CoChat
Sign Up
Cited by 10T-cell and B-cell ImmunologyPsoriasis: Treatment and PathogenesisHematopoietic Stem Cell Transplantation

The Transcription Factor PU.1 Regulates γδ T Cell Homeostasis

Open access
Jabeen, Rukhsana, Chang, Hua-Chen, Goswami, Ritobrata, Nutt, Stephen L., Kaplan, Mark H.·2011·PLOS ONE
DOI 10.1371/journal.pone.0022189PMC PMC3136513
Open this paper in CoChat — free accountOpen PDF

Abstract

BACKGROUND: T cell development results in the generation of both mature αβ and γδ T cells. While αβ T cells predominate in secondary lymphoid organs, γδ T cells are more abundant in mucosal tissues. PU.1, an Ets family transcription factor, also identified as the spleen focus forming virus proviral integration site-1 (Sfpi1) is essential for early stages of T cell development, but is down regulated during the DN T-cell stage. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we show that in mice specifically lacking PU.1 in T cells using an lck-Cre transgene with a conditional Sfpi1 allele (Sfpi1(lck-/-)) there are increased numbers of γδ T cells in spleen, thymus and in the intestine when compared to wild-type mice. The increase in γδ T cell numbers in PU.1-deficient mice is consistent in γδ T cell subsets identified by TCR variable regions. PU.1-deficient γδ T cells demonstrate greater proliferation in vivo and in vitro. CONCLUSIONS/SIGNIFICANCE: The increase of γδ T cell numbers in Lck-Cre deleter strains, where deletion occurs after PU.1 expression is diminished, as well as the observation that PU.1-deficient γδ T cells have greater proliferative responses than wild type cells, suggests that PU.1 effects are not developmental but rather at the level of homeostasis. Thus, our data shows that PU.1 has a negative influence on γδ T cell expansion.

AI snapshot

Written from the paper’s full text.

TL;DR

Conditional deletion of PU.1 in T cells increases γδ T cell numbers in spleen, thymus, and intestinal intraepithelial lymphocytes without affecting αβ T cells or NKT cells. The expanded γδ T cells show greater proliferation in vivo and in vitro, suggesting PU.1 negatively regulates γδ T cell homeostasis rather than development.

Key findings

  • Sfpi1 lck−/− mice had significantly increased percentages and absolute numbers of γδ T cells in spleen, thymus, and intestinal IEL, but not skin; αβ T cells and NKT cells were similar to wild-type (p>0.05).
  • The increase occurred across γδ T cell subsets identified by Vγ2 and Vδ4 TCR expression in all organs examined.
  • PU.1-deficient γδ T cells showed significantly more BrdU incorporation in vivo and greater CFSE dilution/proliferation after anti-CD3/anti-CD28 stimulation for 72 h; apoptosis was only modestly affected.
  • IFNγ production was similar between wild-type and PU.1-deficient γδ T cells, IL-17 showed a non-significant trend higher in PU.1-deficient cells, and CCR6+ γδ T cells were significantly increased.

Methods

  • Mice: wild-type C57BL/6 and Sfpi1 lck−/− mice with conditional PU.1 deletion via Lck-Cre, used at 6–8 weeks; spleen, thymus, intestinal IEL, and skin were examined.
  • Flow cytometry and cell sorting measured γδ TCR, CD3, Vγ2, Vδ4, CD62L, CD44, CCR6, BrdU, CFSE, and annexin V; qPCR measured Sfpi1 and transcription factors; ELISA measured IFNγ and IL-17.
  • Comparisons used Student’s t test with p<0.05 considered significant; data are reported as mean ± SEM from 3–12 mice depending on experiment.

Limitations

  • The paper states that Lck-Cre deletion was not complete in γδ T cells, with at least 50% deletion observed, and that the point at which PU.1-deficiency affects γδ T cell homeostasis is difficult to define.
  • The authors state they cannot distinguish whether increased proliferation occurs during or after γδ T cell development, and that precise PU.1-regulated functions require further study.
  • Evident from the text: some supporting data are described as “data not shown,” and the IL-17 increase was not statistically significant.

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

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0022189

Cites

32 of 32 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

22 of 39 cited sources checked · 8 supported · 1 flagged · accuracy not yet measured

  • Interpretationcitation

    A fetal thymus does not result in the absence of PU.1 expression in γδ T cells.

    “PU.1 expression was observed in γδ T cells derived from fetal thymic organ culture suggesting that a fetal thymus does not result in the absence of PU.1 expression in γδ T cells [18].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [18] MK Anderson, 1999, Precise developmental regulation of Ets family transcription factors during specification and commitment to the T cell lineage., Development source

      The Cell sorting section says the authors cultured fetal thymuses and isolated γδ T-cell subsets. The expression results report PU.1 in immature thymocytes, but the passages do not establish that the cultured γδ T cells expressed PU.1. The claimed observation—and the inference about a fetal thymus—is therefore not supported by the available text. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
  • Premisecitation

    PU.1 expression is observed in γδ T cells derived from fetal thymic organ culture.

    “These results are consistent with a previous report that detected PU.1 expression in γδ T cells derived from fetal thymic organ culture [18].”
    • Also stated in: § Discussion · hedged
      “PU.1 expression was observed in γδ T cells derived from fetal thymic organ culture suggesting that a fetal thymus does not result in the absence of PU.1 expression in γδ T cells [18].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [18] MK Anderson, 1999, Precise developmental regulation of Ets family transcription factors during specification and commitment to the T cell lineage., Development source

      The Cell sorting section says the authors cultured fetal mouse thymuses and isolated γδ T cells. The expression results report PU.1 in immature thymocytes, but the passages do not establish that it was detected in the cultured γδ T-cell subset specifically. (LLM, conf=0.78)

      Experimental — accuracy not yet measured
  • Findingcitation

    αβ T cell development and numbers in various lymphoid organs are unaltered by PU.1-deficiency.

    “Flow cytometry of splenocytes and thymocytes showed similar profiles of CD3+ αβTCR and CD3+ NKT cells between wild-type and Sfpi1 lck−/− mice (Fig. 1A).”
    • Also stated in: § Discussion · plain
      “αβ T cell development and numbers in various lymphoid organs is unaltered by PU.1-deficiency.”
    • Also stated in: § Introduction · boosted
      “Our lab has previously shown that αβ T cell development is normal in mice that conditionally lack PU.1 in T cells [15].”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [15] HC Chang, 2009, PU.1 regulates TCR expression by modulating GATA-3 activity., J Immunol source
  • Study design and conductown data

    Lck-Cre-mediated deletion of the Sfpi1 allele in γδ T cell populations is at least 50% complete.

    “Although deletion was not complete as previously observed in double-positive thymocytes [15], there was at least 50% deletion in γδ T cell populations (Fig. 2A).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [15] HC Chang, 2009, PU.1 regulates TCR expression by modulating GATA-3 activity., J Immunol source

      The Results section (page unavailable) says Lck-Cre-mediated Sfpi1 deletion begins at the DN3 stage and is not complete until later in thymocyte development. It does not report a deletion percentage for γδ T cells. In the citing sentence, the citation follows the statement about incomplete deletion; the at-least-50% figure is attributed to the citing paper’s Fig. 2A. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
  • Findingown data

    There is a significant increase in the numbers of γδ T cells that are CCR6+ in the absence of PU.1.

    “we did observe a significant increase in the numbers of γδ T cells that were CCR6+, supporting a selective increase in the populations of some cytokine secreting γδ T cells in the absence of PU.1 (Fig. 4H and data not shown).”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 21
  • May combine two claims. 14
  • The quote only partly states this claim. 3

  • Probably not a substantial claim: may not pass S1 (asserted), S3 (load-bearing), S4 (based). May combine two claims. The quote only partly states this claim.

    Premisecitation

    PU.1 regulates TCR expression by modulating GATA-3 activity, decreasing the threshold of activation.

    “we have shown that it regulates the expression of TCR in T cells, decreasing the threshold of activation [15].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [15] HC Chang, 2009, PU.1 regulates TCR expression by modulating GATA-3 activity., J Immunol source

      The Discussion says PU.1 regulates TCR expression by modulating GATA-3, but it gives the opposite direction for the activation threshold: CD4 T cells expressing PU.1 have a higher threshold. It is PU.1 deficiency that the paper associates with a reduced threshold (Discussion, passages 7 and 3; page numbers not provided). (LLM, conf=0.90)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured

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

    Premisecitation

    Numerous signals impact γδ T cell lineage including TCR signal strength, notch signaling, IL-7R signaling, and the presence of DP thymocytes.

    “Numerous signals impact on the γδ T cell lineage including, TCR signal strength, notch signaling, IL-7R signaling and the presence of DP thymocytes [6]–[10].”
    Quote in the paperNot a substantial claim3 of 5 sources checked
    • cites [6] SM Hayes, 2005, TCR signal strength influences alphabeta/gammadelta lineage fate., Immunity source
    • cites [7] M Ciofani, 2004, Obligatory role for cooperative signaling by pre-TCR and Notch during thymocyte differentiation., J Immunol source

      The introductory passage in our copy (labeled Abstract; page not provided) reports that Notch1 has been suggested to influence the αβ-versus-γδ T-cell lineage decision. The paper’s own findings concern pre-TCR and Notch signaling during αβ thymocyte differentiation; they do not establish the claimed roles of TCR signal strength, IL-7R signaling, or the presence of DP thymocytes in the γδ lineage. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [8] I Van de Walle, 2009, An early decrease in Notch activation is required for human TCR-alphabeta lineage differentiation at the expense of TCR-gammadelta T cells., Blood source

      The Introduction and Discussion say that TCR signal strength and Notch signaling influence the αβ/γδ lineage decision. The section “IL-7 cannot compensate Notch-dependent γδ T-cell development” shows that changing IL-7 levels affects γδ T-cell numbers, although it does not directly test IL-7R signaling. The paper reports DP thymocytes as an outcome of altered Notch signaling, but does not show that their presence itself affects the γδ lineage. Page numbers were not provided for these passages. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [9] J Kang, 2001, Evidence that gammadelta versus alphabeta T cell fate determination is initiated independently of T cell receptor signaling., J Exp Med source

      The Introduction discusses competing views of how TCR signals affect γδ versus αβ lineage development, and the Discussion states that IL-7R signaling plays a role in γδ cell development. Our copy does not identify Notch signaling or the presence of double-positive (DP) thymocytes as factors affecting the γδ lineage; it describes DP thymocytes chiefly as an αβ-lineage outcome. It also discusses the quality or role of TCR signals rather than establishing an effect of TCR signal strength. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [10] B Silva-Santos, 2005, Lymphotoxin-mediated regulation of gammadelta cell differentiation by alphabeta T cell progenitors., Science source

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

    Premisecitation

    γδ T cells function in immunosurveillance, playing a significant role in innate immunity, autoimmunity, and allergic responses.

    “γδ T cells function in immunosurveillance playing a significant role in innate immunity, autoimmunity and allergic responses [1].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [1] WK Born, 2010, Analysis of gammadelta T cell functions in the mouse., J Immunol source

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

    Premisecitation

    Both αβ and γδ T cells arise from common multipotent DN precursors in the thymus.

    “Both αβ and γδ T cells arise from common multipotent DN precursors in the thymus that can be further separated into four DN subsets based on CD44 and CD25 expression [2], [3].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [2] HE Porritt, 2004, Heterogeneity among DN1 prothymocytes reveals multiple progenitors with different capacities to generate T cell and non-T cell lineages., Immunity source

      The Results and Discussion (page numbers unavailable) identify DN1a and DN1b as early thymic T-cell progenitors, and the Results report that they can also generate cells expressing an NK-lineage marker. Our copy does not establish that both αβ and γδ T cells arise from a common DN precursor; the paper discusses T-lineage output without demonstrating that shared origin. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites [3] D Allman, 2003, Thymopoiesis independent of common lymphoid progenitors., Nat Immunol source

  • May combine two claims.

    Interpretationcitation

    PU.1 appears to have distinct effects in various types of T cells: negatively regulating γδ T cell numbers, negatively regulating Th2 cytokine production and TCR expression, while promoting IL-9 expression.

    “Thus, PU.1 appears to have distinct effects in various types of T cells, negatively regulating γδ T cell numbers, negatively regulating Th2 cytokine production and TCR expression, while promoting IL-9 expression [15]–[17].”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [15] HC Chang, 2009, PU.1 regulates TCR expression by modulating GATA-3 activity., J Immunol source
    • cites [16] HC Chang, 2005, PU.1 expression delineates heterogeneity in primary Th2 cells., Immunity source

      The Results (page not provided) report that PU.1 expression decreased Th2 cytokines, including IL-4, IL-5, IL-10, and IL-13. Our copy does not establish the attributed effects on γδ T cell numbers, TCR expression, or IL-9 expression. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [17] HC Chang, 2010, The transcription factor PU.1 is required for the development of IL-9-producing T cells and allergic inflammation., Nat Immunol source

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

    Premisecitation

    PU.1 expression in T cells and subpopulations of Th2 cells contributes to heterogeneity in Th2 cytokine expression and TCR expression.

    “However the expression of PU.1 in T cells and subpopulations of Th2 cells contributes to heterogeneity in Th2 cytokine expression and TCR expression [16].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [16] HC Chang, 2005, PU.1 expression delineates heterogeneity in primary Th2 cells., Immunity source

      The Results and Discussion (pages unavailable) show that PU.1 is expressed in a subset of Th2 cells and contributes to variation in their cytokine profiles. They do not show that PU.1 contributes to heterogeneity in TCR expression; the paper’s references to anti-CD3 stimulation do not establish that point. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

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

    Premisecitation

    PU.1 promotes development of Th9 cells.

    “Importantly, PU.1 promotes the development of Th9 cells [17].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [17] HC Chang, 2010, The transcription factor PU.1 is required for the development of IL-9-producing T cells and allergic inflammation., Nat Immunol source

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

    Premisecitation

    Dermal γδ T cells have a fetal thymic origin.

    “Since dermal γδ T cells have a fetal thymic origin [1]”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [1] WK Born, 2010, Analysis of gammadelta T cell functions in the mouse., J Immunol source

      The ‘Early functional commitment in the thymus’ section (page unavailable) says γδ T cells develop in the thymus and discusses thymic differentiation of dendritic epidermal T cells. The ‘Clonal selection in the periphery’ section (page unavailable) likewise describes thymic selection of epidermal cells, not dermal cells. Neither passage establishes a fetal thymic origin for dermal γδ T cells. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

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

    Premisecitation

    In several cell types PU.1 can function as a tumor suppressor.

    “In several cell types PU.1 can function as a tumor suppressor [25], [26].”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [25] TC Somervaille, 2006, PU.1 and Junb: suppressing the formation of acute myeloid leukemia stem cells., Cancer Cell source

      The Abstract (page not provided) describes PU.1 as a tumor suppressor and later identifies PU.1 and JUNB as myeloid-lineage tumor suppressors. It does not establish tumor-suppressor activity in several cell types; its discussion is confined to myeloid leukemogenesis. (LLM, conf=0.83)

      Experimental — accuracy not yet measured
    • cites [26] MP Tschan, 2008, PU.1 binding to the p53 family of tumor suppressors impairs their transcriptional activity., Oncogene source

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

    Premisecitation

    The cytokine secretion pattern of γδ T cells may vary with antigen exposure.

    “the cytokine secretion pattern may vary with antigen exposure [23].”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [23] KD Jensen, 2008, Thymic selection determines gammadelta T cell effector fate: antigen-naive cells make interleukin-17 and antigen-experienced cells make interferon gamma., Immunity source

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

    Premisecitation

    PU.1 limits expression of Th2 cytokines and contributes to Th2 heterogeneity.

    “We have also demonstrated that PU.1 limits the expression of Th2 cytokines and contributes to Th2 heterogeneity [15], [16].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [15] HC Chang, 2009, PU.1 regulates TCR expression by modulating GATA-3 activity., J Immunol source
    • cites [16] HC Chang, 2005, PU.1 expression delineates heterogeneity in primary Th2 cells., Immunity source

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

    Premisecitation

    In various cell types PU.1 induces expression of TRAIL and Ink4b, which respectively induce apoptosis and inhibit cell proliferation.

    “In various cell types PU.1 has also been shown to induce expression of TRAIL and Ink4b, which respectively induce apoptosis and inhibit cell proliferation [28], [29].”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [28] S Ueno, 2009, PU.1 induces apoptosis in myeloma cells through direct transactivation of TRAIL., Oncogene source

      The Discussion (page unavailable) reports that PU.1 directly increases TRAIL expression and that TRAIL contributes to apoptosis in two myeloma cell lines. It explicitly says TRAIL induction in other hematopoietic lineages remains to be investigated, so this paper does not establish the broader claim about various cell types. Ink4b appears to be associated with the sentence’s other citation. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites [29] M Schmidt, 2004, The interferon regulatory factor ICSBP/IRF-8 in combination with PU.1 up-regulates expression of tumor suppressor p15(Ink4b) in murine myeloid cells., Blood source

      The Discussion reports that PU.1 and ICSBP regulate p15 Ink4b expression in IFNβ-treated myeloid cells, and the Introduction identifies p15 Ink4b as a cell-cycle inhibitor. This supports the Ink4b portion for this source, but not the broader implication that PU.1 induces it across various cell types or independently of ICSBP. The separate TRAIL claim appears to be assigned to the other citation. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingown data

    There is a trend toward more IL-17 production from PU.1-deficient γδ T cells than wild-type, but it is not statistically significant.

    “There was a trend towards more IL-17 production from PU.1-deficient γδ T cells than from wild type cells, though this was not statistically significant (Fig. 4G).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Among γδ T cell subsets defined by CD62L and CD44, only the CD62Lhi CD44hi population is significantly increased in the absence of PU.1, though all populations show a trend toward increased numbers.

    “though all populations demonstrated a trend towards increased numbers in the absence of PU.1, only the CD62Lhi, CD44hi population was significantly increased (Fig. 2C).”
    Quote in the paperSubstantial claim

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

    Findingown data

    Greater than 90% of splenic γδ T cells are CD5-positive regardless of PU.1 expression.

    “Greater than 90% of the cells were CD5-positive regardless of PU.1 expression”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Sfpi1 expression is lower in γδ T cells isolated from thymus or intraepithelial lymphocytes and undetectable in γδ T cells isolated from skin.

    “Sfpi1 expression was lower in γδ T cells isolated from thymus, or intraepithelial lymphocytes, and was undetectable in γδ T cells isolated from skin (Fig. 1C).”
    • Also stated in: § Discussion · plain
      “Sfpi1 is expressed in lower amounts by thymic and intraepithelial intestinal γδ T cells, and not expressed in dermal γδ T cells.”
    Quote in the paperNot a substantial claim

  • The quote only partly states this claim.

    Findingown data

    The increase in γδ T cell numbers in PU.1-deficient mice occurs across γδ T cell subsets identified by TCR variable regions (Vγ2 and Vδ4).

    “The increase in γδ T cell numbers in PU.1-deficient mice is consistent in γδ T cell subsets identified by TCR variable regions.”
    • Also stated in: § Sfpi1 lck−/− mice contain enhanced numbers of γδ T cell subsets · plain
      “We observed significantly increased percentages of Vγ2+ and Vδ4+ T cells in all organs examined (Fig. 3).”
    • Also stated in: § Sfpi1 lck−/− mice contain enhanced numbers of γδ T cell subsets · plain
      “there was an increase in cell numbers of each of these γδ T cell subsets (Fig. 3).”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    PU.1-deficient γδ T cells show greater proliferation than wild-type γδ T cells in vitro after TCR stimulation.

    “PU.1-deficient γδ T cells demonstrate greater proliferation in vivo and in vitro.”
    • Also stated in: § The absence of PU.1 results in enhanced γδ T cells proliferation · plain
      “γδ T cells from Sfpi1 lck−/− mice had greater proliferation than cells from wild-type mice, determined by staining profile, mean fluorescence intensity of CFSE staining, and proliferative index (Fig. 4B–C).”
    • Also stated in: § Discussion · plain
      “and in vitro TCR-induced proliferation of γδ T cells.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Decreased expression of PU.1 is necessary for continued progress through T cell differentiation.

    “Decreased expression of PU.1 is necessary for continued progress through T cell differentiation [14].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [14] MK Anderson, 2002, Constitutive expression of PU.1 in fetal hematopoietic progenitors blocks T cell development at the pro-T cell stage., Immunity source

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

    Premisecitation

    Commitment to the T cell lineage is complete at the DN3 stage, where cells undergo DNA rearrangements at the β, γ, and δ TCR loci and choose between αβ and γδ developmental programs.

    “Commitment to the T cell lineage is complete at the DN3 stage where the cells undergo extensive DNA rearrangements at the β, γ and δ TCR loci in order to express functional TCR chains and make a choice between two developmental programs, one generating αβ T cell subsets and one generating the distinct characteristics and functions of γδ T cells [4], [5].”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [4] DB Burtrum, 1996, TCR gene recombination and alpha beta-gamma delta lineage divergence: productive TCR-beta rearrangement is neither exclusive nor preclusive of gamma delta cell development., J Immunol source
    • cites [5] T Taghon, 2006, Developmental and molecular characterization of emerging beta- and gammadelta-selected pre-T cells in the adult mouse thymus., Immunity source

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

    Premisecitation

    Expression of Sox13 promotes γδ T cell development while opposing αβ T cell differentiation, and Sox13-deficient mice exhibit impaired γδ T cell development but not αβ T cells.

    “Expression of the transcription factor Sox13 promotes γδ T cell development while opposing αβ T cell differentiation. Mice deficient in Sox13 exhibit impaired γδ T cell development but not αβ T cells [11].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [11] HJ Melichar, 2007, Regulation of gammadelta versus alphabeta T lymphocyte differentiation by the transcription factor SOX13., Science source

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

    Premisecitation

    Expression of c-Jun is not required for, and may antagonize, γδ T cell development.

    “The expression of c-Jun is not required for, and may antagonize, γδ T cell development [12].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [12] L Riera-Sans, 2007, Regulation of alphabeta/gammadelta T cell development by the activator protein 1 transcription factor c-Jun., J Immunol source

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

    Premisecitation

    PU.1 is essential for lymphoid and myeloid development.

    “PU.1 is an Ets family transcription factor that is essential for lymphoid and myeloid development [13].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [13] J Lloberas, 1999, The key role of PU.1/SPI-1 in B cells, myeloid cells and macrophages., Immunol Today source

  • May combine two claims.

    Interpretationown data

    The phenotype of increased γδ T cells in PU.1-deficient mice is likely cell-intrinsic and not due to effects of αβ T cells on γδ T cell development.

    “it is likely that the phenotype observed is cell-intrinsic, and not due to the reported effects of αβ T cells on γδ T cell development [10] because deletion of Sfpi1 by CD4-Cre, where Sfpi1 is deleted in all αβ T cells did not effect γδ T cell numbers in spleen or thymus (data not shown).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [10] B Silva-Santos, 2005, Lymphotoxin-mediated regulation of gammadelta cell differentiation by alphabeta T cell progenitors., Science source

  • May combine two claims.

    Interpretationown data

    The increase in γδ T cell numbers in Lck-Cre deleter strains, where deletion occurs after PU.1 expression is diminished, together with greater proliferative responses of PU.1-deficient γδ T cells, suggests that PU.1 effects are not developmental but at the level of homeostasis.

    “The increase of γδ T cell numbers in Lck-Cre deleter strains, where deletion occurs after PU.1 expression is diminished, as well as the observation that PU.1-deficient γδ T cells have greater proliferative responses than wild type cells, suggests that PU.1 effects are not developmental but rather at the level of homeostasis.”
    • Also stated in: § Discussion · hedged
      “deletion by Lck-Cre, which occurs after Sfpi1 expression is diminished between DN1 and DN2 and extinguished by DN3, is unlikely to affect γδ T cell development.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Lck-Cre-mediated deletion of Sfpi1 occurs after PU.1 expression is diminished between DN1 and DN2 and extinguished by DN3.

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

    Premisecitation

    γδ T cells can secrete IFNγ and IL-17 in response to TCR stimulation.

    “Previous reports have demonstrated that γδ T cells can secrete IFNγ and IL-17 in response to TCR stimulation [20]–[22]”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [20] DA Ferrick, 1995, Differential production of interferon-gamma and interleukin-4 in response to Th1- and Th2-stimulating pathogens by gamma delta T cells in vivo., Nature source
    • cites [21] CL Roark, 2008, gammadelta T cells: an important source of IL-17., Curr Opin Immunol source
    • cites [22] CE Sutton, 2009, Interleukin-1 and IL-23 induce innate IL-17 production from gammadelta T cells, amplifying Th17 responses and autoimmunity., Immunity source

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

    Premisecitation

    γδ T cell commitment during DN T cell development may occur at several stages.

    “Although γδ T cell commitment during DN T cell development may occur at several stages [24]”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [24] K Narayan, 2007, Molecular events that regulate alphabeta versus gammadelta T cell lineage commitment: old suspects, new players and different game plans., Curr Opin Immunol source

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

    Premisecitation

    Development of myeloid leukemia in mice with decreased PU.1 expression is dependent on Jun.

    “The development of myeloid leukemia in mice that have decreased expression of PU.1 is dependent on Jun [27].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [27] U Steidl, 2006, Essential role of Jun family transcription factors in PU.1 knockdown-induced leukemic stem cells., Nat Genet source

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

    Premisecitation

    In initial descriptions, PU.1 was thought to be largely restricted to myeloid lineages.

    “In the initial descriptions of PU.1, it was thought to be largely restricted to myeloid lineages [30].”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [30] T Nagamura-Inoue, 2001, Transcription factors that regulate growth and differentiation of myeloid cells., Int Rev Immunol source
29 of 29

  • The paper relies on this without support.

    The role of other transcription factors in regulating γδ T cell development and peripheral homeostasis has not been examined.

    “The role of other transcription factors in regulating γδ T cell development and peripheral homeostasis has not been examined.”

  • The paper relies on this without support.

    Lck-Cre-mediated deletion of Sfpi1 occurs after PU.1 expression is diminished between DN1 and DN2 and extinguished by DN3.

    “deletion by Lck-Cre, which occurs after Sfpi1 expression is diminished between DN1 and DN2 and extinguished by DN3”
2 of 2

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Conditional deletion of Spi1 in T-lineage cells after DN2 has only modest effects on T cells as a whole, limited to selective reduction of IL-9-producing T-cells and some loss of restraint on γδ T cells and Tfh activity.

    “if Spi1 is conditionally deleted in T-lineage cells only after the cells have passed the DN2 stage, there are very modest effects on T cells as a whole, apparently limited to selective reduction of IL-9-producing T-cells (74), and some loss of restraint on γδ T cells and T follicular helper cell activity (75, 76).”

    From Mechanisms of Action of Hematopoietic Transcription Factor PU.1 in Initiation of T-Cell Development (2019) · cites this paper as [75]

    The Results report that Lck-Cre deletion of Sfpi1 leaves αβ T-cell profiles largely unchanged but increases γδ T-cell numbers, supporting the portion attributed to this paper (Results, ‘γδ T cell numbers are expanded in the absence of PU.1’; page unavailable). The Discussion places deletion after Sfpi1 expression declines between DN1 and DN2, but does not establish that deletion occurs only after cells have passed DN2. The IL-9 and Tfh portions carry other citations in the citing sentence. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

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 cell source for the in vitro proliferation assay

    Minor

    The Results say thymic cells were used for the CFSE proliferation assay, but the Figure 4 caption says splenocytes were used. The Methods also specify sorted γδ T cells from spleen for CFSE labeling.

1 of 1

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Jabeen, R., Chang, H.-C., Goswami, R., Nutt, S. L., & Kaplan, M. H.
Affiliations
  • Indiana University Indianapolis (US)
  • Walter and Eliza Hall Institute of Medical Research (AU)
Corresponding author
Kaplan, Mark H.
Venue
PLOS ONE
Publisher
Public Library of Science (PLoS)
Published
2011-07-14
Volume
6
Issue
7
Pages
e22189
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0022189
PMC
PMC3136513
OpenAlex
W2012402075
Semantic Scholar
cfe909de51e76f739bf4ed4ae721690586eac2dd
Cited by
10
References
32
Citations by year
2019: 2 · 2018: 1 · 2015: 2 · 2014: 2 · 2013: 2 · 2012: 1
FWCI
0.62
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • IUScholarWorks (Indiana University) (submittedVersion, cc-by)
  • Minerva Access (University of Melbourne) (submittedVersion, cc-by)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • U.S. Public Health Service
  • National Institute of Allergy and Infectious Diseases (R01 AI057459)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
  • Life Sciences › Immunology and Microbiology › Immunology › Psoriasis: Treatment and Pathogenesis
  • Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
Keywords
PU.1, γδ T cells, T cell homeostasis, ETS transcription factors, T cell development, T cell proliferation, conditional knockout, spleen, thymus
MeSH terms
Proto-Oncogene Protein Spi-1, Animals, Cells, Cultured, Enzyme-Linked Immunosorbent Assay, Female, Flow Cytometry, Interferon-gamma, Mice, Inbred C57BL, Mice, Mutant Strains, Proto-Oncogene Proteins, T-Lymphocytes, Trans-Activators, Polymerase Chain Reaction, Receptors, Antigen, T-Cell, gamma-delta, Apoptosis, Interleukin-17, Cell Proliferation, 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 2026-10-21.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-21.

Cited sources are checked by gpt-6-sol.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review