Natural Agglutinin Research Articles

Effect of Some Water Supplements on the Performance and Immune System of Chronically Heat-Stressed Broiler Chicks

Effects of water supplements, sodium bicarbonate (NaHCO ), potassium chloride (KCl) and acetic 3 acid were evaluated on the performance and immune system of chronically heat-stressed broiler chicks. Two hundred day old broiler chicks were allotted to one of the four groups (n = 50); 1) control (no supplements), 2) NaHC0 0.5%, 3) KCl, 0.15% and 4) acetic acid, 1.5 mL/Liter. All chicks were kept in a controlled 3, environmental chamber maintained at 33±2 C from day one to 6 weeks (wks) of age. Significantly higher o weight gains coincided with decreased feed conversion ratios for all the supplemented groups as compared to control group at 2, 4 and 6 wks of age. Bursal index, percentage weights of thymus and spleen in relation to body weight and natural agglutinin levels, an indicator of humoral immunity were higher but the heterophil:lymphocyte ratio, an indicator of stress was lower for the supplemented groups as compared to control group. Total aerobic spore formers and Enterobactraecae counts in the intestinal swab samples were higher in control group than supplemented groups. Intestinal pH was lower at 2, 4 and 6 weeks of age but water consumption at 5 and 6 wks of age tended to be higher in acetic acid treated group as compared to other groups. Overall, the results indicated significant improvement in the performance and immune response of chronically heat-stressed broiler chicks given the water supplements, acetic acid being slightly superior to NaHCO and KCl. 3

Isolation and characterization of an acetyl group-recognizing agglutinin from the serum of the Indian white shrimp Fenneropenaeus indicus

A natural agglutinin from the serum of the Indian white shrimp Fenneropenaeus ( Penaeus) indicus was purified to electrophoretic homogeneity by a single-step affinity chromatography on N-acetylglucosamine-Sepharose 6B. The expression of hemagglutinating (HA) activity of F. indicus agglutinin (FIA) was independent of the presence of divalent cations and insensitive to their chelators. FIA gave a single symmetrical peak in its native form with a molecular mass estimate of 200 kDa on gel filtration in HPLC, and SDS-PAGE under reducing conditions revealed that it is a homo-oligomer of a 27-kDa subunit protein. The pattern of reactivity of FIA against anti-FIA rabbit serum in immunodiffusion and immunoelectrophoretic analysis provided additional evidence for its purity and homogeneity. HA-inhibition studies documented exclusive specificity of FIA for acetyl groups in carbohydrates independently of the presence of these groups at the C-2 or C-5 position and its stereochemical arrangement in the axial or equatorial orientation. The unique ability of FIA to recognize acetyl groups was also explicitly demonstrated with sialo- and asialo-glycoproteins. Strikingly, FIA also interacted equally with amino acids and chemicals containing acetyl groups, thereby unambiguously demonstrating the exquisite specificity of FIA for an acetyl group, irrespective of the presence of this group in carbohydrate or noncarbohydrate ligands. The susceptibility of HA activity of FIA to inhibition by lipopolysaccharides from diverse gram-negative bacteria as well as its ability to selectively agglutinate several bacterial species isolated from infected shrimps implicate a potential role of this humoral agglutinin of F. indicus in the host immunodefense reactions against microbial invaders.

Expression of Functional Region of Peanut (Arachis hypogaea) Agglutinin Gene as an Active Lectin in Insect Cells

Natural peanut agglutinin (PNA) gene is expressed with a signal sequence of 23 amino acids and a C terminal peptide of 14 amino acids. Functionally active recombinant PNA having apparent subunit molecular weight of 29kD was obtained when expressed without signal peptide and non-essential C terminal peptide sequences in insect cells. Expression in insect cells (Sf9) was driven by a 129bp Spodoptera litura nucleopolyhedrosis virus (S/NPV) sequence containing its polyhedrin promoter.

Purification and Characterization of a Natural Agglutinin in the Hemolymph of the Prawn Penaeus indicus H. Milne Edwards

A natural agglutinin in the hemolymph of the marine prawn Penaeus indicus was isolated by gel filtration chromatography, purified using polyacrylamide gel electrophoresis, and characterized. Prawn agglutinin has a native molecular mass of 181 kDa and consists of two monomeric units (97 and 84 kDa), maintains some agglutinating activity over a wide pH range (7–9), and is inactivated at 85°C. The agglutinin was denatured upon mixing with trichloroacetic acid, phenol, chloroform, and 45% ammonium sulfate. It was also sensitive to trypsin digestion. The results indicate that prawn agglutinin is proteinaceous in nature, with agglutinating, hemolytic, and antibacterial properties against marine bacteria and erythrocytes with carbohydrate binding sites.

Biological properties of a natural agglutinin in the hemolymph of Indian white prawn, Penaeus indicus H. Milne Edwards

Biological properties of a natural agglutinin in the hemolymph of Penaeus indicus were established. Agglutination was observed against different strains of bacteria and erythrocytes and strong agglutination against a Vibrio sp. isolated from a diseased prawn gut (titer 256). Minimum concentration of agglutinin required for agglutination was 0.8 μg/ml. The agglutination reaction was affected by various sugars and ions. Agglutination and cross-linking process were inhibited by simple sugars like xylose, raffinose and trehalose, but stimulated by maltose, lactose, glucose, mannose and cellobiose. Ca 2+ and Mg 2+ ion supplementations were not essential for agglutination. Natural agglutinin was found to be a specific recognition molecule, which functions as defense protein with simultaneous agglutinating, hemolytic and antibacterial properties. Monitoring hemolymph levels of agglutinin may be helpful in disease detection.

Purification and characterization of a natural agglutinin from the serum of the hermit crab Diogenes affinis

A natural agglutinin from the serum of the hermit crab Diogenes affinis was purified to homogeneity by a single-step affinity chromatography using N-acetylglucosamine-coupled Sepharose 6B. The purified serum agglutinin (PSA) showed a strong affinity for rat RBC, and its hemagglutinating (HA) activity was specifically dependent on Ca 2+ and reversibly sensitive to EDTA. PSA in active form has a molecular mass estimate of 185 kDa and is composed of four non-identical subunits (51, 49, 42 and 39 kDa) cross-linked by interchain disulfide bonds. The homogeneity of PSA was corroborated by immunodiffusion and immunoelectrophoretic analyses using rabbit antiserum raised against the agglutinin. The antibodies in this antiserum appear to be specific for RBC-binding sites of the agglutinin molecules as revealed by the ability of the antiserum to neutralize HA activities of both whole serum and PSA of D. affinis. In HA-inhibition assays performed with several carbohydrates and glycoproteins, PSA showed a distinct and unique specificity for acetyl group in carbohydrates independently of the presence of this group on C-2 or C-5 and its stereochemical arrangement in the axial or equatorial orientation. Besides, this agglutinin appears to recognize the terminal N- and O- acetyl groups in the oligosaccharide chain of glycoconjugates. The HA activity of D. affinis agglutinin was also susceptible to inhibition by lipopolysaccharides from diverse Gram-negative bacteria, which might indicate a significant in vivo role of this humoral agglutinin in the host immune response against bacterial infections.

Characterization of a natural agglutinin present in the body surface mucus of the land slug

Characterization of a natural agglutinin present in the body surface mucus of the land slug

The Effect of the Sex-Linked Dwarfing Gene, dw, on the Immune Responses of Broiler Breeder Chickens

Two generations of a normal (AG) strain and a dwarf (D2) strain and one generation of an F1 (AG × D2) cross-strain of broiler breeder chickens were evaluated to determine if the sex-linked dwarfing gene, dw, affects specific immune competence parameters. The lymphocyte blast transformation response using the mitogen phytohemagglutinin-M was significantly stronger among second generation dwarfs compared to either normal or cross-strain birds.A delayed-type hypersensitivity response was induced by intradermal injection of tuberculin into the wattles of sensitized birds. Dwarf and cross-strain birds had significantly larger wattle swelling indices than normal strain birds. Dwarf males and cross-strain females had significantly stronger tuberculin responses than normal strain birds of either sex at 24 and 48 hr postchallenge. In addition, the tuberculin response of normal strain birds decreased significantly between 24 and 48 hr following antigenic challenge, whereas the tuberculin responses of cross-strain birds increased and dwarf birds remained the same.The dwarfing gene had no significant effect on natural agglutinin titers to 1% rabbit red blood cell (RRBC) in either the parent or second generation. There was also no significant difference in the response of parent generation birds to sheep red blood cells (SRBC) following intravenous sensitization with 1 ml of a 1% SRBC suspension. However, second generation dwarfs had significantly lower SRBC titers than normal or cross-strain birds.Assays of cellular and humoral immunity indicated that sex-linked dwarfs may have more competent T-cell subpopulations and weaker B-cell reactivity compared to normal strain birds.

Resistance of Salmonids to Aeromonas salmonicida: Relation between Agglutinins and Neutralizing Activities

Abstract Serum of furunculosis-resistant rainbow trout Salmo gairdneri, when injected into brook trout Salvelinus fontinalis, increased resistance of the brook trout to virulent challenges of Aeromonas salmonicida, the furunculosis pathogen. Serum from susceptible Atlantic salmon Salmo salar did not confer passive resistance on brook trout although natural agglutinin titers in Atlantic salmon serum were equal to those in rainbow trout. Further analyses indicated that rainbow trout serum neutralized toxic components produced by A. salmonicida, but that Atlantic salmon serum did not. When 11 strains of rainbow trout were challenged with 109 virulent cells of A. salmonicida, a range of mortality from 0 to 83% resulted. Although no correlation was observed between the natural serum agglutinin level of a particular strain and its relative resistance to experimental pathogen challenge, an inverse relation was apparent between cytotoxic serum-neutralization activity and resistance to furunculosis.

Age dependent differential effects of cyclophosphamide on natural antibody levels in chickens

Administration of 5 mg of cyclophosphamide per day for 4 days into neonatal chicks resulted in suppression of natural agglutinin titers up to at least 12 weeks of age. When the administration of cyclophosphamide was delayed until 7 days post-hatching suppression was found up to 8 weeks but titers rose to normal by 10 weeks after initial injections. Injections of cyclophosphamide after 21 days of age did not suppress natural agglutinin titers unless the drug was administered daily for 2 or more weeks.

マガキ組織における凝集素と殺菌素の活性

High natural agglutinin activities against sheep erythrocytes were observed in the extracts of the gill (mean titer: 141) and digestive diverticula (DD, mean titer: 128) from adult oyster, Crassostrea gigas, under hanging culture. Lower but considerable activities were found in the extracts of the mantle and adductor muscle. The natural bactericidal activity against HS 29 strain (Arthrobacter sp.) was conspicuously high in the DD extracts (mean titer: 231) and detectable also in the mantle and gill. The natural agglutinin activites of DD and gill declined sharply with progressive develop ment of the gonad and was minima just before and during the spawning, However, these activities showed a rapid rising trend after the spawning season. The agglutinin activities of DD and gill were enhanced by the injection of 0.1ml/oyster of 10% sheep erythrocyte suspension into the connective tissue surrounding DD. The bactericidal activity of DD was observed to increase after the injection of formalin-killed HS 29 strain vaccine which was given into the connective tissue surrounding DD (dose: 1.8×108 cells/g fresh soft-body wt.) or the adductor muscle (8.3×105 cells/g fresh soft-body wt.).

軟体動物の生体防御機構 Ｉ ホタテガイ組織における殺菌素と凝集素の活性

The extracts of the digestive diverticula (DD) from the normal two-year-old scallop, Patinopecten yessoensis, in hanging cultures were found to possess a low level of natural bactericidal activeity (bactericidin titer: 5 to 10) against HS 29 strain, Arthrobacter sp. No activity, however, was observed in the extracts of the kidney, mantle and gill. The natural agglutinin activity against sheep erythrocytes was extremely high (agglutinin titer: 128) in the DD extracts, as compared with that in the extracts of the gonad, giil and adductor muscle. The natural bactericidal activity of DD which was maximal from March to May declined with the rise of water temperature in the vicinity where scallops were cultivated, and was minimal in hot summer. It exhibited a recovery trend after summer and attained its maximum level in December. The seasonal changes paralleled those in the natural agglutinin activity of DD. The baseline killing power of DD for HS 29 strain was enhanced conspicuously by admi-nistration of HS 29 strain vaccine (formalin-killed, ca. 1×107cells/ml). The duration of the enhancement was approximately 5 days at 10°C.

Etude thermodynamique des allohémagglutinines du système ABO

Many thermodynamic assays on natural allohemagglutinins ABO were undertaken by different authors. Our experiment concerns anti-A and anti-B. The allohemagglutinins level was determined by the Probit Method, using bromelin red blood cells and methyl cellulose, with an autoanalyser. This thermodynamic assay was carried out in almost the same conditions as those used by Wurmser. The data reveal an intraphenotypic (anti-A) or intra genotypic heterogeneity (anti-B) in the repartition of affinities. Average enthalpy changes are, in this methodology, interphenotypic (anti-A) and intergenotypic (anti-B) features. For Wurmser every genotype has a typical feature: so this author concludes a natural agglutinin homogeneity. Heterogeneity showed by our experimentation seems bound to the specific IgM and IgG repartition of each serum. Specific anti-A or anti-B IgM are more exothermic than specific IgG. The apparent disagreement must be interpreted owing to the fact that in N4 Wurmser's method, the only agglutinins determined are specific IgM.

A Natural Anti-A Agglutinin in the Serum of the Brown Trout (Salmo trutta)

A Natural Anti-A Agglutinin in the Serum of the Brown Trout (Salmo trutta)

Protochordate immunity: I. Primary immune response of the tunicate Ciona intestinalis to vertebrate erythrocytes

Serum from the tunicate Ciona intestinalis contains a low titer, natural agglutinin for a variety of vertebrate erythrocytes. Primary injections of 8.5 × 10 3 human erythrocytes or 7.0 × 10 2 duck erythrocytes into the tunic tissues or perivisceral cavity were agglutinated and then cleared by phagocytosis within 24 hr. Higher concentrations of human or duck erythrocytes injected into the tunic tissues were encapsulated. Concomitant with clearance or encapsulation, serum agglutinin levels decreased and then returned to normal within 24 hr. Tunicates are classified taxonomically with the vertebrates in the phylum Chordata. Since phagocytosis and encapsulation reactions are typical of most invertebrates, the results of this study suggest that tunicate internal defense mechanisms are more closely allied to invertebrates than to vertebrates.

Partial characterization of a natural agglutinin in the hemolymph of the lobster, Homarus americanus

Partial characterization of a natural agglutinin in the hemolymph of the lobster showed: (1) Calcium ion stabilized the agglutinin against inactivation at moderate (35–45°C) temperatures as well as against a reversible increase in activity at nonphysiological pH levels ( pH 6 and 9). (2) Serum frozen at −20°C in the absence of calcium showed an increase in agglutinin activity. (3) Heat inactivation curves indicated a single component for the agglutinin. (4) Heat inactivation destroyed the ability of the agglutinin to be adsorbed to the erythrocyte antigen. (5) The chemical nature of the antigenic reactive site differed with the species of erythrocyte tested; D-glucosamine blocked or inhibited adsorption of the agglutinin to erythrocytes of most species tested. (6) In vivo the agglutinin is located in the hemocytes.

Characterization of a natural agglutinin present in the hemolymph of the California sea hare, Aplysia californica

A substance in the serum of the marine gastropod, Aplysia californica, capable of agglutinating marine bacteria and vertebrate red blood cells was subjected to physicochemical analysis in order to ascertain its possible nature. Our studies indicate that agglutinating activity is due to a heterogeneous group of high molecular weight molecules with two activity peaks exhibiting sedimentation coefficients centering ∼18.5 S and ∼31.0 S. This material has a protein component associated with the active site since it is sensitive to heat, pH extremes, and extraction with 2-mercaptoethanol, phenol, chloroform, and trichloracetic acid. Its physicochemical characteristics are different from other known invertebrate agglutinating substances and from classical vertebrate antibody.

Humoral immune factors in the paddlefish, Polyodon spathula

1.1. Natural agglutinin titers to Salmonella typhosa “O” (STO), human group A erythrocytes (HuA) and sheep erythrocytes (SRBC) were studied in twenty-three paddlefish.2.2. Natural agglutinin titers to SRBC were low, with high titers observed to STO and HuA. Cold agglutinin titers (4°C) to STO markedly exceeded titers recorded at room temperature.3.3. Serum immunoglobulins were shown to account for over forty per cent of the total serum proteins in this species.

A Study of Arbovirus Non-Specific Inhibitors and Natural Agglutinins in Bovine Serum

Summary In bovine serum, the natural agglutinin for goose red blood cells was shown to be an IgM antibody which differed from immune IgM antibody in its partial inactivation during acetone-ether extraction. Agglutinins active at pH 6.2 were not readily adsorbed at pH 9, they were partially eluted from the red cells at room temperature. Both natural and immune IgM antibodies were inactivated by 64° C. for 30 min., although “7S” antibodies were not affected. Non-specific inhibitors of arbovirus haemagglutination in bovine serum had properties suggesting they were β-lipoprotein. Both low density or less) and higher density (between l-01 and 1-04) fractions caused inhibition. Kaolin adsorption of bovine serum did not remove all non-specific inhibitors. Acetone-ether extraction was the preferred technique, because it removed non-specific inhibitors more efficiently and because it caused partial inactivation of natural agglutinins without affecting immune IgM antibody.

Serological Aspects of the Suppression of Normal Bactericidal Action of Rabbit Serum Following Whole Body X-Irradiation

Summary The loss or suppression of the normal bactericidal activity of rabbit serum following 650 to 725 r of whole body x-irradiation was not demonstrated to be due to a substance in the sera of irradiated rabbits which inhibits normal bactericidal activity. No decrease in the bactericidal action of normal serum was observed following the addition of serum from x-irradiated rabbits to normal serum. Titrations were carried out with sera from nonimmunized x-irradiated and control rabbits to determine natural agglutinin levels. Suspensions of sheep red blood cells and Bacillus subtilis were employed as antigens. The amount of circulating natural agglutinin was not significantly altered in the sera of x-irradiated rabbits with suppressed bactericidal activity. Complement concentrations determined at a time when the sera from irradiated rabbits showed suppressed bactericidal activity were of the same magnitude in x-irradiated and control rabbits. Addition of either E or M fractions of guinea pig complement failed to restore the bactericidal activity of sera from x-irradiated rabbits.