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T Aebischer

Publications and source records attributed to T Aebischer.

34 records · Page 2Linked to original sources
PubMed1995

The role of macrophage receptors in adhesion and uptake of Leishmania mexicana amastigotes.

Amastigotes of the protozoan parasite Leishmania proliferate in phagolysosomes of mammalian macrophages. Propagation of the infection is considered to occur by host-cell rupture and uptake of released parasites by uninfected macrophages. In this study, the kinetics of binding of L mexicana mexicana amastigotes to COS cells and to COS cells transfected with three different macrophage receptors (FcRII-B2, receptor for the Fc-domain of immunoglobulins; CR3, complement type 3 receptor and the mannose receptor) is compared to the rate of adhesion to peritoneal macrophages. Amastigotes isolated from macrophages cultivated in vitro bind with slow, sigmoid kinetics to COS cells expressing either of the three receptors, or to peritoneal macrophages. In contrast, amastigotes isolated from mouse lesions bind with rapid, hyperbolic kinetics to COS cells expressing the Fc receptor or to peritoneal macrophages but with slow, sigmoid kinetics to COS cells expressing the CR3 or the mannose receptor. As shown by immunofluorescence experiments, lesion-derived amastigotes contain host-derived immunoglobulins (Ig) but no complement component 3 at their surface. It is concluded that amastigotes contain no intrinsic ligand at their surface, which enables high-affinity interactions with macrophages. Opsonization by specific Ig may be of relevance in vivo because firstly, in cryosections of mouse lesions extracellular amastigotes containing surface Ig can be detected and, secondly, B cell-deficient mice reconstituted with parasite-specific Ig show a modest increase in the rate of lesion development. In addition, it is shown that amastigotes are internalized by COS cells and grow in large parasitophorous vacuoles similar to those observed in macrophages.

Adhesiveness↗
PubMed1994

Recurrent cutaneous leishmaniasis: a role for persistent parasites?

Leishmaniosis is, with increasing frequency, reported as an opportunistic infection of immunosuppressed individuals. Re-activation of persistent parasites may be responsible for the disease in a number of these patients. Here, Toni Aebischer reviews some of the evidence for the implication of persistent Leishmania infections in recurrent disease with the emphasis on cutaneous leishmoniasis in humans and in the mouse model. The data suggest that parasite persistence is a common feature in Leishmania infections. The availability of on excellent laboratory model provides on opportunity to study this phenomenon in detail. The findings of these analyses are likely to be important for the identification of people at risk of developing recurrent disease and for the assessment of new therapies for relapsing leishmaniasis and might also have implications for the design of a future anti-Leishmania vaccine.

Journal Article↗
PubMed1994

Ricin-resistant mutants of Leishmania major which express modified lipophosphoglycan remain infective for mice.

Glycosylation variants of the virulent Leishmania major clone V121 were generated by mutagenesis with N-methyl-N-nitroso-N-nitroguanidine and selected using the galactose-specific lectin Ricinus communis II (RCA II). Three mutants, 4B9, 1D1 and 1C12, which failed to bind RCA II, were found to have an altered expression of lipophosphoglycan (LPG), a molecule implicated in the attachment to host macrophages and survival within the phagolysosome. There were differences in the antigenicity, molecular weight and localization of LPG from mutant parasites as compared to V121. Expression of gp63, a surface molecule also implicated in attachment to macrophages, was unaltered. All 3 mutants caused disease when injected into genetically susceptible BALB/c mice but lesions developed at a much slower rate than those caused by the virulent V121 clone. This slow rate of lesion development did not correlate with promastigotes' ability to invade macrophages in vitro. Karyotype analysis showed that there was a reduction in the size of chromosome band number 2 in all 3 mutants. The differences in LPG and chromosome band 2 were retained by mutant clones following passage through mice, suggesting that these phenotypes are stable. Although the mutant parasites were infective and caused lesions, the changed structure of the LPG appeared to influence the virulence of the parasites.

Agglutination↗
PubMed1994

Intravenous injection of irradiated Leishmania major into susceptible BALB/c mice: immunization or protective tolerance.

It is well established that BALB/c mice can be protected from fatal infection with Leishmania major by prophylactic intravenous (i.v.) immunization with irradiated parasites. Protection is critically dependent on the route of injection with i.v. injection being protective and subcutaneous injection not protective. We used this BALB/c-L. major model system to investigate this phenomenon. We analyzed quantitatively the parasite-specific, CD4+ T cell mediated immune responses by limiting dilution. Subcutaneous vaccination resulted in priming of CD4+ precursor T cells, whereas i.v. vaccination was ineffectual. Moreover, i.v. injection prevented the increase in the number of specific precursor cells induced by infection of normal mice during the first weeks post-challenge with virulent parasites. We show here that this was not due to the elimination of the virulent challenge parasites as a result of immunity nor to inefficient antigen presentation of the irradiated organisms after i.v. injection. The data presented here suggest that i.v. injection results in tolerization rather than immunization. Tolerization as a mechanism of host protection is consistent with earlier observations that transient immunosuppression results in cure of L. major infection in BALB/c mice. Transfer of antigen presenting cells (APC) isolated from spleens of mice injected previously with irradiated parasites mimicked to some extent the effect of i.v. immunization with irradiated parasites. The possible involvement of these APC in decreasing the parasite-specific T cell response is discussed.

Animals↗
PubMed1993

Virus-specific cytotoxic T cell-mediated lysis of lymphocytes in vitro and in vivo.

Virus-specific CTL play a major role in early antiviral protection against lymphocytic choriomeningitis virus (LCMV). When mice are infected with high doses of certain LCMV isolates, the initiated CTL response may vanish before the virus is eliminated completely. To evaluate the possibility that this may be because of CTL lysing CTL, we studied the susceptibility to lysis of LCMV-specific CTL clones and of primary immune spleen cells in vitro and in vivo. Confirming earlier reports, CTL were lysed in vitro when incubated with their specific Ag peptide; lysis was MHC restricted because T cell clones derived from allogeneic bone marrow chimeras that do not express the correct restriction element were not susceptible to mutual CTL lysis. The density of CTL cultures correlated with the degree of inactivation of CTL, indicating that CTL lysed each other mutually rather than committed suicide. To assess CTL-mediated lysis of CTL in vivo, mice were infected s.c. into the footpad with LCMV, and shortly before the CTL-dependent footpad swelling developed by day 6 to 7, specific peptide was injected locally. The regional lymph node (LN) was reduced in size, contained about 10 times fewer cells, and only 2% to 3% of the lytic activity when compared with several control LN. This drastic local effect was observed within a few hours and was accompanied by DNA fragmentation. Thus, in this model system, peptide loading of lymph node cells may lead to CTL-mediated cytotoxicity and death of T cells (including CD8+ effector cells and other LN cells) in vivo. These results may suggest that, during overwhelming virus infections, lysis of CTL passively loaded with relevant released peptides possibly may contribute to the impairment of immune responses.

Animals↗
PubMed1993

Resistance of BALB/c mice to Leishmania major infection is associated with a decrease in the precursor frequency of antigen-specific CD4+ cells secreting interleukin-4.

BALB/c mice are highly susceptible to infection with the protozoan parasite Leishmania major and develop a chronic fatal disease. They can, however, be manipulated to resist disease and this has been shown to correlate with increased expression of IFN-gamma mRNA and the absence of IL-4 mRNA in the draining lymph nodes and spleens of these animals. Here we show that anti-IL-4 or anti-CD4 treatment of BALB/c mice resulted in a reduction in the size of the lesion and in the number of parasites in the draining lymph nodes compared with untreated mice. The precursor frequency of CD4+ T cells proliferating in response to Leishmania antigens in vitro in the treated animals was not significantly different from untreated animals. Analysis of the lymphokines secreted by the clonal progeny of these cells showed that the precursor frequency of IL-4 secreting clones was at least 10-fold lower in animals treated with either mAb. However, there was no reciprocal increase in the precursor frequencies of IFN-gamma secreting clones. Comparisons of the total number of precursors of specific CD4+ cells secreting IFN-gamma showed that anti-CD4-treated animals, which are resistant to disease, had considerably fewer for the first 6 weeks than untreated mice with chronic disease. Protection of BALB/c mice was therefore associated with a reduction in the numbers of precursors of cells secreting IL-4 without a concomitant increase in the number of precursors of IFN-gamma secreting cells.

Animals↗
PubMed1993

Persistence of virulent Leishmania major in murine cutaneous leishmaniasis: a possible hazard for the host.

The persistence of Leishmania major parasites in mice resistant to infection was investigated by the polymerase chain reaction and in vitro culture methods. Parasite-specific DNA was detected in the lymph nodes, spleens, bone marrow, and livers of C57BL/6 mice 1 year after their recovery from infection. Live parasites were also recovered from these tissues (except liver tissues) and were used to establish in vitro isolates. Pulsed-field gel electrophoresis, Southern blotting, and Western blot (immunoblot) analyses showed that these isolates retained the karyotype and the phenotype of the original inoculum, including the levels of expression of gp63 and lipophosphoglycan, the two major surface molecules of Leishmania species. More importantly, these isolates were virulent and induced fatal disease when injected into susceptible BALB/c mice. Persistence was shown to be a more general phenomenon, since several different strains of mice which were resistant to L. major infection also harbored persistent parasites. The implications for the etiology of human leishmaniasis in immunocompromised individuals such as AIDS patients are discussed.

Animals↗
PubMed1993

Interleukin-4 but not gamma interferon production correlates with the severity of murine cutaneous leishmaniasis.

For murine cutaneous leishmaniasis, data to date suggest a correlation between the presence of gamma interferon (IFN-gamma) and resistance in C57BL/6 mice and the presence of interleukin-4 (IL-4) and disease in BALB/c mice. In this study, 13 inbred strains of mice covering the range of susceptibility to disease were infected with Leishmania major to determine whether the subsequent expression of IFN-gamma or IL-4 is a reliable indicator of cure or progressive disease. The presence of IL-4 and IFN-gamma mRNAs in the draining lymph nodes was examined 9 weeks after infection, when differences in disease severity became obvious. There were large differences in the levels of IL-4 mRNA among the different strains, whereas IFN-gamma mRNA was detected at similar levels in all strains. The levels of IL-4 mRNA correlated with lesion score, with susceptible and intermediate strains containing up to 100-fold more than any of the resistant strains. Differences in the levels of IFN-gamma mRNA were within only a fourfold range, with significant overlap among susceptible, intermediate, and resistant strains. Similarly, the levels of IFN-gamma secreted in vitro by lymph node cells from infected mice in response to L. major antigens were within a 10-fold range for most strains, and there was no correlation with lesion score. Analysis of Leishmania-specific antibody levels revealed a correlation between immunoglobulin G1 (IgG1) titers and lesion score, consistent with the role of IL-4 as a switch factor for IgG1. In contrast, there was no correlation between IgG2a titers and lesion score, supporting the notion that IFN-gamma synthesis (which promotes IgG2a production) is not correlated with disease state. These data suggest that along the spectrum of murine cutaneous leishmaniasis, IL-4 is a reliable indicator of disease, but IFN-gamma is not prognostic for resistance.

Animals↗
PubMed1991

In vitro selection of lymphocytic choriomeningitis virus escape mutants by cytotoxic T lymphocytes.

Cytotoxic T lymphocyte (CTL)-mediated cytolysis is induced via the interaction of the specific T-cell antigen receptor and the peptidic viral antigen associated with the major histocompatibility complex class I antigen. Here we demonstrate in vitro that lymphocytic choriomeningitis virus (LCMV) can escape the cytotoxic activity of LCMV-specific cloned CTLs by single amino acid changes within the recognized T-cell epitope defined by residues 275-289 of the LCMV glycoprotein [LCMV-GP-(275-289)]. LCMV-infected fibroblasts at a multiplicity of infection of 10(-3) exposed to virus-specific CTL at an effector-to-target cell ratio of 4:1 4 hr after infection was optimal for virus mutant selection. The selections were carried out with three LCMV-GP-(275-289)-specific CTL clones expressing T-cell antigen receptors containing the identical variable gene segments V alpha 4 and V beta 10 but different junctional regions; selection was also possible with LCMV-GP-(275-289)-specific cytotoxic polyclonal T cells. The most common escape mutation was an amino acid change of asparagine (AAT) to aspartic acid (GAT) at position 280; an additional mutation was glycine (GGT) to aspartic acid (GAT) at position 282. The results presented show that relevant point mutations within the T-cell epitope of LCMV-GP-(275-289) occur frequently and that they are selectable in vitro by CTLs.

Amino Acid Sequence↗
PubMed1990

Preferential usage of V alpha 4 and V beta 10 T cell receptor genes by lymphocytic choriomeningitis virus glycoprotein-specific H-2Db-restricted cytotoxic T cells.

Correlations between the T cell receptor (TcR) V gene usage and the specificity of T cells have been primarily described for major histocompatibility complex (MHC) class II-restricted helper T cell responses. In the present study the TcR genes expressed by MHC class I-restricted murine cytotoxic T cells (CTL) specific for a major epitope of the lymphocytic choriomeningitis virus (LCMV), LCMV-GP2(275-289), were investigated. The TcR primary structure of an LCMV-GP2(275-289) specific H-2Db-restricted CTL clone has been determined. It uses a member of the V alpha 4 family joined to J alpha AN14.4 for the alpha chain and V beta 10 rearranged to D beta 2.1 and J beta 2.4 for its beta chain. Four other independent LCMV-GP2(275-289) specific H-2Db-restricted CTL clones also expressed V alpha 4 and V beta 10 gene elements. Furthermore, V alpha 4 and V beta 10 were preferentially expressed by polyclonal CTL of C57BL/6 origin specific for LCMV. These results suggest that both TcR V alpha and V beta regions are important for the recognition of the LCMV-GP2(275-289) epitope on H-2Db molecules.

Amino Acid Sequence↗
PubMed1990

Expression of cellular effector functions and production of reactive nitrogen intermediates: a comparative study including T lymphocytes, T-like cells, neutrophil granulocytes, and mononuclear phagocytes.

The ability of various cell types to secrete reactive nitrogen intermediates (RNI) upon functional activation was comparatively assessed. Neither in T lymphocyte clones mediating MHC class I or class II antigen-restricted killing via alpha/beta T cell receptor (TcR) or MHC-unrestricted killing via gamma/delta TcR, nor in peripheral blood mononuclear cells expressing natural killer or lymphokine-activated killer activity, target cell lysis was associated with detectable RNI production. Also, activated neutrophil granulocytes did not secrete RNI. In contrast, bone marrow-derived mononuclear phagocytes, activated to express tumoricidal activity, secreted marked RNI activity.

Animals↗
PubMed1989

Detection of perforin and granzyme A mRNA in infiltrating cells during infection of mice with lymphocytic choriomeningitis virus.

The analysis of gene expression in cytotoxic T cells by in situ hybridization of serial liver and brain sections from mice infected with lymphocytic choriomeningitis virus (LCMV) and immunostaining with T cell marker- and virus-specific antibodies revealed a close histological association of infiltrating lymphocytes expressing the perforin and granzyme A genes with virally infected cells. Maximal frequency of perforin and granzyme A mRNA-containing cells on liver sections preceded by about 2 days maximal LCMV-specific cytotoxicity of the lymphoid liver infiltrating cells. These results are most consistent with an involvement of perforin and granzyme A in cell-mediated cytotoxicity in vivo.

Animals↗
PubMed1989

Cloning, analysis, and expression of murine perforin 1 cDNA, a component of cytolytic T-cell granules with homology to complement component C9.

The nucleotide sequence coding for the cytotoxic T-lymphocyte (CTL) protein perforin 1 (P1) has been determined and the corresponding protein sequence has been derived. Murine CTL cDNA libraries contained in the vector lambda gt11 were screened by using a monospecific antiserum to purified P1. Three recombinant phages were isolated and their cDNA inserts were sequenced. The derived protein sequence contains 554 amino acids and displays, as expected, considerable homology with certain functional domains in the complement components C9, C8 alpha, C8 beta, and C7. The identity of P1 cDNA clones was verified by prokaryotic expression and the reactivities of antisera produced to the expressed proteins. In addition, antisera were produced to two synthetic peptides located in the center and C-terminal portions of P1. All antisera reacted with purified P1. In Northern blot analyses, P1 cDNA probes recognized a 2.9-kilobase mRNA only in CTL. Perforin mRNA was found in all cloned CTL and in all mixed lymphocyte reactions that gave rise to cytotoxic cells. Perforin mRNA was also detected in virus-specific CTL that had been generated in vivo and isolated from liver tissue of mice infected with lymphocytic choriomeningitis virus. The cell-specific expression of perforin is consistent with its postulated role in cytolysis.

Amino Acid Sequence↗
PubMedDate not supplied

Topical treatment with hexadecylphosphocholine (Miltex) efficiently reduces parasite burden in experimental cutaneous leishmaniasis.

Ether-lipids and alkylphosphocholines have been found to have anti-leishmanial activity. Oral treatment with hexadecylphosphocholine (HePC) efficiently reduces parasite burden in murine visceral leishmaniasis. Drugs for the treatment of cutaneous leishmaniasis are most commonly administered parenterally, whereas efficient drugs for topical treatment are not in current use. Here we investigate the efficacy of topical treatment with HePC in mice infected with Leishmania mexicana or L. major, causative agents of cutaneous leishmaniasis in the New and Old World, respectively. BALB/c, CBA/J and C57BL/6 inbred mice do not control infection with L. mexicana because they do not mount an efficient Th1-type anti-parasitic lymphocyte response. In contrast, C57BL/6 mice are resistant to an infection with L. major, developing only transient lesions that heal spontaneously owing to an efficient Th1 response. BALB/c, CBA/J and C57BL/6 mice were infected subcutaneously with L. mexicana amastigotes, causing nodular lesions after 5 months. Topical treatment with HePC (Miltex) was highly effective in reducing parasite burden and healed established lesions. The treatment did not induce a Th1 response in L. mexicana-infected susceptible mice and most of the mice relapsed. In resistant C57BL/6 mice infected subcutaneously with 2 x 10(6) L. major promastigotes at the tail base, nodular lesions developed after 2 weeks. Topical treatment with Miltex reduced the parasite load and the mice healed their lesions much faster than the untreated infected controls. The clinical application of Miltex for treatment of cutaneous leishmaniasis may be highly efficient because humans, similarly to resistant mice, in general do not relapse after healing. Clinical trials should be straightforward considering that Miltex is an approved drug for the treatment of breast cancer metastases.

Administration, Topical↗