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A chronic model of arthritis supported by a strain-specific periarticular lymph node in BALB/c mice.

Baddack U, Hartmann S, Bang H, Grobe J, Loddenkemper C, Lipp M, Müller G - Nat Commun (2013)

Bottom Line: However, BALB/c mice generate significantly higher titres of anticollagen and anticitrullinated peptide antibodies, show a stronger progressive joint destruction, and in the chronic phase the disease spreads between joints.Concomitant to the observation of a more severe pathology, we discovered a previously undescribed small periarticular lymph node in close proximity to the knee joint of BALB/c mice, which acts as the primary draining lymph node for the synovial cavity.Our model more closely reflects the pathology of rheumatoid arthritis than classical models of arthritis and is hence particularly suitable for further studies of disease pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Max-Delbrück-Center of Molecular Medicine, Department of Tumor Genetics and Immunogenetics, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.

ABSTRACT
Current animal models of arthritis only partially reflect the complexity of rheumatoid arthritis and typically lack either chronicity or autoantibody formation. Here we describe a model that combines features of antigen-induced arthritis and collagen-induced arthritis, which can be efficiently induced in BALB/c and C57BL/6 mice. However, BALB/c mice generate significantly higher titres of anticollagen and anticitrullinated peptide antibodies, show a stronger progressive joint destruction, and in the chronic phase the disease spreads between joints. Concomitant to the observation of a more severe pathology, we discovered a previously undescribed small periarticular lymph node in close proximity to the knee joint of BALB/c mice, which acts as the primary draining lymph node for the synovial cavity. Our model more closely reflects the pathology of rheumatoid arthritis than classical models of arthritis and is hence particularly suitable for further studies of disease pathogenesis.

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ACIA leads to chronic autoimmune arthritis in BALB/c mice.(a) Time line of immunization procedure and development of arthritis. CFA, Freund’s complete adjuvant, ICFA, Freund’s incomplete adjuvant, mBSA, methylated bovine serum albumine, CII, bovine collagen type II. (b) Disease chronicity (left) and joint destruction (right) were assessed by scoring histology sections of BALB/c and C57BL/6 mice 5–8 weeks after arthritis induction. Statistics: Mann–Whitney-U-test, n=12–21, White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. (c) HE-stained sections of paraffin-embedded knee joints from healthy (left) and arthritic BALB/c mice in the acute (middle) or chronic (right) inflammation phase. Black bars, 200 μm. (d) Arthritis-specific antibody production was higher in BALB/c than in C57BL/6 mice. Serum was tested at different time points in disease development for (auto)antibodies directed against the inducing agent mBSA and murine Collagen II using ELISA. Time points: healthy untreated, 4 days after first immunization (−17 days), 4 days after second immunization (−10 days), acute phase (4 days after induction), transition phase (10 days after induction), early chronic phase (3 weeks after induction) and late chronic phase (8–9 weeks after induction). White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per time point and group. Statistics: Mann–Whitney-U-test. (e) Generation of mBSA- and Collagen II-specific CD4+ T cells by BALB/c and C57BL/6 mice in ACIA. Cells from the draining lymph nodes of immunized (+) or control (−) mice were stained with E-Fluor 670 and stimulated with mBSA or CII for 3 days. Cell proliferation was analysed by flow cytometry. Proliferating antigen-specific cells are indicated as percentages of CD4+ lymphocytes. White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per group. Statistics: Mann–Whitney-U-test (n.s., not significant, OD, optical density, *P<0.05, **P<0.01, ***P<0.001).
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f1: ACIA leads to chronic autoimmune arthritis in BALB/c mice.(a) Time line of immunization procedure and development of arthritis. CFA, Freund’s complete adjuvant, ICFA, Freund’s incomplete adjuvant, mBSA, methylated bovine serum albumine, CII, bovine collagen type II. (b) Disease chronicity (left) and joint destruction (right) were assessed by scoring histology sections of BALB/c and C57BL/6 mice 5–8 weeks after arthritis induction. Statistics: Mann–Whitney-U-test, n=12–21, White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. (c) HE-stained sections of paraffin-embedded knee joints from healthy (left) and arthritic BALB/c mice in the acute (middle) or chronic (right) inflammation phase. Black bars, 200 μm. (d) Arthritis-specific antibody production was higher in BALB/c than in C57BL/6 mice. Serum was tested at different time points in disease development for (auto)antibodies directed against the inducing agent mBSA and murine Collagen II using ELISA. Time points: healthy untreated, 4 days after first immunization (−17 days), 4 days after second immunization (−10 days), acute phase (4 days after induction), transition phase (10 days after induction), early chronic phase (3 weeks after induction) and late chronic phase (8–9 weeks after induction). White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per time point and group. Statistics: Mann–Whitney-U-test. (e) Generation of mBSA- and Collagen II-specific CD4+ T cells by BALB/c and C57BL/6 mice in ACIA. Cells from the draining lymph nodes of immunized (+) or control (−) mice were stained with E-Fluor 670 and stimulated with mBSA or CII for 3 days. Cell proliferation was analysed by flow cytometry. Proliferating antigen-specific cells are indicated as percentages of CD4+ lymphocytes. White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per group. Statistics: Mann–Whitney-U-test (n.s., not significant, OD, optical density, *P<0.05, **P<0.01, ***P<0.001).

Mentions: BALB/c and C57BL/6 mice were immunized with mBSA in CFA and 1 week later with mBSA and bovine CII in Freund’s incomplete adjuvant. In addition, mice received Bordetella pertussis toxin (PTx) intraperitoneally at both time points to enhance the immune response. Arthritis was induced by a single injection of mBSA into the left knee-joint cavity (Fig. 1a). This combined AIA/CIA (ACIA) protocol induces a strong inflammation of the knee joint characterized by fibrinous exudates and an influx of neutrophils into the synovial cavity within the first 4 days after arthritis induction (Fig. 1c). After 4–7 days, fibrin exudation ceases and synovial hyperplasia with a massive infiltration of mononuclear cells can be observed. Eventually, arthritis passes into the chronic phase 3–4 weeks after induction. Chronic inflammation was characterized by dense clusters of lymphocyte infiltrates in the synovial and parasynovial tissue, chronic fibrosis and cartilage, as well as bone erosion (Fig. 1c, right panel). The severity of chronic inflammation and joint destruction was evaluated by semiquantitative scoring of hematoxylin and eosin (HE)-stained paraffin sections of the left, that is, induced knee joint. The score for chronic inflammation reflects synovial hyperplasia, infiltration of the synovium by mononuclear cells and fibrosis. Joint destruction scores are based on pannus formation and cartilage and bone erosion. The histological analysis revealed no significant differences between BALB/c and C57BL/6 mice during the acute phase of inflammation; however, BALB/c mice showed significantly elevated scores for chronic arthritis compared with C57BL/6 animals. In particular, we observed joint destruction with massive bone erosion and calcification of cartilage tissue only in BALB/c but not in C57BL/6 mice (Fig. 1b).


A chronic model of arthritis supported by a strain-specific periarticular lymph node in BALB/c mice.

Baddack U, Hartmann S, Bang H, Grobe J, Loddenkemper C, Lipp M, Müller G - Nat Commun (2013)

ACIA leads to chronic autoimmune arthritis in BALB/c mice.(a) Time line of immunization procedure and development of arthritis. CFA, Freund’s complete adjuvant, ICFA, Freund’s incomplete adjuvant, mBSA, methylated bovine serum albumine, CII, bovine collagen type II. (b) Disease chronicity (left) and joint destruction (right) were assessed by scoring histology sections of BALB/c and C57BL/6 mice 5–8 weeks after arthritis induction. Statistics: Mann–Whitney-U-test, n=12–21, White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. (c) HE-stained sections of paraffin-embedded knee joints from healthy (left) and arthritic BALB/c mice in the acute (middle) or chronic (right) inflammation phase. Black bars, 200 μm. (d) Arthritis-specific antibody production was higher in BALB/c than in C57BL/6 mice. Serum was tested at different time points in disease development for (auto)antibodies directed against the inducing agent mBSA and murine Collagen II using ELISA. Time points: healthy untreated, 4 days after first immunization (−17 days), 4 days after second immunization (−10 days), acute phase (4 days after induction), transition phase (10 days after induction), early chronic phase (3 weeks after induction) and late chronic phase (8–9 weeks after induction). White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per time point and group. Statistics: Mann–Whitney-U-test. (e) Generation of mBSA- and Collagen II-specific CD4+ T cells by BALB/c and C57BL/6 mice in ACIA. Cells from the draining lymph nodes of immunized (+) or control (−) mice were stained with E-Fluor 670 and stimulated with mBSA or CII for 3 days. Cell proliferation was analysed by flow cytometry. Proliferating antigen-specific cells are indicated as percentages of CD4+ lymphocytes. White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per group. Statistics: Mann–Whitney-U-test (n.s., not significant, OD, optical density, *P<0.05, **P<0.01, ***P<0.001).
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f1: ACIA leads to chronic autoimmune arthritis in BALB/c mice.(a) Time line of immunization procedure and development of arthritis. CFA, Freund’s complete adjuvant, ICFA, Freund’s incomplete adjuvant, mBSA, methylated bovine serum albumine, CII, bovine collagen type II. (b) Disease chronicity (left) and joint destruction (right) were assessed by scoring histology sections of BALB/c and C57BL/6 mice 5–8 weeks after arthritis induction. Statistics: Mann–Whitney-U-test, n=12–21, White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. (c) HE-stained sections of paraffin-embedded knee joints from healthy (left) and arthritic BALB/c mice in the acute (middle) or chronic (right) inflammation phase. Black bars, 200 μm. (d) Arthritis-specific antibody production was higher in BALB/c than in C57BL/6 mice. Serum was tested at different time points in disease development for (auto)antibodies directed against the inducing agent mBSA and murine Collagen II using ELISA. Time points: healthy untreated, 4 days after first immunization (−17 days), 4 days after second immunization (−10 days), acute phase (4 days after induction), transition phase (10 days after induction), early chronic phase (3 weeks after induction) and late chronic phase (8–9 weeks after induction). White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per time point and group. Statistics: Mann–Whitney-U-test. (e) Generation of mBSA- and Collagen II-specific CD4+ T cells by BALB/c and C57BL/6 mice in ACIA. Cells from the draining lymph nodes of immunized (+) or control (−) mice were stained with E-Fluor 670 and stimulated with mBSA or CII for 3 days. Cell proliferation was analysed by flow cytometry. Proliferating antigen-specific cells are indicated as percentages of CD4+ lymphocytes. White bars indicate the mean value for BALB/c, grey bars for C57BL/6 samples. Data is representative of two independent experiments with 8–15 animals per group. Statistics: Mann–Whitney-U-test (n.s., not significant, OD, optical density, *P<0.05, **P<0.01, ***P<0.001).
Mentions: BALB/c and C57BL/6 mice were immunized with mBSA in CFA and 1 week later with mBSA and bovine CII in Freund’s incomplete adjuvant. In addition, mice received Bordetella pertussis toxin (PTx) intraperitoneally at both time points to enhance the immune response. Arthritis was induced by a single injection of mBSA into the left knee-joint cavity (Fig. 1a). This combined AIA/CIA (ACIA) protocol induces a strong inflammation of the knee joint characterized by fibrinous exudates and an influx of neutrophils into the synovial cavity within the first 4 days after arthritis induction (Fig. 1c). After 4–7 days, fibrin exudation ceases and synovial hyperplasia with a massive infiltration of mononuclear cells can be observed. Eventually, arthritis passes into the chronic phase 3–4 weeks after induction. Chronic inflammation was characterized by dense clusters of lymphocyte infiltrates in the synovial and parasynovial tissue, chronic fibrosis and cartilage, as well as bone erosion (Fig. 1c, right panel). The severity of chronic inflammation and joint destruction was evaluated by semiquantitative scoring of hematoxylin and eosin (HE)-stained paraffin sections of the left, that is, induced knee joint. The score for chronic inflammation reflects synovial hyperplasia, infiltration of the synovium by mononuclear cells and fibrosis. Joint destruction scores are based on pannus formation and cartilage and bone erosion. The histological analysis revealed no significant differences between BALB/c and C57BL/6 mice during the acute phase of inflammation; however, BALB/c mice showed significantly elevated scores for chronic arthritis compared with C57BL/6 animals. In particular, we observed joint destruction with massive bone erosion and calcification of cartilage tissue only in BALB/c but not in C57BL/6 mice (Fig. 1b).

Bottom Line: However, BALB/c mice generate significantly higher titres of anticollagen and anticitrullinated peptide antibodies, show a stronger progressive joint destruction, and in the chronic phase the disease spreads between joints.Concomitant to the observation of a more severe pathology, we discovered a previously undescribed small periarticular lymph node in close proximity to the knee joint of BALB/c mice, which acts as the primary draining lymph node for the synovial cavity.Our model more closely reflects the pathology of rheumatoid arthritis than classical models of arthritis and is hence particularly suitable for further studies of disease pathogenesis.

View Article: PubMed Central - PubMed

Affiliation: Max-Delbrück-Center of Molecular Medicine, Department of Tumor Genetics and Immunogenetics, Robert-Rössle-Strasse 10, 13125 Berlin, Germany.

ABSTRACT
Current animal models of arthritis only partially reflect the complexity of rheumatoid arthritis and typically lack either chronicity or autoantibody formation. Here we describe a model that combines features of antigen-induced arthritis and collagen-induced arthritis, which can be efficiently induced in BALB/c and C57BL/6 mice. However, BALB/c mice generate significantly higher titres of anticollagen and anticitrullinated peptide antibodies, show a stronger progressive joint destruction, and in the chronic phase the disease spreads between joints. Concomitant to the observation of a more severe pathology, we discovered a previously undescribed small periarticular lymph node in close proximity to the knee joint of BALB/c mice, which acts as the primary draining lymph node for the synovial cavity. Our model more closely reflects the pathology of rheumatoid arthritis than classical models of arthritis and is hence particularly suitable for further studies of disease pathogenesis.

Show MeSH
Related in: MedlinePlus