scholarly journals Tandem mass spectrometry, but not T-cell receptor excision circle analysis, identifies newborns with late-onset adenosine deaminase deficiency

2013 ◽  
Vol 131 (6) ◽  
pp. 1604-1610 ◽  
Author(s):  
Giancarlo la Marca ◽  
Clementina Canessa ◽  
Elisa Giocaliere ◽  
Francesca Romano ◽  
Marzia Duse ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
T Cell ◽  
Tandem Mass Spectrometry ◽  
Adenosine Deaminase ◽  
T Cell Receptor ◽  
Late Onset ◽  
Cell Receptor ◽  
Tandem Mass ◽  
Adenosine Deaminase Deficiency

scholarly journals Adenosine deaminase deficiency increases thymic apoptosis and causes defective T cell receptor signaling

2001 ◽  
Vol 108 (1) ◽  
pp. 131-141 ◽  
Author(s):  
Sergey G. Apasov ◽  
Michael R. Blackburn ◽  
Rodney E. Kellems ◽  
Patrick T. Smith ◽  
Michail V. Sitkovsky
Keyword(s):  
T Cell ◽  
Adenosine Deaminase ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Receptor Signaling ◽  
Adenosine Deaminase Deficiency ◽  
T Cell Receptor Signaling ◽  
Cell Receptor Signaling

Long-Term Culture and T Cell Receptor Analysis of T Cell Clones Isolated from a Patient with Adenosine Deaminase Deficiency and Type I Diabetes

1994 ◽  
Vol 73 (3) ◽  
pp. 362-366 ◽  
Author(s):  
Piergiuseppe De Berardinis ◽  
Maria Neve Ombra ◽  
Catello Buono ◽  
Roberto Toraldo ◽  
Francesco Vetrano ◽  
...  
Keyword(s):  
T Cell ◽  
Adenosine Deaminase ◽  
T Cell Receptor ◽  
Type I Diabetes ◽  
Cell Receptor ◽  
Type I ◽  
Adenosine Deaminase Deficiency ◽  
T Cell Clones ◽  
Cell Clones

Newborn screening and molecular features of patients with multiple acyl-CoA dehydrogenase deficiency in Quanzhou, China

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Yiming Lin ◽  
Weifeng Zhang ◽  
Zhixu Chen ◽  
Chunmei Lin ◽  
Weihua Lin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Genetic Analysis ◽  
Tandem Mass Spectrometry ◽  
Newborn Screening ◽  
Dehydrogenase Deficiency ◽  
Late Onset ◽  
False Negative ◽  
Tandem Mass ◽  
Lipid Storage Myopathy ◽  
Molecular Features

Abstract Objectives Multiple acyl-CoA dehydrogenase deficiency (MADD) is an autosomal recessive disorder of fatty acid, amino acid and choline metabolism. Late-onset MADD is caused by ETFDH mutations and is the most common lipid storage myopathy in China. However, few patients with MADD have been identified through newborn screening (NBS). This study assessed the acylcarnitine profiles and molecular features of patients with MADD identified through NBS. Methods From January 2014 to June 2020, 479,786 newborns screened via tandem mass spectrometry were recruited for this study. Newborns with elevated levels of multiple acylcarnitines were recalled, those who tested positive in the reassessment were referred for genetic analysis. Results Of 479,786 newborns screened, six were diagnosed with MADD. The MADD incidence in the Chinese population was estimated to be 1:79,964. Initial NBS revealed five patients with typical elevations in the levels of multiple acylcarnitines; however, in one patient, acylcarnitine levels were in the normal reference range during recall. Notably, one patient only exhibited a mildly increased isovalerylcarnitine (C5) level at NBS. The patient with an atypical acylcarnitine profile was diagnosed with MADD by targeted gene sequencing. Six distinct ETFDH missense variants were identified, with the most common variant being c.250G>A (p.A84T), with an allelic frequency of 58.35 (7/12). Conclusions These findings revealed that it is easy for patients with MADD to go unidentified, as they may have atypical acylcarnitine profiles at NBS and the recall stage, indicating the value of genetic analysis for confirming suspected inherited metabolic disorders in the NBS program. Therefore, false-negative (FN) results may be reduced by combining tandem mass spectrometry (MS/MS) with genetic testing in NBS for MADD.

Skewing of the T-Cell Receptor Repertoire in Patients Receiving Rituximab after Allogeneic Hematopoietic Cell Transplantation: What Lies Beneath?

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3962-3962
Author(s):  
Apostolia Papalexandri ◽  
Maria Karypidou ◽  
Evangelia Stalika ◽  
Michail Iskas ◽  
Anna Vardi ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Transplantation ◽  
T Cell Receptor ◽  
Hematopoietic Cell Transplantation ◽  
Late Onset ◽  
Cell Receptor ◽  
Hematopoietic Cell ◽  
Cytotoxic T Cell ◽  
Gene Repertoire ◽  
Lgl Leukemia

Abstract The development of CD3+/CD8+/CD57+ cytotoxic cell expansions after allogeneic hematopoietic cell transplantation (allo-HCT) driven by antigenic stimulation, viral or associated with chronic graft-versus-host disease (cGvHD), has been suggested as related with favorable outcome. Rituximab, an anti-CD20 humanized monoclonal antibody, has been linked to the development of oligo- or even monoclonal expansions of CD3+/CD8+/CD57+ T-large granular lymphocytes (T-LGLs) that can manifest with neutropenia of delayed origin in relation to Rituximab administration. We have recently reported remarkable skewing of the T-cell receptor (TR) gene repertoire in two allo-HCT transplanted patients with delayed neutropenia associated with T-LGL expansions developing in a context of GvHD and Rituximab administration for EBV reactivation. Prompted by these preliminary findings, we here extend our immunogenetic studies of the TR repertoire in patients receiving Rituximab post allo-HCT. The study group was comprised of 9 patients (including the two previously reported) aged 14-50 years (median 41) who were subjected to myeloablative allo-HCT (4 from matched related, 3 from matched unrelated donors), haplo-identical transplantation (1) or Reduced Intensity Conditioning-allo-HCT from sibling donor (1), all for hematological malignancies. All patients received Rituximab consecutively between 2010-2013 either as pre-emptive treatment for EBV reactivation or against refractory cGvHD. In all patients TR gene repertoire analysis was performed at least one year after the transplantation (range 12-72 months), when immune reconstitution normally would have been achieved, and 5-24 months after the first treatment with Rituximab. Each patient received a mean of 7 cycles of Rituximab (range, 1-14). TRBV-TRBD-TRDJ gene rearrangements were PCR-amplified on genomic DNA isolated from bone marrow samples using the BIOMED2 protocol and subjected to classic subcloning/Sanger sequencing. Sequence data was interpreted using the IMGT/V-QUEST tool. A total of 164 sequences were analyzed (9-25/case, median=18) revealing 106 productive TRBV-TRBD-TRBJ rearrangements. Among the 29 TRBV functional genes identified only three accounted for 48% of cases: (i) TRBV27*01 (25%), (ii) TRBV6-5*01 (13%), (iii) TRBV6-2*01 (10%). Of note, TRBV27*01 has been reported as the most frequent TRBV gene in Rituximab-related late-onset neutropenia in CLL. All cases were found to carry clusters of identical (>=2) rearrangements corresponding to clonotypes. In the majority of cases (5/9), 2-4 (median 3) immunodominant clonotypes accounted for over 30% of the analyzed sequences (frequency of immunodominant clonotype/case 13-40%). Lymphocyte subpopulation analysis by flow cytometry in 6 patients revealed T-LGL expansion. Samples from additional time points (spanning a period of 10 years), pre- and post- Rituximab, were studied in one patient. Analysis of 71 sequences demonstrated progressive expansion of a certain clonotype overtime, associated with the emergence of steroid-refractory autoimmune hemolytic anemia in a context of CD3+CD8+CD57+ lymphoproliferation. This particular clonotype dominated the repertoire by far, thus establishing a diagnosis of T-LGL leukemia. which, remarkably, proved to be of donor origin (97% and 30% donor chimerism in T lymphocytes and total hematopoeisis, respectively). No association of oligoclonality to stronger GvL effect could be found among the rest of the patients. However, a strong correlation with cGvHD (100% vs 25% among polyclonal cases) was identified. Late-onset neutropenia was documented in 4/9 patients, regardless of the composition of the repertoire i.e whether it was polyclonal or oligo(mono)clonal. In conclusion, we report frequent development of oligoclonal cytotoxic T-cell populations after Rituximab treatment post allo-HCT likely of multifactorial evidence. Direct evidence of the anti-leukemic effect of this phenomenon could not be provided, however, the observed association of oligoclonality with GvHD and the development of a possible “T-LGL leukemia vs leukemia” effect in one patient is noteworthy and merits further investigation. Finally, the observed skewing of the TR gene repertoire strongly implicates antigen selection in the development of cytotoxic T-cell expansions after allo-HCT. Disclosures No relevant conflicts of interest to declare.

scholarly journals Somatic mutations and T-cell clonality in patients with immunodeficiency

Haematologica ◽  
2019 ◽  
Vol 105 (12) ◽  
pp. 2757-2768 ◽  
Author(s):  
Paula Savola ◽  
Timi Martelius ◽  
Matti Kankainen ◽  
Jani Huuhtanen ◽  
Sofie Lundgren ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Receptor ◽  
Common Variable Immunodeficiency ◽  
Somatic Mutations ◽  
Late Onset ◽  
Cell Receptor ◽  
Cd8 Cells ◽  
Receptor Repertoire ◽  
Common Variable

Common variable immunodeficiency and other late-onset immunodeficiencies often co-manifest with autoimmunity and lymphoproliferation. The pathogenesis of most cases is elusive, as only a minor subset harbors known monogenic germline causes. The involvement of both B and T cells is however implicated. To study whether somatic mutations in CD4+ and CD8+ T cells associate with immunodeficiency, we recruited 17 patients and 21 healthy controls. Eight patients had late-onset common variable immunodeficiency and nine patients other immunodeficiency and/or severe autoimmunity. In total, autoimmunity occurred in 94% and lymphoproliferation in 65%. We performed deep sequencing of 2533 immune-associated genes from CD4+ and CD8+ cells. Deep T-cell receptor beta sequencing was used to characterize CD4+ and CD8+ T-cell receptor repertoires. The prevalence of somatic mutations was 65% in all immunodeficiency patients, 75% in common variable immunodeficiency and 48% in controls. Clonal hematopoiesis-associated variants in both CD4+ and CD8+ cells occurred in 24% of immunodeficiency patients. Results demonstrated mutations in known tumor suppressors, oncogenes, and genes that are critical for immune- and proliferative functions, such as STAT5B (two patients), C5AR1 (two patients), KRAS (one patient), and NOD2 (one patient). Additionally, as a marker of T-cell receptor repertoire perturbation, common variable immunodeficiency patients harbored increased frequencies of clones with identical complementarity determining region 3 sequences despite unique nucleotide sequences when compared to controls. In conclusion, somatic mutations in genes implicated for autoimmunity and lymphoproliferation are common in CD4+ and CD8+ cells of patients with immunodeficiency. They may contribute to immune dysregulation in a subset of immunodeficiency patients.

scholarly journals Phosphorylation of CrkL S114 induced by common gamma chain cytokines and T-cell receptor signal transduction

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Armando Estrada ◽  
Alejandro C. Rodriguez ◽  
Georgialina Rodriguez ◽  
Alice H. Grant ◽  
Yoshira M. Ayala-Marin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
T Cell ◽  
T Cell Receptor ◽  
T Cell Activation ◽  
Interleukin 2 ◽  
Cell Receptor ◽  
Serine Phosphorylation ◽  
Mass Spectrometry Analysis ◽  
Gamma Chain ◽  
Common Gamma Chain

AbstractT-cell activation and cellular expansion by common gamma chain cytokines such as Interleukin-2 is necessary for adaptive immunity. However, when unregulated these same pathways promote pathologies ranging from autoimmune disorders to cancer. While the functional role of Interleukin-2 and downstream effector molecules is relatively clear, the repertoire of phosphoregulatory proteins downstream of this pathway is incomplete. To identify phosphoproteins downstream of common gamma chain receptor, YT cells were radiolabeled with [32P]-orthophosphate and stimulated with Interleukin-2. Subsequently, tyrosine phosphorylated proteins were immunopurified and subjected to tandem mass spectrometry—leading to the identification of CrkL. Phosphoamino acid analysis revealed concurrent serine phosphorylation of CrkL and was later identified as S114 by mass spectrometry analysis. S114 was inducible through stimulation with Interleukin-2 or T-cell receptor stimulation. Polyclonal antibodies were generated against CrkL phospho-S114, and used to show its inducibility by multiple stimuli. These findings confirm CrkL as an Interleukin-2 responsive protein that becomes phosphorylated at S114 by a kinase/s downstream of PI3K and MEK/ERK signaling.

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