Sorting of BHRF1-specific CD4 T cells with DR*0401/BHRF1 multimers
. | . | Sorting with DR4/PYY multimers . | . | . | . | |||
|---|---|---|---|---|---|---|---|---|
| . | . | TNF production upon stimulation with PYY peptide, pg/mL . | . | % of DR4/PYY tetramer + cells . | . | |||
| Donors . | HLA-DR4 subtype . | Presorted cells . | Postsorted cells . | Presorted cells . | Postsorted cells . | |||
| PBL-derived T-cell lines | ||||||||
| D3 | DR*0401 | <1 | >200 | 0.7 | 95.7 | |||
| D5 | DR*0401 | NT | >200 | NT | NT | |||
| KR1 | DR*0401 | <1 | >200 | NT | NT | |||
| KR7 | DR*0407 | <1 | <1 | NT | NT | |||
| SF-derived T-cell lines | ||||||||
| RA1 | DR*0401 | 85 | >200 | 2.2 | 99 | |||
| RA3 | DR*0401 | <1 | >200 | 0.26 | 40 | |||
| RA5 | DR*0401 | 15 | >200 | 0.22 | 99.2 | |||
| RA6 | DR*0401 | <1 | 60 | NT | NT | |||
| RA14 | DR*0401 | 150 | >200 | NT | 97.5 | |||
| RS2 | DR*0401 | 12 | >200 | 0.26 | 97 | |||
| RA2 | DR*0404 | <1 | <1 | NT | NT | |||
| RA7 | DR*0404 | <1 | <1 | NT | NT | |||
| RA9 | DR*0404 | <1 | <1 | NT | NT | |||
| RA19 | DR*0404 | <1 | <1 | NT | NT | |||
| AS1 | DR*0404 | <1 | <1 | NT | NT | |||
. | . | Sorting with DR4/PYY multimers . | . | . | . | |||
|---|---|---|---|---|---|---|---|---|
| . | . | TNF production upon stimulation with PYY peptide, pg/mL . | . | % of DR4/PYY tetramer + cells . | . | |||
| Donors . | HLA-DR4 subtype . | Presorted cells . | Postsorted cells . | Presorted cells . | Postsorted cells . | |||
| PBL-derived T-cell lines | ||||||||
| D3 | DR*0401 | <1 | >200 | 0.7 | 95.7 | |||
| D5 | DR*0401 | NT | >200 | NT | NT | |||
| KR1 | DR*0401 | <1 | >200 | NT | NT | |||
| KR7 | DR*0407 | <1 | <1 | NT | NT | |||
| SF-derived T-cell lines | ||||||||
| RA1 | DR*0401 | 85 | >200 | 2.2 | 99 | |||
| RA3 | DR*0401 | <1 | >200 | 0.26 | 40 | |||
| RA5 | DR*0401 | 15 | >200 | 0.22 | 99.2 | |||
| RA6 | DR*0401 | <1 | 60 | NT | NT | |||
| RA14 | DR*0401 | 150 | >200 | NT | 97.5 | |||
| RS2 | DR*0401 | 12 | >200 | 0.26 | 97 | |||
| RA2 | DR*0404 | <1 | <1 | NT | NT | |||
| RA7 | DR*0404 | <1 | <1 | NT | NT | |||
| RA9 | DR*0404 | <1 | <1 | NT | NT | |||
| RA19 | DR*0404 | <1 | <1 | NT | NT | |||
| AS1 | DR*0404 | <1 | <1 | NT | NT | |||
The efficiency of sorting of BHRF1-specific CD4 T cells using DR*0401/BHRF1 monomers coated on magnetic beads was evaluated (1) by staining presorted and postsorted cells with the DR4/BHRF1 tetramer, and (2) by measuring in presorted cells and in postsorted cells, the TNF production (pg/mL) upon stimulation with the BHRF1122-133 peptide in an autopresentation assay.
NT indicates not tested.