To deplete CD8+ T cells, anti-CD8α (BioXCell; clone YTS169.4; 500 μg) or an isotype control (BioXCell; clone LTF-2; 500 μg) was administered to immunized mice by intraperitoneal injection at days −7, −3, +1, +5 and +12. Mice then were challenged with 3 × 105 FFU of mouse-adapted ZIKV Dakar (41525) at day 0 that was preceded by intraperitoneal inoculation of anti-ifnar1 blocking mAb. To confirm immune cell depletion, peripheral blood was collected at 7 dpi followed by erythrocyte lysis with ACK lysis buffer (GIBCO) and resuspension in RPMI supplemented with 10% heat-inactivated FBS. Cells were blocked for FcγR binding and stained with CD45 BUV395 (BD BioSciences clone30-F11), CD3 Alexa488 (BioLegend clone1452C11), CD4 PE-Cy7 (BioLegend clone GK1.5), CD8b PreCP-Cy5.5 (BioLegend clone YTS156.7.7), and Fixable Aqua Dead Cell Stain (Invitrogen, L34966). Subsequently, cells were fixed by Fix/Lyse solution (eBioSciences 00–5333). Datasets were acquired on a LSRII flow cytometer and analyzed using FlowJo software X 10.0.7.
Cd8b precp cy5
Manufactured by BioLegend
CD8b PreCP-Cy5.5 is a fluorescently-labeled antibody that binds to the CD8b cell surface protein. It can be used in flow cytometry applications to identify and characterize CD8+ T cells.
Automatically generated - may contain errors
Lab products found in correlation
Cd4 pe cy7, by BioLegend (4 mentions)
Cd45 buv395, by BD (4 mentions)
Fixable aqua dead cell stain, by Thermo Fisher Scientific (2 mentions)
Cd3 alexa 488, by BioLegend (2 mentions)
Fix lyse solution, by Thermo Fisher Scientific (2 mentions)
Ack lysis buffer, by Thermo Fisher Scientific (2 mentions)
Ltf 2, by BioXCell (2 mentions)
Efluor 506, by Thermo Fisher Scientific (2 mentions)
Cd19 apc cy7, by BioLegend (2 mentions)
Cd62l pacific blue, by BioLegend (2 mentions)
Klrg1 fitc, by BioLegend (2 mentions)
Cd127 alexa700, by Thermo Fisher Scientific (2 mentions)
Lsrii flow cytometer, by FlowJo (2 mentions)
Cd44 pe, by BioLegend (2 mentions)
Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (2 mentions)
Brefeldin a, by BioLegend (2 mentions)
Anti tnf α bv605, by BD (2 mentions)
Anti ifn γ alexa 647, by BD (2 mentions)
4 protocols using cd8b precp cy5
1
Depletion of CD8+ T Cells in ZIKV Challenge
2
Assessing ZIKV-specific T-cell responses in mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Freshly-isolated mouse splenocytes were stimulated with an H-2Db-restricted immunodominant ZIKV peptide (amino acids 294–302), with rat anti-mouse CD3 used as a positive control and medium as negative control, for 12 h at 37°C before brefeldin A (BioLegend, 420601) was added for an additional 4 h. Subsequently, single-cell suspensions were blocked for FcγR binding (BioLegend; clone 93) and stained with the following antibodies: CD45 BUV395 (BD BioSciences clone30-F11), CD62L Pacific Blue, KLRG1 FITC, CD44 PE, CD4 PE-Cy7, CD8b PreCP-Cy5.5, CD19 APC-Cy7 (BioLegend clones MEL-14, 2F1/KLRG1, IM7, GK1.5, YTS156.7.7, 6D5, respectively), CD127 Alexa700 (eBioScience clone A7R34), and fixable viability dye (eFluor 506, eBioscience). Subsequently, cells were fixed and permeabilized with Foxp3/Transcription Factor Staining Buffer Set (eBiosciences, 00–5523-00) followed by intracellular staining with anti-TNF-α BV605 and anti-IFN-γ Alexa 647 (BD Biosceinces clones MP6-XT22 and XMG1.2, respectively). Datasets were acquired on a LSRII flow cytometer and analyzed using FlowJo software X 10.0.7.
3
Depletion of CD8+ T Cells in ZIKV Challenge
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To deplete CD8+ T cells, anti-CD8α (BioXCell; clone YTS169.4; 500 μg) or an isotype control (BioXCell; clone LTF-2; 500 μg) was administered to immunized mice by intraperitoneal injection at days −7, −3, +1, +5 and +12. Mice then were challenged with 3 × 105 FFU of mouse-adapted ZIKV Dakar (41525) at day 0 that was preceded by intraperitoneal inoculation of anti-ifnar1 blocking mAb. To confirm immune cell depletion, peripheral blood was collected at 7 dpi followed by erythrocyte lysis with ACK lysis buffer (GIBCO) and resuspension in RPMI supplemented with 10% heat-inactivated FBS. Cells were blocked for FcγR binding and stained with CD45 BUV395 (BD BioSciences clone30-F11), CD3 Alexa488 (BioLegend clone1452C11), CD4 PE-Cy7 (BioLegend clone GK1.5), CD8b PreCP-Cy5.5 (BioLegend clone YTS156.7.7), and Fixable Aqua Dead Cell Stain (Invitrogen, L34966). Subsequently, cells were fixed by Fix/Lyse solution (eBioSciences 00–5333). Datasets were acquired on a LSRII flow cytometer and analyzed using FlowJo software X 10.0.7.
4
Assessing ZIKV-specific T-cell responses in mice
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