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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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BALB/c mice develop an additional paLN in close vicinity to the knee joint.(a) Section of paraffin-embedded hind leg from BALB/c mouse in the early chronic phase of RA with hematoxylin eosin staining. Arrows: periarticular lymph node (paLN) close to the knee joint and popliteal lymph node (pLN) embedded in fat tissue. Black bars, 400 μm. (b) paLN and popliteal lymph node stained for B cells (B220, blue) and T cells (CD3, red). (c) HE staining of cryosections of paLN in healthy untreated BALB/c mice (top) and 10 days after induction of arthritis (bottom). Black bars, 200 μm. (d) paLNs are connected to the vasculature. Biotinylated splenocytes were injected intravenously into arthritic mice. Eighteen hours after transfer, biotinylated cells (in red) could be detected in the spleen (top) and the paLN (bottom). Magnification X40. (e) paLNs are fully functional and develop GCs following arthritis induction. Characterization of the microarchitecture of the paLN from healthy and arthritic BALB/c mice: T cells were stained with peroxidase-labelled anti-CD3 antibody (in red) and several other lymph node and activation markers (in blue). FDC-M2 is a marker for FDCs, Lyve for lymphatic vessels, PNAd for high endothelial venules, PNA for GCs and Ki67 for proliferating cells. Only after arthritis induction, GCs are formed that harbour proliferating cells. The tissue sections represent two independent experiments with four animals per time point. Magnification: X40 for lymph nodes isolated 10 days after induction of arthritis and X100 for lymph nodes from healthy untreated mice. Black bars, 200 μm.
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f2: BALB/c mice develop an additional paLN in close vicinity to the knee joint.(a) Section of paraffin-embedded hind leg from BALB/c mouse in the early chronic phase of RA with hematoxylin eosin staining. Arrows: periarticular lymph node (paLN) close to the knee joint and popliteal lymph node (pLN) embedded in fat tissue. Black bars, 400 μm. (b) paLN and popliteal lymph node stained for B cells (B220, blue) and T cells (CD3, red). (c) HE staining of cryosections of paLN in healthy untreated BALB/c mice (top) and 10 days after induction of arthritis (bottom). Black bars, 200 μm. (d) paLNs are connected to the vasculature. Biotinylated splenocytes were injected intravenously into arthritic mice. Eighteen hours after transfer, biotinylated cells (in red) could be detected in the spleen (top) and the paLN (bottom). Magnification X40. (e) paLNs are fully functional and develop GCs following arthritis induction. Characterization of the microarchitecture of the paLN from healthy and arthritic BALB/c mice: T cells were stained with peroxidase-labelled anti-CD3 antibody (in red) and several other lymph node and activation markers (in blue). FDC-M2 is a marker for FDCs, Lyve for lymphatic vessels, PNAd for high endothelial venules, PNA for GCs and Ki67 for proliferating cells. Only after arthritis induction, GCs are formed that harbour proliferating cells. The tissue sections represent two independent experiments with four animals per time point. Magnification: X40 for lymph nodes isolated 10 days after induction of arthritis and X100 for lymph nodes from healthy untreated mice. Black bars, 200 μm.

Mentions: A histological analysis of the chronically inflamed knees with their surrounding tissue revealed the existence of a lymph node-like structure in close proximity to knee joints in BALB/c mice (Fig. 2a) that was not detectable in C57BL/6 mice. These periarticular structures were smaller in size than the popliteal lymph node and also present in healthy BALB/c animals, albeit at a smaller size (Fig. 2c). We next asked whether these structures are fully accessible to lymphocytes via the circulation. To test this, we transferred biotin-labelled splenocytes intravenously into arthritic animals and analysed the spleens and periarticular structures 18 h later for the presence of the transferred cells. Splenocytes were able to access not only the spleen but also the periarticular nodes, indicating that these follicles are fully functional lymph nodes (Fig. 2d). Finally, we characterized these structures for typical features of secondary lymphoid organs such as T/B compartmentalization and GC formation. Double staining of paraffin sections for T cells (CD3, red) and B cells (B220, blue) revealed a clear T-cell—B-cell segregation both in naive and arthritic animals (Fig. 2e). Both groups of mice showed networks of follicular dendritic cells (FDCs) within the B cell zones (FDC-M2, blue), lymphatic vessels (LYVE-1, blue) and high endothelial venules (PNAd, blue) in these periarticular structures (Fig. 2e). In contrast, GC formation could only be detected after arthritis induction, by staining with peanut agglutinin (PNA) and for Ki67, a marker for proliferating cells. The number of proliferating cells increased from day 4 on and both GCs and proliferative activity persisted until the end of the period examined. Given the full functionality of a regular lymph node we term the newly discovered periarticular structures paLN.


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)

BALB/c mice develop an additional paLN in close vicinity to the knee joint.(a) Section of paraffin-embedded hind leg from BALB/c mouse in the early chronic phase of RA with hematoxylin eosin staining. Arrows: periarticular lymph node (paLN) close to the knee joint and popliteal lymph node (pLN) embedded in fat tissue. Black bars, 400 μm. (b) paLN and popliteal lymph node stained for B cells (B220, blue) and T cells (CD3, red). (c) HE staining of cryosections of paLN in healthy untreated BALB/c mice (top) and 10 days after induction of arthritis (bottom). Black bars, 200 μm. (d) paLNs are connected to the vasculature. Biotinylated splenocytes were injected intravenously into arthritic mice. Eighteen hours after transfer, biotinylated cells (in red) could be detected in the spleen (top) and the paLN (bottom). Magnification X40. (e) paLNs are fully functional and develop GCs following arthritis induction. Characterization of the microarchitecture of the paLN from healthy and arthritic BALB/c mice: T cells were stained with peroxidase-labelled anti-CD3 antibody (in red) and several other lymph node and activation markers (in blue). FDC-M2 is a marker for FDCs, Lyve for lymphatic vessels, PNAd for high endothelial venules, PNA for GCs and Ki67 for proliferating cells. Only after arthritis induction, GCs are formed that harbour proliferating cells. The tissue sections represent two independent experiments with four animals per time point. Magnification: X40 for lymph nodes isolated 10 days after induction of arthritis and X100 for lymph nodes from healthy untreated mice. Black bars, 200 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3644064&req=5

f2: BALB/c mice develop an additional paLN in close vicinity to the knee joint.(a) Section of paraffin-embedded hind leg from BALB/c mouse in the early chronic phase of RA with hematoxylin eosin staining. Arrows: periarticular lymph node (paLN) close to the knee joint and popliteal lymph node (pLN) embedded in fat tissue. Black bars, 400 μm. (b) paLN and popliteal lymph node stained for B cells (B220, blue) and T cells (CD3, red). (c) HE staining of cryosections of paLN in healthy untreated BALB/c mice (top) and 10 days after induction of arthritis (bottom). Black bars, 200 μm. (d) paLNs are connected to the vasculature. Biotinylated splenocytes were injected intravenously into arthritic mice. Eighteen hours after transfer, biotinylated cells (in red) could be detected in the spleen (top) and the paLN (bottom). Magnification X40. (e) paLNs are fully functional and develop GCs following arthritis induction. Characterization of the microarchitecture of the paLN from healthy and arthritic BALB/c mice: T cells were stained with peroxidase-labelled anti-CD3 antibody (in red) and several other lymph node and activation markers (in blue). FDC-M2 is a marker for FDCs, Lyve for lymphatic vessels, PNAd for high endothelial venules, PNA for GCs and Ki67 for proliferating cells. Only after arthritis induction, GCs are formed that harbour proliferating cells. The tissue sections represent two independent experiments with four animals per time point. Magnification: X40 for lymph nodes isolated 10 days after induction of arthritis and X100 for lymph nodes from healthy untreated mice. Black bars, 200 μm.
Mentions: A histological analysis of the chronically inflamed knees with their surrounding tissue revealed the existence of a lymph node-like structure in close proximity to knee joints in BALB/c mice (Fig. 2a) that was not detectable in C57BL/6 mice. These periarticular structures were smaller in size than the popliteal lymph node and also present in healthy BALB/c animals, albeit at a smaller size (Fig. 2c). We next asked whether these structures are fully accessible to lymphocytes via the circulation. To test this, we transferred biotin-labelled splenocytes intravenously into arthritic animals and analysed the spleens and periarticular structures 18 h later for the presence of the transferred cells. Splenocytes were able to access not only the spleen but also the periarticular nodes, indicating that these follicles are fully functional lymph nodes (Fig. 2d). Finally, we characterized these structures for typical features of secondary lymphoid organs such as T/B compartmentalization and GC formation. Double staining of paraffin sections for T cells (CD3, red) and B cells (B220, blue) revealed a clear T-cell—B-cell segregation both in naive and arthritic animals (Fig. 2e). Both groups of mice showed networks of follicular dendritic cells (FDCs) within the B cell zones (FDC-M2, blue), lymphatic vessels (LYVE-1, blue) and high endothelial venules (PNAd, blue) in these periarticular structures (Fig. 2e). In contrast, GC formation could only be detected after arthritis induction, by staining with peanut agglutinin (PNA) and for Ki67, a marker for proliferating cells. The number of proliferating cells increased from day 4 on and both GCs and proliferative activity persisted until the end of the period examined. Given the full functionality of a regular lymph node we term the newly discovered periarticular structures paLN.

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