RESUMO
BACKGROUND: Simian immunodeficiency virus (SIV) infection and persistent CD8(+) lymphocyte depletion rapidly leads to encephalitis and neuronal injury. The objective of this study is to confirm that CD8 depletion alone does not induce brain lesions in the absence of SIV infection. METHODS: Four rhesus macaques were monitored by proton magnetic resonance spectroscopy ((1) H-MRS) before and biweekly after anti-CD8 antibody treatment for 8 weeks and compared with four SIV-infected animals. Post-mortem immunohistochemistry was performed on these eight animals and compared with six uninfected, non-CD8-depleted controls. RESULTS: CD8-depleted animals showed stable metabolite levels and revealed no neuronal injury, astrogliosis or microglial activation in contrast to SIV-infected animals. CONCLUSIONS: Alterations observed in MRS and lesions in this accelerated model of neuroAIDS result from unrestricted viral expansion in the setting of immunodeficiency rather than from CD8(+) lymphocyte depletion alone.
Assuntos
Encéfalo/patologia , Linfócitos T CD8-Positivos/patologia , Depleção Linfocítica/veterinária , Macaca mulatta , Síndrome de Imunodeficiência Adquirida dos Símios/patologia , Animais , Anticorpos Monoclonais/metabolismo , Astrócitos/metabolismo , Astrócitos/patologia , Astrócitos/virologia , Encéfalo/metabolismo , Encéfalo/virologia , Linfócitos T CD8-Positivos/metabolismo , Linfócitos T CD8-Positivos/virologia , Modelos Animais de Doenças , Encefalite Viral/imunologia , Encefalite Viral/metabolismo , Encefalite Viral/patologia , Encefalite Viral/veterinária , Citometria de Fluxo/veterinária , Proteína Glial Fibrilar Ácida/metabolismo , Imuno-Histoquímica/veterinária , Espectroscopia de Ressonância Magnética , Proteínas dos Microfilamentos/metabolismo , Microglia/metabolismo , Microglia/patologia , Microglia/virologia , Proteínas Associadas aos Microtúbulos/metabolismo , Doenças dos Macacos/imunologia , Doenças dos Macacos/patologia , Doenças dos Macacos/virologia , Neurônios/metabolismo , Neurônios/patologia , Neurônios/virologia , Prótons , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/virologia , Vírus da Imunodeficiência Símia/fisiologia , Sinaptofisina/metabolismoRESUMO
Six cytokines of the interleukin (IL)-6 family involved in various inflammatory or tumoral diseases share the same gp130 signal transducer chain. We made a panel of anti-gp130 monoclonal antibodies (mAb) to study the structure and function of the gp130 molecule. These mAb recognized different epitopes of the gp130 that we called A to J. Most of the mAb were found to be inhibitors and we studied whether some of them could also induce gp130 activation. When used alone, none of them was able to initiate the proliferation of IL-6-dependent cell lines. However, some particular associations of the mAb were able to induce a proliferative response. mAb B1 could activate the lines in association with F1 or with I2 but not with I1, which in ELISA was similar to I2. In contrast mAb B2, which in ELISA appeared to be very similar to B1, was able to activate the cells in association with I1 but not with F1 or I2. Two other mAb belonging to specificities A and C were found to be activators either in association with I1 only, or with I1 or B2, respectively. These associations of mAb appeared to be nearly as potent activators as IL-6 itself. Although we still have no precise idea of the mechanisms involved, they are interesting tools to study the molecular interactions leading to gp130 activation and, from a practical point of view, valuable growth factors of hematopoietic stem cells.
Assuntos
Anticorpos Monoclonais/imunologia , Antígenos CD/imunologia , Glicoproteínas de Membrana/imunologia , Reações Antígeno-Anticorpo , Divisão Celular , Células Cultivadas , Receptor gp130 de Citocina , Mapeamento de Epitopos , Humanos , Interleucina-6/fisiologia , Transdução de SinaisRESUMO
Agonist antihuman gp130 transducer monoclonal antibodies (MoAbs) were used in SCID mice to grow myeloma cells whose survival and proliferation is dependent on gp130 transducer activation. The agonist anti-gp130 MoAbs neither bound to murine gp130 nor activated murine cells and, as a consequence, did not induce interleukin-6 (IL-6)-related toxicities in mice. They have a 2-week half-life in vivo when injected in the peritoneum. The agonist antibodies made possible the in vivo growth of exogenous IL-6-dependent human myeloma cells as well as that of freshly explanted myeloma cells from 1 patient with secondary plasma cell leukemia. Tumors occurred 4 to 10 weeks after myeloma cell graft and weighed 3 to 5 g. They grew as solid tumors in the peritoneal cavity and metastasized to the different peritoneal organs: liver, pancreas, spleen, and intestine. Tumoral cells were detected in blood and bone marrow of mice grafted with the XG-2 myeloma cells. Tumoral cells grown in SCID mice had kept the phenotypic characteristics of the original tumoral cells and their in vitro growth required the presence of IL-6 or agonist anti-gp130 MoAbs. Myeloma cells from 4 patients with medullary involvement persisted for more than 1 year as judged by detectable circulating human Ig. However, no tumors were detected, suggesting a long-term survival of human myeloma cells without major proliferation. These observations paralleled those made in in vitro cultures as well as the tumor growth pattern in these patients. This gp130 transducer-dependent SCID model of multiple myeloma should be useful to study various therapeutical approaches in multiple myeloma in vivo.
Assuntos
Anticorpos Monoclonais/imunologia , Interleucina-6/imunologia , Mieloma Múltiplo , Neoplasias Experimentais , Animais , Anticorpos Monoclonais/administração & dosagem , Divisão Celular/efeitos dos fármacos , Divisão Celular/imunologia , Modelos Animais de Doenças , Humanos , Interleucina-6/administração & dosagem , Camundongos , Camundongos SCID , Mieloma Múltiplo/imunologia , Mieloma Múltiplo/patologia , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/patologiaRESUMO
The objective of our research was to study the mechanisms of activation of mAb against the gp130 transducer chain common to the IL-6 cytokine family. It has been found that among the 56 anti-gp130 available worldwide, none was able to activate the growth of IL-6-dependent myeloma cell lines. When certain of them were associated in pairs they allowed the cells to grow; alone, they were inhibitory. The same activation was also obtained by cross-linking certain anti-gp130 mAb on the cell membrane with a goat anti-mouse Ig antiserum. A bispecific mAb was prepared by the somatic fusion of two hybridomas secreting two mAb whose association was able to activate gp130 signaling; the bispecific mAb was inactive. The activating mAb were able to support long-term proliferation of the IL-6-dependent myeloma cell lines, which indicates that they are potential valuable growth factors of tumor cells and hematopoietic stem cells. When they were injected into SCID mice, they allowed human IL-6-dependent myeloma cell lines to grow, develop tumors and metastasize. By studying the functional epitopes of the cell membrane gp130 receptors, it was shown that the activating mAb induced gp130 dimerization and STAT3 activation, as did IL-6.