Gerardo R. Vasta

Characterization of interleukin-1 activity in tunicates☆

1. Eight North American species of tunicates were examined for the presence of interleukin-1 (IL-1) like activity. 2. The tunicates studied produce molecules with readily detectable lymphocyte activation factor (LAF) activity. 3. G50 column chromatography separated molecular species that were directly mitogenic for thymocytes (mol. wt greater than 50,000) from those that were comitogenic in an IL-1 assay (mol. wt 20,000). 4. Tunicate fractions with LAF activity induced increased vascular permeability in rabbit skin. 5. Tunicate LAF activity was neutralized by polyclonal anti-human IL-1 antisera. 6. These data further support the conclusion that IL-1 is an ancient and functionally conserved molecule.

A lectin on the hemocyte membrane of the oyster (Crassostrea virginica)

Using antisera produced against a serum lectin we have shown by employing immunocytofluorescence that hemocytes from the oyster, Crassostrea virginica, possess a lectin which is situated on the external surface of the cell membrane. The antisera block the binding of hemocyte microsomes to protease-treated vertebrate erythrocytes, thus confirming that the hemocyte membrane lectin is serologically related to the serum lectin. The major serum lectin has an apparent mass of 34,000. Flow cytometry has revealed that the distribution of the surface lectin on hemocytes represents a heterogeneous expression on a population basis, but no discrete cell subpopulations can be identified.

A cell membrane-associated lectin of the oyster hemocyte

Abstract The presence of a lectin in association with hemocytes of the American oyster, Crassostrea virginica, has been demonstrated by utilizing a microhemagglutination assay. The plasma membrane association of this lectin is shown by its copurification with the plasma membrane fraction of disrupted hemocytes, using sucrose density gradient centrifugation, and also by the binding of 125I-labeled glycoproteins to intact hemocytes at 4°C. Based upon agglutinating spcificity for a range of vertebrate erythrocytes, both untreated and enzyme-treated, along with hemagglutination-inhibition assays and crossed-absorption tests, it is apparent that there are also two serum (soluble) lectins, each having a distinct serological agglutination specificity, and that the hemocyte membrane-associated lectin has a specificity that is identical with one of these two serum lectins. It is proposed that the hemocyte membrane-associated lectin may be a true integral membrane protein, and therefore may function as a membrane receptor in nonself recognition by molluscan hemocytes.

Tunicate lectins: distribution and specificity

Abstract 1. 1. The plasma lectins of ten species of North American tunicate were studied by agglutination, crossed absorption and hemagglutination-inhibition with a panel of vertebrate erythrocytes. 2. 2. All species possessed detectable lectins. 3. 3. All but one of the species possessed multiple lectins. 4. 4. The specificities of the lectins which included d -galactose, sialic acid and complex carbohydrate structures, were not correlated with taxonomic relationships of the species studied.

Serological characterization of humoral lectins from the freshwater prawn Macrobrachiumrosenbergii

We have detected and partially characterized multiple lectins present in the serum of the freshwater prawn Macrobrachium rosenbergii. Since agglutination of erythrocytes (RBC) is not abolished by treatment with Vibrio cholerae neuraminidase (VCN), Macrobrachium shows an agglutination pattern different from that of other sialic acid-specific lectins such as Limulus polyphemus lectin. However, after absorption with primate and bird VCN-treated RBC, Macrobrachium serum exhibits high titers with untreated and pronase-treated RBC and no agglutination of VCN-treated RBC, suggesting a typical sialic acid specific lectin agglutination profile. Hemagglutination-inhibition tests indicate that sialic acid containing compounds are the best inhibitors for Macrobrachium lectins. Subterminal sugars and type of linkage are probably important for the lectin binding since bovine submaxillary mucin (containing mainly terminal NANA-alpha-2 6-GalNAc-) is a better inhibitor than fetuin (containing mainly terminal NANA-alpha-2 leads to 3-Gal-) and colominic acid (-NANA-alpha-2 leads to 8-NANA-) is a weak inhibitor.

Heterogeneous humoral and hemocyteassociated lectins with N-acylaminosugar specificities from the blue crab, Callinectes sapidus rathbun☆

Callinectes sapidus serum and hemocyte microsomal fraction agglutinated a panel of untreated and enzyme treated vertebrate erythrocytes and cultured lymphoid cell lines. Crossed absorption experiments suggested the presence of multiple specific lectins in the serum. The microsomal fraction showed a 35-fold increase in specific activity when compared to the hemocyte lysate suggesting that hemocyte lectins are membrane-associated. Agglutination by serum and hemocyte lectins was inhibited by low concentrations of N-acylamino compounds including sialic, N-acetylmuramic and N-acetylglutamic acids, GalNAc, GlcNAc, ManNAc, and glycoproteins and polysaccharides which contain these carbohydrates: bovine submaxillary mucin, human orosomucoid, porcine stomach mucin and colominic acid. Hemagglutination by lectins of both serum and hemocyte microsomal fraction required divalent cations as suggested by the reduction in hemagglutination titer in the presence of the chelators EDTA, EGTA, CDTA and citrate.

Humoral and cell membrane-associated lectins from invertebrates and lower chordates: Specificity, molecular characterization and their structural relationships with putative recognition molecules from vertebrates

Based on their carbohydrate-binding properties and their ubiquitous presence in pro- and eukaryotes, recognition functions have been hypothesized for many humoral and tissue lectins. In this review three main topics relevant to the possible biological roles and evolution of invertebrate and chordate lectins are discussed. They include the broad carbohydrate-binding spectrum of lectins from chelicerata, the distribution and specificity of cell membrane-associated lectins in mollusks and the serological and biochemical characterization of lectins from tunicates and their structural relationships with putative non-self recognition molecules from vertebrates.

Sialic acid-binding lectins in the whip scorpion (Mastigoproctus giganteus) serum

Abstract Sialic acid-specific lectins have been detected in the serum of the “whip scorpion,” Mastigoproctus giganteus . When compared to Limulus lectins, Mastigoproctus agglutination profiles for a panel of untreated and enzyme-treated vertebrate erythrocytes were almost identical except for the agglutination of nonhuman primate erythrocytes. However, both chelicerate species exhibited heterogeneous serum lectins which showed some differences in their serological reactivity. At least three distinct specific fractions could be demonstrated in Mastigoproctus serum by crossed absorption and hemagglutination-inhibition experiments. These fractions are specific for sialic acids and/or sialoconjugates but also bind substances such as N -acetylglutamic acid, N -acetylmuramic acid, chitobiose, and chitotriose. These adjunct specificities are important clues in the interpretation of the possible biological role of chelicerate lectins.

Sialic acid binding lectins in the serum of american spiders of the genus Aphonopelma

Multiple lectins were detected in the serum of three American species of the genus Aphonopelma (A. chalcodes, A. cochise and A. chiricawa) through tests of agglutination and crossed absorption with a panel of untreated and enzyme treated of vertebrate erythrocytes. Hemagglutination-inhibition experiments showed that a fraction of A. chalcodes serum lectins bind sialic acids and sialoconjugates although other related carbohydrates as N-acetyl-D-glucosamine and 2-keto-3-deoxyoctonate also are recognized. Results of these studies extend our observations about the presence of sialoconjugate binding lectins in members of the Chelicerata to species from the Order Araneae. At the present time the Chelicerata (horseshoe crabs, scorpions, whip scorpions and spiders), would constitute the only group of high taxonomic rank which includes species that exhibit certain common patterns in the specificity of their humoral lectins.

Some T-cell leukemia lines express surface markers related to a restricted set of human VH determinants

Serological studies were carried out to obtain information regarding the relationship of the VH-related determinants expressed by certain permanent in vitro T-cell leukemia lines and corresponding determinants expressed by characterized human serum immunoglobulins. A panel of conventional (goat and rabbit) antisera, produced against various Fab-related fragments of monoclonal human Waldenstrom macroglobulins and polyclonal IgG molecules, bound to certain in vitro T-cell leukemia lines, notably, 70-N2, MT-1, YT4E, and HUT78, as shown by microhemagglutination. Inhibition studies using characterized myeloma proteins to inhibit this agglutination indicated the expression of a restricted VH-related determinant by these T-cell lines. Parallel studies performed using conventional (rabbit) and murine monoclonal/hybridoma antibodies produced against the isolated 68,000-Da VH-related product synthesized by the 70-N2 line showed that the determinant expressed by this molecule was restricted in expression, comprising 2-3% of the normal, polyclonal human Fab pool, and that the determinants found on the other positive T-cell leukemias were cross-reactive rather than identical. The inhibition studies suggest that the determinant resides between residue 22 and the end of the VH region. These results further define the antigenic nature of the VH-related marker found on the surfaces of certain normal and neoplastic T-cell lines.