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Cited by 4Immunodeficiency and Autoimmune DisordersT-cell and B-cell ImmunologyBlood disorders and treatments

A Novel Homozygous RHOH Variant Associated with T Cell Dysfunction and Recurrent Opportunistic Infections

Open access
Zhou, Jingyu, Qian, Mengqing, Jiang, Ning, Wu, Jing, Feng, Xiaoqian, Yu, Meiping, Min, Qing, Xu, Haoxin +9 more·2024·Journal of Clinical Immunology
DOI 10.1007/s10875-024-01735-4

Published version of a preprint

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Abstract

Abstract RHOH, an atypical small GTPase predominantly expressed in hematopoietic cells, plays a vital role in immune function. A deficiency in RHOH has been linked to epidermodysplasia verruciformis, lung disease, Burkitt lymphoma and T cell defects. Here, we report a novel germline homozygous RHOH c.245G > A (p.Cys82Tyr) variant in a 21-year-old male suffering from recurrent, invasive, opportunistic infections affecting the lungs, eyes, and brain. His sister also succumbed to a lung infection during early adulthood. The patient exhibited a persistent decrease in CD4+ T, B, and NK cell counts, and hypoimmunoglobulinemia. Despite normal RHOH mRNA levels in his family, the patient’s T cell showed impaired activation upon in vitro TCR stimulation. In Jurkat T cells transduced with RHOHC82Y, a similar reduction in CD69 activation marker up-regulation was observed. However, ectopic expression of the C82Y variant did not exhibit a negative dominance over wild type RHOH. Furthermore, the C82Y variant showed reduced RHOH protein expression and impaired interaction with the TCR signaling molecule ZAP70. Together, these data suggest that the newly identified autosomal-recessive RHOH variant is associated with T cell dysfunction and recurrent opportunistic infections, functioning as a hypomorph by disrupting ZAP70-mediated TCR signaling.

AI snapshot

Written from the paper’s full text.

TL;DR

This paper reports a novel homozygous germline RHOH c.245G>A (p.Cys82Tyr) missense variant in a 21-year-old male with recurrent invasive opportunistic infections of the lungs, eyes, and brain, plus persistent CD4+ T, B, and NK cell decreases and hypoimmunoglobulinemia. Functional data indicate the C82Y variant acts as a hypomorph, reducing RHOH protein expression and disrupting ZAP70-mediated TCR signaling without dominant-negative effect.

Key findings

  • The patient, from a consanguineous family, had recurrent infections involving lungs, retina, and brain caused by pathogens including CMV, rubella virus, *Pneumocystis jirovecii*, helminth, and multiple bacteria; his sister also had recurrent severe lung infections and died of severe respiratory infection.
  • Immunological testing during 2018–2019 showed persistent CD4+ T and B cell reductions, sometimes decreased NK cells, and profound hypoimmunoglobulinemia; for example, serum IgG was 0.8 g/L (reference 7.0–16.0 g/L), B cells were 0.9% (reference 6.48–16.64%), and NK cells were 3.5% (reference 5.17–24.65%).
  • Whole-exome sequencing identified the RHOH c.245G>A (p.Cys82Tyr) variant as the only homozygous rare variant in the patient; both parents were heterozygous carriers, and in silico tools predicted pathogenicity (CADD 26.4; REVEL 0.831; SIFT 0.001; PolyPhen-2 1.0; MutationTaster 1).
  • RHOH mRNA was comparable to parents and controls, but patient T cells showed impaired activation upon TCR stimulation; Jurkat cells expressing RHOH C82Y showed reduced CD69 up-regulation and lower IL-2 production, and C82Y showed reduced RHOH protein expression and undetectable binding to ZAP70 before and after anti-CD3 stimulation.

Methods

  • Clinical and family study of a 21-year-old male proband, his consanguineous parents, and an affected sibling; whole-exome sequencing and Sanger sequencing were performed on the patient and parents, with variant filtering against population databases and in silico prediction tools.
  • Functional assays used PBMCs from the proband and parents, HEK293 cells transfected with wild-type or C82Y RHOH, and Jurkat cells transduced with HA-tagged RHOH WT, RHOH C82Y, or empty vector; analyses included qPCR, immunoblot, co-immunoprecipitation, flow cytometry for CD69, and ELISA for IL-2.
  • Statistical analysis used one-way ANOVA with Bonferroni’s multiple comparisons test or two-tailed unpaired Student’s t tests in GraphPad Prism 8, with P < 0.05 considered significant.

Limitations

  • Stated: detailed immunological analysis of the patient could not be conducted because he succumbed to infection and no more biological specimens were available; genetic testing of the sibling could not be performed due to unavailability of a specimen; exome sequencing data are not publicly available due to privacy restrictions.
  • Stated: further investigation is required to understand the role of RHOH C82Y in pre-TCR/TCR signaling and T-cell function, and a mouse model is needed to explain B and NK cell deficiency.
  • Evident from the text: the findings rely on a single family/case, and much of the mechanistic work uses ectopic expression in Jurkat and HEK293 cell lines; the authors also note they could not determine ZAP70 phosphorylation/activation because RHOH-bound ZAP70 was barely detected.

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

Integrity signals

Registry

Crossref · 10.1007/s10875-024-01735-4

Cites

24 of 24 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

18 of 28 cited sources checked · 10 supported · 0 flagged · accuracy not yet measured

  • Interpretationcitation

    The impaired response to TCR stimulation of the patient T cell was consistent with previous RHOH deficiency cases.

    “The impaired response to TCR stimulation of the patient T cell was consistent with previous RHOH de ciency cases.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Crequer A, Troeger A, Patin E, Ma CS, Picard C, Pedergnana V, et al. Human RHOH deficiency causes T cell defects and susceptibility to EV-HPV infections. J Clin Invest. 2012;122(9):3239–47. source
  • Premisecitation

    Previously, a homozygous nonsense Y38X mutation in RHOH was reported in siblings suffering from epidermodysplasia verruciformis, increased susceptibility to certain HPV, lung disease, Burkitt lymphoma and impaired T-cell function.

    “Previously, a homozygous nonsense Y38X mutation in the RHOH gene (GenBank: NM_004310.5:c.114C > G, p.Tyr38Ter) was reported in siblings suffering from epidermodysplasia verruciformis, increased susceptibility to certain human papillomaviruses (HPV), lung disease, Burkitt lymphoma and impaired T-cell function (MIM: 618307) (6).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Crequer A, Troeger A, Patin E, Ma CS, Picard C, Pedergnana V, et al. Human RHOH deficiency causes T cell defects and susceptibility to EV-HPV infections. J Clin Invest. 2012;122(9):3239–47. source

      The mutation-identification section reports a homozygous RHOH Y38X nonsense mutation in two siblings with persistent EV-HPV infections and T-cell defects. The Introduction and Methods specify that bronchopulmonary disease and Burkitt lymphoma occurred in P1, not both siblings; the Introduction also describes their phenotype as related to, but distinct from, epidermodysplasia verruciformis. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
  • Premisecitation

    RHOH exerts a critical role in the activation, development and differentiation of T lymphocytes.

    “As a membrane-bound adaptor for proximal protein kinases involved in TCR signal transduction, RHOH exerts a critical role in the activation, development and differentiation of T lymphocytes (1,4).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [1] Gu Y, Jasti AC, Jansen M, Siefring JE. RhoH, a hematopoietic-specific rho GTPase, regulates proliferation, survival, migration, and engraftment of hematopoietic progenitor cells. Blood. 2005;105(4):1467–75. source
    • cites [4] Ahmad Mokhtar AM, Hashim IF, Mohd Zaini Makhtar M, Salikin NH, Amin-Nordin S. The role of RhoH in TCR Signalling and its involvement in diseases. Cells. 2021;10(4). source

      Section 2 describes RhoH as an adaptor that recruits kinases for TCR signalling and calls that signalling crucial for T-cell activation and development; it also describes RhoH’s membrane targeting. Section 3.2.2 links loss of RhoH to increased differentiation into TH17 cells, but says development of some unconventional T-cell subsets is not critically dependent on RhoH-mediated signalling. The source therefore supports a role in differentiation, not an unqualified critical role across T lymphocytes. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
  • Premisecitation

    The Y38X variant led to a complete loss of RHOH protein expression.

    “The Y38X variant led to a complete loss of RHOH protein expression.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Crequer A, Troeger A, Patin E, Ma CS, Picard C, Pedergnana V, et al. Human RHOH deficiency causes T cell defects and susceptibility to EV-HPV infections. J Clin Invest. 2012;122(9):3239–47. source

      In ‘Identification of a nonsense mutation in the RHOH gene,’ the authors report that the 21-kDa RHOH protein was absent from both patients’ T cells and that no protein was detected from Y38X-transfected cells. They also state that translation reinitiation cannot be formally excluded, so the evidence supports loss of detectable full-length protein but not an unqualified complete loss of all RHOH protein expression. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Premisecitation

    A germline mutation in RHOH has been identified as the cause of an autosomal recessive IEI.

    “A germline mutation in RHOH has been identi ed as the cause of an autosomal recessive IEI (6).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Crequer A, Troeger A, Patin E, Ma CS, Picard C, Pedergnana V, et al. Human RHOH deficiency causes T cell defects and susceptibility to EV-HPV infections. J Clin Invest. 2012;122(9):3239–47. source
  • Classified as probably not a substantial claim. 14
  • May combine two claims. 36

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

    Premisecitation

    RHOH is expressed in T, B, NK cells, and eosinophils in peripheral blood, but not in monocytes and neutrophils.

    “Physiologically, RHOH is expressed in T, B, NK cells, and eosinophils in peripheral blood, but not in monocytes and neutrophils (2,3).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [2] Schmidt-Mende J, Geering B, Yousefi S, Simon HU. Lysosomal degradation of RhoH protein upon antigen receptor activation in T but not b cells. Eur J Immunol. 2010;40(2):525–9. source
    • cites [3] Stoeckle C, Geering B, Yousefi S, Rožman S, Andina N, Benarafa C, et al. RhoH is a negative regulator of eosinophilopoiesis. Cell Death Differ. 2016;23(12):1961–72. source

  • May combine two claims.

    Findingcitation

    The patient showed increased number of CD8+ T cells, which showed a more activated and differentiated phenotype as indicated by an elevated proportion of HLA-DR+ or CD38+ cells.

    “The patient showed increased number of CD8 + T cells, which showed a more activated and differentiated phenotype as indicated by an elevated proportion of HLA-DR + or CD38 + cells (7).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [7] Mahnke YD, Brodie TM, Sallusto F, Roederer M, Lugli E. The who’s who of T-cell differentiation: human memory T-cell subsets. Eur J Immunol. 2013;43(11):2797–809. source

      Table 1 (Concluding remarks; page unavailable) lists HLA-DR and CD38 under activation, supporting their use as activation markers. It does not support treating a higher proportion of cells positive for either marker as evidence of greater differentiation; the table shows CD38 expression declining across the listed differentiation stages. The paper does not report the citing authors’ patient observation. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Disruption of RHOH gene has been associated with various types of B-cell malignancies and inborn errors of immunity.

    “Disruption of RHOH (MIM: 602037) gene has been associated with various types of B-cell malignancies (4) and inborn errors of immunity (IEI) (6).”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [4] Ahmad Mokhtar AM, Hashim IF, Mohd Zaini Makhtar M, Salikin NH, Amin-Nordin S. The role of RhoH in TCR Signalling and its involvement in diseases. Cells. 2021;10(4). source
    • cites [6] Crequer A, Troeger A, Patin E, Ma CS, Picard C, Pedergnana V, et al. Human RHOH deficiency causes T cell defects and susceptibility to EV-HPV infections. J Clin Invest. 2012;122(9):3239–47. source

  • May combine two claims.

    Findingcitation

    Both the CD4+ and CD8+ T-cell subsets displayed a significantly lower proportion of CD28+ cells, suggesting a decrease in naïve T cells.

    “Both the CD4 + and CD8 + T-cell subsets displayed a signi cantly lower proportion of CD28 + cells, suggesting a decrease in naïve T cell which express a high level of differentiation marker CD28 (7).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [7] Mahnke YD, Brodie TM, Sallusto F, Roederer M, Lugli E. The who’s who of T-cell differentiation: human memory T-cell subsets. Eur J Immunol. 2013;43(11):2797–809. source

  • May combine two claims.

    Interpretationcitation

    Our patient's immunophenotype shared similarities with previously reported RHOH deficiency cases in terms of reduced CD4+ T cells, increased CD8+ T cells, and an elevated proportion of activated, differentiated effector and memory CD8+ T cells.

    “Our patient's immunophenotype shared similarities with previously reported RHOH de ciency cases in terms of reduced CD4 + T cells, increased CD8 + T cells, and an elevated proportion of activated, differentiated effector and memory CD8 + T cells.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [6] Crequer A, Troeger A, Patin E, Ma CS, Picard C, Pedergnana V, et al. Human RHOH deficiency causes T cell defects and susceptibility to EV-HPV infections. J Clin Invest. 2012;122(9):3239–47. source

      In ‘Impact of human RHOH deficiency on T cells’ (page unavailable), the paper reports slightly lower CD4+ and higher CD8+ T-cell frequencies in both patients, along with increased effector-memory and revertant-memory CD8+ subsets. It describes the expanded memory cells as showing signs of exhaustion, but does not establish an elevated proportion of activated CD8+ T cells. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    RHOH is involved in T-cell receptor signaling through the recruitment and activation of ZAP70 and LCK.

    “RHOH is involved in T-cell receptor signaling through the recruitment and activation of ZAP70 and LCK (4,5).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [4] Ahmad Mokhtar AM, Hashim IF, Mohd Zaini Makhtar M, Salikin NH, Amin-Nordin S. The role of RhoH in TCR Signalling and its involvement in diseases. Cells. 2021;10(4). source
    • cites [5] Gu Y, Chae HD, Siefring JE, Jasti AC, Hildeman DA, Williams DA. RhoH GTPase recruits and activates Zap70 required for T cell receptor signaling and thymocyte development. Nat Immunol. 2006;7(11):1182–90. source

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

    Premisecitation

    Somatic mutations in RHOH gene have been linked to certain hematological malignancies.

    “Somatic mutations in RHOH gene have been linked to certain hematological malignancies (4).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [4] Ahmad Mokhtar AM, Hashim IF, Mohd Zaini Makhtar M, Salikin NH, Amin-Nordin S. The role of RhoH in TCR Signalling and its involvement in diseases. Cells. 2021;10(4). source

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

    Premisecitation

    A recent study in patients with CD28 deficiency strongly suggests that the poor antibody responses seen in those patients after vaccination result from a deficiency of CD4 T cell response, due to the absence of CD28 co-stimulation signaling.

    “A recent study in patients with CD28 de ciency strongly suggest that the poor antibody responses seen in those patients after vaccination results from a de ciency of CD4 T cell response, due to the absence of CD28 co-stimulation signaling (16).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [16] Béziat V, Rapaport F, Hu J, Titeux M, Bonnet des Claustres M, Bourgey M, et al. Humans with inherited T cell CD28 deficiency are susceptible to skin papillomaviruses but are otherwise healthy. Cell. 2021;184(14):3812–e2830. source

      Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Studies on RhoH-/- mice have observed a defect in thymocyte selection, maturation, lower T-cell cytotoxicity and cytokine release, and demonstrated that it is a result of reduced activation of ZAP70-mediated signaling and impaired translocation of ZAP70 to the immunological synapse.

    “Studies on RhoH -/-mice have observed a defect in thymocyte selection, maturation, lower T-cell cytotoxicity and cytokine release, and have demonstrated that it is a result of reduced activation of ZAP70-mediated signaling and impaired translocation of ZAP70 to the immunological synapse (5,9).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [5] Gu Y, Chae HD, Siefring JE, Jasti AC, Hildeman DA, Williams DA. RhoH GTPase recruits and activates Zap70 required for T cell receptor signaling and thymocyte development. Nat Immunol. 2006;7(11):1182–90. source
    • cites [9] Chae HD, Siefring JE, Hildeman DA, Gu Y, Williams DA. RhoH regulates subcellular localization of ZAP-70 and Lck in T cell receptor signaling. PLoS ONE. 2010;5(11):e13970. source

      Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

      Experimental — accuracy not yet measured

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

    Premisecitation

    BCR signaling has been found intact in Rhoh-deficient mice.

    “However, BCR signaling has also been found intact in Rhoh-de cient mice (15).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [15] Sanchez-Aguilera A, Rattmann I, Drew DZ, Müller LU, Summey V, Lucas DM, et al. Involvement of RhoH GTPase in the development of B-cell chronic lymphocytic leukemia. Leukemia. 2010;24(1):97–104. source

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

    Findingown data

    HIV serology was negative.

    “His HIV serology was negative.”
    Quote in the paperNot a substantial claim

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

    Findingown data

    HPV serology test results of our patient and his parents were negative.

    “HPV infection was found in previous reported RHOH de ciency cases, but HPV serology test results of our patient and his parents were negative.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The patient exhibited recurrent infections affecting multiple organs, including the lungs, retina, and brain, caused by various opportunistic pathogens such as CMV, RV, P. jirovecii, helminth, and multiple bacteria.

    “Thirdly, the patient exhibited recurrent infections affecting multiple organs, including the lungs, retina, and brain, caused by various opportunistic pathogens such as CMV, RV, P. jirovecii, helminth, and multiple bacteria.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingcitation

    In RHOH WT-expressing Jurkat cells, ZAP70 could bind with RHOH in a resting state, and RHOH-bound ZAP70 increased after anti-CD3 stimulation.

    “In RHOH WTexpressing Jurkat cells, ZAP70 could bind with RHOH in a resting state, and RHOH-bound ZAP70 increased after anti-CD3 stimulation, consistent with previous publications (5).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [5] Gu Y, Chae HD, Siefring JE, Jasti AC, Hildeman DA, Williams DA. RhoH GTPase recruits and activates Zap70 required for T cell receptor signaling and thymocyte development. Nat Immunol. 2006;7(11):1182–90. source

  • May combine two claims.

    Findingown data

    Throughout this period, serial immunological tests revealed persistent CD4+ T cell lymphopenia and hypoimmunoglobulinemia.

    “Throughout this period, a series of immunological tests revealed persistent CD4 + T cell lymphopenia and hypoimmunoglobulinemia (Table 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Genetic analysis identified a novel germline homozygous missense mutation in RHOH (p.Cys82Tyr), which represents the first reported case of a missense variant in RHOH.

    “Genetic analysis identi ed a novel germline homozygous missense mutation in RHOH (p.Cys82Tyr), which, to the best of our knowledge, represents the rst reported case of a missense variant in RHOH.”
    Quote in the paperSubstantial claim

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

    Findingown data

    Brain MRI revealed neurocysticercosis of the right cerebellar hemisphere.

    “He developed a headache during this period, and a brain MRI revealed neurocysticercosis of the right cerebellar hemisphere.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The proportions of B cells and NK cells were both below average.

    “The proportions of B cells (0.9%, reference range: 6.48-16.64%)and NK cells (3.5%, reference range: 5.17-24.65%)were both below average.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Immunological tests indicated a normal CD4+ T cell number, but a decreased CD4:CD8 ratio.

    “Immunological tests indicated a normal CD4 + T cell number, but a decreased CD4:CD8 ratio (Table 1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingnone given

    The patient is from a consanguineous family, with both parents being healthy.

    “The patient is from a consanguineous family, with both parents being healthy.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingnone given

    The patient is a 21-year-old male with recurrent infections affecting multiple organs, yet without any skin lesions.

    “Here we present a unique case of a 21-yearold male patient with recurrent infections affecting multiple organs, yet without any skin lesions.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The clinical possibility of acquired immunodeficiency was considered but later ruled out as tests for anti-IFN-γ autoantibody and HIV RNA were both negative.

    “The clinical possibility of acquired immunode ciency was considered but later ruled out as tests for anti-IFN-γ autoantibody and HIV RNA were both negative.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingnone given

    The C82Y variant allows for normal transcription of the RHOH mutant, but disrupted its protein expression.

    “In our case, the C82Y variant allows for normal transcription of the RHOH mutant, but disrupted its protein expression.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The typical skin lesion associated with epidermodysplasia verruciformis was absent, and no evidence of HPV infection was found.

    “Secondly, the typical skin lesion associated with epidermodysplasia verruciformis was absent, and no evidence of HPV infection was found.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The patient had been suffering from recurrent lung infections and deterioration of eyesight for over a year.

    “Upon admission to our hospital, the patient had been suffering from recurrent lung infections and deterioration of eyesight for over a year.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The patient remained well for two years, but succumbed to an unmanageable central nerve system infection in 2021.

    “Following discharge, the patient remained well for two years, but unfortunately succumbed to an unmanageable central nerve system infection in 2021.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Both parents are heterozygous carriers of this variant, supporting an autosomal-recessive inheritance pattern for this novel germline RHOH mutation.

    “Both parents are heterozygous carriers of this variant, supporting an autosomal-recessive inheritance pattern for this novel germline RHOH mutation (Fig. 2A).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Serial immunological tests revealed persistently reduced numbers of CD4+ T and B cells, profound hypoimmunoglobulinemia, and sometimes a decreased number of NK cells.

    “As shown in Table 1, serial immunological tests conducted during 2018-2019 revealed that the patient exhibited persistently reduced numbers of CD4 + T and B cells, profound hypoimmunoglobulinemia, and sometimes a decreased number of NK cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The patient's T cells failed to enlarge, as indicated by the significantly lower FSC of viable cells compared to his parents, suggesting a general defect in TCR-mediated T cell activation in the patient.

    “The patient's T cells failed to enlarge, as indicated by the signi cantly lower FSC of viable cells compared to his parents, suggesting a general defect in TCR-mediated T cell activation in the patient.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    His sibling also suffered from recurrent severe lung infections and eventually died of severe respiratory infection.

    “His sibling also suffered from recurrent severe lung infections.”
    • Also stated in: § Clinical features of the patient · plain
      “Despite the implementation of IVIg and mechanical ventilation, she kept deteriorating, and eventually died of severe respiratory infection.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Our patient presented a persistent decrease in B and NK cell numbers, and profound hypoimmunoglobulinemia, indicating a combined defect in both cellular and humoral immune responses.

    “In contrast, our patient presented a persistent decrease in B and NK cell numbers, and profound hypoimmunoglobulinemia, indicating a combined defect in both cellular and humoral immune responses.”
    Quote in the paperSubstantial claim

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

    Findingnone given

    His sister also succumbed to a lung infection during early adulthood.

    “His sister also succumbed to a lung infection during early adulthood.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Results from the immunoprecipitation analysis support our hypothesis, as RHOH-bound ZAP70 was undetectable in RHOH C82Y Jurkat cells.

    “Results from the immunoprecipitation analysis support our hypothesis, as RHOH-bound ZAP70 was undetectable in RHOH C82Y Jurkat cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingnone given

    The patient's parents were consanguineous cousins, and both were in good health.

    “The patient's parents were consanguineous cousins, and both were in good health.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The patient exhibited a persistent decrease in CD4+ T, B, and NK cell counts, and hypoimmunoglobulinemia.

    “The patient exhibited a persistent decrease in CD4 + T, B, and NK cell counts, and hypoimmunoglobulinemia.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The C82Y variant showed reduced RHOH protein expression and impaired interaction with the TCR signaling molecule ZAP70.

    “Furthermore, the C82Y variant showed reduced RHOH protein expression and impaired interaction with the TCR signaling molecule ZAP70.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    RHOH is an atypical small GTPase predominantly expressed in hematopoietic cells and plays a vital role in immune function.

    “RHOH, an atypical small GTPase predominantly expressed in hematopoietic cells, plays a vital role in immune function.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [1] Gu Y, Jasti AC, Jansen M, Siefring JE. RhoH, a hematopoietic-specific rho GTPase, regulates proliferation, survival, migration, and engraftment of hematopoietic progenitor cells. Blood. 2005;105(4):1467–75. source
    • cites [2] Schmidt-Mende J, Geering B, Yousefi S, Simon HU. Lysosomal degradation of RhoH protein upon antigen receptor activation in T but not b cells. Eur J Immunol. 2010;40(2):525–9. source

  • May combine two claims.

    Interpretationown data

    The C82Y is a loss-of-function variant but does not exert a dominant negative effect over the endogenous WT RHOH.

    “These observations suggest that the C82Y is a loss-of-function variant but does not exert a dominant negative effect over the endogenous WT RHOH.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    RHOH is specifically expressed in the hematopoietic system, with variable expression among different leukocyte subsets.

    “It is speci cally expressed in the hematopoietic system, with variable expression observed among different leukocyte subsets (2).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [2] Schmidt-Mende J, Geering B, Yousefi S, Simon HU. Lysosomal degradation of RhoH protein upon antigen receptor activation in T but not b cells. Eur J Immunol. 2010;40(2):525–9. source

  • May combine two claims.

    Interpretationown data

    The C82Y variant is a hypomorphic mutation which hampers RHOH protein expression and impairs its function in TCR-mediated T cell activation.

    “Collectively, these results indicate that the C82Y variant is a hypomorphic mutation which hampers RHOH protein expression and impairs its function in TCR-mediated T cell activation.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The functional effects of RHOH C82Y observed in HEK293 and Jurkat cell lines reflect the effects of the endogenous RHOH C82Y variant in the patient's T cells.

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

    Study design and conductnone given

    Written informed consent was obtained from the patient and his parents.

    “Written informed consent was obtained from the patient and his parents.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Together, these data suggest that the newly identified autosomal-recessive RHOH variant is associated with T cell dysfunction and recurrent opportunistic infections, functioning as a hypomorph by disrupting ZAP70-mediated TCR signaling.

    “Together, these data suggest that the newly identi ed autosomal-recessive RHOH variant is associated with T cell dysfunction and recurrent opportunistic infections, functioning as a hypomorph by disrupting ZAP70-mediated TCR signaling.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Dorn et al. found that RHOH is not required for TCR-induced activation of ZAP70, but is indispensable for the efficient interaction of ZAP70 with the LAT signalosome.

    “Nevertheless, Dorn, T., et al. found that RHOH is not required for TCR-induced activation of ZAP70, but is indispensable for the e cient interaction of ZAP70 with the LAT signalosome (10).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [10] Dorn T, Kuhn U, Bungartz G, Stiller S, Bauer M, Ellwart J, et al. RhoH is important for positive thymocyte selection and T-cell receptor signaling. Blood. 2007;109(6):2346–55. source

  • May combine two claims.

    Interpretationown data

    In conclusion, our case expands the genetic and phenotypic spectrum of RHOH deficiency and provides new insights into the development and regulation of T, B and NK cells.

    “In conclusion, our case expands the genetic and phenotypic spectrum of RHOH de ciency and provides new insights into the development and regulation of T, B and NK cells.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    The study received approval from the institutional ethics review board of Huashan Hospital, Fudan University, and was conducted in accordance with the principles outlined in the Declaration of Helsinki.

    “The study received approval from the institutional ethics review board of Huashan Hospital, Fudan University, and was conducted in accordance with the principles outlined in the Declaration of Helsinki.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    The C82Y variant led to a major defect in the interaction between RHOH and ZAP70, and impaired subsequent TCR signaling, at an extent similarly to what has been observed in cases with stop-gain RHOH mutation.

    “The C82Y variant led to a major defect in the interaction between RHOH and ZAP70, and impaired subsequent TCR signaling, at an extent similarly to what has been observed in cases with stop-gain RHOH mutation.”
    Quote in the paperSubstantial claim
46 of 46

  • The paper relies on this without support.

    The functional effects of RHOH C82Y observed in HEK293 and Jurkat cell lines reflect the effects of the endogenous RHOH C82Y variant in the patient's T cells.

    “We therefore evaluate the expression and function of the C82Y RHOH variant using cell lines generated to express RHOH WT or RHOH C82Y .”
    “Impaired TCR-mediated T-cell activation was observed in both the patient and in RHOH C82Yexpressing Jurkat cells”

  • The paper relies on this without support.

    The patient's sibling's severe lung infection and death were due to the same RHOH C82Y variant.

    “His sibling also suffered from recurrent severe lung infections.”
    “Unfortunately, genetic assays of the patient's sibling could not be performed due to the unavailability of a specimen.”

  • The paper relies on this without support.

    The in silico pathogenicity predictions are reliable indicators that the C82Y variant is pathogenic.

    “It is predicted to be pathogenic by multiple in silico pathogenicity prediction tools, including CADD (score = 26.4),REVEL (0.831, pathogenic), SIFT (0.001, deleterious), PolyPhen-2 (1.0, probably damaging), and MutationTaster (1, disease causing).”
3 of 3

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.

  • September 2019 CD4 count and percentage disagree

    Minor

    The September 2019 CD8 count and percentage imply about 2,210 lymphocytes; 464 CD4 cells would therefore be about 21.0%, not the reported 20.1%.

  • November 2019 NK count and percentage disagree

    Minor

    The November CD3 count and percentage imply about 991 lymphocytes. An NK count of 244 would be about 24.6% of that total, rather than the reported 14.19%.

  • September 2018 white blood cell counts disagree

    Minor

    The narrative gives a white blood cell count of 7.41×109/L for the initial episode, equivalent to 7,410 cells/μl. Table 1 lists 8,880 cells/μl for September 2018.

  • CD4 count is called persistently reduced despite a normal initial result

    Minor

    The paper describes the CD4 count as normal at the initial assessment, but later says the 2018-2019 tests showed persistently reduced CD4 counts.

4 of 4

Cites

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Authors
Zhou, J., Qian, M., Jiang, N., Wu, J., Feng, X., Yu, M., Min, Q., Xu, H., Yang, Y., Yang, Q., Zhou, F., Shao, L., Zhu, H., Yang, Y., Wang, J.-Y., Ruan, Q., & Zhang, W.
Affiliations
  • Shanghai Medical College of Fudan University (CN)
  • Children's Hospital of Fudan University (CN)
Corresponding author
Ruan, Qiaoling
ORCID
Venue
Journal of Clinical Immunology
Publisher
Springer Science and Business Media LLC
Published
2024-05-22
Volume
44
Issue
6
Pages
131-131
ISSN
  • 0271-9142 (ISSN-L)
  • 1573-2592
DOI
10.1007/s10875-024-01735-4
OpenAlex
W4398222262
Semantic Scholar
0493a8f9d4034ddf802f84357ea537921912d385
Cited by
4
References
23
Citations by year
2026: 1 · 2025: 3
FWCI
0.67
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Clinical Immunology (publishedVersion)
  • Research Square (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
National Natural Science Foundation of China (32330033, 82271794)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Immunodeficiency and Autoimmune Disorders
  • Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Blood disorders and treatments
Keywords
RhoH, T cell dysfunction, ZAP-70, T cell receptor signaling, CD69, CD4+ T cell deficiency, NK cell deficiency, autosomal recessive immunodeficiency, epidermodysplasia verruciformis
MeSH terms
Homozygote, Humans, Lymphocyte Activation, Male, Opportunistic Infections, Pedigree, Receptors, Antigen, T-Cell, Recurrence, T-Lymphocytes, Jurkat Cells, ZAP-70 Protein-Tyrosine Kinase, Young Adult

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 2026-11-02.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-11-02.

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