MHC I Tetramer Profiling of T. cruzi-specific CD8+ T Cells

Mouse peripheral blood was obtained, processed and analyzed as previously described [15 (link),17 (link)]. Whole blood was incubated with a major histocompatibility complex I (MHC I) tetramer containing the T. cruzi transialidase TSKB20 peptide (ANYKFTLV/Kb) labeled with BV421 (Tetramer Core Facility at Emory University, Atlanta, GA) and the following labeled antibodies: anti-CD8 FITC, anti-CD4 APC EF780, anti-CD127 PE (BD Bioscience, San Jose, CA). At least 500,000 cells were acquired using a CyAn ADP flow cytometer (Beckman Coulter, Hialeah, Florida) and analyzed with FlowJo software v10.6.1 (Treestar, Inc., Ashland, OR).

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Bustamante J.M., Padilla A.M., White B., Auckland L.D., Busselman R.E., Collins S., Malcolm E.L., Wilson B.F., Saunders A.B., Hamer S.A, & Tarleton R.L. (2022). Prophylactic low-dose, bi-weekly benznidazole treatment fails to prevent Trypanosoma cruzi infection in dogs under intense transmission pressure. PLOS Neglected Tropical Diseases, 16(10), e0010688.

Publication 2022

anti-CD8 FITC anti-CD4 APC EF780 anti-CD127 PE CyAn ADP

Antibodies Blood Cells Fitc Major histocompatibility complex Mouse Peptide T cruzi Tetramer

Corresponding Organization : Texas A&M University

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Variable analysis

independent variables

T. cruzi transialidase TSKB20 peptide (ANYKFTLV/Kb) labeled with BV421

dependent variables

Number of cells expressing CD8, CD4, and CD127 markers

control variables

Whole blood incubation conditions
Flow cytometry acquisition settings
Antibodies used for staining (anti-CD8 FITC, anti-CD4 APC EF780, anti-CD127 PE)

controls

Positive control: None specified
Negative control: None specified

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