The immunophenotyping of MAIT, iNKT, and γδ T cells was performed as previously described8 (link) on cryopreserved PBMCs from P1 prepared from a sample collected at the age of three years; and as previously described10 (link) on cryopreserved PBMCs from P2 prepared from a sample collected at the age of six years. Both patients were receiving broad-spectrum antimycobacterial drugs. Briefly, staining was performed in the presence of Fcblock (Miltenyi Biotec), with Zombie-NIR live-dead exclusion dye (#423105, BioLegend), anti-CD3-Alexa532 (Clone UCHT1, # 58-0038-42, Thermo Fisher Scientific), anti-γδTCR-FITC (#11-9959-42, Thermo Fisher Scientific), anti-Vδ2-APC-Fire750 (#331420, BioLegend), anti-CD56-BV605 (clone 5.1H11, #362538, BioLegend), anti-CD4-BV750 (#5663656, BD Biosciences), anti-CD8a-BV510 (clone RPA-T8, #301047, BioLegend), anti-Vα7.2-BV711 (clone 3C10, #351731, BioLegend), anti-Vα24-Jα18-PE-Cy7 (clone 6B11, #342912, BioLegend), anti-Vδ1-Vioblue (#30-100-555, Miltenyi Biotec), anti-CD161-PE (clone HP-3G10, #339938, BioLegend) and anti-Vβ11-APC (Miltenyi Biotec) antibodies. Cells were analyzed with an Aurora cytometer (Cytek). The gating strategy for MAIT cells, iNKT cells, γδ1+ T cells, and γδ2+ T cells has been described elsewhere8 (link),69 (link).
Rosain J., Neehus A.L., Manry J., Yang R., Le Pen J., Daher W., Liu Z., Chan Y.H., Tahuil N., Türel Ö., Bourgey M., Ogishi M., Doisne J.M., Izquierdo H.M., Shirasaki T., Le Voyer T., Guérin A., Bastard P., Moncada-Velez M., Han J.E., Khan T., Rapaport F., Hong S.H., Cheung A., Haake K., Mindt B.C., Perez L., Philippot Q., Lee D., Zhang P., Rinchai D., Al Ali F., Ata M.M., Rahman M., Peel J.N., Heissel S., Molina H., Kendir-Demirkol Y., Bailey R., Zhao S., Bohlen J., Mancini M., Seeleuthner Y., Roelens M., Lorenzo L., Soudée C., Paz M.E., Gonzalez M.L., Jeljeli M., Soulier J., Romana S., L’Honneur A.S., Materna M., Martínez-Barricarte R., Pochon M., Oleaga-Quintas C., Michev A., Migaud M., Lévy R., Alyanakian M.A., Rozenberg F., Croft C.A., Vogt G., Emile J.F., Kremer L., Ma C.S., Fritz J.H., Lemon S.M., Spaan A.N., Manel N., Abel L., MacDonald M.R., Boisson-Dupuis S., Marr N., Tangye S.G., Di Santo J.P., Zhang Q., Zhang S.Y., Rice C.M., Béziat V., Lachmann N., Langlais D., Casanova J.L., Gros P, & Bustamante J. (2023). Human IRF1 governs macrophagic IFN-γ immunity to mycobacteria. Cell, 186(3), 621-645.e33.
Anti cd3Antibodies Antimycobacterial Cd161Cells Clone Drugs Fitc Inkt cells Mait cellsPatients T cells
Corresponding Organization : Howard Hughes Medical Institute
Other organizations :
Medizinische Hochschule Hannover, Institut de Recherche en Infectiologie de Montpellier, Centre National de la Recherche Scientifique, Université de Montpellier, Bezmiâlem Vakıf Üniversitesi, Ontario Genomics, McGill University, Institut Pasteur, Université Paris Sciences et Lettres, Institut Curie, University of North Carolina at Chapel Hill, St Vincent's Clinic, Garvan Institute of Medical Research, Garrahan Hospital, Ümraniye Eğitim ve Araştırma Hastanesi, Centro Científico Tecnológico - Tucumán, Hôpital Cochin, Hôpital Robert-Debré, Hôpital Saint-Louis, Department of Virology, Vanderbilt University Medical Center, Hôpital Necker-Enfants Malades, European Genomic Institute for Diabetes, Hôpital Ambroise-Paré, University Medical Center Utrecht, Utrecht University, Hamad bin Khalifa University, German Center for Lung Research
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
