Bacteria isolated from the gut of different developmental stages of Phlebotomus duboscqi Neveu-Lemaire, 1906 belonged almost all to aerobic or facultatively anaerobic gram-negative rods. In females, the highest bacterial counts were observed two days after bloodfeeding; seven days after bloodfeeding the bacterial counts returned to pre-feeding levels. Most isolates were identified phenotypically as Ochrobactrum sp. The distinctiveness and homogeneity of the phenotypic and genotypic characteristics of Ochrobactrum isolates indicated that they belonged to a single strain (designated AK). This strain was acquired by larvae from food and passaged transtadially: it was isolated from the guts of fourth-instar larvae shortly before pupation, from pupae as well from newly emerged females. Most other bacteria found in females were acquired from the sugar solution fed to adults. To determine if the midgut lectin activity may serve as antibacterial agent females were membrane-fed on blood with addition of inhibitory carbohydrates. No significant differences in bacterial infections were found between experimental and control groups and we suppose that the lectin activity has no effect on gram-negative bacteria present in sandfly gut.
Physico-chemical properties and carbohydrate-binding specificity of hemagglutination activity (HA) were compared in tissue lysates and haemolymph of unfed and bloodied females of five sandfly species. Sandfly gut lectins were found to be heat-labile, sensitive to dithiotreitol treatment, freezing/thawing procedures and were affected by divalent cations. The pH optimum of HA ranged between 7.0-7.5. Specificity of gut HA of all species studied was directed towards aminosugars and some glycoconjugates, mainly lipopolysaccharide from Escherichia coli K-235, heparin and fetuin. Gut HA of Phlebotomus papatasi (Scopoli, 1786) was strongly inhibited by lipophosphoglycan (LPG) from Leishmania major promastigotes. In females, that took blood, the HA was higher but the carbohydrate-binding specificity remained the same; this suggests that the same lectin molecule was present, at different levels, both in unfed and fed flies. High HA was found in ovaries of fed females of Lutzomyia longipalpis (Lutz et Nieva, 1912), P. papatasi and P. duhoscqi Neveu-Lemaire, 1906. In P. papatasi and P. duboscqi the HA was present also in the haemolymph and head lysates of both fed and unfed females. Carbohydrate-binding specificity of HA present in these tissues was similar with the gut lectin.
F.xcept other functions, surface saccharide residues on trematode larvae are supposed either to be the targets of the intermediate (molluscan) and final host immune systems, or to represent candidates for molecular mimicry. Therefore, changes in surface saccharide patterns during the development of the avian schistosome Trichobilharzia szidati were characterized. Whole parasite larval stages and their tissue sections were examined using FITC-conjugated lectins. Marked surface differences were found among larval stages (miracidia, mother sporocysts, daughter sporocysts, cercariae, schistosomula). Staining by some lectins reflected known ultrastructural changes of the outer tegument. Reaction of lectins with cercarial embryos was almost negative. In case of other developmental stages, binding of at least one member from each carbohydrate-specificity group of lectins (Man/Glc-, GIcNAc-, Gal/GalNAc- and Fuc-specific) occurred. One exception is represented by mother and daughter sporocysts which practically failed to react with Fuc-specific lectins. Besides other lectins which recognized larval surfaces, a-L-fucose-specific lectins (LTA, UEA-I) and (GlcNAcfll —>4)„-spccific WGA bound very strong to certain stages. The comparison of mature intrasporocystic cercariae with those emerged from snails brought the indication that some snail glycosylated molecules adhere to the surface of schistosome larvae or that emerged cercariae express some new carbohydrate epitopes under changed environmental conditions. The result partially supports the theory of parasite mimicry/masking strategies and immune evasion in the host.
The glycophenotyping of mammalian cells with plant lectins maps aspects of the glycomic profile and disease-associated alterations. A salient step toward delineating their functional dimension is the detection of endogenous lectins. They can translate sugar-encoded changes into cellular responses. Among them, the members of the lectin family of galectins are emerging regulators of cell adhesion, migration and proliferation. Focusing on galectins-1, -3 and -7, we addressed the issue whether their expression is regulated during wound healing in porcine skin as model. A conspicuous upregulation is detected for galectin-1 in the dermis and a neoexpression in the epidermis, where an increased level of galectin-7 was also found. Applying biotinylated tissue lectins as probes, the signal intensities for accessible binding sites decreased, intimating an interaction of the cell lectin with reactive sites. In contrast, galectin-3 parameters remained rather constant. Of note, epidermal cells in culture also showed an increase in expression/presence of galectin-1, measured on the levels of mRNA and protein, in this case by Western blotting and quantitative immunocytochemistry. Used as matrix, galectin-1 conferred resistance to trypsin treatment to attached human keratinocytes and reduced migration into scratch-wound areas in vitro. This report thus presents new information on endogenous lectins in wound healing and differential regulation among the three tested cases., J. Klíma ... [et al.]., and Obsahuje seznam literatury
Transgenic tobacco plants expressing the Allium porrum L. (leek) agglutinin (called APA) under the control of the 35S constitutive promoter were tested for their insecticidal activity against the cotton leafworm, Spodoptera littoralis (Lepidoptera: Noctuidae). Southern blot and PCR analysis confirmed that the APA gene was integrated into the plant genome. Northern and Western blots as well as semi-quantitative agglutination assays revealed lectin expression at various levels in the transgenic lines. Biochemical analyses indicated that the recombinant APA has the same molecular structure as the native lectin. Native and recombinant lectin have the same N-terminal amino acid sequence. Bioassays using detached leaves from transgenic tobacco plants demonstrated that the ectopically expressed APA significantly (p < 0.05) reduced the weight gain of 2nd-instar larvae of S. littoralis. This inhibitory effect was observed throughout the experiment and on day 11 the caterpillars fed on transgenic tobacco leaves were 25-30% lighter in weight than the control caterpillars fed on wild type plants. In addition the lectin retarded the development of the larvae and metamorphosis, reduced pupal weight and increased mortality rate. These findings suggest that APA is a suitable insect resistance protein for integrating into plant genomes for controlling S. littoralis.
Using degenerative primers designed on the basis of known sequences of lectin genes from different sources a fragment of genomic DNA of Borrelia burgdorferi ( strain B31) that contained a lectin-like sequence was isolated, cloned and sequenced. The presence of an open reading frame of 268 amino acids (position 1501-2304 bp) and the computer analysis of the predicted amino acid sequence showed 37% of identity and 75% of homology over region of 25 amino acids with the legume lectin proteins, including erythroagglutinating phytohcmagglutinin (РНЛ-Е) and leucoagglutinating phylohemagglutinin (PHA-L). The further analysis of the predicted amino acid sequence showed the presence of another two domains (positions 198-211 and 215-226 aa) consisting of the characteristic conserved sequence motifs for legume lectin proteins. Hemagglutinating activity was detected in lysate of В burgdorferi (strain B31) spirochete and the affinity to fetuin was determined in a hemagglutination inhibition test. Hemagglutinating activity was also found in a crude lysate of the recombinant clones carrying the fragment of B. burgdorferi genomic DNA. The inhibition of agglutinating activity by fetuin, D-galactosamine and D-mannosamine was determined using the standard procedure of hemagglutination inhibition test with native rabbit red blood cells (RBC).
Lectins and their glycosylated receptors in a system of the tick-transmitted pathogen are the addressed topics which the minireview is dealing with. They participate in the reciprocal protein-saccharide interactions in the transmission of the causative agents of the tick-borne encephalitis and Lyme borreliosis by the ticks. Functional significance of the tick tissue specific lectins as well the lectins/aggulutinis of the transmitted pathogens in molecular ecology of the tick borne diseases has been shown.
Lectins as carbohydrate recognition proteins other than enzymes or immunoglobulins play important roles in living systems, e.g., in celi celi recognition. They are considered to be involved in snail-trematode immune interactions, i.e., in a system where antibodies are lacking and lectins might at least partially substitute immunoglobulin functions. From the snail side, lectins can be located on haemocyte surfaces as receptors for foreignness and they can be found freely in plasma. The latter can function as agglutinins/opsonins helping in the recognition of parasites by haemocytes. They may also link immune cells and pathogens by recognition of surface carbohydrates on both. Lectins of parasite origin could also be involved in snail-trematode interactions. They might function as trematode surface receptors for snail glycoconjugates in parasite masking strategies. Functions other than the involvement in the snail's immune response or the parasite’s evasion strategies might be fulfilled by lectins as well. Among these may be host-finding, penetration, orientation in the host, nutrition. It cannot be omitted that lectin-saccharide reactions represent only a part of the snail-trematode interactions and thus, results obtained from lectin experiments are a rough simplification of the actual, very complicated situation. An array of immune and other reactions comprised of yet unknown bioactive molecules certainly exists in snails and, on the other hand, trematode mechanisms to escape or otherwise interact with these, might be involved at the same time. But we can certainly conclude that a more complete view of the complex snail-trematode interactions also necessitates a more profound knowledge of the identity and functioning of lectins and their ligands, in host and parasite.
Lectin activities were studied in five different species of tsetse flies. Different native or enzymatically treated human or animal red blood cells were used to detect hemagglutination activity in midgut extracts. Two inducible lectin systems in the midgut of C. tachinoides were distinguished.