Mapping and Characterization of HCMV-Specific Unconventional HLA-E-Restricted CD8 T Cell Populations and Associated NK and T Cell Responses Using HLA/Peptide Tetramers and Spectral Flow Cytometry
| Autor: | Amélie Rousselière, Laurence Delbos, Céline Bressollette, Maïlys Berthaume, Béatrice Charreau |
|---|---|
| Přispěvatelé: | Le Bihan, Sylvie, Centre de Recherche en Transplantation et Immunologie - Center for Research in Transplantation and Translational Immunology (U1064 Inserm - CR2TI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Nantes Université - UFR de Médecine et des Techniques Médicales (Nantes Univ - UFR MEDECINE), Nantes Université - pôle Santé, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Nantes Université - pôle Santé, Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ), Centre hospitalier universitaire de Nantes (CHU Nantes) |
| Rok vydání: | 2021 |
| Předmět: |
HLA-E
CD8 T cells NK γδT HCMV pHLA tetramers spectral flow cytometry [SDV]Life Sciences [q-bio] Receptors Antigen T-Cell alpha-beta viruses Cytomegalovirus CD8-Positive T-Lymphocytes MESH: Cytomegalovirus Infections / virology Biology (General) Spectroscopy MESH: Cytomegalovirus / immunology virus diseases MESH: Histocompatibility Antigens Class I / immunology Cell Differentiation General Medicine Flow Cytometry MESH: Flow Cytometry Computer Science Applications [SDV] Life Sciences [q-bio] Killer Cells Natural Chemistry Cytomegalovirus Infections MESH: Cell Differentiation MESH: Killer Cells Natural / immunology MESH: Lymphocyte Count MESH: Immunophenotyping MESH: Peptides / metabolism QH301-705.5 chemical and pharmacologic phenomena MESH: CD8-Positive T-Lymphocytes / immunology Article Catalysis Immunophenotyping Inorganic Chemistry MESH: Receptors Antigen T-Cell alpha-beta / metabolism Humans Lymphocyte Count Physical and Theoretical Chemistry QD1-999 Molecular Biology B. Mapping and Characterization of HCMV-Specific HLA-E MESH: Humans Histocompatibility Antigens Class I Organic Chemistry biochemical phenomena metabolism and nutrition MESH: Cytomegalovirus Infections / pathology Peptides |
| Zdroj: | International Journal of Molecular Sciences, Vol 23, Iss 263, p 263 (2022) International Journal of Molecular Sciences; Volume 23; Issue 1; Pages: 263 International Journal of Molecular Sciences International Journal of Molecular Sciences, 2021, 23 (1), pp.263. ⟨10.3390/ijms23010263⟩ |
| ISSN: | 1422-0067 1661-6596 |
| DOI: | 10.3390/ijms23010263 |
| Popis: | HCMV drives complex and multiple cellular immune responses, which causes a persistent immune imprint in hosts. This study aimed to achieve both a quantitative determination of the frequency for various anti-HCMV immune cell subsets, including CD8 T, γδT, NK cells, and a qualitative analysis of their phenotype. To map the various anti-HCMV cellular responses, we used a combination of three HLApeptide tetramer complexes (HLA-EVMAPRTLIL, HLA-EVMAPRSLLL, and HLA-A2NLVPMVATV) and antibodies for 18 surface markers (CD3, CD4, CD8, CD16, CD19, CD45RA, CD56, CD57, CD158, NKG2A, NKG2C, CCR7, TCRγδ, TCRγδ2, CX3CR1, KLRG1, 2B4, and PD-1) in a 20-color spectral flow cytometry analysis. This immunostaining protocol was applied to PBMCs isolated from HCMV− and HCMV+ individuals. Our workflow allows the efficient determination of events featuring HCMV infection such as CD4/CD8 ratio, CD8 inflation and differentiation, HCMV peptide-specific HLA-EUL40 and HLA-A2pp65CD8 T cells, and expansion of γδT and NK subsets including δ2−γT and memory-like NKG2C+CD57+ NK cells. Each subset can be further characterized by the expression of 2B4, PD-1, KLRG1, CD45RA, CCR7, CD158, and NKG2A to achieve a fine-tuned mapping of HCMV immune responses. This assay should be useful for the analysis and monitoring of T-and NK cell responses to HCMV infection or vaccines. |
| Databáze: | OpenAIRE |
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