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Biomedical subjects

M A Leon

Publications and source records attributed to M A Leon.

At least 19 recordsLinked to original sources
PubMed1991

Environmental signals induce major changes in virulence of Shigella spp.

Growth conditions play a major role in expression of virulence by Shigella spp. both in vitro (adherence and internalization in eukaryotic host cells) and in vivo (keratoconjunctivitis). Optimized expression of virulence required anaerobic growth to log phase in particular media such as brain heart infusion broth. Kinetic studies of guinea pig eye infections showed that as few as 2 x 10(5) S. dysenteriae CG097 or S. flexneri M90T, grown under these optimized conditions, produced keratoconjunctivitis in 15 h. In vitro studies demonstrated that adherence to and invasion of Henle 407 cells, at 37 degrees C, by organisms grown under these optimized conditions, were significantly greater than when organisms were grown aerobically under the same conditions.

Anaerobiosis↗
PubMed1991

Shigella dysenteriae 60R strain adheres to and invades tissue culture cells in the absence of virulence plasmid.

Evidence is presented that a high level Shiga toxin-producing strain Shigella dysenteriae 60R adheres to and invades the epithelial cell lines Hct8 and Henle 407. The invasive phenotype of S. dysenteriae 60R differs in four ways from the heretofore studied invasive Shigella phenotypes. First, S. dysenteriae 60R lacks the virulence plasmid characteristic of other invasive Shigella spp. and enteroinvasive Escherichia coli. Second, hybridization studies show that the known ipa genes are neither present in the chromosome nor in the small plasmid of 60R. Third, 60R adheres to and invades Hct8 and Henle 407 cells at 37 degrees C as well as at 30 degrees C. Fourth, the phenotype of adherence and invasion of 60R is remarkably stable, even during prolonged growth in laboratory media and storage.

Antigens, Bacterial↗
PubMed1989

Cloning and DNA sequence of plasmid determinant iss, coding for increased serum survival and surface exclusion, which has homology with lambda DNA.

Escherichia coli K12 cells carrying a cloned 1.4 kb HindIII fragment from plasmid ColV2-K94, showed increased survival in guinea pig serum. The recombinant plasmid also conferred group II surface exclusion, i.e. the cells were reduced in recipient ability towards the incoming plasmid R538drd in conjugation experiments. Southern blotting suggested homology with bacteriophage lambda DNA and to the insertion element IS2. Determination of the DNA sequence of the fragment demonstrated the presence of a truncated IS2 (165 bp), separated by 250 bp from a 900 bp stretch of homology with lambda DNA, beginning within the Rz gene and continuing in the rightward direction on the lambda map. A 97 amino acid open reading frame (ORF) adjacent to Rz and on the opposite strand, remained intact in iss, with several amino acid changes. The ORF in iss is preceded by sequences resembling prokaryotic ribosome binding sites and promoters.

Amino Acid Sequence↗
PubMed1987

The carbohydrate specificity of conglutinin and its homology to proteins in the hepatic lectin family.

Inhibition experiments with D-mannose oligosaccharides establish that conglutinin recognises terminal alpha 1----2 mannobiosyl units present in the glycopeptide of the alpha-chain of the complement component C3b. On the basis of its three domain structure and the homology of its N-terminal amino-acid sequence to that of the dog pulmonary surfactant protein, it is proposed that conglutinin is a member of the hepatic lectin family.

Animals↗
PubMed1987

Roles of IL-2 and antigen in the later stages of the primary antibody response.

Spleen cells, obtained 2-5 days after in vivo priming with sheep erythrocytes (SRBC), were cultured to determine the presence of plaque-forming cell (PFC) precursors capable of developing into mature PFC under the influence of various stimulants. Lipopolysaccharide (LPS), added together with SRBC at the initiation of a 48-hr in vitro culture, enhanced the PFC response of primed spleen cells. In vivo priming for a minimum of 3 days was required, and maximal numbers of PFC were obtained from spleen cells primed for 4 days. Depletion of T lymphocytes from Day 3-primed spleen cells abrogated LPS-mediated enhancement, and addition of concanavalin A supernatants to the T-cell depleted system restored the enhancement, suggesting that LPS action required co-operation with a product(s) of activated T cells. Addition of various interleukin-2 preparations including recombinant human IL-2 to the system restored the LPS-mediated enhancement. The response of Day 3 cells from which T cells were eliminated as vigorously as possible was similarly restored by the addition of IL-2, LPS and antigen, suggesting that IL-2 reacts directly with PFC precursors that have developed IL-2 receptors. LPS-mediated enhancement, in the presence or absence of T cells, was also markedly dependent on the presence of SRBC during in vitro culture. These data suggest that, in co-operation with IL-2 and other co-factors, antigen plays a significant role in driving the later stages of differentiation and/or division of PFC precursors to mature PFC.

Animals↗
PubMed1979

Genetic control of the murine corneal response to Pseudomonas aeruginosa.

Inbred mouse strains differ in susceptibility to intracorneal challenge with Pseudomonas aeruginosa. Genetic studies indicate that resistance to corneal infection is dominant over susceptibility and is controlled by autosomal genes, at least one of which is located outside of the H-2 locus. On the basis of genetic complementation studies, the susceptible strains BALB/c and C57BL/6 each bear one resistance gene, since the F1 hybrid (BALB/c X C57BL/6) was uniformly resistant to infection.

Animals↗
PubMed1979

Reversal by insulin of concanavalin A inhibition of myotube formation and evidence for common binding sites.

Concanavalin A (Con A) inhibits fusion of trypsin-treated myoblasts. This inhibition is reversed by the addition of supraphysiological concentrations (4 micrograms/ml) of insulin either during continuous presence in culture or by pulse additions at 36 and 48 h of culture, just before the time that cultures not treated with Con A undergo myoblast fusion. This reversal is not due to the mitogenic effects of insulin. Under reversal conditions, no specific displacement of bound [125I]iodo-Con A was detected nor did insulin stimulate metabolite uptake. Cell surface replicas of hemocyanin-tagged Con A showed that insulin reversal of the inhibition of myotube formation correlated with the alteration of Con A-binding sites from a clustered configuration present in the inhibited cells to a dispersed state correlated with normal myotube formation. Although a causal relationship has yet to be shown, the data suggest that insulin-mediated reversal of Con A inhibition of myoblast fusion may be related to the ability of insulin at supraphysiological levels to alter the translational mobility of cell surface components containing glucose and/or mannose residues capable of binding Con A. Evidence is presented which suggests that insulin and Con A share common binding sites, since in the physiological range of insulin concentrations (1 ng/ml), Con A pretreatments results in an inhibition of specific [125I]iodo-insulin binding, and antagonistic interactions of insulin and Con A on metabolite uptake and cell proliferation occur. Thus, it appears that the insulin receptors of developing skeletal muscle are glycoproteins containing glycopyranosides.

Animals↗
PubMed1977

Suppression of myoblast fusion by concanavalin A: possible involvement of membrane fluidity.

Experimental evidence is presented which is consistent with the involvement of membrane fluidity during myoblast fusion. Treatment of pretrypsinized myoblasts with tetrameric Con A, but not with the dimeric succinyl derivate, inhibits fusion. Inhibition is reversed by treatment with alpha-methyl-D-mannoside or subsequent trypsinization. No inhibition is observed when the lectin is incubated with cells at 4 degrees C unless the incubation is followed by treatment with glycogen, a multivalent Con A cross-linking agent. This effect of glycogen is reversed by subsequent treatment with alpha-amylase. Direct observation of Con A-binding site topography by transmission electron microscopy of membrane replicas of cells labelled with Con A and haemocyanin reveals that inhibition of fusion correlates with a clustered distribution of Con A-binding sites, whereas normal fusion correlates with a dispersed distribution.

Animals↗
PubMed1976

Isolation of a human lymphocyte mitogen from wheat germ with N-acetyl-D-glucosamine specificity.

A lectin, isolated from wheat germ by affinity chromatography on chitin, was mitogenic for purified human peripheral blood lymphocytes. Peak incorporation of 3H-thymidine was observed after incubation of lymphocyte cultures with wheat germ mitogen for 7 to 10 days. When lymphocytes were separated into two fractions based on their ability to form rosettes with unsensitized sheep erythrocytes, the mitogen induced a negligible proliferative response in either fraction. Mixing experiments demonstrated a strong response in the T lymphocyte fraction which required the collaboration, but not proliferation, of cells present in the nonrosetting fraction. Stimulation was specifically abolished by addition of N-acetyl-D-glucosamine at initiation of culture.

Acetylglucosamine↗
PubMed1976

The immune response to dextran in BALB/c mice. I. Modification of "thymus-independent" response by the T cell mitogen concanavalin A.

Normal and congenitally athymic mice respond to immunization with dextran, thus indicating that the response is thymus independent. Athymic mice have lower plaque-forming cell (PFC) response than normal mice 3 days after immunization, but similar PFC response as normal mice by 5 days. Seven to 10 days after immunization, athymic mice maintain a high level of PFC response whereas the PFC response declines sharply in normal mice. Concanavalin A (Con A), added at initiation of in vitro culture, enhances the response of spleen cells from primed normal mice. Treatment with methyl-alpha-D-mannoside (MAM), a competitive inhibitor of binding of Con A, immediately after addition of Con A, abolishes the enhancement. Treatment with MAM, 3 to 6 hr after Con A addition, partially blocks the enhancement. The enhancing effect is dependent on the dose of Con A and the duration of in vivo priming before in vitro culture. Con A does not enhance the in vitro PFC response of cells from primed athymic mice. Supernatants from Con A-treated non-primed normal spleen cell cultures enhance the PFC response of "nude" cells, thereby indicating that enhancement by Con A is mediated through a soluble T cell product(s) with no apparent antigen specificity. Hydroxyurea, an inhibitor of DNA synthesis, blocks the enhancement by Con A of cells primed for 1 to 3 days. Release of hydroxyurea block after 19 hr incubation does not restore the PFC response, thereby indicating the responding anti-dextran PFC precursor cells are engaging actively in DNA synthesis. PFC response of primed cells taken at the peak of anti-dextran response (day 5) shows that they are less susceptible to the hydroxyurea block. Con A enhances the PFC response of cells primed for 5 days even in the absence of DNA synthesis.

Animals↗
PubMed1975

Modification of the biological activities of concanavalin A by anti-concanavalin A.

Concanavalin A (Con A) bound to cell membrane glycoproteins, may be dissociated from the membrane receptors by competitive ligands such as alpha-methyl-D-mannoside. Addition of antibody to Con A to the system forms complexes of antibody and Con A which are still bound to the membrane receptors. Such complexes are not dissociable from the membrane by alpha-methyl-D-mannoside. Presence of the complexes on the membrane is monitored by radioactive label or by passive lysis of the cells with guinea pig complement. Antibody to Con A can completely suppress Con A mediated stimulation of lymphocytes as measured by incorporation of tritiated thymidine. However production of lymphokines involved in migration inhibition, enhancement of plaque forming cells or blastogenesis are differentially suppressed by antibody to Con A.

Animals↗