cd4+ negative selection magnetic bead kit  (STEMCELL Technologies Inc)

Journal: Nature Communications

Article Title: Persistence of viral RNA in lymph nodes in ART-suppressed SIV/SHIV-infected Rhesus Macaques

doi: 10.1038/s41467-021-21724-0

Figure Lengend Snippet: A Tissue distribution of intact proviruses per 10 6 CD4 T cells from rhesus macaques in the Late and Early ART cohorts measured by the SIV Intact Proviral DNA Assay (IPDA). Each colored dot represents one animal. B Correlation between IPDA viruses measured in each lymphoid tissue selected in ( A ) and viral DNA gag copies per 10 6 lymph node mononuclear cells (LNMCs) (Fig. ).

Article Snippet: For DNA proviral sequences from tissue cell suspensions, total genomic DNA was isolated from enriched CD4 cells from each tissue analyzed using a negative CD4 selection magnetic bead kit, following the manufacturer’s instructions (STEMCELL).

Techniques:

Journal: Nature Communications

Article Title: Persistence of viral RNA in lymph nodes in ART-suppressed SIV/SHIV-infected Rhesus Macaques

doi: 10.1038/s41467-021-21724-0

Figure Lengend Snippet: A , B SIV-Gag specific IFN-γ, IL-2, and TNFα responses in CD69 + CD4 + ( A ) and CD8 + ( B ) T cells at necropsy following 1 year of ART for the Late ART monkeys (red dots) and Early ART monkeys (blue dots). Colored dots refer to individual animals. All samples with approximately 2,000 or more CD69 + cells were kept for further analysis. Flow cytometry gating strategy used is illustrated in Supplementary Fig. . IFN-γ, Interferon gamma, IL-2, Interleukin 2, TNFα, Tumor necrosis factor alpha.

Article Snippet: For DNA proviral sequences from tissue cell suspensions, total genomic DNA was isolated from enriched CD4 cells from each tissue analyzed using a negative CD4 selection magnetic bead kit, following the manufacturer’s instructions (STEMCELL).

Techniques: Flow Cytometry

Journal: Cells

Article Title: CERI, CEFX, and CPI: Largely Improved Positive Controls for Testing Antigen-Specific T Cell Function in PBMC Compared to CEF

doi: 10.3390/cells10020248

Figure Lengend Snippet: Identifying the CD4/CD8 lineage of T cells responding to antigens CEF, CERI, CEFX, and CPI. The SFU counts in the unseparated PBMC was set as 100%, to which the SFU counts in the CD4 cell-depleted PBMC (PBMC-CD4 that still contain the CD8+ T cells) and the CD8 cell-depleted PBMC (PBMC-CD8 that sill contain the CD4+ T cells) are compared. PBMC, PBMC-CD4, and PBMC-CD8 were all adjusted to 300,000 cells/well. The PBMC tested were from Donor ID 162 and the 100% SFU counts for the CEF, CERI, CEFX, and CPI-responses in the unseparated PBMC were 195, 266, 400, 486, SFU, respectively. For each experimental condition, means and standard deviations calculated from triplicate wells are shown, and an independent samples student’s T -test was done to detect significant differences between the specified conditions.

Article Snippet: CD4+ and CD8+ T cell subsets were depleted from PBMC using magnetic bead-based CD4 and CD8 negative selection kits (Stem Cell Technologies, Vancouver, Canada).

Techniques: