibody NLDC 145. Such cells are numerous in the developing inflammatory foci but are not detected individually at any other site in the infected liver. This finding suggests that mature tissue DC are not recruited during infection and that expression of this antigen is under precise microenvironmental control.

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PubMed1986

Regulation of macrophage accessory cell activity by mycobacteria. II. In vitro inhibition of Ia expression by Mycobacterium microti.

In our preceding study, we showed that infection of mice with Mycobacterium microti leads to a dramatic increase in Ia expression on local inflammatory macrophage populations. However, the majority of these cells did not contain intracellular organisms. To evaluate the effect of parasitism of macrophages by M. microti, Ia-induction experiments were performed in vitro. We show here that Ia expression is increased on peritoneal macrophages treated with either crude lymphokine preparations or recombinant gamma-interferon (gamma-IFN) and that this expression is suppressed by M. microti in a dose dependent fashion. The degree of suppression varied between macrophage populations and could be achieved to a lesser extent with killed organisms. It was partially reversed with indomethacin but only poorly so at high infection levels. Inhibition of Ia expression may be of importance in the generation and maintenance of chronic infection.

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PubMed1986

Regulation of macrophage accessory cell activity by mycobacteria. I. Ia expression in normal and irradiated mice infected with Mycobacterium microti.

CBA/Ca mice were infected by either the intravenous or intraperitoneal route with Mycobacterium microti and the subsequent changes in local macrophage populations examined. Following infection, the number of macrophages increased and they showed greater expression of both MHC Class II molecules. This response was not dependent on viability of the mycobacteria, in contrast to reports with other microorganisms such as Listeria. Studies in sublethally irradiated mice indicated that persistent antigen could give rise to a response after a period of host recovery which was radiation dose dependent. This procedure also highlighted differences in the regulation of different murine class II antigens in vivo, as seen by delayed re-expression of I-E antigens. Macrophage accessory cell function, as assessed by an in vitro T cell proliferation assay, correlated with Ia expression after fixation, but not after indomethacin treatment; this highlights the diverse nature of regulatory molecules produced by these cells.

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