Repositorio
Institucional

Latorre, Irene

Diaz-Fernandez, Sergio

Villar-Hernandez, Raquel

Stojanovic, Zoran

Fernandez, Marco

Galvao, Maria Luiza De Souza

Tolosa, Guillermo

Sanchez-Montalva, Adrian

Abad, Jorge

Jimenez-Fuentes, Maria Angeles

Safont, Guillem

Romero, Iris

Sabria, Josefina

Prat, Cristina

Dominguez, Jose

Study of CD27, CD38, HLA-DR and Ki-67 immune profiles for the characterization of active tuberculosis, latent infection and end of treatment

FRONTIERS MEDIA SA

FRONTIERS IN MICROBIOLOGY

en

BackgroundCurrent blood-based diagnostic tools for TB are insufficient to properly characterize the distinct stages of TB, from the latent infection (LTBI) to its active form (aTB), nor can they assess treatment efficacy. Several immune cell biomarkers have been proposed as potential candidates for the development of improved diagnostic tools. ObjectiveTo compare the capacity of CD27, HLA-DR, CD38 and Ki-67 markers to characterize LTBI, active TB and patients who ended treatment and resolved TB. MethodsBlood was collected from 45 patients defined according to clinical and microbiological criteria as: LTBI, aTB with less than 1 month of treatment and aTB after completing treatment. Peripheral blood mononuclear cells were stimulated with ESAT-6/CFP-10 or PPD antigens and acquired for flow cytometry after labelling with conjugated antibodies against CD3, CD4, CD8, CD27, IFN-gamma, TNF-alpha, CD38, HLA-DR, and Ki-67. Conventional and multiparametric analyses were done with FlowJo and OMIQ, respectively. ResultsThe expression of CD27, CD38, HLA-DR and Ki-67 markers was analyzed in CD4(+) T-cells producing IFN-gamma and/or TNF-alpha cytokines after ESAT-6/CFP-10 or PPD stimulation. Within antigen-responsive CD4(+) T-cells, CD27(-) and CD38(+) (ESAT-6/CFP-10-specific), and HLA-DR+ and Ki-67(+) (PPD- and ESAT-6/CFP-10-specific) populations were significantly increased in aTB compared to LTBI. Ki-67 demonstrated the best discriminative performance as evaluated by ROC analyses (AUC > 0.9 after PPD stimulation). Data also points to a significant change in the expression of CD38 (ESAT-6/CFP-10-specific) and Ki-67 (PPD- and ESAT-6/CFP-10-specific) after ending the anti-TB treatment regimen. Furthermore, ratio based on the CD27 median fluorescence intensity in CD4(+) T-cells over Mtb-specific CD4(+) T-cells showed a positive association with aTB over LTBI (ESAT-6/CFP-10-specific). Additionally, multiparametric FlowSOM analyses revealed an increase in CD27 cell clusters and a decrease in HLA-DR cell clusters within Mtb-specific populations after the end of treatment. ConclusionOur study independently confirms that CD27(-), CD38(+), HLA-DR+ and Ki-67(+) populations on Mtb-specific CD4(+) T-cells are increased during active TB disease. Multiparametric analyses unbiasedly identify clusters based on CD27 or HLA-DR whose abundance can be related to treatment efficacy. Further studies are necessary to pinpoint the convergence between conventional and multiparametric approaches.

artículo original

10.3389/fmicb.2022.885312

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Mycobacterium tuberculosis

T-cells

immune response

activation markers

treatment

flow cytometry

multiparametric analysis

cluster

IFN-GAMMA

EXPRESSION

DIAGNOSIS

RESPONSES

CHILDREN

ASSAY

MODEL

Microbiología

Mycobacterium tuberculosis

células T

respuesta inmune

marcadores de activación

tratamiento

citometría de flujo

análisis multiparamétrico

cluster

IFN-GAMMA

EXPRESIÓN

DIAGNOSIS

RESPUESTAS

NIÑOS

ENSAYO

MODELO

Enfermedades Infecciosas

Inmunología

Tecnología Médica

Study of CD27, CD38, HLA-DR and Ki-67 immune profiles for the characterization of active tuberculosis, latent infection and end of treatment

artículo original

version publicada

CC BY 4.0

CC BY 4.0

acceso abierto

acceso abierto

WOS:000837101100001