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The Fasciola hepatica surface and whole body lectins (FHLs) were separated partially purified and tentatively identified .They are of protein nature and of glucose ,lactose and mannose binding potentials .As well as they agglutinate sheep red corpuscles 2% suspension. FHLs are T lymphocyte mitogens ,but the whole body lectins was more potent than surface lectins .Thus ,whole body lectin was selected for further study .The whole body lectin stimulate specific antibody formation at both mucosal and systemic level ,post to an oral immunization program in rabbits .The immunodominant epitope can be of B cell or Th2 dependent nature and in combination with E. coli "O" antigen induces mucosal tolerance . E. coli induce significant leukocyte migration inhibitory factor ( LIF ) and the combination of E .coli -FHLs induced both significant macrophage migration inhibitory factor (MIF) and LIF in the immunoprimed rabbits .FHLs posses augumenting potential for the tuberculin skin DTH reaction indicating its cellular immunopotentiating ability .Bilateral humoral and cellular shared immunogenicity .Such shared character may be helpful in immunoprotection model of E.coli infection in one hand and source of confusion in immunomodiagnosts of Fasciola infection ,on the other hand. The conclusion of this study FHL functional epitope mapping ,maps the following possible epitope ,B or Th2 dependent .Th1 , Tdth and or shared immunodominant epitopes. Introduction Lectins are carbohydrate containing conjugate substances other than immunoglobulins or enzymes.Their presence are mapped into plants ,animals ,fungi and bacteria (Hirabayashi and Kasai,1997).Lectins are capable of specific recogniation of reversible binding to carbohydrate moieties of complex glycoconjugates without altering the covelent structure of any of the recognized ligands (Horak ,1996).Among the known lectins that are abiqitous in the natural biological systems ,animal lectins mediate biological processes ranging from protein folding and trafficking to the modulation of cell – cell and cell matrix interaction s (Hirabayashi and Kasai ,1997).In general ,lectins can be grouped in eight groups .Animal lectins .however ,occupied the first six are namely,Galectins,C type, selectins,collectins, annexins and the invertebrate body protection factors (Drickamer and Tylor,1998; Hirabayashi and Kasai,1997).Although lectins are considered to be ubiquitous and important molecules in organisms,but data on lectins in helminth are scarce (Horak and Vanderknaop,1997; Hirabayashi and Kasai,1997,Guteirrez,2000). These glycolprotein have been found mainly in nematodes .In trermatodes however data are even more scarce (Horak ,1996; Horak and Vanderknaop,1997).The mapped termatodes for presence of lectins were ; Schistosoma mansoni ,Tricobilharzia szidati,Diplostomum pseudospathaceum and Acanthostomum brauni (Horak ,1996) Fasciola hepatica The trematod helminth ,was wide,wide shoulders and large cephalic cone as well as branched seca .Ventral and dorsal suckers ,the ventral one located at the base of cephalic cone .The size of the fluke was of up to cm in length by 1.3 cm in width and 0.1 cm in thickness (Guteirrez ,2000),studies have been shown ;screted thioredoxin peroxidase product and secretory excretory (Donnely et al .; 2005) tegument (Hillyer et al .; 1980) Thus the aim of the present work were to 1studies on Fasciola hepatica lectin ,2study lectin immunogenicity, modulating ,and their allerginicity effect, 3study their shared allerginicity and immunogenicity with E.coli AL-Qadisiya Journal of Vet.Med.Sci. Vol./9 No./1 2010  72 Materials and methods ILectins & antigens I-1 Helminth : Faschiola hepatica was identified I2 Escherichia coli The isolate was Gram negative facultative anaerobic rod. lactose fermenting. catalase positive,oxidase negative ,produced metallic sheen on eosin methylene blue medium by conventional and Api 20 methods (Macffadin ,2000). such characters are consistent with E .coli. I-3lectin: Faschiola hepatica was collected from the infected liver tissue of the sheep brought from the Babylon slugther house during the period (March– June2008).Rinsed three times with sterile saline,then each helminth was sliced then mortared with 10 ml sterile saline , the crud mortared materials was collected into centrifuge tube and centrifuged at 5000 rpm for 15 min .supernate was saved into sterile plane tube,then equal volume of poly ethylene glycol (PEG) 6%(Mw.6000) was added and mixed followed by 1 hr standing at 4 C ◦ . The supernatant PEG mixture was centrifuged at 5000 rpm for 15 min .Pellet was reconstuted with saline to 1 ml (this solution was ready to use as whole body. Lectin surface of flukes were scraped by scalpel in 5 ml saline per fluke and processed as above to prepare surface lectins)as the concept of mucosal scrapping (Shnawa and Abd,2005)and tegument antigen of F .hepatica(Hillyer , 1980). I-4 lectin characterization I-4 -1 ; sugar binding ability : Direct microhaemagglutination between tanned ovine erythrocytes with lectin and three types of sugar . Erythrocytes were tittered against serial twofold dilution of membrane filter sugar stock of 10% of manose ,glucose and lactose.(Garveyetal.,1977). I-4-2:Lectin skin test in chicken Sixty bird of one week age were used in this test,0.1 ml of lectin( surface or whole lectin) were injected intradermaly (Smit and Willams ,1999)in the total of three bird for each lectin solution. II-laboratory animals II-1Animals Rabbits of (1-1.5)Kg body weight were selected as test experimental animals .They were brought from local market and of local breed (Orcyctalagus cuniculus) among which 12 were checked and found to be free of pathogenic agents and grouped into four groups each of three and kept at libitum. II-2 : Dose preparation Lectin dose was determined by using Biurete method (Bishop et al .,1985) as 3 mg/ml of lectin was used.E.coli (heat killed antigen 10 IU) was prepared according to (Garvey et al,1977). IIIImmunization program Table (1) The animal groups were displayed in the following items Site of administration Dose Group Oral doses weekly ( for four week ) 5 ml of saline ISaline control Oral doses weekly ( for four week ) 3mg /ml (1.5 ml completed to 5 ml for each animal ) IIFasciola hepatica lectin Oral doses weekly (for four week) 10 Iu(1.5 ml completed to 5 ml for each animal l) IIIE.coli Oral doses weekly (for four week) Volume to volume of these antigens (1.5 ml completed to 5 ml for each animal l) IV-Fasciola hepatica lectin + E.coli antigen On the fifth week all groups were left for one week then were ready for investigation IV -Immune sera and immunoprimed cells. Macrophages were stimulated by using casein digest (1.2g for 100ml D. W. and was sterilized by autoclave) 5ml of casein digest was injected in peritoneal cavity of rabbits (Bloom and Bennett, 1966). This AL-Qadisiya Journal of Vet.Med.Sci. Vol./9 No./1 2010  72 stimulation performed in all groups of animals and after 72hrs the peritoneal fluid was collected and used in macrophage migration inhibitory study rabbit spun sera were made (Garvey et al ,1977). V-Immune function tests Tube agglutination ,microhaemagglutination ,MIF ,LIF,and skin DTH tests were done as in (Garvey et al., 1977; Soborg ,1968 ,Burel,1979).

maǧallaẗ al-qādisiyyaẗ li-ʻulūm al-ṭibb al-bayṭarī

The immunogenicity,allerginicity and immunomodulating potentials as well as shared antigenicity of Fasciola hepatica (whole body lectin FHL) with E .coli in rabbit