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Cytokine regulation of angiogenesis in breast tumours
Abstracts
I 581
P8 October4: LymphokinedCytokines in Disease Al 6aa
A171c HUMAN INTERFERON ALPHA BLOCKS RELEASE OF NON-ENVELOPED HUMAN IMMUNODEFICIENCY VIRUS PARTICLES Lilia M Babe and Charles S. Craik Khepri Pharmaceuticals Inc. San Francisco, CA 94080, University of California, San Francisco, CA 94143 The effect of human inferferon alpha (hulFNa) on the maturation process of the human immunodeficiency virus has been studied using a mammalian cell hne that produces non-enveloped particles. This monkey cell line efficiently assembles and secretes particles devoid of viral env proteins gp120 and gp41 because the gp/160 gene was deleted from the viral genome. Active viral protease is produced and correctly processes its natural substrates p55gag and pl80gag. These particles contain reverse transcriptase activity and packaged RNA, but are non-infectious. A block in the secretion of these particles is observed in cells treated with hulFNu at concentrations between 100 and 1000 U/ml. This concentration-dependent block in particle secretion is also observed in cells treated with a viral protease inhibitor that block polyprotein processing. Analysis of the intracellular content of capsid proteins in hulFNa treated cells shows no decrease in protein synthesis at levels below 1000 U/ml. Electron microscopy shows hulFNn treated cells are decorated with assembled particles at the cell surface. These particles are either mature or immature in morphology depending on whether or not they also received treatment with a protease inhibitor. These results suggest that the observed block in particle release is independent of capsid precursor processing, and viral envelope protein.
CHEMOKINE EXPRESSION IN RHEUMATOID ARTHRITIS E.N. Fish, E. Robinson, E.C. Keystone, T.Schall. University of Toronto, Canada & DNAX Res. Inst. A central question in rheumatoid arthritis (RA) concerns the mechanism of lymphocyte trafficking by which monocytes and specific T cell subpopulations are recruited into synovial tissues (ST) and fluids (SF). The factors responsible for lymphocyte trafficking to the affected joint may be crucial in orchestrating inflammation. The selective chemoattractant and activation effects of chemokines on leukocytes identifies them as potentially ideal candidates in mediating inflammatory processes in RA. The preferential accumulation of memory T cells in the rheumatoid joint suggests that their recruitment may contribute to the persistent synovitis. Earlier studies from this laboratory demonstrated restricted cytokine expression in the T cell population in RA tissues: IL-2, IL-4, IL-6 and IFN-ygene expression levels were low. Accordingly, we undertook studies to examine gene expression for RANTES, MCP-1 and MIP-IO, in both the T- and non-T cell populations in RA tissues. Our results identified elevated levels of both RANTES and MIP-IB gene expression in circulating RA PB and SF T-cells. By contrast, elevated MCP-1 mRNA levels were detected primarily in the non-T cell populations of the SF and ST samples. These data suggest that specific chemokines may have a central role in the trafficking of reactive molecules involved in immunoregulation and the inflammatory processes in RA.
A171a
Al 73a CYTOKINE BETWEEN MVRIS.
DEPENDANT SCHISTOSOMA
IMMUNE MANSONI
INTERACTIONS AND TRICHVRIS
Curry AJ,. Else K.J *, Jones F., Bancroft A.*, Grencis R.K*, Dunne D. W. Depatt. Pathology, University of Cambridge, Cambridge, U.K. CBZlOP. *Deuatt. Biological Sciences, University of Manchester, Manch&ter, U.-K. Ml3 9F?. Polyparasitism is prevalent throughout Schistosome endemic areas. We have therefore investigated whether cytokine production during infection with S.mansoni influences a concurrent infection with another parasite. The characteristic production of Th2 associated cytokines to both parasite specific and non specific antigens in response to egg deposition has been utilised to address this question. Initial experimentation has focused on the murine T.mwis-infection model were expulsion of T.muris worms is dependant on a dominant Th2 response. We demonstrate that susceptible AKR mice (normally eliciting Thl type responses) acquire the ability to expel T.muris when concurrently infected with S.mun.soni. This is associated with altered cytokine production and modulation of T. murk specific IgG2a production. Secondary experiments confvm this to be a cytokine mediated phenomen independant of disease associated pathology. This data is compared to that from our other co-infection studies with Leishmania major.
COMPLEX APPLICATION OF TUMOR NECROSIS FACTOR AND INTERLEUKINE-1 DURING STAPHYU3COCCUS AUREUS INFECTION. Ganova
L.,
Spivak
N.
Institute of Microbiology and Virology of Academy Sciences, Kiev, Ukraine The destroing of the cell immune response has been found in CBA mice with S.aureus (LD50) Treatment animals with optimal. doses of flNF and rIL-1 at 24 h after infection promotes restoring mutual regulation of these cytokines production by cells, tha% leads to enhance of the phagocytes and natural killer cells effectory functions, essential clearance of the bacteria from the organism.
Al aoa
A171b
CELL MEDIATED RESPONSES OF IMMUNISED VERVET MONKEYS TO DEFINED T CELL EPITOPES. Curry, A.J.“, Jardim, A., Olobo, J.O.*, Olafson, R.O. University of Victoria, B.C. Canada.*Institute of Primate Research, Narobi, Kenya. *Present address, University of Cambridge, U.K Ververt monkevs were immunised with killed Leishmania parasites, infected w&t. major and allowed to cute. The proliferation of PBMC from naive and recovered animals and ability to to produce IL2, IL-5 and IFNg following in vitro stimulation was then compared PBMC were stimulated with T and B cell mitogens, killed parasites and peptides (containing defined T cell epitopes) from the major surface glycoprotein GP63. All animals responded to mitogenic stimuli although responses were reduced with PBMC from infected animals. Control animals did not respond to parasites or synthetic peptides. In conaast, recovered animals produced IL-2 and IFNy but not IL-5 in response to parasites. FACS analysis revealed an increase in CD45Ro+ ‘memory’ cells within this population. Various responses were observed from infected (not control) to the different synthetic peptides. The data supports the consensus that synthetic epitopes are prime candidates for molecular defined vaccines.
CYTOKINE REGULATION OF ANGIOGENESIS IN BREAST TUMOURS C. E. Lewis'.R.D. Leek',A.L. Harris*& JO'D.McGee' 'Nuffield l&t. of Pathology, University of Oxford, .John Radcliffe Hospital, Oxford OX3 9DU., 'ICRF Molecular Oncology Labs., Institute of Molecular Medicine, Oxford. OX3 9DU. UK Cytokines produced by the various cell types present within the microenvironment of solid turnours form a cmplex, dynamic network, in which they have multiple effects on tumxx progression. We have combined immunohistochemistry with single-cell assays to investigate the cellular sources and multifaceted role of several key cytokines in the stimulation of a new blood supply within breast carcinomas. Stromal macrophages and malignant cells were found to be the major producer cells for TNFor and its receptors (~55 and ~75) in breast lesions,whilst focal tumour cell islands, endothelial cells and/or occasional macrophages expressed TGF-p and the specific endothelial cell mitogen, VEGF. IFN-I, an angiogenesis inhibitor, was secreted by tumour-associated lymphocytes, but also some malignant cells and macrophages. Here we also report on our work to correlate the cellular distribution of these cytokines with the degree of angiogenesis and metastasis.
I 581
P8 October4: LymphokinedCytokines in Disease Al 6aa
A171c HUMAN INTERFERON ALPHA BLOCKS RELEASE OF NON-ENVELOPED HUMAN IMMUNODEFICIENCY VIRUS PARTICLES Lilia M Babe and Charles S. Craik Khepri Pharmaceuticals Inc. San Francisco, CA 94080, University of California, San Francisco, CA 94143 The effect of human inferferon alpha (hulFNa) on the maturation process of the human immunodeficiency virus has been studied using a mammalian cell hne that produces non-enveloped particles. This monkey cell line efficiently assembles and secretes particles devoid of viral env proteins gp120 and gp41 because the gp/160 gene was deleted from the viral genome. Active viral protease is produced and correctly processes its natural substrates p55gag and pl80gag. These particles contain reverse transcriptase activity and packaged RNA, but are non-infectious. A block in the secretion of these particles is observed in cells treated with hulFNu at concentrations between 100 and 1000 U/ml. This concentration-dependent block in particle secretion is also observed in cells treated with a viral protease inhibitor that block polyprotein processing. Analysis of the intracellular content of capsid proteins in hulFNa treated cells shows no decrease in protein synthesis at levels below 1000 U/ml. Electron microscopy shows hulFNn treated cells are decorated with assembled particles at the cell surface. These particles are either mature or immature in morphology depending on whether or not they also received treatment with a protease inhibitor. These results suggest that the observed block in particle release is independent of capsid precursor processing, and viral envelope protein.
CHEMOKINE EXPRESSION IN RHEUMATOID ARTHRITIS E.N. Fish, E. Robinson, E.C. Keystone, T.Schall. University of Toronto, Canada & DNAX Res. Inst. A central question in rheumatoid arthritis (RA) concerns the mechanism of lymphocyte trafficking by which monocytes and specific T cell subpopulations are recruited into synovial tissues (ST) and fluids (SF). The factors responsible for lymphocyte trafficking to the affected joint may be crucial in orchestrating inflammation. The selective chemoattractant and activation effects of chemokines on leukocytes identifies them as potentially ideal candidates in mediating inflammatory processes in RA. The preferential accumulation of memory T cells in the rheumatoid joint suggests that their recruitment may contribute to the persistent synovitis. Earlier studies from this laboratory demonstrated restricted cytokine expression in the T cell population in RA tissues: IL-2, IL-4, IL-6 and IFN-ygene expression levels were low. Accordingly, we undertook studies to examine gene expression for RANTES, MCP-1 and MIP-IO, in both the T- and non-T cell populations in RA tissues. Our results identified elevated levels of both RANTES and MIP-IB gene expression in circulating RA PB and SF T-cells. By contrast, elevated MCP-1 mRNA levels were detected primarily in the non-T cell populations of the SF and ST samples. These data suggest that specific chemokines may have a central role in the trafficking of reactive molecules involved in immunoregulation and the inflammatory processes in RA.
A171a
Al 73a CYTOKINE BETWEEN MVRIS.
DEPENDANT SCHISTOSOMA
IMMUNE MANSONI
INTERACTIONS AND TRICHVRIS
Curry AJ,. Else K.J *, Jones F., Bancroft A.*, Grencis R.K*, Dunne D. W. Depatt. Pathology, University of Cambridge, Cambridge, U.K. CBZlOP. *Deuatt. Biological Sciences, University of Manchester, Manch&ter, U.-K. Ml3 9F?. Polyparasitism is prevalent throughout Schistosome endemic areas. We have therefore investigated whether cytokine production during infection with S.mansoni influences a concurrent infection with another parasite. The characteristic production of Th2 associated cytokines to both parasite specific and non specific antigens in response to egg deposition has been utilised to address this question. Initial experimentation has focused on the murine T.mwis-infection model were expulsion of T.muris worms is dependant on a dominant Th2 response. We demonstrate that susceptible AKR mice (normally eliciting Thl type responses) acquire the ability to expel T.muris when concurrently infected with S.mun.soni. This is associated with altered cytokine production and modulation of T. murk specific IgG2a production. Secondary experiments confvm this to be a cytokine mediated phenomen independant of disease associated pathology. This data is compared to that from our other co-infection studies with Leishmania major.
COMPLEX APPLICATION OF TUMOR NECROSIS FACTOR AND INTERLEUKINE-1 DURING STAPHYU3COCCUS AUREUS INFECTION. Ganova
L.,
Spivak
N.
Institute of Microbiology and Virology of Academy Sciences, Kiev, Ukraine The destroing of the cell immune response has been found in CBA mice with S.aureus (LD50) Treatment animals with optimal. doses of flNF and rIL-1 at 24 h after infection promotes restoring mutual regulation of these cytokines production by cells, tha% leads to enhance of the phagocytes and natural killer cells effectory functions, essential clearance of the bacteria from the organism.
Al aoa
A171b
CELL MEDIATED RESPONSES OF IMMUNISED VERVET MONKEYS TO DEFINED T CELL EPITOPES. Curry, A.J.“, Jardim, A., Olobo, J.O.*, Olafson, R.O. University of Victoria, B.C. Canada.*Institute of Primate Research, Narobi, Kenya. *Present address, University of Cambridge, U.K Ververt monkevs were immunised with killed Leishmania parasites, infected w&t. major and allowed to cute. The proliferation of PBMC from naive and recovered animals and ability to to produce IL2, IL-5 and IFNg following in vitro stimulation was then compared PBMC were stimulated with T and B cell mitogens, killed parasites and peptides (containing defined T cell epitopes) from the major surface glycoprotein GP63. All animals responded to mitogenic stimuli although responses were reduced with PBMC from infected animals. Control animals did not respond to parasites or synthetic peptides. In conaast, recovered animals produced IL-2 and IFNy but not IL-5 in response to parasites. FACS analysis revealed an increase in CD45Ro+ ‘memory’ cells within this population. Various responses were observed from infected (not control) to the different synthetic peptides. The data supports the consensus that synthetic epitopes are prime candidates for molecular defined vaccines.
CYTOKINE REGULATION OF ANGIOGENESIS IN BREAST TUMOURS C. E. Lewis'.R.D. Leek',A.L. Harris*& JO'D.McGee' 'Nuffield l&t. of Pathology, University of Oxford, .John Radcliffe Hospital, Oxford OX3 9DU., 'ICRF Molecular Oncology Labs., Institute of Molecular Medicine, Oxford. OX3 9DU. UK Cytokines produced by the various cell types present within the microenvironment of solid turnours form a cmplex, dynamic network, in which they have multiple effects on tumxx progression. We have combined immunohistochemistry with single-cell assays to investigate the cellular sources and multifaceted role of several key cytokines in the stimulation of a new blood supply within breast carcinomas. Stromal macrophages and malignant cells were found to be the major producer cells for TNFor and its receptors (~55 and ~75) in breast lesions,whilst focal tumour cell islands, endothelial cells and/or occasional macrophages expressed TGF-p and the specific endothelial cell mitogen, VEGF. IFN-I, an angiogenesis inhibitor, was secreted by tumour-associated lymphocytes, but also some malignant cells and macrophages. Here we also report on our work to correlate the cellular distribution of these cytokines with the degree of angiogenesis and metastasis.