Assessment of activated gut-homing CD8+ T cells in blood by flow cytometry during a 3-day gluten challenge

Núñez, Concepción; Gómez-Aguililla, Sara; Corzo, María; Fernández‐Bañares, Fernando; Bodas, Andrés; Farrais, Sergio; López-Palacios, Natalia; Rubio, Mercedes

doi:10.1016/bs.mcb.2022.09.020

Cited by 2 publications

(2 citation statements)

References 26 publications

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“…Most studies evaluating T cell responses to gluten challenge used gluten-containing sliced white bread ( 6 , 50 – 52 , 54 , 61 ), although other gluten vehicles have been also successfully employed ( 55 , 62 , 63 ). We started using a bread-challenge, but replaced it by pure gluten, after ascertaining that the activated gut-homing CD8 + T cells were also detected ( 64 ), aimed to reduce ATIs and FODMAPs and the possible consequently associated clinical symptoms.…”

Section: Blood Lymphogrammentioning

confidence: 99%

See 1 more Smart Citation

Intestinal and blood lymphograms as new diagnostic tests for celiac disease

Roy

Fernández‐Bañares²,

Corzo³

et al. 2023

Front. Immunol.

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Accurate celiac disease (CD) diagnosis is still challenging for some specific patients or circumstances. Thus, much effort has been expended last decades focused on seronegative or low grade enteropathy CD and, especially, on enable early diagnosis of individuals on a gluten-free diet (GFD). We discuss here two diagnostic approaches based on immunophenotyping by flow cytometry that we expect to reduce the persistent low diagnostic rates and the common diagnostic delay. The intraepithelial lymphogram is based on determining the percentage of γδ+ and surface CD3- lymphocytes in the intestinal epithelium. The concomitant increase in γδ+ and decrease in surface CD3- intraepithelial lymphocytes has been termed the celiac lymphogram and has been proved to be discriminative in seronegative, low grade enteropathy and potential CD, as well as in most CD patients on a GFD. A blood lymphogram based on the analysis of activated gut-homing CD8+ T cells combined with a 3-day gluten challenge is also considered, which has shown high sensitivity and specificity to diagnose seropositive Marsh 1 and Marsh 3 CD in individuals following a GFD. In addition, flow cytometry can be extremely useful in cases of refractory CD type II to identify aberrant cells. Those approaches represent highly accurate methods for CD diagnosis, being simple, fast, highly reproducible and of easy implementation in clinical practice.

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Section: Blood Lymphogrammentioning

confidence: 99%

“…High reproducibility exists also when using different monoclonal antibodies. Finally, it must be underlined the speed to get the results: less than 3 hours from the second blood extraction ( 64 , 65 ).…”

Section: Blood Lymphogrammentioning

confidence: 99%

Intestinal and blood lymphograms as new diagnostic tests for celiac disease

Roy

Fernández‐Bañares²,

Corzo³

et al. 2023

Front. Immunol.

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Diagnosis of celiac disease on a gluten-free diet: a multicenter prospective quasi-experimental clinical study

Gómez-Aguililla,

Farrais,

López-Palacios

et al. 2024

Preprint

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BackgroundDiagnosing celiac disease (CD) in individuals adhering to a gluten-free diet (GFD) presents significant challenges. Current guidelines recommend a gluten challenge (GC) lasting at least 6-8 weeks, which has several limitations.ObjectivesThis study compares four approaches previously proposed for diagnosing CD on a GFD: IL-2 serum levels, gut-homing CD8+T cells, %TCRγδ+intraepithelial lymphocytes (IELs), andUBE2L3gene expression. Additionally, we evaluated the CD8+T-cell based method with a 3-day GC against the standard GC protocol.MethodsWe conducted a multicenter prospective quasi-experimental clinical study. Two subsets of individuals were considered: 1) 20 patients with CD and 15 non-CD controls previously diagnosed, to evaluate the first aim; 2) 45 individuals with uncertain diagnosis who were on a GFD and required GC following current clinical guidelines, to assess the second aim. All participants underwent a 3-day GC (10 g gluten/day).ResultsAmong CD patients and non-CD controls, the sensitivity and specificity of IL-2, gut-homing CD8+T cells, andUBE2L3were 82.4% and 83.3%, 88.2% and 100%, and 52.9% and 100%, respectively. The percentage of TCRγδ+IELs showed 88.2% sensitivity. In the uncertain diagnosis group, a CD8+T-cell positive response was observed in 8 of the 45 subjects.ConclusionThe percentage of TCRγδ+IELs and the gut-homing CD8+T-cell assay are promising diagnostic methods for CD on a GFD. Notably, the CD8+T-cell assay provides a consistent and reliable alternative to the extended GC, eliminating the need for the invasive procedures to obtain duodenal samples and the prolonged gluten ingestion.

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Assessment of activated gut-homing CD8+ T cells in blood by flow cytometry during a 3-day gluten challenge

Cited by 2 publications

References 26 publications

Intestinal and blood lymphograms as new diagnostic tests for celiac disease

Intestinal and blood lymphograms as new diagnostic tests for celiac disease

Diagnosis of celiac disease on a gluten-free diet: a multicenter prospective quasi-experimental clinical study

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