UTS site search

Emeritus Professor Robert Raison

Emeritus Professor, The ithree Institute
Associate Member, CHT - Centre for Health Technologies
BSc (Syd), PhD (Monash)
 

Research Interests

A long term interest and involvement in the study of the evolutionary development of the immune system provides the background for current research projects in Professor Raison’s laboratory that address the relationship between mucosal and systemic immune responses in fish, and the development of targeted vaccines for the aquaculture industry. In addition, his laboratory continues research in the area of antibody engineering that led to the development of the first recombinant, membrane active immunotoxin. Current research in this area addresses the unique anti-tumour activity of a monoclonal antibody targeting human kappa light chains. Commercial development of a therapy for multiple myeloma (an incurable blood cancer) based on this antibody, is being undertaken through a UTS spin-off company, PacMab Ltd.

Professor Raison’s expertise lies in the areas of immunology and antibody engineering. He is the subject coordinator for Immunology 2 and presents specialist lectures and tutorials in selected biotechnology subjects.

Journal articles

Hutchinson, A.T., Jones, D.R. & Raison, R.L. 2015, 'Preclinical and clinical development of an anti-kappa free light chain mAb for multiple myeloma.', Molecular immunology, vol. 67, no. 2 Pt A, pp. 89-94.
Monoclonal antibodies (mAb) have had tremendous success in treating a variety of cancers over the past twenty years. Yet despite their widespread clinical use, which includes treatments for haematological malignancies, there are still no approved mAb therapies for multiple myeloma (MM). This is likely to change within the next few years with a number of mAb therapies being assessed in late stage clinical trials, most notably, the anti-CS-1 mAb, elotuzumab, and the anti-CD38 mAb, daratumumab, which are currently being evaluated in Phase III clinical trials for MM. In this review, we will discuss the preclinical and clinical development of MDX-1097, a Phase II candidate which targets cell membrane-associated kappa immunoglobulin free light chains expressed on the surface of MM cells.
Hutchinson, A.T., Jones, D.R., McCauley Winter, P., Tangye, S.G. & Raison, R.L. 2014, 'Cell membrane associated free kappa light chains are found on a subset of tonsil and in vitro-derived plasmablasts.', Human immunology, vol. 75, no. 9, pp. 986-990.
The monoclonal antibody, MDX-1097, is currently progressing through clinical trials as a possible therapy for multiple myeloma. MDX-1097 targets a cell membrane bound form of free immunoglobulin kappa light chain (FLC), termed kappa myeloma antigen (KMA), which is found on the surface of malignant plasma cells. The clinical potential of MDX-1097 highlights the need to characterise the expression of its cognate antigen, KMA, in normal tissue. In this study, we have analysed the expression of KMA on B cell subsets found in tonsils, peripheral blood and bone marrow. We found KMA expression on a small population of tonsillar and in vitro derived plasmablasts. In contrast, no KMA expression was observed on peripheral blood or bone marrow resident B cell subsets. This study yields important insights into the possible subsets of B cells that might be depleted as a result of an immunotherapy targeting KMA.
Hutchinson, A.T., Malik, A., Berkahn, M.B., Agostino, M., To, J., Tacchi, J.L., Djordjevic, S.P., Turnbull, L., Whitchurch, C.B., Edmundson, A.B., Jones, P.M., Raison, R.L. & Ramsland, P.A. 2013, 'Formation of Assemblies on Cell Membranes by Secreted Proteins: Molecular Studies of Free Lambda Light Chain Aggregates Found on the Surface of Myeloma Cells.', Biochemical Journal, vol. 454, no. 3, pp. 479-489.
View/Download from: UTS OPUS or Publisher's site
We have described the presence of cell membrane-associated ? free immunoglobulin light chains (FLC) on the surface of myeloma cells. Notably, the anti-?FLC mAb, MDX-1097, is being assessed in clinical trials as a therapy for ? light chain isotype multiple myeloma. Despite the clinical potential of anti-FLC mAbs, there have been limited studies on characterizing membrane-associated FLCs at a molecular level. Furthermore, it is not known if ?FLCs can associate with cell membranes of myeloma cells. In this study, we describe the presence of ?FLCs on the surface of myeloma cells. We found that cell surface-associated ?FLC are bound directly to the membrane and in an aggregated form. Subsequently, membrane interaction studies revealed that ?FLCs interact with saturated zwitterionic lipids such as phosphatidylcholine and phosphatidylethanolamine, and using automated docking, we characterize a potential recognition site for these lipids. Atomic force microscopy confirmed that membrane-associated ?FLCs are aggregated. Given our findings, we propose a model whereby individual FLCs show modest affinity for zwitterionic lipids, with aggregation stabilizing the interaction due to multivalency. Notably, this is the first study to image FLCs bound to phospholipids and provides important insights into the possible mechanisms of membrane association by this unique myeloma surface antigen.
Hutchinson, A.T., Jones, D. & Raison, R.L. 2012, 'The Ability To Interact With Cell Membranes Suggests Possible Biological Roles For Free Light Chain', Immunology Letters, vol. 142, no. 1-2, pp. 75-77.
View/Download from: Publisher's site
During antibody synthesis, immunoglobulin light chains are produced in excess of heavy chains and, as a consequence, can be secreted by plasma cells as free light chains (FLC). Thus, FLC were considered to be a by-product of immunoglobulin synthesis, lac
Hutchinson, A.T., Alexova, R., Bockhorni, V., Ramsland, P.A., Jones, D.R., Jennings, C.V., Broady, K.W., Edmundson, A.B. & Raison, R.L. 2011, 'Characterization of a unique conformational epitope on free immunoglobulin kappa light chains that is recognized by an antibody with therapeutic potential', Molecular Immunology, vol. 48, no. 9-10, pp. 1245-1252.
View/Download from: UTS OPUS or Publisher's site
The murine mAb, K-1-21, recognizes a conformational epitope expressed on free Ig kappa light chains (FkLCs) and also on cell membrane-associated FkLCs found on kappa myeloma cells. This has led to the development of a chimeric version of K-1-21, MDX-1097, which is being assessed in a Phase II clinical trial for the treatment of multiple myeloma. The epitope recognized by K-1-21 is of particular interest, especially in the context that it is not expressed on heavy chain-associated light chains such as in an intact Ig molecule. Using epitope excision techniques we have localized the K-1-21 epitope to a region spanning residues 104-110 of FkLC. This short strand of residues links the variable and constant domains, and is a flexible region that adopts different conformations in FkLC and heavy chain-associated light chain. We tested this region using site-directed mutations and found that the reactivity of K-1-21 for FkLC was markedly reduced. Finally, we applied in silico molecular docking to generate a model that satisfied the experimental data. Given the clinical potential of the Ag, this study may aid the development of next generation compounds that target the membrane form of FkLC expressed on the surface of myeloma plasma cells.
Villavedra Sierra, M., To, J., Lemke, S.J., Birch, D., Crosbie, P., Adams, M.A., Broady, K.W., Nowak, B., Raison, R.L. & Wallach, M. 2010, 'Characterisation Of An Immunodominant, High Molecular Weight Glycoprotein On The Surface Of Infectious Neoparamoeba Spp., Causative Agent Of Amoebic Gill Disease (Agd) In Atlantic Salmon', Fish and Shellfish Immunology, vol. 29, no. 6, pp. 946-955.
View/Download from: UTS OPUS or Publisher's site
Amoebic gill disease can be experimentally induced by the exposure of salmonids to Neoparamoeba spp. freshly isolated from infected fish, while cultured amoebae are non-infective.
Hutchinson, A.T., Ramsland, P.A., Jones, D., Agostino, M., Lund, M.E., Jennings, C., Bockhorni, V., Yuriev, E., Edmundson, A.B. & Raison, R.L. 2010, 'Free Ig light chains interact with Sphingomyelin and are found on the surface of Myeloma plasma Cells in an aggregated form', Journal of Immunology, vol. 185, no. 7, pp. 4179-4188.
View/Download from: UTS OPUS or Publisher's site
Free kappa L chains (FkappaLCs) are expressed on the surface of myeloma cells and are being assessed as a therapeutic target for the treatment of multiple myeloma. Despite its clinical potential, the mechanism by which FkappaLCs interact with membranes remains unresolved. In this study, we show that FkappaLCs associate with sphingomyelin on the plasma membrane of myeloma cells. Moreover, membrane-bound FkappaLCs are aggregated, suggesting that aggregation is required for intercalation with membranes. Finally, we propose a model where the binding of FkappaLCs with sphingomyelin on secretory vesicle membranes is stabilized by self-aggregation, with aggregated FkappaLCs exposed on the plasma membrane after exocytosis. Although it is well known that protein aggregates bind membranes, this is only the second example of an aggregate being found on the surface of cells that also secrete the protein in its native form. We postulate that many other aggregation-prone proteins may associate with cell membranes by similar mechanisms.
Vallas, V., Farrugia, W., Raison, R.L., Edmundson, A.B. & Ramsland, P.A. 2007, 'Dissimilar aggregation processes govern precipitation and gelation of human IgM cryoglobulins', Journal of Molecular Recognition, vol. 20, no. 2, pp. 90-96.
View/Download from: UTS OPUS or Publisher's site
Cryoglobulinemia is associated witha range of diseases including rheumatoid arthritis, B-cell malignancies, and chronic viral infections. This "cold sensitivity" condition is caused by cryoglobulins that precipitate, gel, or occasionally crystallise in the cold. Clinical manifestations vary widely in severity, depending on many factors, including the type of cryoglobulin (monoclonal or mixed immunoglobulins) and the physical nature of the aggregates (precipitate, gel or crystal). Dynamic light scattering (DLS) was used to examine the cold-induced precipitation or gelation of two human cryoglobulins, namely, Pot IgM and Yvo IgM. The DLS assay was highly reproducible, sensitive and had low intra-assay variations for both igM, cryoglobulins. Distinct processes were revealed to contribute to precipitation and gelation of cryoglobulins. The precipitation of Pot IgM displayed a rapid transition from solution to solid phases, with a wide distribution of aggregated sizes. in contrast, the gelation fo Yvo IgM progressed gradually across a broad temperature range to produce a relatively uniform gel matrix. Initial cryoglobulin concentrations determined the kinetics and critical temperatures for both precipitation and gelation. Moreover, the Yvo IgM was observed to have a distinct relationship between concentrations and mean hydrodynamic diameters or particle sizes. Concentration-dependent effects on particle sizes were present but not as pronounced for the pot IgM. Precipitation and gelation of cryoglobulins were also found to be differentially responsive to changes in the aqueous environment. Our results indicate that DLS is a rapid, reliable and sensitive method for characterising the nature of disease-associated cryoglobulins.
Villavedra Sierra, M., Lemke, S.J., To, J.H., Broady, K.W., Wallach, M. & Raison, R.L. 2007, 'Carbohydrate epitopes are immunodominant at the surface of infectious Neoparamoeba spp', Journal Of Fish Diseases, vol. 30, no. 4, pp. 191-199.
View/Download from: UTS OPUS or Publisher's site
Amoebic gill disease, the main disease of concern to the salmon industry is Tasmania is caused by the amoeba Neoparamoeba spp. Experimental infection can onlybe induced by exposure to wild-type (WT) oarasites isolated from the gills of infected fish, as cultured amoeba are non-infective. To characterise the surface antigens of WT parasites, we produced monoclonal antibodies (mAbs) using subtractive immunization. Mice inoculated with non-infective parasites were treated with cyclophosphamide, to deplete reactive lumphocytes, and then immunized with different antigen preparations from infective parasites. When whole parasites were used for boosting the percentage of WT unique mAbs was very high (86%) as was the percentage of mAbs specific for carbohydrate pritopes (89%). When degloycosylated membranes were used, the numbers of mAbs spefic for non-carbohydrate spitopes id not increase, but the total number of WT unique mAbs was reduced (86-40%). Using an untreated membrane preparation, the total number of mAbs to surface molecules was very high, but all recognized carbohydrate epitopes. The total number of MAbs recognising carbohydrate epitopes on the surface of the WT parasites was 97%, suggesting that the dominant epitopes on the surface molecules unique to WT parasites are carbohydrate in nature.
Lynch, G.W., Turville, S., Carter, B., Sloane, A.J., Chan, A., Muljadi, N., Li, S., Low, L., Armati, P., Raison, R.L., Zoellner, H., Williamson, P., Cunningham, A.L. & Church, W.B. 2006, 'Marked differences in the structures and protein associations of lymphocyte and monocyte CD4 Resolution of a novel CD4 isoform', Immunology And Cell Biology, vol. 84, no. 2, pp. 154-165.
View/Download from: UTS OPUS or Publisher's site
The structures, molecular interactions and functions of CD4 in a subset of T lymphocytes have been well characterized. The CD4 receptors of other cell types have, however, been poorly documented. We have previously shown that lymphocytes and monocytes/ma
Villavedra Sierra, M., To, J.H., Lemke, S.J., Broady, K.W., Melrose, J., Birch, D., Wallach, M. & Raison, R.L. 2006, 'Carbohydrate Epitopes Are Immunodominant At The Surface Of Infectious Neoparamoeba Spp', Glycobiology, vol. 16, no. 11, pp. 1143-1143.
NA
Villavedra Sierra, M., McCarthy, K., To, J.H., Morrison, R., Crosbie, P., Broady, K.W. & Raison, R.L. 2005, 'Changes in antigenic profile during culture of Neoparamoeba sp., causative agent of amoebic gill disease in Atlantic salmon', International Journal for Parasitology, vol. 35, pp. 1417-1423.
View/Download from: UTS OPUS or Publisher's site
Amoebic gill diseas (AGD) the most serious infectious disease affecting farmed salmon in Tasmania, is caused by free-living marine amoeba Neoparamoeba sp. The parasites on the gills induce proliferation of epithelial cells initiating a hyperplastic response and reducing the surface area available for gaseous exchange. AGD casn be induced in salmon by exposure to frshly isolated Neoparamoeba from AGD infected fish, however cultured Neoparamoeba are non-infective. We describe here antigenic difference between freshly isolated and in vitro cultured parasites, and within individual isolates of the parasite cultured under different conditions.
Asvadi, P., Jones, D., Dunn, R., Choo, A.B., Raison, M.J. & Raison, R.L. 2004, 'A Monoclonal Antibody Specific For Free Human Kappa Light Chains Induces Apoptosis Of Multiple Myeloma Cells And Exhibits Anti-tumor Activity In Vivo', Blood, vol. 104, no. 11, pp. 1-1.
NA
Choo, A.B., Dunn, R., Broady, K.W. & Raison, R.L. 2002, 'Soluble expression of a functional recombinant cytolytic immunotoxin in insect cells', Protein Expression And Purification, vol. 24, no. 3, pp. 338-347.
View/Download from: Publisher's site
We have previously described the production of a recombinant melittin-based cytolytic immunotoxin (IT), scFv-mel-FLAG, in bacterial cells. While the IT exhibited specific cytotoxicity for a human lymphoblastoid cell line, HMy2, yields from expression were low. Here, we describe a baculovirus expression system for the overexpression and secretion of scFv-mel-FLAG. A novel snake phospholipase A2 inhibitor signal peptide was used to aid in the secretion of the immunotoxin. Sf21 insect cells infected with the recombinant virus secreted soluble scFv-mel-FLAG into the culture medium from which it was purified directly on an affinity column. The final yield of scFv-mel-FLAG was estimated at 3â5 mg/L, which was an improvement of 30-fold compared to expression in the prokaryotic system. The cell binding characteristics of the recombinant IT were assessed by flow cytometry using the antigen expressing cell line HMy2. ScFv-mel-FLAG bound specifically to HMy2 cells in direct binding assays and this binding was completely inhibited in the presence of an excess of soluble antigen. Significant cytotoxicity for HMy2 cells, measured by leakage of cytosolic LDH, was also observed for the IT at a concentration of 60 pmol/104 cells. Cytotoxicity was concentration dependent and was specific for antigen-positive cells. Thus the baculovirus expression system, under the control of a novel secretion signal, can be used for the production of soluble and functional recombinant cytolytic immunotoxins. To our knowledge, this is the first report of expression of a recombinant immunotoxin in the baculovirus expression vector system.
Raison, R.L. & Jorgensen, T.O. 2002, 'Immunology And Sustainable Aquaculture', Developmental And Comparative Immunology, vol. 26, no. 2, pp. 129-130.
View/Download from: Publisher's site
The contribution by aquaculture to total fisheries production has increased significantly over the last decade. Indeed the increase in production of approximately 20 million tonnes over that period has been almost entirely the result of increased aquaculture-based production. Furthermore, impacts of global climate change together with overfishing of capture-based fisheries mean the contribution of aquaculture to this important area of food production will increase in the future.
Cain, K.D., Jones, D. & Raison, R.L. 2002, 'Antibody-antigen kinetics following immunization of rainbow trout (Oncorhynchus mykiss) with a T-cell dependent antigen', Developmental & Comparative Immunology, vol. 26, no. N/A, pp. 181-190.
View/Download from: UTS OPUS
Lovas, J.M., Craig, A.R., Raison, R.L., Weston, K.M., Segal, Y. & Markus, M.R. 2002, 'The effects of massage therapy on the human immune response in healthy adults', Journal of Bodywork and Movement Therapies, vol. 6, no. 30, pp. 143-150.
View/Download from: UTS OPUS
Asvadi, P., Fletcher, A. & Raison, R.L. 2002, 'Expression and functional analysis of recombitant scFv and diabody fragments with specificity for human RhD', Journal of Molecular Recognition, vol. 15, no. N/A, pp. 321-330.
View/Download from: UTS OPUS
n an attempt to generate recombinant anti-D reagents for possible diagnostic and therapeutic use we cloned the genes encoding the variable (V) domains of a human anti-D antibody secreted by the lymphoblastoid cell line BTSN4. A single-chain Fv (scFv) fragment was constructed using a 21 amino acid linker to join the genes encoding the variable domains of the BTSN4 heavy (VH) and light chains (VL). A diabody construct was also generated by reducing the length of the scFv linker from 21 to 10 residues. The scFv and diabody constructs were cloned into the pFLAG-CTS vector, expressed in E. coli host cells and the recombinant proteins were affinity-isolated from bacterial culture medium. Analysis of the recombinant proteins indicated that they retained the D antigen binding specificity of the parental BTSN4 IgG. Furthermore, both fragments mediated agglutination of papain-treated D positive erythrocytes in the absence of a cross-linking second antibody. While the agglutinating property of BTSN4 diabody was readily explained by the noncovalent association of this protein as a bivalent dimer, oligomeric forms of BTSN4 scFv were not detected when the protein was analysed by size exclusion chromatography. Thus, the agglutinating property of the scFv is not the result of the formation of non-covalently associated multimeric forms of the antibody fragment
Cain, K.D., Jones, D.R. & Raison, R.L. 2002, 'Antibody-antigen kinetics following immunization of rainbow trout (Oncorhynchus mykiss) with a T-cell dependent antigen.', Developmental and comparative immunology, vol. 26, no. 2, pp. 181-190.
Enhancement of the immune response through affinity maturation of the antibody response is a feature of the mammalian immune system and has important implications with respect to development of vaccination strategies. However, an absence of germinal centres and apparent lack of somatic hypermutation of immunoglobulin V genes suggests that this phenomenon does not occur in fish. We investigated the question of affinity maturation in rainbow trout (Oncorhynchus mykiss) by measuring antibody-antigen binding kinetics using a BIAcore biosensor. Following immunization with a T-cell dependent antigen (FITC-KLH), relative binding affinities of serum and mucosal antibodies were assessed based on their dissociation rate constants (k(diss.)). A detectable serum anti-FITC response developed by 4 weeks post-immunization, and a consistent shift to higher affinity antibody production (i.e. a decrease in k(diss.)) was observed over the ensuing course of the immune response. An average k(diss.) of 3.5 x 10(-4)+/-0.27 x 10(-4)sec(-1) was observed during early stages of the response (4 weeks), while by 6 weeks this decreased significantly (p<0.05). Further reduction in k(diss.) was observed, with a low of 1.2 x 10(-4)+/-0.06 x 10(-4)sec(-1) being observed by week 12. Analysis of the anti-FITC response in skin-derived mucus revealed a similar pattern of decreasing k(diss.) as the immune response progressed. While these data clearly demonstrate a 2-3 fold increase in antibody-antigen binding during the course of the immune response in trout, the magnitude of this increase is much less than that seen in the mammalian immune response. This may reflect differences in the mechanisms underpinning this phenomenon in divergent species.
Ramsland, P.A., Ollerenshaw, J., Schultz, B.B., DeWitt, C.R., Chissoe, W.F., Raison, R.L. & Edmundson, A.B. 2001, 'Interconversion of Different Crystal Forms of Fabs from Human IgM Cryoglobulins', Journal of Crystal Growth, vol. 232, pp. 204-214.
View/Download from: UTS OPUS or Publisher's site
Nair, S.V., Burandt, M., Hutchinson, A., Raison, R.L. & Raftos, D.A. 2001, 'A C-type lectin from the tunicate, Styela plicta, that modulates cellular activity', Comparative Biochemistry and Physiology Part C: ..., vol. C129, pp. 11-24.
View/Download from: UTS OPUS
Weston, K.M., Tangye, S.G., Dunn, R., Smith, A., Morris, M. & Raison, R.L. 2001, 'IgM Expressed by Leukemic CD5+B Cells Binds Mouse Immunoglobulin Light Chain', Journal of Molecular Recognition, vol. 14, pp. 245-253.
View/Download from: UTS OPUS or Publisher's site
Mouse immunoglobulin (Ig) molecules have previously been shown to bind to the surface of CD5 B cells from patients with B-cell chronic lymphocytic leukemia (B-CLL). The results indicated that surface IgM was involved in the interaction and suggested the phenomenon was an example of the polyreactive binding capacity of the surface Ig (sIg) expressed by these malignant cells. This article describes the further characterization of the interaction between human IgM and mouse Ig molecules and subunits. Mouse Ig molecules of both kappa and lambda light chain classes bound to the B-CLL cell surface. The dissociation constant for the interaction of mouse IgG1 (K121) with the B-CLL cell surface was 3.6 107 M. To confirm the involvement of the human IgM expressed by the B-CLL cells in the interaction, the malignant cells were stimulated in vitro to induce secretion of human IgM. Enzyme immunoassay was used to show that secreted human IgM bound to intact mouse Ig, as occurred with the cell surface analysis. The mouse Ig epitope recognized by the purified secreted human IgM was shown by Western blot analysis to be located on the light chain of the mouse Ig molecule and to be conformationally dependent. K121 light chain was cloned and expressed in E. coli and the recombinant light chain bound to the surface of CLL B cells. The results confirm that human IgM is the reactive ligand in the interaction with mouse Ig and indicate that the interaction of polyreactive IgM with mouse IgG occurs via the light chain component of IgG.
Cain, K.D., Jones, D. & Raison, R.L. 2000, 'Characterisation of Mucosal and Systemic Immune Responses in Rainbow Trout (Oncorhynchus mykiss) Using Surface Plasmon Resonance', Fish and Shellfish Immunology, vol. 10, no. 0, pp. 651-666.
Dos Remedies, N., Ramsland, P.A., Hook, J.M. & Raison, R.L. 1999, 'Identification Of A Homologue Of Cd59 In A Cyclostome: Implications For The Evolutionary Development Of The Complement System', Developmental And Comparative Immunology, vol. 23, no. 1, pp. 1-14.
We have employed a COS cell expression cloning procedure to isolate a full length cDNA clone encoding a hagfish leukocyte-associated membrane protein (HLMP1). The protein. which is identified by a monoclonal antibody (JB3) generated in our laboratory, is
Jones, D., Hannan, C., Russell-jones, G. & Raison, R.L. 1999, 'Selective B Cell Non-responsiveness In The Gut Of The Rainbow Trout (oncorhynchus Mykiss)', Aquaculture, vol. 172, no. 1-2, pp. 29-39.
View/Download from: Publisher's site
In order to assess the potential for the development of economically viable and effective oral vaccines we have examined the humoral immune response resulting from the anal administration of a hapten-carrier conjugate in the posterior intestine of the ra
Ramsland, P.A., Brock, C., Moses, J., Robinson, B., Edmundson, A.B. & Raison, R.L. 1999, 'Structural Aspects Of Human Igm Antibodies Expressed In Chronic B Lymphocytic Leukemia', Immunotechnology, vol. 4, no. 3-4, pp. 217-229.
View/Download from: Publisher's site
Background: Malignant B cells from patients with chronic B lymphocytic leukemia (B CLL) generally express both surface IBM and the pan T cell antigen CD5, a characteristic of the B1 population of B lymphocytes. The IgM on the surface of these B CLL cells
Kehrer, S., Hannan, C. & Raison, R.L. 1998, 'Identification Of A Subpopulation Of Leucocytes From The Rainbow Trout (oncorhynchus Mykiss) Responsive To Pokeweed Mitogen', Fish & Shellfish Immunology, vol. 8, no. 6, pp. 477-487.
View/Download from: Publisher's site
Several compounds which are known to be potent polyclonal activators of lymphocytes in mammals have also been demonstrated to have mitogenic activity for leucocytes from a number of fish species, including rainbow trout. In the present study, the rainbow
Weston, K.M., Mulligan, S. & Raison, R.L. 1998, 'In Vivo Binding Of Mouse Igg Via Polyreactive Surface Igm Abrogates Progressive Lymphocytosis In Prolymphocytic Leukemia', Leukemia & Lymphoma, vol. 29, no. 3-4, pp. 361-373.
View/Download from: Publisher's site
Surface IgM expressed by malignant CD5(+) B-cells from patients with B-chronic lymphocytic leukemia (B-CLL) has previously been shown to bind mouse Ig in what appears to be an example of polyreactive antigen-binding activity. This report demonstrates the
Weston, K.M. & Raison, R.L. 1998, 'Interaction Of Melittin With A Human Lymphoblastoid Cell Line, Hmy2', Journal Of Cellular Biochemistry, vol. 68, no. 2, pp. 164-173.
View/Download from: Publisher's site
We have examined the cytolytic effects of the membrane-active peptide, melittin, on a human lymphoblastoid cell line (HMy2) in the context of the use of melittin as the toxic component of an immunotoxin. The toxicity of melittin for HMy2 cells was linear
Tangye, S.G., Weston, K.M. & Raison, R.L. 1998, 'Interleukin-10 Inhibits The In Vitro Proliferation Of Human Activated Leukemic Cd5(+) B-cells', Leukemia & Lymphoma, vol. 31, no. 1-2, pp. 121-130.
View/Download from: Publisher's site
B-cell chronic lymphocytic leukemia (B-CLL) is characterised by the proliferation and accumulation of sIgM(+)/CD5(+) B-cells that fail to progress to the final stages of B-cell development. Despite their developmental arrest, leukemic CD5(+) B-cells can
Ramsland, P.A., Guddat, L., Edmundson, A.B. & Raison, R.L. 1997, 'Diverse Binding Site Structures Revealed In Homology Models Of Polyreactive Immunoglobulins', Journal Of Computer-aided Molecular Design, vol. 11, no. 5, pp. 453-461.
View/Download from: Publisher's site
We describe here computer-assisted homology models of the combining site structure of three polyreactive immunoglobulins. Template-based models of Fv (V-L-V-H) fragments were derived for the surface IgM expressed by the malignant CD5 positive B cells fro
Tangye, S.G. & Raison, R.L. 1997, 'Human Cytokines Suppress Apoptosis Of Leukaemic Cd5(+) B Cells And Preserve Expression Of Bcl-2', Immunology And Cell Biology, vol. 75, no. 2, pp. 127-135.
View/Download from: Publisher's site
Leukaemic CD5(+) B cells obtained from B cell chronic lymphocytic leukaemia (B-CLL) patients rapidly undergo apoptosis during in vitro culture. This is associated with down-regulation in expression of bcl-2. Spontaneous apoptosis of these cells contrasts
Tangye, S.G., Weston, K.M. & Raison, R.L. 1997, 'Cytokines And Cross-linking Of Sigm Augment Pma-induced Activation Of Human Leukaemic Cd5(+) B Cells', Immunology And Cell Biology, vol. 75, no. 6, pp. 561-567.
View/Download from: Publisher's site
Purified leukaemic CD5(+) B cells obtained from patients with B cell chronic lymphocytic leukaemia (B-CLL) undergo activation and differentiation following in vitro stimulation with optimal concentrations of the phorbol ester PMA. This paper examines the
Shum, B., Azumi, K., McCrimmon, L., Kehrer, S., Raison, R.L., Detrich, H. & Parham, P. 1996, 'Unexpected Beta(2)-microglobulin Sequence Diversity In Individual Rainbow Trout', Proceedings Of The National Academy Of Sciences Of The United States Of America, vol. 93, no. 7, pp. 2779-2784.
View/Download from: Publisher's site
For mammals beta(2)-microglobulin (beta(2)m), the light chain of major histocompatibility complex (MHC) class I molecules, is invariant (or highly conserved) and is encoded by a single gene unlinked to the MHC. We find that beta(2)m of a salmonid fish, t
Dunn, R., Weston, K.M., Longhurst, T.J., Lilley, G.G., Rivett, D., Hudson, P. & Raison, R.L. 1996, 'Antigen Binding And Cytotoxic Properties Of A Recombinant Immunotoxin Incorporating The Lytic Peptide, Melittin', Immunotechnology, vol. 2, no. 3, pp. 229-240.
View/Download from: Publisher's site
Background: The majority of immunotoxins studied to date incorporate toxins that act in the cytosol and thus need to be endocytosed by the target cell. An alternative strategy for immunotoxin development is the use of membrane active toxins, such as the
Tangye, S.G. & Raison, R.L. 1996, 'Leukaemic Cd5(+) B-cell Apoptosis: Co-incidence Of Cell Death And Dna Fragmentation With Reduced Bcl-2 Expression', British Journal Of Haematology, vol. 92, no. 4, pp. 950-953.
View/Download from: Publisher's site
Apoptosis and bcl-2 expression were characterized in leukaemic B cells during in vitro culture. Prior to culture, > 85% of cells from each B-CLL patient expressed high levels of bcl-2. Despite this, all leukaemic B-cell populations underwent apoptosis in
Tangye, S.G., Weston, K.M. & Raison, R.L. 1995, 'Phorbol Ester Activates Cd5(+) Leukemic B-cells Via A T-cell Independent Mechanism', Immunology And Cell Biology, vol. 73, no. 1, pp. 44-51.
View/Download from: Publisher's site
B cell chronic lymphocytic leukaemia (B-CLL) is characterized by the proliferation and accumulation of sIgM(+)CD5(+) lymphocytes that fail to progress to the final stages of B cell development. Stimulation of unfractionated PBL from three patients with B
Newton, R.U., Raftos, D.A., Raison, R.L. & Geczy, C.L. 1994, 'Chemotactic Responses Of Hagfish (vertebrata, Agnatha) Leukocytes', Developmental And Comparative Immunology, vol. 18, no. 4, pp. 295-303.
View/Download from: Publisher's site
The chemotactic responses of hagfish leucocytes were tested using a variety of chemoattractants. Leucocyte migration was significantly enhanced by purified mammalian complement anaphylotoxin (C5a) and LPS-activated hagfish plasma. Checkerboard analyses c
Raison, R.L., Coverley, J., Hook, J.M., Towns, P., Weston, K.M. & Raftos, D.A. 1994, 'A Cell-surface Opsonic Receptor On Leukocytes From The Phylogenetically Primitive Vertebrate, Eptatretus-stouti', Immunology And Cell Biology, vol. 72, no. 4, pp. 326-332.
View/Download from: Publisher's site
A humoral recognition molecule that is homologous to the mammalian complement components C3, C4 and C5 has recently been identified in the Pacific hagfish, Eptatretus stouti. One function of this complement-like protein (CLP) is to opsonize foreign mater
Tangye, S.G., Weston, K.M. & Raison, R.L. 1994, 'Mechanisms For Maintenance Of Leukemic B-cell Survival In-vitro - Role Of Normal T-cells', Journal Of Cellular Biochemistry, vol. 0, pp. 440-440.
NA
Weston, K.M., Alsalami, M. & Raison, R.L. 1994, 'Cell-membrane Changes Induced By The Cytolytic Peptide, Melittin, Are Detectable By 90-degrees Laser Scatter', Cytometry, vol. 15, no. 2, pp. 141-147.
View/Download from: Publisher's site
The 90 degrees light scatter parameter of the now cytometer was used to observe changes in the membrane of human lymphoblastoid cells (HMy2) as a result of the action of the cytolytic peptide, melittin. There was a rapid and concentration-dependent incre
Dunn, R., Hudson, P., Lilley, G. & Raison, R.L. 1994, 'Cloning And Expression Of Antibody Fragments For Construction Of A Recombinant Immunotoxin', Journal Of Cellular Biochemistry, vol. 0, pp. 212-212.
NA
Weston, K.M., Tangye, S.G. & Raison, R.L. 1993, 'Human Polyreactive Igm Binds Mouse Igg Light-chain In A Conformationally Dependent Interaction', Journal Of Leukocyte Biology, vol. 0, pp. 92-92.
NA
Ramsland, P.A., Tangye, S.G. & Raison, R.L. 1993, 'Ag Binding-properties Of A Structurally Defined Human-igm Cryoglobulin', Journal Of Leukocyte Biology, vol. 0, pp. 92-92.
NA
Raftos, D.A. & Raison, R.L. 1993, 'The Echinoid Immune-system Revisited', Immunology Today, vol. 14, no. 2, pp. 92-93.
NA
Tangye, S.G., Weston, K.M. & Raison, R.L. 1993, 'In-vitro Induction Of Proliferation And Ig Class Switch In B-cll Cells', Journal Of Leukocyte Biology, vol. 0, pp. 77-77.
NA
Shan, L., Guddat, L., Raison, R.L. & Edmundson, A.B. 1993, 'Crystallization Of An Fv Fragment From A Human-igm Cryoglobulin By A Microseeding Technique', Journal Of Crystal Growth, vol. 126, no. 2-3, pp. 229-244.
View/Download from: Publisher's site
In contrast to many fragments of human IgM immunoglobulins, V(L)-V(H) heterodimer (Fv) from a patient (Pot) with macroglobulinemia produces large single crystals in relatively short periods of time (days). It was therefore convenient to segregate the cry
Raftos, D.A., Hook, J.M. & Raison, R.L. 1992, 'Complement-like Protein From The Phylogenetically Primitive Vertebrate, Eptatretus-stouti, Is A Humoral Opsonin', Comparative Biochemistry And Physiology B-biochemistry & Molecular Biology, vol. 103, no. 2, pp. 379-384.
1. Serum from the Pacific hagfish, Eptatretus stouti, contains a complement-like protein (CLP). 2. CLP from unfractionated hagfish serum and from affinity-purified preparations binds to yeast cell surfaces. 3. Incubation with CLP enhances the phagocytosi
Hanley, P., Hook, J.M., Raftos, D.A., Gooley, A., Trent, R. & Raison, R.L. 1992, 'Hagfish Humoral Defense Protein Exhibits Structural And Functional Homology With Mammalian Complement Components', Proceedings Of The National Academy Of Sciences Of The United States Of America, vol. 89, no. 17, pp. 7910-7914.
View/Download from: Publisher's site
A genomic clone and cDNA fragment encoding a portion of a humoral recognition molecule from the hagfish were isolated and sequenced. The serum protein has previously been described as having structural features that are immunoglobulin-like. Amino acid se
Fan, Z., Shan, L., Guddat, L., He, X., Gray, W., Raison, R.L. & Edmundson, A.B. 1992, 'Three-dimensional Structure Of An Fv From A Human IgM Immunoglobulin', Journal of Molecular Biology, vol. 228, no. 1, pp. 188-207.
View/Download from: UTS OPUS or Publisher's site
An IgM(?) immunoglobulin from a patient (Pot) with Waldenstrom's macroglobulinemia was hydrolyzed with pepsin to release a fragment consisting of the `variable (V) domains of the light and heavy chains plus eight residue `tails from the `constant (C) domains. The crystal structure of this fragment was determined at 2.3 &Aring; resolution by molecular replacement and crystallographic refinement methods. When examined separately, the light chain component closely resembles another human ? chain (Rei) in both the ?-pleated sheet regions and the `hypervariable loops. The conserved pleated sheets in the heavy chain are similar to those in the human Kol IgG1 protein, but the third hypervariable loop in particular is different from that in any immunoglobulin structure described to date. As in the Kol protein, this loop blocks the access to any internal active site along the light-heavy chain interface. Unlike the Kol protein, however, the loop does not protrude beyond the boundaries of a conventional antigen combining site. Instead, it forms a very compact structure, which fills almost all residual space between the domains. This is an example of one dominant complementarity-determining region (CDR) essentially negating the diversity possible with five other CDRs in the two chains. Ordered water molecules are associated with light chain constituents along the interface, but not with CDR3 of the heavy chain. In screening exercises the Pot IgM failed to bind a wide variety of peptides. Together, the results suggest that ligand binding can only occur on external surfaces of the protein. These surfaces carry a limited number of side chains usually assigned to CDRs in more typical antibodies.
Weston, K.M. & Raison, R.L. 1991, 'Low Affinity Binding Of Mouse Immunoglobulin To Human Cd5+ B-cells', Immunology And Cell Biology, vol. 69, pp. 261-271.
View/Download from: Publisher's site
Polyreactive immunoglobulin (Ig) secreted by CD5-bearing B cells has the capacity to bind a broad range of self and foreign antigens. Flow cytometric analysis was used to detect low-affinity binding of mouse Ig molecules to the surface of CD5-bearing B c
Foley, R., Raison, R.L. & Beh, K. 1991, 'Monoclonal-antibody Against Sheep Kappa-light Chain', Hybridoma, vol. 10, no. 4, pp. 507-515.
View/Download from: Publisher's site
The production of a monoclonal antibody specific for sheep kappa light chain protein is described. The monoclonal antibody was designated McM11 and its specificity was verified using western blots of sheep IgG and slides of efferent lymph cells. The sp
Hanley, P., Seppelt, I., Gooley, A., Hook, J.M. & Raison, R.L. 1990, 'Distinct Ig H Chains In A Primitive Vertebrate, Eptatretus-stouti', Journal Of Immunology, vol. 145, no. 11, pp. 3823-3828.
NA
Nethery, A., Raison, R.L. & Easterbrooksmith, S. 1990, 'Single-step Purification Of Immunoglobulin-m On C1q-sepharose', Journal Of Immunological Methods, vol. 126, no. 1, pp. 57-60.
View/Download from: Publisher's site
NA
Mckenzie, J.A., Raison, R.L. & Rivett, D. 1988, 'Development Of A Bifunctional Crosslinking Agent With Potential For The Preparation Of Immunotoxins', Journal Of Protein Chemistry, vol. 7, no. 5, pp. 581-592.
View/Download from: Publisher's site
NA
Walker, K., Seymourmunn, K., Towson, J., Axiak, S.M., Raison, R.L., Bautovitch, G., Morris, J.M. & Basten, A. 1988, 'Radioimmunolocalization And Selective Delivery Of Radiation In A Rat Model System - Comparison Of Intact And Fragmented Antibody', Nuclear Medicine Communications, vol. 9, no. 7, pp. 517-526.
NA
Wilson, M., Mulligan, S. & Raison, R.L. 1988, 'A New Microsphere-based Immunofluorescence Assay For Antibodies To Membrane-associated Antigens', Journal Of Immunological Methods, vol. 107, no. 2, pp. 231-237.
View/Download from: UTS OPUS or Publisher's site
The screening of panels of hybridoma supernatants for specific secreted monoclonal antibodies is often achieved by cellular immunofluorescent staining and flow cytometric analysis. In some circumstances such assays are difficult because the required antigen-bearing cell population is not suitable for use in flow cytometry, has limited cell cycle expression or poor in vitro growth. A method is presented here that provides a solution to these difficulties. A system was developed, using polyacrylamide microspheres coupled with cell membranes, which permitted the production of an easily stored, standardised antigen source which could be used in subsequent flow cytometric assays. Studies comparing the binding of antibodies to whole cells and cell membrane-coupled microspheres indicate a strong qualitative and quantitative correlation in the expression of surface antigens. It is shown here that membrane antigens can be stored coupled to microspheres for months and still retain good reactivity with the appropriate antibodies. This same technique could be used in studies of antigens other than those of the mammalian cell membrane - for example, membrane antigens of sub-cellular organelles such as the mitochondrion and endoplasmic reticulum, plant or bacterial membrane antigens, or antigens not associated with membranes such as viral protein
Hayden, G., Walker, K., Miller, J., Wotherspoon, J. & Raison, R.L. 1988, 'Simultaneous Cytometric Analysis For The Expression Of Cytoplasmic And Surface-antigens In Activated T-cells', Cytometry, vol. 9, no. 1, pp. 44-51.
View/Download from: UTS OPUS or Publisher's site
A method of two-colour immunofluorescence staining has been developed to allow the simultaneous analysis of both surface and cytoplasmic antigens. This involves the use of direct fluorochrome antibody conjugates for cell-surface antigen staining, followed by cell permeabilization and the staining of cytoplasmic antigens with biotinylated antibodies and streptavidin-fluorochrome conjugates. Fluorochrome-antibody conjugates bound to cell-surface epitopes were found not to be affected by the subsequent permeabilisation and cytoplasmic staining. This method was used to examine the surface phenotype of T cells expressing a cytoplasmic antigen, STA. STA is a unique determinant detected in activated human T cells by the monoclonal antibody K-1-21, which also recognizes a cross-reactive conformation-dependent epitope on human free kappa light chains. Cytometric analysis showed that STA is found in both Leu 2a+ cytotoxic/suppressor T cells and Leu 3a+ helper/inducer T cells but is not induced in the Leu 15+ population which contains suppressor T cells. STA was also shown to be an activation antigen in murine T cells.
Raison, R.L. & Edmundson, A.B. 1987, 'Localization Of An Idiotope On The L-chain Dimer And Intact Igg1 Immunoglobulin From The Patient Mcg', Molecular Immunology, vol. 24, no. 9, pp. 937-943.
View/Download from: Publisher's site
NA
Axiak, S.M., Krishnamoorthy, L., Guinan, J. & Raison, R.L. 1987, 'Quantitation Of Free-kappa-light Chains In Serum And Urine Using A Monoclonal-antibody Based Inhibition Enzyme-linked Immunoassay', Journal Of Immunological Methods, vol. 99, no. 1, pp. 141-147.
View/Download from: Publisher's site
NA
Raison, R.L., Gilbertson, P. & Wotherspoon, J. 1987, 'Cellular-requirements For Mixed Leukocyte Reactivity In The Cyclostome, Eptatretus-stoutii', Immunology And Cell Biology, vol. 65, pp. 183-188.
View/Download from: Publisher's site
NA
Mulligan, S., Walker, K., Axiak, S. & Raison, R.L. 1986, 'Applications Of A Monoclonal-antibody Against Free Kappa Light-chains In B-cell Neoplasms', Australian And New Zealand Journal Of Medicine, vol. 16, no. 1, pp. 129-129.
NA
Gilbertson, P., Wotherspoon, J. & Raison, R.L. 1986, 'Evolutionary Development Of Lymphocyte Heterogeneity - Leukocyte Subpopulations In The Pacific Hagfish', Developmental And Comparative Immunology, vol. 10, no. 1, pp. 1-10.
View/Download from: Publisher's site
NA
Walker, K., Hayden, G., Goodnow, C., Boux, H., Adams, E., Basten, A. & Raison, R.L. 1985, 'Co-expression Of An Epitope On Human Free Kappa-light Chains And On A Cytoplasmic Component In Activated T Cells', Journal Of Immunology, vol. 134, no. 2, pp. 1059-1064.
NA
Raison, R.L. & Boux, H. 1985, 'Conformation Dependence Of A Monoclonal-antibody Defined Epitope On Free Human Kappa-chains', Molecular Immunology, vol. 22, no. 12, pp. 1393-1398.
View/Download from: Publisher's site
NA
Walker, K., Boux, H., Hayden, G., Goodnow, C. & Raison, R.L. 1985, 'A Monoclonal-antibody With Selectivity For Human Kappa-myeloma And Lymphoma-cells Which Has Potential As A Therapeutic Agent', Advances In Experimental Medicine And Biology, vol. 186, pp. 833-841.
NA
Goodnow, C. & Raison, R.L. 1985, 'Structural-analysis Of The Myeloma-associated Membrane Antigen Kma', Journal Of Immunology, vol. 135, no. 2, pp. 1276-1280.
NA
Hambly, B.D., Cameron, S., Raison, R.L. & Dosremedios, C. 1985, 'A Potentially Useful Probe For Structural Studies On Actin - Monoclonal-antibodies Directed Against 2 Isotypes Of Actin', Journal Of Muscle Research And Cell Motility, vol. 6, no. 1, pp. 72-73.
NA
Britton, W.J., Hellqvist, L., Basten, A. & Raison, R.L. 1985, 'Mycobacterium-leprae Antigens Involved In Human Immune-responses .1. Identification Of 4 Antigens By Monoclonal-antibodies', Journal Of Immunology, vol. 135, no. 6, pp. 4171-4177.
NA
Prentice, R., Krilis, S. & Raison, R.L. 1985, 'Purification Of 2 Allergens From Dermatophagoides Pteronyssinus Using Monoclonal-antibodies', Molecular Immunology, vol. 22, no. 9, pp. 1131-1134.
View/Download from: Publisher's site
NA
Raison, R.L., Walker, K., Hayden, G. & Goodnow, C. 1984, 'Tumor Recognition And Targeting With Monoclonal-antibodies', Proceedings Of The Australian Biochemical Society, vol. 16, pp. 1-1.
NA
Walker, K., Boux, H., Goodnow, C., Hayden, G. & Raison, R.L. 1984, 'The Membrane Expression Of Non-heavy Chain Associated Kappa Light-chains In Normal And Malignant B-cells', Immunobiology, vol. 167, no. 1-3, pp. 25-25.
NA
Hayden, G., Raison, R.L., Goodnow, C. & Walker, K. 1984, 'A Cytoplasmic Determinant Present In Certain Transformed Or Activated T-cells Is Recognized By A Monoclonal-antibody To Free Kappa-chains', Immunobiology, vol. 167, no. 1-3, pp. 244-244.
NA
Hambly, B.D., Dosremedios, C. & Raison, R.L. 1984, 'Monoclonal-antibodies To Purified G-actin', Journal Of Anatomy, vol. 139, no. AUG, pp. 191-192.
NA
Boux, H., Raison, R.L., Walker, K., Musgrove, E. & Basten, A. 1984, 'The Surface Expression Of A Tumor-associated Antigen On Human Kappa-myeloma Cells', European Journal Of Immunology, vol. 14, no. 3, pp. 216-222.
View/Download from: Publisher's site
NA
Raison, R.L. & Hildemann, W. 1984, 'Immunoglobulin Bearing Blood Leukocytes In The Pacific Hagfish', Developmental And Comparative Immunology, vol. 8, no. 1, pp. 99-108.
View/Download from: Publisher's site
NA
Hambly, B.D., Raison, R.L. & Dosremedios, C. 1983, 'Monoclonal-antibodies Directed Against Skeletal-muscle Actin', Biochemistry International, vol. 7, no. 6, pp. 739-746.
NA
Reddel, R., Baxter, R.C., Raison, R.L. & Sutherland, R. 1983, 'Immunoreactive Somatomedin-c/igf-1 Production By T47d Human-breast Cancer-cells - A Possible Autocrine System', Clinical And Experimental Pharmacology And Physiology, vol. 10, no. 4, pp. 484-484.
NA
Krilis, S., Baldo, B.A., Raison, R.L., Callard, R. & Basten, A. 1983, 'Standardization Of Antigen-e In Ragweed Pollen Extracts Using A Monoclonal-antibody Based Enzyme-immunoassay', Journal Of Allergy And Clinical Immunology, vol. 71, no. 3, pp. 261-265.
View/Download from: Publisher's site
NA
Boux, H., Raison, R.L., Walker, K., Hayden, G. & Basten, A. 1983, 'A Tumor-associated Antigen Specific For Human Kappa Myeloma Cells', Journal Of Experimental Medicine, vol. 158, no. 5, pp. 1769-1774.
View/Download from: Publisher's site
NA
Raison, R.L., Walker, K., Halnan, C., Briscoe, D. & Basten, A. 1982, 'Loss Of Secretion In Mouse-human Hybrids Need Not Be Due To The Loss Of A Structural Gene', Journal Of Experimental Medicine, vol. 156, no. 5, pp. 1380-1389.
View/Download from: Publisher's site
NA
Mountford, C., Grossman, G., Holmes, K., Osullivan, W., Hampson, A., Raison, R.L. & Webster, R.L. 1982, 'Effect Of Monoclonal Anti-neuraminidase Antibodies On The Kinetic-behavior Of Influenza-virus Neuraminidase', Molecular Immunology, vol. 19, no. 6, pp. 811-816.
View/Download from: Publisher's site
NA
Baxter, R.C., Axiak, S. & Raison, R.L. 1982, 'Monoclonal-antibody Against Human Somatomedin-c Insulin-like Growth Factor-i', Journal Of Clinical Endocrinology And Metabolism, vol. 54, no. 2, pp. 474-476.
View/Download from: Publisher's site
NA
Holmes, K., Hampson, A., Raison, R.L., Webster, R.L., Osullivan, W. & Mountford, C. 1982, 'A Comparison Of 2 Anti-neuraminidase Monoclonal-antibodies By Complement Activation', European Journal Of Immunology, vol. 12, no. 6, pp. 523-526.
View/Download from: Publisher's site
NA