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Cited by 13Rheumatoid Arthritis Research and TherapiesNatural Compounds in Disease TreatmentSystemic Lupus Erythematosus Research

The oncoprotein TBX3 is controlling severity in experimental arthritis

Open access
Sardar, Samra, Kerr, Alish, Vaartjes, Daniëlle, Moltved, Emilie Riis, Karosiene, Edita, Gupta, Ramneek, Andersson, Åsa·2019·Arthritis Research & Therapy
DOI 10.1186/s13075-018-1797-3PMC PMC6329118
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Abstract

BACKGROUND: Development of autoimmune diseases is the result of a complex interplay between hereditary and environmental factors, with multiple genes contributing to the pathogenesis in human disease and in experimental models for disease. The T-box protein 3 is a transcriptional repressor essential during early embryonic development, in the formation of bone and additional organ systems, and in tumorigenesis. METHODS: With the aim to find novel genes important for autoimmune inflammation, we have performed genetic studies of collagen-induced arthritis (CIA), a mouse experimental model for rheumatoid arthritis. RESULTS: We showed that a small genetic fragment on mouse chromosome 5, including Tbx3 and three additional protein-coding genes, is linked to severe arthritis and high titers of anti-collagen antibodies. Gene expression studies have revealed differential expression of Tbx3 in B cells, where low expression was accompanied by a higher B cell response upon B cell receptor stimulation in vitro. Furthermore, we showed that serum TBX3 levels rise concomitantly with increasing severity of CIA. CONCLUSIONS: From these results, we suggest that TBX3 is a novel factor important for the regulation of gene transcription in the immune system and that genetic polymorphisms, resulting in lower expression of Tbx3, are contributing to a more severe form of CIA and high titers of autoantibodies. We also propose TBX3 as a putative diagnostic biomarker for rheumatoid arthritis.

AI snapshot

Written from the paper’s full text.

TL;DR

Fine-mapping of the Eae39r locus on mouse chromosome 5 identified a 1.4-Mbp Eae39r2 fragment containing Tbx3, Tbx5, and Rbm19 that enhances collagen-induced arthritis severity and anti-collagen antibody responses. The authors propose TBX3 as a novel regulator of B cell activation and a putative rheumatoid arthritis biomarker.

Key findings

  • B10.RIIIS/J-Eae39r2 sub-congenic mice developed more severe CIA than littermate controls: mean maximum score 8.30 ± 1.14 vs 4.4 ± 0.8 (p = 0.0118), AUC 67.8 ± 17.6 vs 25.7 ± 7.2 (p = 0.0134), and 5/10 (50%) vs 2/19 (10.5%) euthanized (p = 0.0302). The larger Eae39r congenic line also showed higher severity: mean maximum score 9.7 ± 1.6 vs 3.9 ± 1.3 (p = 0.0041).
  • Anti-CII antibody titers were significantly higher in Eae39r2 sub-congenic mice at day 15 and day 72 for IgG1, IgG2C, IgG3, and IgM (early only); for example, day 72 IgG1 was 5044 ± 1363 vs 1852 ± 486 AU/ml (p = 0.0002), and IgG2C was 822 ± 113 vs 323 ± 45 AU/ml (p < 0.0001).
  • Tbx3 mRNA was significantly lower in spleens (p = 0.0036) and purified splenic B lymphocytes (p = 0.0286) of Eae39r2 sub-congenic mice versus controls; Tbx5 was not detectable in spleen, and joint Tbx3/Tbx5 expression did not differ.
  • B cells from sub-congenic mice proliferated more upon anti-IgM stimulation (p = 0.0317) but not upon LPS or anti-CD40/IL-4; active TBX3 fell after BCR stimulation in vitro (p = 0.0286), and serum TBX3 rose in clinical CIA, with arthritic mice higher than immunized non-arthritic mice (p = 0.0087) and naïve mice (p = 0.0095); serum TBX3 correlated with clinical score (rs = 0.62, n = 24, p = 0.0012).

Methods

  • Genetic fine-mapping used B10.RIIIS/J-Eae39r congenic and Eae39r2 sub-congenic mice on the CIA-susceptible B10.RIII background, with littermate controls; CIA was induced with bovine type II collagen in incomplete Freund’s adjuvant and scored clinically. Sample sizes included Eae39r congenic n = 11 vs controls n = 15, and Eae39r2 sub-congenic n = 10 vs controls n = 19.
  • A separate time-course CIA study used 24 male B10.RIII mice (20 immunized, 4 unimmunized), with sampling at pre-clinical, onset/progressive, and late phases.
  • Analyses included Sanger sequencing and bioinformatic prediction of regulatory variants, anti-CII isotype ELISAs, B and T cell isolation and proliferation assays, RT-PCR for Tbx3/Tbx5, a TBX3 DNA-binding activity assay, and serum TBX3 ELISA; statistics used Mann-Whitney U, chi-square/Fisher’s exact, and two-way Student’s t tests.

Limitations

  • The paper states that the biomarker study is limited by group size and by being performed in an experimental model, and that future studies in human cohorts are needed to prove TBX3’s biomarker potential.
  • The paper notes that involvement of non-coding genes within Eae39r2 cannot be disregarded, and that the mechanism linking reduced Tbx3 expression to increased B cell responses was not directly demonstrated.
  • Evident from the text: the Eae39r2 fragment contains multiple protein-coding genes (Tbx3, Tbx5, Rbm19) plus non-coding elements, so the findings support Tbx3 as a candidate but do not establish it as the sole causal gene; several in vitro and cellular experiments also used small numbers of mice.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1186/s13075-018-1797-3

Cites

81 of 81 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

23 of 41 cited sources checked · 7 supported · 1 flagged · accuracy not yet measured

  • Findingown data

    The SNP rs260388829 is predicted to disrupt a transcription factor binding site for c-Rel and Sp1.

    “Disruption of TFBS for c-Rel (member of NF-ĸB TF family, linked to CIA development, and lymphoid cell growth and development [86]) and Sp1 (involved in cellular processes including growth, differentiation and apoptosis)”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [86] Campbell IK, Gerondakis S, O’Donnell K, Wicks IP. Distinct roles for the NF-kappaB1 (p50) and c-Rel transcription factors in inflammatory arthritis. J Clin Invest. 2000;105(12):1799–806. source

      The Introduction identifies c-Rel as a Rel/NF-κB transcription factor and reports that c-Rel-deficient mice largely failed to develop collagen-induced arthritis. It also describes defects in mature B- and T-cell activation, but does not establish a role in lymphoid-cell development. The citation is attached to this description of c-Rel, not to the SNP prediction or the statement about Sp1. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
  • Premisecitation

    B10.RIIIS/J-Eae39r1 sub-congenic mice (with only Med13l in the sub-congenic fragment) develop arthritis to the same extent as wild-type B10.RIII mice.

    “We have previously shown that B10.RIIIS/J-Eae39r1 sub-congenic mice (with only Med13l in the sub-congenic fragment) develop arthritis to the same extent as the wild-type B10.RIII mice [46].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [46] Sardar S, Kanne K, Andersson Å. Analysis of polymorphisms in the Mediator complex subunit 13-like (Med13L) gene in the context of immune function and development of experimental arthritis. Arch Immunol Ther Exp. 2018;66(5):365–77. source
  • Premisecitation

    The time point of lowest TBX3 activity corresponds to the time point reported for the highest B cell activation and proliferation of CII-specific IgM-positive B cells in the CIA model.

    “which corresponds to the time point reported for the highest B cell activation and proliferation of CII-specific IgM-positive B cells in the CIA model [64].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [64] Palm A-KE, Friedrich HC, Mezger A, Salomonsson M, Myers LK, Kleinau S. Function and regulation of self-reactive marginal zone B cells in autoimmune arthritis. Cell Mol Immunol. 2015;12(4):493–504. source

      The Results (page unavailable) report that CII-reactive IgM-positive marginal-zone B cells expanded after immunization and peaked in number on day 12. That supports a peak in the B-cell response, but the paper does not report that activation and proliferation themselves were highest at that time point. The comparison with TBX3 activity is the citing authors’ own framing, not a finding attributed to this source. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
  • Premisecitation

    Tissue-specific regulation of Tbx3 has been previously documented in mice and humans.

    “This can be explained by tissue-specific regulation of Tbx3 that has been previously documented in mice [18] and humans [18, 25].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [18] Washkowitz AJ, Gavrilov S, Begum S, Papaioannou VE. Diverse functional networks of Tbx3 in development and disease. Wiley Interdiscip Rev Syst Biol Med. 2012;4(3):273–83. source

      The Abstract (page not available) describes Tbx3 expression in specific mouse tissues, and the limb-development section describes spatial regulation of that expression. The paper also reports tissue-specific effects of human TBX3 mutations, but those effects do not establish tissue-specific regulation of TBX3 in humans. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [25] Willmer T, Cooper A, Sims D, Govender D, Prince S. The T-box transcription factor 3 is a promising biomarker and a key regulator of the oncogenic phenotype of a diverse range of sarcoma subtypes. Oncogene. 2016;5(2):e199. source

      The Results and Discussion (page unavailable) show that TBX3 has different effects across human sarcoma cell types, with opposite effects on tumour growth in mouse xenografts. They do not document tissue-specific regulation of Tbx3 expression in mice and humans generally; the mouse experiments test implanted tumour cells. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
  • Findingown data

    Computational analysis predicted that some of the identified variations in Tbx3 and Tbx5 might affect promoter function, transcription factor binding, or microRNA binding.

    “Computational analysis of these variations predicted that some of these might affect promoter function, transcription factor binding, or microRNA (miRNA) binding (Table 4).”
    Quote in the paperNot a substantial claim
  • May combine two claims. 27
  • Classified as probably not a substantial claim. 14

  • May combine two claims.

    Premisecitation

    The Eae39 locus on mouse chromosome 5 was previously linked to experimental autoimmune encephalomyelitis and also influences collagen-induced arthritis development and severity.

    “The genetic locus Eae39, located on mouse chromosome 5, was previously linked to experimental autoimmune encephalomyelitis (EAE), an experimental model for multiple sclerosis. Subsequently, it was also shown to influence CIA development and severity [8, 10].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [8] Lindvall T, Karlsson J, Holmdahl R, Andersson Å. Dissection of a locus on mouse chromosome 5 reveals arthritis promoting and inhibitory genes. Arthritis Res Ther. 2009;11(1):R10. source
    • cites [10] Karlsson J, Zhao X, Lonskaya I, Neptin M, Holmdahl R, Andersson Å. Novel quantitative trait loci controlling development of experimental autoimmune encephalomyelitis and proportion of lymphocyte subpopulations. J Immunol. 2003;170(2):1019–26. source

      The Linkage analysis section identifies the chromosome 5 EAE locus as Eae26, not Eae39, and the Discussion says no Eae locus had previously been reported on mouse chromosome 5. The Discussion mentions other arthritis loci on chromosome 5 but does not link Eae26 to collagen-induced arthritis development or severity. Page numbers are unavailable in our copy. (LLM, conf=0.88)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TBX3 functions predominantly as a transcriptional repressor.

    “TBX3 functions predominantly as a transcriptional repressor.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [17] Carlson H, Ota S, Campbell CE, Hurlin PJ. A dominant repression domain in Tbx3 mediates transcriptional repression and cell immortalization: relevance to mutations in Tbx3 that cause ulnar-mammary syndrome. Hum Mol Genet. 2001;10(21):2403–13. source
    • cites [18] Washkowitz AJ, Gavrilov S, Begum S, Papaioannou VE. Diverse functional networks of Tbx3 in development and disease. Wiley Interdiscip Rev Syst Biol Med. 2012;4(3):273–83. source

      The Conclusion (page unavailable) describes a transcriptional repression module, and the cancer section gives examples of genes TBX3 represses. But the Abstract and Conclusion also say TBX3 can activate transcription depending on context; the paper does not establish that repression is its predominant function. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    The B10.RIII mouse strain develops collagen-induced arthritis after immunization with bovine collagen type II, whereas the MHC-matched RIIIS/J mouse strain is resistant to arthritis development.

    “After immunization with bovine collagen type II (CII), the B10.RIII mouse strain develops CIA, whereas the MHC-matched RIIIS/J mouse strain is resistant to arthritis development.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [7] Jirholt J, Cook A, Emahazion T, Sundvall M, Jansson L, Nordquist N, et al. Genetic linkage analysis of collagen-induced arthritis in the mouse. Eur J Immunol. 1998;28:3321–8. source
    • cites [8] Lindvall T, Karlsson J, Holmdahl R, Andersson Å. Dissection of a locus on mouse chromosome 5 reveals arthritis promoting and inhibitory genes. Arthritis Res Ther. 2009;11(1):R10. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Both TBX3 and TBX5 are involved in bone development and remodeling.

    “Among other functions, both proteins are involved in bone development [18–20] and remodeling [21].”
    Quote in the paperNot a substantial claim3 of 4 sources checked
    • cites [18] Washkowitz AJ, Gavrilov S, Begum S, Papaioannou VE. Diverse functional networks of Tbx3 in development and disease. Wiley Interdiscip Rev Syst Biol Med. 2012;4(3):273–83. source

      The 'Tbx3 in limb development' section reports that Tbx3-null mice have missing or abnormal forelimb elements, supporting a role for TBX3 in skeletal development. Our copy does not establish a comparable role for TBX5 or address bone remodeling; its reference to remodeling concerns the heart. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [19] Chapman DL, Garvey N, Hancock S, Alexiou M, Agulnik SI, Gibson-Brown JJ, et al. Expression of the T-box family genes, Tbx1-Tbx5, during early mouse development. Dev Dyn. 1996;206(4):379–90. source
    • cites [20] Bickley SRB, Logan MPO. Regulatory modulation of the T-box gene Tbx5 links development, evolution, and adaptation of the sternum. Proc Natl Acad Sci U S A. 2014;111(50):17917–22. source

      The paper’s section on the early requirement for Tbx5 in sternum development reports that mice lacking Tbx5 fail to form a sternum, supporting a role for TBX5 in skeletal development. It does not establish a role for TBX3. In the citing sentence, remodeling has a separate citation [21], so it is not attributed to this source. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [21] Denninger KCM, Litman T, Marstrand T, Moller K, Svensson L, Labuda T, et al. Kinetics of gene expression and bone remodelling in the clinical phase of collagen-induced arthritis. Arthritis Res Ther. 2015;17:43. source

      The Results and Discussion discuss bone formation and remodelling in collagen-induced arthritis, but the passages found do not address TBX3 or TBX5. Searches for “TBX3 TBX5 bone development remodeling” and “T-box transcription factors Tbx3 Tbx5” found no passage linking either protein to those processes. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Mutations in the human TBX3 and TBX5 genes cause ulnar-mammary syndrome and Holt-Oram syndrome, respectively.

    “Mutations in the human TBX3 and TBX5 genes cause ulnar-mammary syndrome (UMS, OMIM 181450) and Holt-Oram syndrome (HOS, OMIM 142900), respectively.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [16] Ghosh TK, Packham E, Bonser J, Robinson TE, Cross SJ, Brook JD. Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome. Hum Mol Genet. 2001;10(18):1983–94. source
    • cites [22] Frank DU, Emechebe U, Thomas KR, Moon AM. Mouse Tbx3 mutants suggest novel molecular mechanisms for ulnar-mammary syndrome. PLoS One. 2013;8(7):1–7. source

      The Introduction states that people with TBX3 mutations have ulnar-mammary syndrome, and the Abstract says TBX3 mutations cause it. The paper’s References link TBX5 with Holt-Oram syndrome, but no passage in our copy states that human TBX5 mutations cause the syndrome. Page numbers are unavailable in our copy. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    B lymphocytes and autoantibodies are key in aspects of RA pathogenesis, and the clinical success of B cell targeted therapies further strengthens this.

    “It has recently been acknowledged that B lymphocytes and autoantibodies are key in aspects of RA pathogenesis [48, 49] and the clinical success of B cell targeted therapies further strengthens this [6, 50].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [48] Panayi GS. B cells: a fundamental role in the pathogenesis of rheumatoid arthritis? Rheumatology (Oxford). 2005;44(Suppl 2):ii3–7. source
    • cites [49] Bugatti S, Vitolo B, Caporali R. B cells in rheumatoid arthritis: from pathogenic players to disease biomarkers. Biomed Res. 2014;2014:681678. source
    • cites [6] Sardar S, Andersson Å. Old and new therapeutics for rheumatoid arthritis: in vivo models and drug development. Immunopharmacol Immunotoxicol. 2016;38(1):2–13. source

  • May combine two claims.

    Interpretationown data

    This is the first study to suggest the involvement of TBX3 in B cell proliferation and development of CIA, and adds mechanistic links to three independent GWAS that linked TBX3 to RA.

    “To our knowledge, this is the first study to suggest the involvement of TBX3 in B cell proliferation and development of CIA, and adds mechanistic links to three independent genome-wide association studies (GWAS) that linked TBX3 to RA [65–67].”
    Quote in the paperSubstantial claim1 of 3 sources checked
    • cites [65] Robert P, Seielstad M, Lee T, D P, Remmers EF, Ding B, et al. TRAF1-C5 as a risk locus for rheumatoid arthritis — a genomewide study. N Engl J Med 2007;357(12):1199–1209. source
    • cites [66] Freudenberg J, Lee HS, Han BG, Shin H, Do KYM, Sung YK, et al. Genome-wide association study of rheumatoid arthritis in Koreans: population-specific loci as well as overlap with European susceptibility loci. Arthritis Rheum. 2011;63(4):884–93. source
    • cites [67] Julia A, Ballina J, Caete JD, Balsa A, Tornero-Molina J, Naranjo A, et al. Genome-wide association study of rheumatoid arthritis in the Spanish population: KLF12 as a risk locus for rheumatoid arthritis susceptibility. Arthritis Rheum. 2008;58(8):2275–86. source

      The introduction says that three earlier RA genome-wide association studies had been performed, but it does not say they linked TBX3 to RA. The section on genomic regions shared with earlier studies discusses other loci; searches for TBX3 and T-box 3 found no passage addressing a TBX3–RA association. The sentence’s claim about a first mechanistic study is the citing authors’ own framing, not a finding attributed to this source. (LLM, conf=0.75)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The importance of TBX3 in relation to proliferation is underpinned by studies of its role in oncogenesis.

    “The importance of TBX3 in relation to proliferation is underpinned by studies of its role in oncogenesis [24, 25, 58, 59].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [24] Lomnytska M, Dubrovska A, Hellman U, Volodko N, Souchelnytskyi S. Increased expression of cSHMT, Tbx3 and utrophin in plasma of ovarian and breast cancer patients. Int J Cancer. 2006;118(2):412–21. source
    • cites [25] Willmer T, Cooper A, Sims D, Govender D, Prince S. The T-box transcription factor 3 is a promising biomarker and a key regulator of the oncogenic phenotype of a diverse range of sarcoma subtypes. Oncogene. 2016;5(2):e199. source
    • cites [58] Ito A, Asamoto M, Hokaiwado N, Takahashi S, Shirai T. Tbx3 expression is related to apoptosis and cell proliferation in rat bladder both hyperplastic epithelial cells and carcinoma cells. Cancer Lett. 2005;219(1):105–12. source

  • May combine two claims.

    Premisecitation

    Although TBX3 promotes migration and invasiveness of cancer cells, it negatively regulates cell proliferation.

    “More recently, it was recognized that although TBX3 promotes migration and invasiveness of cancer cells, it negatively regulates cell proliferation [37, 63].”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [37] Li J, Weinberg MS, Zerbini L, Prince S. The oncogenic TBX3 is a downstream target and mediator of the TGF-β1 signaling pathway. Mol Biol Cell. 2013;24:3569–76. source
    • cites [63] Li J, Ballim D, Rodriguez M, Cui R, Goding CR, Teng H, et al. The anti-proliferative function of the tgf-β1 signaling pathway involves the repression of the oncogenic tbx2 by its homologue tbx3. J Biol Chem. 2014;289(51):35633–43. source

  • Classified as probably not a substantial claim.

    Premisecitation

    TBX3 has previously been shown to directly interact with these signaling cascades in other cell types to regulate cell survival and proliferation.

    “TBX3 has previously been shown to directly interact with these signaling cascades in other cell types to regulate cell survival and proliferation [54–57]”
    Quote in the paperSubstantial claim2 of 4 sources checked
    • cites [54] Chang F, Lee JT, Navolanic PM, Steelman LS, Shelton JG, Blalock WL, et al. Involvement of PI3K/Akt pathway in cell cycle progression, apoptosis, and neoplastic transformation: a target for cancer chemotherapy. Leukemia. 2003;17(3):590–603. source
    • cites [55] Woyach JA, Johnson AJ, Byrd JC. The B-cell receptor signaling pathway as a therapeutic target in CLL. Blood. 2012;120(6):1175–85. source

      The section ‘Downstream activation pathways influenced by BCR’ (page unavailable) links MAPK and Akt signaling to cell survival and proliferation. It does not say that TBX3 directly interacts with those pathways in other cell types, which is the central interaction claimed here. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [56] Peres J, Mowla S, Prince S. The T-box transcription factor, TBX3, is a key substrate of AKT3 in melanomagenesis. Oncotarget. 2014;6(3):1821–33. source

      The Discussion (page unavailable) reports TBX3 interactions with PI3K/AKT signaling in mouse embryonic stem cells and with p38 MAPK in COS cells, supporting the reference to other cell types. It also describes PI3K/AKT as promoting survival and proliferation in melanoma, but does not establish that TBX3’s interactions with those cascades regulate both outcomes in other cell types. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [57] Niwa H, Ogawa K, Shimosato D, Adachi K. A parallel circuit of LIF signalling pathways maintains pluripotency of mouse ES cells. Nature. 2009;460(7251):118–22. source

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    Anti-IgM antibodies stimulate B cells through the B cell receptor (BCR) thereby activating downstream signaling cascades like PI3K, AKT and MAPK pathways.

    “Anti-IgM antibodies stimulate B cells through the B cell receptor (BCR) thereby activating further downstream signaling cascades like the phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and mitogen-activated protein kinase (MAPK) pathways [51–53].”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [51] Scharenberg AM, Humphries LA, Rawlings DJ. Calcium signalling and cell-fate choice in B cells. Nat Rev Immunol. 2007;7:778. source

      The Introduction describes BCR engagement activating a signalling complex and identifies PI3K as part of BCR signalling. It does not establish that anti-IgM antibodies activate the claimed AKT and MAPK pathways; MAPK appears only in the title of a cited reference. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [52] Kurosaki T. Regulation of B-cell signal transduction by adaptor proteins. Nat Rev Immunol. 2002;2:354. source
    • cites [53] Satpathy S, Wagner SA, Beli P, Gupta R, Kristiansen TA, Malinova D, et al. Systems-wide analysis of BCR signalosomes and downstream phosphorylation and ubiquitylation. Mol Syst Biol. 2015;11(6):810. source

      The Results (BCR-regulated phosphoproteome; page not provided) identify PI3K–AKT and RAS–MAPK as pathways activated by BCR signaling. However, the study stimulated BCRs with anti-IgG, not anti-IgM, as specified in the Results (Strategy for analysis of BCR-regulated signaling networks) and Methods (Cell stimulation). It therefore supports the downstream-pathway claim but not the stated antibody condition. (LLM, conf=0.90)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    TBX3 functions primarily as a transcriptional repressor and initial reports suggested that it can promote proliferation by repressing the cell cycle regulators p14/p19ARF and p21.

    “TBX3 functions primarily as a transcriptional repressor and initial reports suggested that it can promote proliferation by repressing the cell cycle regulators p14/p19ARF and p21 [60–62].”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [60] Carlson H, Ota S, Song Y, Chen Y, Hurlin PJ. Tbx3 impinges on the p53 pathway to suppress apoptosis, facilitate cell transformation and block myogenic differentiation. Oncogene. 2002;21(24):3827–35. source
    • cites [61] Brummelkamp TR, Kortlever RM, Lingbeek M, Trettel F, MacDonald ME, Van Lohuizen M, et al. TBX-3, the gene mutated in ulnar-mammary syndrome, is a negative regulator of p19ARF and inhibits senescence. J Biol Chem. 2002;277(8):6567–72. source

      The Results section ('TBX-3 Down-regulates p19 ARF'; page unavailable) reports TBX-3-mediated repression of the human p14 ARF promoter and reduced p19 ARF expression. The Discussion says TBX-3 drives proliferation and inhibits senescence, consistent with that part of the claim. The passages found do not show that TBX-3 represses p21, so this source does not support the claim in full. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [62] Wensing LA, Campos AH. TBX3, a downstream target of TGF-beta1, inhibits mesangial cell apoptosis. Exp Cell Res. 2014;328(2):340–50. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    The overall incidence of disease was 75%.

    “The overall incidence of disease was 75%, which is in accordance with previous reports on the B10.RIII mouse strain [42].”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The Eae39r fragment spans over 1.8 Mbp between rs33731225 (5:118442102 bp) and rs32160309 (5:120248221 bp), and Eae39r2 spans 1.4 Mbp between rs51153450 (5:118811970 bp) and rs32160309 (5:120248221 bp).

    “The Eae39r fragment spanned over 1.8 Mbp, between rs33731225 (5:118442102 bp) and rs32160309 (5:120248221 bp); and Eae39r2 was defined to be 1.4 Mbp, between rs51153450 (5:118811970 bp) and rs32160309 (5:120248221 bp) (Fig. 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In the time-course study, mice started to show signs of arthritis on day 37 post immunization, which peaked around day 50 and slightly declined in the late phase.

    “mice started to show signs of arthritis on day 37 post immunization, which peaked around day 50 and slightly declined in the late phase, as shown in Fig. 5b.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Tbx3 is differentially expressed in B cells, and low Tbx3 expression is accompanied by a higher B cell response upon B cell receptor stimulation in vitro.

    “Gene expression studies have revealed differential expression of Tbx3 in B cells, where low expression was accompanied by a higher B cell response upon B cell receptor stimulation in vitro.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Arthritic mice (sick mice from days 42 and 68; n = 6) had significantly higher TBX3 levels in serum than immunized, non-arthritic mice (n = 14) (p = 0.0087) and naïve control mice (p = 0.0095).

    “arthritic mice (sick mice from days 42 and 68 samples; n = 6) had significantly higher TBX3 levels in serum than immunized, non-arthritic mice (days 7, 15, 25, and non-arthritic mice from days 42 and 68; n = 14) (p = 0.0087) and naïve control mice (p = 0.0095) Fig. 6b.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The Eae39 region contains a smaller QTL (Eae39r) that encompasses 1.8 Mbp and has an enhancing effect on CIA severity.

    “we have shown that the Eae39 region contains a smaller QTL (Eae39r), which encompasses 1.8 Mbp and has an enhancing effect on CIA severity.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductown data

    Cell purity was >90% for CD19+ B cells and >92% for pan T and CD4+ T cells, as determined by flow cytometry.

    “Cell purity was > 90% for CD19+ B cells and > 92% for pan T and CD4+ T cells, as determined by flow cytometry.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    TBX5 acts as a transcription activator.

    “TBX5 acts as a transcription activator [16]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Ghosh TK, Packham E, Bonser J, Robinson TE, Cross SJ, Brook JD. Characterization of the TBX5 binding site and analysis of mutations that cause Holt-Oram syndrome. Hum Mol Genet. 2001;10(18):1983–94. source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Clinical disease was monitored in a blinded manner.

    “Clinical disease was monitored in a blinded manner three times a week, whereby arthritis was evaluated based on inflammation (erythema and swelling) of affected joints.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Genetic polymorphisms resulting in lower expression of Tbx3 contribute to a more severe form of collagen-induced arthritis and high titers of autoantibodies.

    “and that genetic polymorphisms, resulting in lower expression of Tbx3, are contributing to a more severe form of CIA and high titers of autoantibodies.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The identified regulatory polymorphisms in Tbx3 cause lower Tbx3 expression.
    • Rests on an assumption: The Eae39r2 locus effect on collagen-induced arthritis severity is mediated by Tbx3 rather than by other genes or elements in the fragment.

  • May combine two claims.

    Interpretationown data

    TBX3 is involved in BCR downstream pathways and might contribute to humoral immune responses.

    “These data suggested that TBX3 is involved in BCR downstream pathways and might contribute to humoral immune responses.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Study design and conductcitation

    The TBX3 ELISA had a sensitivity limit of 0.091 ng/ml and inter-assay and intra-assay coefficient of variation ≤10%, as reported by the manufacturer.

    “The sensitivity limit of the assay was 0.091 ng/ml, and the inter-assay and intra-assay coefficient of variation (CV) was ≤ 10%, as reported by the manufacturer.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    TBX3 is associated with proliferation of B cells, but not the number.

    “indicating that TBX3 is associated with proliferation of B cells, but not the number.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    TBX3 is a player in CIA pathogenesis by modulating B cell proliferative responses, but it does not affect the development of B lymphocytes.

    “This suggests that TBX3 is a player in CIA pathogenesis by modulating B cell proliferative responses, but that it does not affect the development of B lymphocytes.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    TBX3 interacts with signaling pathways downstream of the IgM-BCR, thereby being modulated by, or modulating, the activation signal, resulting in consequences for the outcome of the B cell response.

    “This, again, suggests that TBX3 interacts with signaling pathways downstream of the IgM-BCR, thereby being modulated by, or modulating, the activation signal, resulting in consequences for the outcome of the B cell response.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    A similar mechanism might operate in B cells.

    “and a similar mechanism might operate in B cells.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    The CIA enhancing Eae39r locus is fine-mapped and the therein located Tbx3 gene is a disease candidate gene for experimental arthritis in the mouse.

    “We have fine-mapped the CIA enhancing Eae39r locus and suggest that the therein located Tbx3 gene is a disease candidate gene for experimental arthritis in the mouse.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Decreased expression of Tbx3 in B lymphocytes from mice with a 1.4-Mbp fragment, in addition to increased in vitro proliferation of these cells in response to anti-IgM stimulation, can explain the enhanced antibody response and more severe disease in these mice.

    “Although not directly demonstrated, we believe that decreased expression of Tbx3 in B lymphocytes from mice with a 1.4-Mbp fragment derived from a genetically different mouse strain, in addition to the increased in vitro proliferation of these cells in response to anti-IgM stimulation, is a finding that can explain the enhanced antibody response and more severe disease in these mice.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    TBX3 is involved in RA pathogenesis and possibly also in other autoimmune diseases dependent on B cell activation and autoantibody production.

    “Taken together, these results indicate that TBX3 is involved in RA pathogenesis and possibly also in other autoimmune diseases dependent on B cell activation and autoantibody production.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Lack of, or low expression of Tbx3, may lead to increased activation of B lymphocytes, manifested by proliferation and increased autoantibody production, which subsequently would cause severe arthritis in the CIA model.

    “Lack of, or low expression of Tbx3, may thus lead to increased activation of B lymphocytes, manifested by proliferation and increased autoantibody production, which subsequently would cause severe arthritis in the CIA model.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    TBX3 is a candidate biomarker that can potentially aid in diagnosis and assessment of RA disease severity.

    “We therefore propose TBX3 as a candidate biomarker that can potentially aid in diagnosis and assessment of RA disease severity.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    One or more of these variants are responsible for the allele-specific differences in Tbx3 transcript levels observed in total spleen and splenic B lymphocytes, but not in joints.

    “We propose that one or more of these variants are responsible for the allele-specific differences in Tbx3 transcript levels observed in total spleen and splenic B lymphocytes, but not in joints.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    TBX3 impacts CIA development and autoantibody production by regulating B lymphocyte activation.

    “From studies of gene transcription and B cell proliferation, we propose that TBX3 impacts CIA development and autoantibody production by regulating B lymphocyte activation.”
    Quote in the paperSubstantial claim
35 of 35

  • The paper relies on this without support.

    The role of TBX3 and TBX5 proteins in the immune system, their link to rheumatoid arthritis, and their biomarker potential remains unexplored.

    “However, the role of the TBX3 and TBX5 proteins in the immune system, their link to RA, and their biomarker potential, remains unexplored.”

  • The paper relies on this without support.

    The identified regulatory polymorphisms in Tbx3 cause lower Tbx3 expression.

    “and that genetic polymorphisms, resulting in lower expression of Tbx3, are contributing to a more severe form of CIA and high titers of autoantibodies.”

  • The paper relies on this without support.

    The Eae39r2 locus effect on collagen-induced arthritis severity is mediated by Tbx3 rather than by other genes or elements in the fragment.

    “we suggest that TBX3 is a novel factor important for the regulation of gene transcription in the immune system”

  • The paper relies on this without support.

    The B cell response upon B cell receptor stimulation in vitro reflects the B cell response in vivo.

    “low expression was accompanied by a higher B cell response upon B cell receptor stimulation in vitro.”

  • The paper relies on this without support.

    Serum TBX3 levels reflect TBX3 activity relevant to rheumatoid arthritis.

    “We also propose TBX3 as a putative diagnostic biomarker for rheumatoid arthritis.”
5 of 5

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Conflicting tests reported for ethical survival

    Minor

    The Methods specify Fisher's exact test for ethical survival, but the Table 1 note says the number of mice euthanized was analyzed with a chi-squared test. These are different tests for the same outcome.

1 of 1

Cites

Flagged first, then by citations

Research works printed but not linked: 4

  1. PubMed database. Available from: https://www.ncbi.nlm.nih.gov/pubmed /. Accessed 18 Nov 2018.
  2. Google scholar. Available from: https://scholar.google.com /. Accessed 18 Nov 2018.
  3. NCBI SNP database. Available from: https://www.ncbi.nlm.nih.gov/SNP /. Accessed 2 June 2018.
  4. Freudenberg J, Lee H-S, Han B-G, Do SH, Kang YM, Sung Y-K, et al. Genome-wide association study of rheumatoid arthritis in Koreans: population-specific loci as well as overlap with European susceptibility loci. Arthritis Rheum. 2011;63(4):884–93. · DOI · Paper page

Cited by

Sorted by citations

Related

Authors
Sardar, S., Kerr, A., Vaartjes, D., Moltved, E. R., Karosiene, E., Gupta, R., & Andersson, Å.
Affiliations
  • University of Copenhagen (DK)
  • Novo Nordisk (Denmark) (DK)
  • Technical University of Denmark (DK)
Corresponding author
Andersson, Åsa
ORCID
Venue
Arthritis Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2019-01-10
Volume
21
Issue
1
Pages
16-16
ISSN
  • 1478-6354 (ISSN-L)
  • 1478-6362
DOI
10.1186/s13075-018-1797-3
PMC
PMC6329118
OpenAlex
W2941376028
Semantic Scholar
00dcd2d9c686cbc35794d3399dc7d6911d283470
Cited by
13
References
81
Citations by year
2025: 1 · 2024: 2 · 2023: 1 · 2022: 2 · 2021: 2 · 2020: 3 · 2019: 2
FWCI
1.47
Type
journal article
Language
en
Open access
Licence
Locations
  • Arthritis Research & Therapy (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (submittedVersion, other-oa)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Technical University of Denmark, DTU Orbit (Technical University of Denmark, DTU) (submittedVersion, other-oa)
Funders
  • Gigtforeningen
  • Oticon Fonden
Topics
  • Health Sciences › Medicine › Rheumatology › Rheumatoid Arthritis Research and Therapies
  • Health Sciences › Medicine › Complementary and alternative medicine › Natural Compounds in Disease Treatment
  • Health Sciences › Medicine › Rheumatology › Systemic Lupus Erythematosus Research
Keywords
TBX3, collagen-induced arthritis, rheumatoid arthritis, autoimmune arthritis, disease severity, anti-collagen antibodies, B cells, B cell receptor signaling, transcriptional regulation, genetic polymorphism, serum biomarkers, mouse chromosome 5
MeSH terms
Animals, Arthritis, Experimental, Cattle, Cells, Cultured, Male, Mice, Transgenic, Severity of Illness Index, Mice, Congenic, T-Box Domain Proteins, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-23.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-23.

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