Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [88]-114).

Thesis--University of Copenhagen. / Summary in Danish. Bibliography: p. 71-80.

MALA-2 (Murine Lymphocyte Activated Antigen-2) is a murine cell surface antigen that is detected at high concentration on activated, proliferating lymphocytes, but only weakly on resting lymphocytes. It is thought to play an important role in lymphocyte activation since the rat monoclonal antibody YN1/1.7.4 which recognizes MALA-2 is capable of inhibiting the mixed lymphocyte reaction. Considering the central role of lymphocyte activation to the generation and maintenance of the immune response, I undertook the purification and biochemical characterization of MALA-2. In these studies, MALA-2 was isolated and purified to homogeneity using immobilized YN1/1.7.4 monoclonal antibody and sodium docecylsulphate-polyacrylamide gel electrophoresis. Biochemical characterization studies revealed that MALA-2 is a Mr 95-100 kD glycoprotein containing a protein backbone of approximately 66 kD, and N-linked carbohydrate chains amounting to a Mr of approximately 35 kD. Two dimensional gel electrophoresis suggested that MALA-2 has an isoelectric point of 4.9. Although it was previously suspected that MALA-2 might be associated with the transferrin receptor on the cell surface, this was shown not to be the case on NS-1 cells. Additionally, ³²P-orthophosphate labelling of MALA-2 on NS-1 or MBL-2 cells could not be detected. Finally, the partial amino acid sequence of MALA-2 was determined by sequencing trypsin-generated peptides from purified MALA-2. Computer-assisted homology comparisons of the MALA-2 partial amino acid sequences with other known sequences showed that MALA-2 shared its most consistent homology with a class of proteins known as the immunoglobulin superfamily. Subsequent to this study, the partial amino acid sequences obtained within this study were used to construct oligonucleotide probes. These probes were used for the screening of cDNA libraries, facilitating the successful cloning of the MALA-2 gene. This, in turn, resulted in the identification of MALA-2 as the murine counterpart of the human ICAM-1 molecule, a protein known to play a significant role in intercellular adhesion and lymphocyte activation within the immune system. Significantly, results obtained from the biochemical characterization of MALA-2 carried out in this thesis have been confirmed by the subsequent nucleotide sequence data from the cloning of MALA-2. / Medicine, Faculty of / Pathology and Laboratory Medicine, Department of / Graduate

No description available.

No description available.

Tumour necrosis factor receptor (TNFR) family members are widely expressed in cells of the immune system and are essential for the development and function of many immune cell types. The TNFR associated factor (TRAF) family are signal adapter molecules that are recruited to various members of the TNFR family and are important for the transduction of signals downstream of these receptors. In these studies, gene targeting was used to create a mouse capable of undergoing conditional inactivation of the Traf3 gene. These mice were studied alongside previously generated mice that were similarly genetically modified with respect to the Traf2 gene. The mice produced lacked expression of either TRAF2 or TRAF3 in either B or T cells. In resting B cells TRAF2 and TRAF3 were shown to cooperate to negatively regulate the signalling of B cell activating factor of the TNF family (BAFF) and its receptor (BAFF-R), the TNF ligand and receptor pair that provide obligate survival signals to B cells. Thus, TRAF2- and TRAF3-deficient B cells displayed hyperactive NF-kB2 signalling, an increased ability to survive, and almost identical gene expression profiles, emphasizing the cooperative nature of their roles in resting B cells. Importantly, the survival of these B cells was completely independent of BAFF. In normal B cells, BAFF signalling was shown to lift the negative regulation of survival mediated by TRAF2 and TRAF3, by depleting TRAF3 from the cell. This process was shown to require TRAF2. T cells deficient in TRAF2 or TRAF3 also displayed hyperactivity of the NF-kB2 pathway, but they did not accumulate in vivo or show extended survival in vitro. Mice lacking TRAF2 or TRAF3 in their T cells did however display a decrease in the number of memory phenotype CD8+ T cells. These studies indicate that some of the roles of TRAF2 and TRAF3 are common between B and T cells. However, the consequences of loss of TRAF2 or TRAF3 in B and T cells differs considerably, presumably due to the differential TNFR expression and usage by each cell type.

Tumour necrosis factor receptor (TNFR) family members are widely expressed in cells of the immune system and are essential for the development and function of many immune cell types. The TNFR associated factor (TRAF) family are signal adapter molecules that are recruited to various members of the TNFR family and are important for the transduction of signals downstream of these receptors. In these studies, gene targeting was used to create a mouse capable of undergoing conditional inactivation of the Traf3 gene. These mice were studied alongside previously generated mice that were similarly genetically modified with respect to the Traf2 gene. The mice produced lacked expression of either TRAF2 or TRAF3 in either B or T cells. In resting B cells TRAF2 and TRAF3 were shown to cooperate to negatively regulate the signalling of B cell activating factor of the TNF family (BAFF) and its receptor (BAFF-R), the TNF ligand and receptor pair that provide obligate survival signals to B cells. Thus, TRAF2- and TRAF3-deficient B cells displayed hyperactive NF-kB2 signalling, an increased ability to survive, and almost identical gene expression profiles, emphasizing the cooperative nature of their roles in resting B cells. Importantly, the survival of these B cells was completely independent of BAFF. In normal B cells, BAFF signalling was shown to lift the negative regulation of survival mediated by TRAF2 and TRAF3, by depleting TRAF3 from the cell. This process was shown to require TRAF2. T cells deficient in TRAF2 or TRAF3 also displayed hyperactivity of the NF-kB2 pathway, but they did not accumulate in vivo or show extended survival in vitro. Mice lacking TRAF2 or TRAF3 in their T cells did however display a decrease in the number of memory phenotype CD8+ T cells. These studies indicate that some of the roles of TRAF2 and TRAF3 are common between B and T cells. However, the consequences of loss of TRAF2 or TRAF3 in B and T cells differs considerably, presumably due to the differential TNFR expression and usage by each cell type.

Errata & addenda tipped in behind back end paper. / Copies of author's previously published articles in pocket on back end-paper. / Bibliography: leaves 147-194. / xi, 194, [69] leaves, [68] leaves of plates : ill. (chiefly col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Explores the hypothesis that structures similar to lymphocyte-filled villi in rats and mice are present in the small human intestine. / Thesis (Ph.D.)--University of Adelaide, Dept. of Microbiology and Immunology, 1999

It has been shown that when implanted individually, both cells and biomaterials elicit biological responses. Implanted cells are often destroyed by the host's immune system, while biomaterials activate foreign body reactions which can result in inflammation and fibrotic encapsulation. However, when implanted simultaneously, the inflammatory responses to the biomaterial component can alter the immune responses to the cellular component. The experiments described in this thesis were designed to characterize the effect of different biomaterials on adaptive immune responses towards an allogeneic challenge. Balb/c splenocytes were challenged with irradiated allogeneic L929 cells, and treated with different biomaterials. Alterations in adaptive immune responses were quantified by T cell proliferation and cytokine release (i.e. IL-1(beta), IL-4, IL-12, and IFN-(gamma)). The roles various cell types played in first set responses were investigated. Experimental results indicated that biomaterials had a significant influence on nonspecific proliferation of splenocytes. In particular, analysis of the degree to which biomaterials affected specific proliferation indicated that the soluble alginate treatment significantly increased proliferation differences when compared to the control. However, biomaterials neither significantly affected specific splenocyte proliferation to an allogeneic challenge, nor the profile of secreted cytokines. To elucidate this response, alginate-treated splenocytes were depleted of adherent macrophages, CD4+ cells or CD8+ cells. Within non-challenged mixtures, CD4+ depletion had the most obvious effect. These results were supported by the non-depleted challenges, and indicated the direct influence biomaterials on CD4+ T cell proliferation. / Thesis / Master of Applied Science (MASc)

Commercial equipment used by the turkey industry at hatch sterilizes the germinal tissue of the claw with microwave energy and the beak tissue with infrared energy. This effectively claw and beak trims the birds. To test this technique on chickens, one-half of two strains of 1,200 Leghorn chicks were each subjected to the claw reduction (RC) technique at hatch, while one-half retained intact claws (IC). The beaks of one-third of these treatments were reduced at hatch using the infrared technique (1-day), one-third were precision trimmed at 7 d of age (7-day), and one-third were not trimmed (IB). Body weight, weight gain, feed intake, feed conversion, mortality, and fearfulness were measured. Rearing followed standard commercial feeding and husbandry procedures. During the preliminary experiment, heterophil to lymphocyte ratios did not consistently differ significantly between treatments. The RC birds had significantly lower body weight, except from 3 to 6 wk and had significantly lower feed consumption from 8 to 18 wk. The 1-day beak trimmed (BT) birds had significantly lower body weight from 3 to 14 wk and ate less total feed by 4 wk. Subjective evaluation showed that the RC birds exhibited less fearfulness during the growing period than the IC birds. Throughout lay, the body weight of RC and BT birds was significantly affected. Feed consumption was not lessened for RC birds, but was for BT birds throughout lay. Egg production, egg quality, and mortality were not affected by either treatment. / Master of Science