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Recombinant galectin-1 and its genetic delivery suppress collagen-induced arthritis via T cell apoptosis.

Rabinovich GA, Daly G, Dreja H, Tailor H, Riera CM, Hirabayashi J, Chernajovsky Y - J. Exp. Med. (1999)

Bottom Line: This effect was reproduced by daily administration of recombinant GAL-1.GAL-1 treatment resulted in reduction in anticollagen immunoglobulin (Ig)G levels.Moreover, GAL-1-expressing fibroblasts and recombinant GAL-1 revealed a specific dose-dependent inhibitory effect in vitro in antigen-dependent interleukin 2 production to an A(q)-restricted, collagen type 2-specific T cell hybridoma clone.

View Article: PubMed Central - PubMed

Affiliation: Immunology Department, Faculty of Chemical Sciences, National University of Córdoba 5000, Argentina.

ABSTRACT
Galectin-1 (GAL-1), a member of a family of conserved beta-galactoside-binding proteins, has been shown to induce in vitro apoptosis of activated T cells and immature thymocytes. We assessed the therapeutic effects and mechanisms of action of delivery of GAL-1 in a collagen-induced arthritis model. A single injection of syngeneic DBA/1 fibroblasts engineered to secrete GAL-1 at the day of disease onset was able to abrogate clinical and histopathological manifestations of arthritis. This effect was reproduced by daily administration of recombinant GAL-1. GAL-1 treatment resulted in reduction in anticollagen immunoglobulin (Ig)G levels. The cytokine profile in draining lymph node cells and the anticollagen IgG isotypes in mice sera at the end of the treatment clearly showed inhibition of the proinflammatory response and skewing towards a type 2-polarized immune reaction. Lymph node cells from mice engaged in the gene therapy protocol increased their susceptibility to antigen-induced apoptosis. Moreover, GAL-1-expressing fibroblasts and recombinant GAL-1 revealed a specific dose-dependent inhibitory effect in vitro in antigen-dependent interleukin 2 production to an A(q)-restricted, collagen type 2-specific T cell hybridoma clone. Thus, a correlation between the apoptotic properties of GAL-1 in vitro and its immunomodulatory properties in vivo supports its therapeutic potential in the treatment of T helper cell type 1-mediated autoimmune disorders.

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Cytokine profile in draining lymph node cells after gene therapy with mGAL-1. Arthritic DBA/1 mice treated with mGAL-1 transfected cells (white bars) or control transfectants (black bars) were killed on day 12 after onset. Inguinal lymph nodes from two mice with compatible clinical scores were excised and pooled together. Cells (5 × 106 cells/ml) were cultured in triplicate in 96-well plates in the presence of CII (100 μg/ml). After 72 h, supernatants were collected and analyzed for IFN-γ (a) and IL-5 (b) detection by a capture ELISA. Controls included cells cultured in medium alone or with 5 μg/ml Con A (data not shown). Mean values of different groups are indicated (mean ± SEM) as a combination of two independent experiments.
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Figure 5: Cytokine profile in draining lymph node cells after gene therapy with mGAL-1. Arthritic DBA/1 mice treated with mGAL-1 transfected cells (white bars) or control transfectants (black bars) were killed on day 12 after onset. Inguinal lymph nodes from two mice with compatible clinical scores were excised and pooled together. Cells (5 × 106 cells/ml) were cultured in triplicate in 96-well plates in the presence of CII (100 μg/ml). After 72 h, supernatants were collected and analyzed for IFN-γ (a) and IL-5 (b) detection by a capture ELISA. Controls included cells cultured in medium alone or with 5 μg/ml Con A (data not shown). Mean values of different groups are indicated (mean ± SEM) as a combination of two independent experiments.

Mentions: To elucidate whether class switching to IgG1 was correlated with changes in the cytokine secretion pattern, inguinal lymph nodes and spleens were excised at the end of the treatment from GAL-1–treated or untreated mice. Cells were cultured in the presence of CII and supernatants were analyzed after 72 h for IFN-γ (Fig. 5 a) and IL-5 (Fig. 5 b) production. The main differences were found at the level of draining lymph nodes of mice engaged in the gene therapy protocol with GAL-1, where IL-5 raised to mean high levels of 1,800 pg/ml (P < 0.005), in comparison to the lower levels exhibited by arthritic mice, treated with control vector alone (<250 pg/ml). Consistently, gene therapy with GAL-1 resulted in a severe decline in IFN-γ to background levels of <50 pg/ml (P < 0.005), whereas the control group secreted large amounts of this proinflammatory cytokine (>500 pg/ml). No significant differences in cytokine secretion could be detected at the level of spleens between experimental and control groups (data not shown).


Recombinant galectin-1 and its genetic delivery suppress collagen-induced arthritis via T cell apoptosis.

Rabinovich GA, Daly G, Dreja H, Tailor H, Riera CM, Hirabayashi J, Chernajovsky Y - J. Exp. Med. (1999)

Cytokine profile in draining lymph node cells after gene therapy with mGAL-1. Arthritic DBA/1 mice treated with mGAL-1 transfected cells (white bars) or control transfectants (black bars) were killed on day 12 after onset. Inguinal lymph nodes from two mice with compatible clinical scores were excised and pooled together. Cells (5 × 106 cells/ml) were cultured in triplicate in 96-well plates in the presence of CII (100 μg/ml). After 72 h, supernatants were collected and analyzed for IFN-γ (a) and IL-5 (b) detection by a capture ELISA. Controls included cells cultured in medium alone or with 5 μg/ml Con A (data not shown). Mean values of different groups are indicated (mean ± SEM) as a combination of two independent experiments.
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Related In: Results  -  Collection

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Figure 5: Cytokine profile in draining lymph node cells after gene therapy with mGAL-1. Arthritic DBA/1 mice treated with mGAL-1 transfected cells (white bars) or control transfectants (black bars) were killed on day 12 after onset. Inguinal lymph nodes from two mice with compatible clinical scores were excised and pooled together. Cells (5 × 106 cells/ml) were cultured in triplicate in 96-well plates in the presence of CII (100 μg/ml). After 72 h, supernatants were collected and analyzed for IFN-γ (a) and IL-5 (b) detection by a capture ELISA. Controls included cells cultured in medium alone or with 5 μg/ml Con A (data not shown). Mean values of different groups are indicated (mean ± SEM) as a combination of two independent experiments.
Mentions: To elucidate whether class switching to IgG1 was correlated with changes in the cytokine secretion pattern, inguinal lymph nodes and spleens were excised at the end of the treatment from GAL-1–treated or untreated mice. Cells were cultured in the presence of CII and supernatants were analyzed after 72 h for IFN-γ (Fig. 5 a) and IL-5 (Fig. 5 b) production. The main differences were found at the level of draining lymph nodes of mice engaged in the gene therapy protocol with GAL-1, where IL-5 raised to mean high levels of 1,800 pg/ml (P < 0.005), in comparison to the lower levels exhibited by arthritic mice, treated with control vector alone (<250 pg/ml). Consistently, gene therapy with GAL-1 resulted in a severe decline in IFN-γ to background levels of <50 pg/ml (P < 0.005), whereas the control group secreted large amounts of this proinflammatory cytokine (>500 pg/ml). No significant differences in cytokine secretion could be detected at the level of spleens between experimental and control groups (data not shown).

Bottom Line: This effect was reproduced by daily administration of recombinant GAL-1.GAL-1 treatment resulted in reduction in anticollagen immunoglobulin (Ig)G levels.Moreover, GAL-1-expressing fibroblasts and recombinant GAL-1 revealed a specific dose-dependent inhibitory effect in vitro in antigen-dependent interleukin 2 production to an A(q)-restricted, collagen type 2-specific T cell hybridoma clone.

View Article: PubMed Central - PubMed

Affiliation: Immunology Department, Faculty of Chemical Sciences, National University of Córdoba 5000, Argentina.

ABSTRACT
Galectin-1 (GAL-1), a member of a family of conserved beta-galactoside-binding proteins, has been shown to induce in vitro apoptosis of activated T cells and immature thymocytes. We assessed the therapeutic effects and mechanisms of action of delivery of GAL-1 in a collagen-induced arthritis model. A single injection of syngeneic DBA/1 fibroblasts engineered to secrete GAL-1 at the day of disease onset was able to abrogate clinical and histopathological manifestations of arthritis. This effect was reproduced by daily administration of recombinant GAL-1. GAL-1 treatment resulted in reduction in anticollagen immunoglobulin (Ig)G levels. The cytokine profile in draining lymph node cells and the anticollagen IgG isotypes in mice sera at the end of the treatment clearly showed inhibition of the proinflammatory response and skewing towards a type 2-polarized immune reaction. Lymph node cells from mice engaged in the gene therapy protocol increased their susceptibility to antigen-induced apoptosis. Moreover, GAL-1-expressing fibroblasts and recombinant GAL-1 revealed a specific dose-dependent inhibitory effect in vitro in antigen-dependent interleukin 2 production to an A(q)-restricted, collagen type 2-specific T cell hybridoma clone. Thus, a correlation between the apoptotic properties of GAL-1 in vitro and its immunomodulatory properties in vivo supports its therapeutic potential in the treatment of T helper cell type 1-mediated autoimmune disorders.

Show MeSH
Related in: MedlinePlus