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G Mayrhofer

Publications and source records attributed to G Mayrhofer.

At least 37 records · Page 2Linked to original sources
PubMed1996

Recruitment of lymphoblasts derived from peripheral and intestinal lymph to synovium and other tissues in normal rats and rats with adjuvant arthritis.

Recruitment of [125I]iododeoxyuridine-labeled syngeneic lymphoblasts from thoracic duct (TD) lymph into periarticular tissues has been examined after intravenous administration to normal rats and to rats with adjuvant-induced arthritis. Uptake of label was observed in the inflamed paws of arthritic rats and cells were located in synovium and periarticular bone marrow by autoradiography. Uptake was greater with lymphoblasts from donors in the late prodromal phase of adjuvant-induced arthritis (arthritic donors) than from normal donors. With arthritic donors, recruitment of lymphoblasts from TD lymph was greater than from mesenteric duct lymph, suggesting that most of the joint-seeking lymphoblasts in arthritic rats arose in peripheral lymphoid tissues. Lymphoblasts from arthritic donors were also detected in the synovium of paws from normal rats. Recovery of lymphoblasts was monitored in other tissues; this revealed, in arthritic recipients, competition among extra-articular sites of inflammation (adjuvant injection site, draining lymph nodes, and lymph nodes draining affected joints), the lungs, and the inflamed synovium for recruitment of lymphoblasts from arthritic donors. In contrast, while some lymphoblasts from normal donors were recruited to inflamed joints, the small intestine was the main site of recruitment. The results reflect the known propensity of T lymphoblasts generated in peripheral lymphoid tissues to enter inflamed tissues. However, some mesenteric duct lymphoblasts also entered inflamed synovium. The observed pattern of recruitment of lymphoblasts to synovium is pertinent to the pathogenesis of arthritis, the potential roles of arthritogenic and "bystander" lymphocytes and the known links between the joints and inflammation in the intestine.

Adoptive Transfer↗
PubMed1996

Complete prevention of the clinical expression of adjuvant-induced arthritis in rats by cyclosporin-A and lobenzarit: the regulation of lymph node cell populations and cytokine production.

A single dose of either cyclosporin-A (CsA) or lobenzarit (CCA) given with an arthrogenic adjuvant completely prevented expression of experimental adjuvant arthritis in rats. The aim of this study was to understand how these drugs prevented the arthritis expression by studying the popliteal lymph nodes draining the arthritic joints at various times after adjuvant injection. Neither drug affected the proliferation in popliteal lymph nodes at the time arthritis was normally expressed, however, there was a marked change in the types of cells present. Immunofluorescence assays showed a reduction in the proportion of CD4+ cells, while the proportion of B-lymphocytes was almost doubled. This coincided with a marked elevation in the ability of these cells to produce interleukin (IL)-6. At the same time production of other cytokines (IL-2, tumour necrosis factor (TNF) and interferon (IFN)-gamma) was not greatly affected. However, one day after adjuvant injection IL-2 and IFN-gamma production was reduced. In vitro experiments showed that IL-6 production by lymphoid cells was relatively unaffected by CsA and CCA but IL-2, TNF and IFN-gamma were suppressed by CsA. The results indicate that CsA and CCA may modify the response to the arthritic adjuvant by specifically inhibiting IL-2, TNF and IFN-gamma production at the time of adjuvant injection. The lack of inhibition of IL-6 by these drugs reveals it may not play a key role in the initiation of this model of chronic inflammation.

Animals↗
PubMed1996

Molecular genetic analysis of Giardia intestinalis isolates at the glutamate dehydrogenase locus.

Samples of DNA from a panel of Giardia isolated from humans and animals in Europe and shown previously to consist of 2 major genotypes--'Polish' and 'Belgian'--have been compared with human-derived Australian isolates chosen to represent distinct genotypes (genetic groups I-IV) defined previously by allozymic analysis. Homologous 0.52 kilobase (kb) segments of 2 trophozoite surface protein genes (tsa417 and tsp11, both present in isolates belonging to genetic groups I and II) and a 1.2 kb segment of the glutamate dehydrogenase (gdh) gene were amplified by the polymerase chain reaction (PCR) and examined for restriction fragment length polymorphisms (RFLPs). Of 21 'Polish' isolates that were tested, all yielded tsa417-like and tsp11-like PCR products that are characteristic of genetic groups I or II (15 and 6 isolates respectively) in a distinct assemblage of G. intestinalis from Australia (Assemblage A). Conversely, most of the 19 'Belgian' isolates resembled a second assemblage of genotypes defined in Australia (Assemblage B) which contains genetic groups III and IV. RFLP analysis of gdh amplification products showed also that 'Polish' isolates were equivalent to Australian Assemblage A isolates (this analysis does not distinguish between genetic groups I and II) and that 'Belgian' isolates were equivalent to Australian Assemblage B isolates. Comparison of nucleotide sequences determined for a 690 base-pair portion of the gdh PCR products revealed > or = 99.0% identity between group I and group II (Assemblage A/'Polish') genotypes, 88.3-89.7% identity between Assemblage A and Assemblage B genotypes, and > or = 98.4% identity between various Assemblage B/'Belgian' genotypes. The results confirm that the G. duodenalis isolates examined in this study (inclusive of G. intestinalis from humans) can be divided into 2 major genetic clusters: Assemblage A (= 'Polish' genotype) containing allozymically defined groups I and II, and Assemblage B (= 'Belgian' genotype) containing allozymically defined groups III and IV and other related genotypes.

Amino Acid Sequence↗
PubMed1996

Dependence of epithelial growth of the small intestine on T-cell activation during weaning in the rat.

BACKGROUND & AIMS: Intestinal crypt hyperplasia is associated with local T-cell activation in both clinical and experimental examples of immunologically mediated enteropathy. This suggests that T cell-derived factors may be trophic for epithelial proliferation in the intestine postnatally. The purpose of this study was to investigate T-cell activity during weaning in the rat and to investigate immune dependence of intestinal growth on T-cell activation. METHODS: The expression of interleukin-2 receptor (IL-2R) by mesenteric lymph node T cells was investigated from days 14 to 160 of life. Rats were treated with monoclonal antibodies against the IL-2R that were nonblocking (control) or blocking (experimental) from day 7, and intestinal growth was assessed at days 19, 25, and 29 of life. RESULTS: The mean +/- SEM of T cells expressing the IL-2R during weaning (days 15-28) was 6.1% +/- 0.3% compared with 3.3% +/- 0.3% at other ages (P < 0.001). The small intestine in rats treated with blocking antibody had reduced crypt length and mitotic count compared with control animals. CONCLUSIONS: Weaning is associated with activation of T cells and blockade of the IL-2R reduces intestinal growth.

Animals↗
PubMed1996

Ovarian steroid hormones regulate granulocyte-macrophage colony-stimulating factor synthesis by uterine epithelial cells in the mouse.

Uterine epithelial cells have been shown by in vitro studies to be a potent source of the inflammatory cytokine granulocyte-macrophage colony-stimulating factor (GM-CSF), and the luminal and glandular epithelium has been confirmed as the predominant site of GM-CSF expression in the intact endometrium by in situ hybridization. To examine the role of ovarian steroid hormones in GM-CSF synthesis, GM-CSF bioactivity has been measured in the supernatants of short-term primary cultures of endometrial cells prepared from mice in which steroid levels were perturbed by ovariectomy and steroid replacement or by steroid antagonists. GM-CSF production was found to fluctuate in cells harvested at different times during the estrous cycle, peaking at estrus. Endometrial cells derived from ovariectomized mice produced 25-fold less GM-CSF than did cells from estrous mice, and production was increased if ovariectomized mice were pretreated with estrogen, but not progesterone, 3 h or more before harvest. This estrogen-induced increase was inhibited by coadministration of progesterone or by induction of a decidual response and was blocked by the estrogen antagonist ZK 119,010. By contrast, pretreatment of mice with the anti-progestin RU486 significantly elevated GM-CSF output in cells from ovariectomized mice given estrogen and progesterone in combination and antagonized the inhibition of GM-CSF release seen in cells harvested from mice treated with hCG. These studies demonstrate that GM-CSF synthesis and/or release by uterine epithelial cells is stimulated by estrogen, with progesterone having a moderate inhibitory effect. Analysis of GM-CSF mRNA expression in uterine epithelial cell cultures and in intact uteri from steroid hormone-treated ovariectomized mice by quantitative reverse transcription-polymerase chain reaction indicated that the effects of estrogen and progesterone on GM-CSF release are mediated at least in part at the transcriptional level. These findings implicate GM-CSF as a local mediator of steroid-driven remodeling events in the cycling and preimplantation endometrium, possibly acting through the recruitment and behavioral regulation of granulocytes and macrophages.

Animals↗
PubMed1996

Measurement of virulence of aeromonads using a suckling mouse model of infection.

Virulent and avirulent strains of Aeromonas spp. were identified and virulence quantified using an animal model. Virulence was measured by determining a 50% lethal dose (LD50) 43 h after oral administration of live bacteria. The LD50 of virulent Aeromonas isolates ranged from log10 7.53 (mean) organisms to log10 8.88 (mean). Some isolates were avirulent in this model. Detection of cytotoxic activity in culture supernatants correlated with virulence (Fisher exact test, P = 0.0029). There was no correlation between LD50 and the source of the isolate, beta-haemolysis or lipopolysaccharide (LPS) banding profile on SDS-PAGE. In this animal model, virulence was multifactorial in that: (i) bacterial multiplication in the gut was associated with fatal infection; (ii) the increase in bacterial numbers in the gut of mice administered a lethal dose of bacteria was accompanied by accumulation of fluid; and (iii) there was evidence of extraintestinal spread of infection. Protection of suckling mice by rabbit antiserum to Aeromonas cell envelopes was observed.

Aeromonas↗
PubMed1995

Division of Giardia isolates from humans into two genetically distinct assemblages by electrophoretic analysis of enzymes encoded at 27 loci and comparison with Giardia muris.

Giardia that infect humans are known to be heterogeneous but they are assigned currently to a single species, Giardia intestinalis (syn. G. lamblia). The genetic differences that exist within G. intestinalis have not yet been assessed quantitatively and neither have they been compared in magnitude with those that exist between G. intestinalis and species that are morphologically similar (G. duodenalis) or morphologically distinct (e.g. G. muris). In this study, 60 Australian isolates of G. intestinalis were analysed electrophoretically at 27 enzyme loci and compared with G. muris and a feline isolate of G. duodenalis. Isolates of G. intestinalis were distinct genetically from both G. muris (approximately 80% fixed allelic differences) and the feline G. duodenalis isolate (approximately 75% fixed allelic differences). The G. intestinalis isolates were extremely heterogeneous but they fell into 2 major genetic assemblages, separated by fixed allelic differences at approximately 60% of loci examined. The magnitude of the genetic differences between the G. intestinalis assemblages approached the level that distinguished the G. duodenalis isolate from the morphologically distinct G. muris. This raises important questions about the evolutionary relationships of the assemblages with Homo sapiens, the possibility of ancient or contemporary transmission from animal hosts to humans and the biogeographical origins of the two clusters.

Alleles↗
PubMed1995

Absorption and presentation of antigens by epithelial cells of the small intestine: hypotheses and predictions relating to the pathogenesis of coeliac disease.

Effects of the route of antigen absorption and presentation in the small intestine are discussed in relation to outcomes from antigen exposure. It is proposed that antigens presented by epithelial cells to intrinsic T cell populations in the mucosa induce anti-inflammatory immune responses, while antigens presented to conventional T cells by lamina propria APC predisposes to inflammation. The pathogenesis of coeliac disease is attributed to three factors: a defect in antigen processing by epithelial cells; the intrinsic properties of the gliadins; and the HLA-D haplotype of the individual. It is suggested that MHC class II molecules play a role in antigen uptake by enterocytes, in addition to their function in antigen presentation to T cells.

Antigen Presentation↗
PubMed1994

Origin of the oocyte shell membrane of a dasyurid marsupial: an immunohistochemical study.

Polyclonal antibodies were raised against the extracellular coats (mucoid and shell membrane) of unfertilized oocytes and early embryos of a dasyurid marsupial, the fat-tailed dunnart (Sminthopsis crassicaudata). Indirect immunofluorescence was used to test the specificity of the antibodies to the shell membrane. Streptavidin/biotin immunoperoxidase cytochemistry revealed that precursors of the tertiary egg membranes are secreted by the luminal epithelium of the ampulla, isthmus (including the crypts), utero-tubal junction, and adjacent endometrial glands. Histochemistry distinguished between the sites of mucoid and shell membrane precursor secretion. The mucoid coat stained positively with Alcian blue at pH 1.0 and 2.5, with Alcian blue at pH 0.2 after performic acid oxidation, and with PAS which was amylase resistant. Some of the luminal epithelial cells of the ampulla and isthmus, as well as its crypts, also stained positively by these histochemical methods but the luminal epithelium of the utero-tubal junction and endometrial glands were negative for all methods. The shell membrane did not stain with any of the above methods nor with dihydroxy-dinapthyl-disulphide (DDD) or ferric ferricyanide but it was eosinophilic and stained positively with the red cytoplasmic stain of Masson's trichrome. Therefore, it is concluded that shell membrane precursors are secreted by the luminal epithelium of the utero-tubal junction, adjacent glands, and by scattered glands in the anterior region of the uterus but not by any cell population of the oviduct.

Animals↗
PubMed1993

Two genes encoding homologous 70-kDa surface proteins are present within individual trophozoites of the binucleate protozoan parasite Giardia intestinalis.

A 0.52-kb DNA sequence encoding part of a major surface antigen of Giardia intestinalis trophozoites has been amplified by the polymerase chain reaction (PCR) using primers specific for nucleotide sequences that are conserved between two apparently related genes, tsp11 (cloned previously from the Australian G. intestinalis isolate, Ad-1) and tsa417 (cloned from the Afghanistani isolate, WB). Restriction analysis revealed that the DNA amplified from each of seven axenic isolates of G. intestinalis was not homogeneous, even though the template DNA had been purified from cultures that had been established from single trophozoites. Every isolate yielded two PCR products, whose respective cleavage fragments corresponded to tsp11 (HindIII+, PstI-, KpnI-) and to tsa417 (HindIII-, PstI+, KpnI+). This was confirmed by cloning individual amplification products into the plasmid vector, pGEM-7Zf(+). The sequence of one cloned fragment (1-P4), derived from the Ad-1 isolate, but possessing restriction sites characteristic of tsa417, exhibited 98.6% identity over 425 bp with tsa417 and only 63.8% identity with the corresponding region of tsp11. The data indicate that individual trophozoites of all the isolates examined contain a copy of each of these homologous genes.

Amino Acid Sequence↗
PubMed1993

Giardia intestinalis: detection of major genotypes by restriction analysis of gene amplification products.

The polymerase chain reaction (PCR) has been used to amplify a 0.52 kb segment of Giardia intestinalis DNA, using primers specific for nucleotide sequences conserved within two genes (tsp11 and tsa417) that encode homologous, cysteine-rich trophozoite surface proteins. Using products amplified from axenic isolates belonging to genetic groups I and II (defined on the basis of allozyme electrophoresis data), restriction endonuclease analysis revealed both tsp11-like and tsa417-like fragments within all samples. The study also identified among the amplification products of group II organisms an additional fragment, containing a novel PstI site, that is not detected in the reaction products of group I isolates. The recovery of three distinct PCR products from each group II isolate was verified by cloning the fragments into the plasmid vector pGEM-7. Fragments containing the new PstI site possess the ClaI site common to both tsp11 and tsa417-like fragments, but they lack the HindIII site which characterizes tsp11-like fragments and also lack the PstI and KpnI sites which characterize tsa417-like fragments. Spot-blot analyses using cloned fragments of all three types as probes showed strong homologous hybridization but weak heterologous hybridization, indicating that each type differs substantially in nucleotide sequence from the others. Because the samples of Giardia DNA used in the PCR were purified from cultures that had been established from single trophozoites, the data indicate that individual trophozoites belonging to genetic group II possess three homologous genes defined by these related fragments. The presence of a PstI site in the amplified segment of the newly-discovered third gene of group II organisms provides a simple diagnostic means of differentiating group I and II isolates.

Animals↗
PubMed1993

A gene encoding a 69-kilodalton major surface protein of Giardia intestinalis trophozoites.

A gene encoding a 68.5-kDa trophozoite surface protein (TSP11) of the Australian Giardia intestinalis (syn. G. lamblia) isolate, Ad-1, has been cloned from a genomic expression library screened with an antiserum specific for 3 major surface antigens. Sequence analysis of two overlapping genomic fragments identified a single open reading frame that contained no introns and predicted a cysteine-rich, 667-residue polypeptide with features common to other trophozoite surface proteins. These include the presence of 27 copies of the 4-amino acid Cys-X-X-Cys motif, an N-terminal signal sequence and a highly conserved, hydrophobic C-terminal segment. Transcripts from the tsp11 gene were detected as a single band on Northern blots using total RNA extracted from Ad-1 trophozoites. Primer extension analysis indicated that the mRNA has a 5' untranslated region of only 5 nt, similar to the very short (1-6 nt) leader sequences reported for other Giardia mRNAs. A large portion of the promoter distal segment of tsp11 has homology with tsa417, a gene encoding a 72.5-kDa trophozoite surface antigen of the Afghanistan-derived G. intestinalis isolate, WB [13].

Amino Acid Sequence↗
PubMed1993

Differentiation of major genotypes of Giardia intestinalis by polymerase chain reaction analysis of a gene encoding a trophozoite surface antigen.

The polymerase chain reaction (PCR) has been used to amplify in vitro a semi-conserved region of a gene encoding an M(r) 68-72,000 surface antigen of Giardia intestinalis trophozoites. Using primers specific for conserved nucleotide sequences identified within the promoter-distal portion of two homologous genes (tsp11 and tsa417) cloned previously from the G. intestinalis isolates Ad-1 (from Australia) and WB (from Afghanistan), a single PCR-amplified DNA fragment of the expected size (0.52 kilobases) was obtained in high yield from either purified DNA or whole trophozoites of the Ad-1 isolate and from every 1 to 9 other axenic G. intestinalis isolates belonging to genetic groups I and II (defined previously on the basis of allozyme electrophoresis data--Andrews et al. 1989). Discernible product was recovered from as few as 2-4 trophozoites. In contrast, 6 G. intestinalis isolates that were assigned by allozymic analysis to genetic groups III/IV yielded small amounts of a 0.37-kilobase (kb) amplification product (with evidence in some samples of an additional 0.4 or 0.18 kb fragment) but no 0.52 kb product. Two animal-derived isolates of G. duodenalis (one from an Australian native rodent, Notomys alexis, the other from a domestic cat) also yielded a single 0.37 kb PCR-amplified fragment, whereas an isolate from another cat produced a 0.34 kb fragment. No product was recovered from G. muris, a morphologically distinct species of Giardia. The results demonstrate that different genotypes of G. duodenalis can be distinguished using this assay and that it is diagnostic for isolates belonging to two major clusters (groups I/II and III/IV) of G. intestinalis. The amplified DNA segment appears to be relatively conserved among group I and group II isolates of G. intestinalis. A related but clearly distinct sequence seems to be conserved among group III/IV isolates of G. intestinalis and some isolates of G. duodenalis.

Animals↗
PubMed1993

Localization of leukocyte subsets in the rat ovary during the periovulatory period.

The ovulatory process has been compared with inflammation because several classical inflammatory mediators appear to participate in this process. One component of the inflammatory reaction is the migration of leukocytes to the site of inflammation and the subsequent activation of these cells. We have reported recently that perfusion of leukocytes into the rat ovary in vitro enhances the number of LH-induced ovulations, which suggests an active role of leukocytes in ovulation. In the present study we characterize immunohistochemically the distribution of macrophages, T lymphocytes, and granulocytes in the ovaries of untreated immature rats and of eCG-hCG-primed rats killed prior to hCG injection, at ovulation, and at 33-36 h post-ovulation. Macrophages, identified with monoclonal antibodies ED1 and ED2, were the major leukocyte population and were found primarily in the medullary region surrounding the blood vessels. The density of the cells in this region increased continuously during development to sexual maturity and until after ovulation. Macrophages were also present in the thecal layer of the preovulatory follicles, and the numbers of these cells increased about 5-fold in this area in ovulating follicles (12 h after hCG) compared to preovulatory follicles (before hCG). A portion of macrophages in both areas expressed major histocompatibility complex (MHC) class II antigens (OX6+); these cells were present mostly in the medullary region, with no apparent change in density during the periovulatory period. Neutrophilic granulocytes comprised a lesser proportion of the total leukocyte population in the medullary region but were abundant in the thecal layer. The density of neutrophils increased 3-fold in the medullary region and 8-fold in the thecal region in ovulatory compared to preovulatory follicles. T lymphocytes (OX52+) were evenly distributed at relatively low density in the medulla and the stroma of the cortex. Most T lymphocytes expressed the CD8 antigen (OX8+) and hence were of the MHC class I-restricted phenotype. Few T lymphocytes were present in the thecal layer. In summary, macrophages, neutrophilic granulocytes, and T lymphocytes are present in the ovary at ovulation. There is a selective increase in the numbers of macrophages and neutrophilic granulocytes in the medullary region and in the thecal layer as the ovulatory period progresses, indicating that these cells may actively be involved in the tissue remodeling occurring at ovulation.

Analysis of Variance↗
PubMed1992

Identification of a broad-specificity nucleoside transporter with affinity for the sugar moiety in Giardia intestinalis trophozoites.

A broad-specificity nucleoside transporter has been identified in Giardia intestinalis trophozoites, using a rapid sampling assay to measure influx of [3H]deoxycytidine, [3H]adenosine and [3H]guanosine at 0 degrees C. The influx of each labelled nucleoside was inhibited strongly by all common, naturally-occurring nucleosides but only poorly or not at all by nucleobases, indicating that the transporter recognizes structural features on the furanosyl moiety of ribo- and 2'-deoxyribonucleosides. Both 2'- and 5'-deoxyadenosine were potent inhibitors of influx (greater than 95% inhibition at 2 mM), whereas 3'-deoxyadenosine was significantly less effective (approx. 70% inhibition), and 2',3'-dideoxycytidine and cytosine arabinoside were virtually inactive (0-20% inhibition). The data reveal that the 2'- and 5'-hydroxyl groups are not necessary for the recognition of nucleosides by this transporter. However, the 3'-hydroxyl appears to be important. Michaelis-Menten constants (Km) were calculated for the influx at 0 degrees C of deoxycytidine (220 +/- 116 microM) and adenosine (45 +/- 24 microM), with respective Vmax values of 13 +/- 4 and 11 +/- 2 pmol min-1 (10(6) cells)-1. Only 12-26% of [3H]thymidine influx occurred through this transporter, the remainder entering the cells through a thymine/uracil-specific transporter described previously. Thymidine exhibited a Ki of 205 +/- 90 microM against [3H]deoxycytidine influx.

Adenosine↗
PubMed1992

Changes in allozyme pattern of the protozoan parasite Giardia intestinalis.

The present study compares the allelic profiles of Giardia intestinalis grown in vivo and in vitro. Three clinical isolates of G. intestinalis were established in suckling mice and subsequently adapted to in vitro culture to test the null hypothesis that samples of the same clinical isolate grown in different culture conditions have identical allelic profiles. For each isolate, a mouse-derived and an axenically cultured sample were analysed electrophoretically at 11 enzyme loci. In each case, the axenically cultured sample of each isolate showed marked allelic differences from its corresponding in vivo sample. These data suggest that there may be either regulated expression of alternative genes encoding distinct isozymes (i.e. gene switching) or selection by different growth conditions of specific genotypes from a mixture present within the original clinical isolate. Although these hypotheses are not tested in this study, the data highlight the importance of confirming that allozymes (or isozymes) are stable genetic characters for the identification and characterization of protozoan taxa.

Alleles↗
PubMed1992

The use of suckling mice to isolate and grow Giardia from mammalian faecal specimens for genetic analysis.

A simple technique is described for preparation of Giardia cysts from faecal samples, the growth of trophozoites in suckling mice and the isolation of trophozoites for genetic analysis by allozyme electrophoresis. In total, 125 new isolates of Giardia have been collected from human and animal sources over a wide geographical area of South Australia and the Northern Territory of Australia. A number of long-established axenized isolates of G. intestinalis belonging to Groups I and II also adapted to grow in suckling mice. These findings indicate that suckling mice are permissive hosts for a variety of genetically dissimilar but morphologically similar organisms of the G. duodenalis type and that this in vivo technique may be less selective than isolation by in vitro culture. The use of suckling mice has revealed that infections can be composed of mixed genotypes and that isolation and purification techniques can be selective. Allozymic interpretation is essential to reveal the genetic complexity of such mixtures.

Alleles↗