Elucidating Immune Cell Changes in Celiac Disease: Revealing New Insights from Spectral Flow Cytometry
Abstract
1. Introduction
2. Results
2.1. Immunological Cell Patterns
2.2. Characterization of CD4+, CD8+, and TCRγδ+ T Lymphocytes
3. Discussion
4. Materials and Methods
4.1. Study Design
4.2. Participants, Intervention, and Sample Collection
4.3. Panel Design
4.4. Sample Processing
4.5. Quality Control and Reproducibility: Panel Validation
4.6. Unsupervised Analysis
4.7. Supervised Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
Appendix B
Appendix C
Product | Company | Reference |
---|---|---|
RBC lysis buffer (10X) | Biolegend | 420302 |
LIVE/DEAD™ Fixable Blue Stain Fluorescence | Invitrogen | L34961 |
True-Stain Monocyte Blocker™ | Biolegend | 426102 |
Brilliant Stain Buffer | BD Biosciences | 563794 |
anti-TCRγδ PerCP-eFluor 710 (clone: B1.1) | Invitrogen | 46-9959-42 |
anti-CX3CR1 in BV711 (clone: 2A9-1) | Biolegend | 341630 |
anti-CCR7 in BV421 (clone: G043H7) | Biolegend | 353208 |
anti-CCR9 in PE/Dazzle (clone: L053E8) | Biolegend | 358918 |
anti-CCR4 in BV750 (clone: 1G1) | BD Biosciences | 746980 |
anti-CD11c in e-Fluor® 450 (clone: 3.9) | Invitrogen | 48-0116-42 |
anti-CD127 in APC-R700 (clone: HIL-7R-M21) | BD Biosciences | 565185 |
anti-CD3 in BV510 (clone: SK7) | Biolegend | 344828 |
anti-CXCR3 in PE/Cy7 (clone: G025H7) | Biolegend | 353720 |
anti-CD20 in Pacific Orange (clone: HI47) | Invitrogen | MHCD2030 |
anti-CD39 in BUV615 (clone: TU66) | BD Biosciences | 751269 |
anti-PD-1 in BV785 (clone: EH12.2H7) | Biolegend | 329930 |
anti-CD1c in AF647 (clone: L161) | Biolegend | 331510 |
anti-PD-L1 in BV650 (clone: 29E.2A3) | Biolegend | 329740 |
anti-integrin β7 in PerCP/Cy5.5 (clone: FIB27) | Biolegend | 121008 |
anti-CD38 in APC/Fire 810 (clone: HB-7) | Biolegend | 356644 |
anti-CD25 in PE/AF-700 (clone: 3G10) | Invitrogen | MHCD2524 |
anti-CD123 in SuperBright 436 (clone: 6H6) | Invitrogen | 62-1239-42 |
anti-CD14 in Spark Blue 550 (clone: 63D3) | Biolegend | 367148 |
anti-CD45 in PerCP (clone: 2D1) | Biolegend | 368506 |
Anti-CLA in BV605 (HECA-452) | BD Biosciences | 563960 |
anti-CD103 in PE (clone: Ber-ACT8) | Biolegend | 350206 |
anti-CD141 in BB515 (clone: 1A4) | BD Biosciences | 566017 |
anti-CD45RA in BUV395 (clone: 5H9) | BD Biosciences | 740315 |
anti-CD49d in BUV615 (clone: 9F10) | BD Biosciences | 751596 |
anti-CD19 in Spark NIR 685 (clone: HIB19) | Biolegend | 302270 |
anti-CD69 in PE/Cy5 (clone: FN50) | Biolegend | 310908 |
anti-HLA-DR in BUV563 (clone: G46-6) | BD Biosciences | 748340 |
anti-CD56 in BUV737 (clone: NCAM16.2) | BD Biosciences | 612766 |
anti-IgD in BUV480 (clone: IA6-2) | BD Biosciences | 566138 |
anti-CD16 in BUV496 (clone: 3G8) | BD Biosciences | 612944 |
anti-CD8 in BUV805 (clone: SK1) | BD Biosciences | 612889 |
anti-CD4 in cFLUOR® YG584 (clone: SK3) | Cytek | SKU R7-20041 |
anti-CD27 in APC/Cy7 (clone: M-T271) | Biolegend | 356424 |
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Analysis | Cluster | Lineage | Phenotypic Markers |
---|---|---|---|
CD45+ | Clu-29 | CD8+ CD3+ | CD127+ CD27+ CD45RA+ CCR7+ |
Clu-68 | CD20+ CD19+ | CD27+ CD45RA+ HLA-DR+ CD49d+ | |
Clu-04 | CD4+ CD3+ | CD38+ CD49d+ β7+ | |
Clu-05 | CD4+ CD3+ | CD127+ CD27+ CD45RA+ CCR7+ | |
Clu-69 | CD20+ CD19+ | CCR7+ CD45RA+ HLA-DR+ IgD+ | |
CD3+ | Clu-21 | CD4+ | CD38+ CD49d+ β7+ |
Clu-22 | CD8+ | CD38+ CD103+ β7+ |
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Gómez-Aguililla, S.; Farrais, S.; Senosiain, C.; López-Palacios, N.; Arau, B.; Ruiz-Carnicer, Á.; Sánchez-Domínguez, R.; Corzo, M.; Casado, I.; Pujals, M.; et al. Elucidating Immune Cell Changes in Celiac Disease: Revealing New Insights from Spectral Flow Cytometry. Int. J. Mol. Sci. 2025, 26, 2877. https://doi.org/10.3390/ijms26072877
Gómez-Aguililla S, Farrais S, Senosiain C, López-Palacios N, Arau B, Ruiz-Carnicer Á, Sánchez-Domínguez R, Corzo M, Casado I, Pujals M, et al. Elucidating Immune Cell Changes in Celiac Disease: Revealing New Insights from Spectral Flow Cytometry. International Journal of Molecular Sciences. 2025; 26(7):2877. https://doi.org/10.3390/ijms26072877
Chicago/Turabian StyleGómez-Aguililla, Sara, Sergio Farrais, Carla Senosiain, Natalia López-Palacios, Beatriz Arau, Ángela Ruiz-Carnicer, Rebeca Sánchez-Domínguez, María Corzo, Isabel Casado, Mar Pujals, and et al. 2025. "Elucidating Immune Cell Changes in Celiac Disease: Revealing New Insights from Spectral Flow Cytometry" International Journal of Molecular Sciences 26, no. 7: 2877. https://doi.org/10.3390/ijms26072877
APA StyleGómez-Aguililla, S., Farrais, S., Senosiain, C., López-Palacios, N., Arau, B., Ruiz-Carnicer, Á., Sánchez-Domínguez, R., Corzo, M., Casado, I., Pujals, M., Bodas, A., Sousa, C., & Núñez, C. (2025). Elucidating Immune Cell Changes in Celiac Disease: Revealing New Insights from Spectral Flow Cytometry. International Journal of Molecular Sciences, 26(7), 2877. https://doi.org/10.3390/ijms26072877