CD8ACD8 Antigen, Alpha Polypeptide
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[12] Human Genes: WDR20, WD repeatdomain 20; CD4, CD4 molecule; CD8A, CD8a molecule; FYN, FYN protooncogene, Src family tyrosine kinase; BCL11B, B-cell CLL/lymphoma 11B; WIPI2, WD repeat domain, phosphoinositide interacting 2; SLC15A4, solute carrier family 15 member 4; CENPA, centromere protein A; LSM14B, LSM family member 14B; ZC3H3, zinc finger CCCH-type containing 3.
We analyzed the correlation between PD-L1 and CD8A expression at the mRNA level.
The following monoclonal antibodies were used for staining: (1) Live/Dead Fixable Far Red Dead Cell Stain Kit (Invitrogen); (2) PE-Cy5 Anti-Mouse CD8a (BD Pharmingen); (3) Rat Anti-Mouse CD4/L3T4a-FITC (Beckman Coulter); and (4) PE Rat Anti-Mouse Gamma Interferon (BD Pharmingen).
Cells were then labeled with the following fluorescence-conjugated monoclonal antibodies: anti-Rat CD45 PE, anti-Rat CD8a APC, anti-Rat CD44 PE, and anti-Rat CD62L (eBioscience, San Diego, CA, USA).
Transcription analysis of 44 CD genes was also carried out, indicating 24 CD genes (CD3E, CD244, CD69, CD247, CD40L, CD38, CD5L, CD44, CD5, CD83, CD163L1, CD36, CD14, CD28, CD200R1A, CD40, CD59, CD200, CD4, CD2, CD72, CD180, CD79B, and CD93) to be markedly increased by 2.1- to 8.4-fold, and 4 CD genes (CD163, CD8A, CD34, and CD7) to be significantly decreased by 2.1- to 8.2-fold in chicken line 6.3 with p < 0.01 and fold [greater than or equal to] 2.
We also measured mRNA genes derived from stem cells [CD34 molecule (CD34)], T cells [CD3e molecule, epsilon (CD3-TCR complex) (CD3E)], CD4 molecule (CD4), CD8a molecule (CD8A), and B cells [CD19 molecule (CD19)], but these genes were not amplified (data not shown).
Fluorescein isothiocyanate (FITC)-conjugated CD3, phycoerythrin-conjugated CD4, and FITC-conjugated CD8a were used to detect specific cell populations, which were counted on a Becton Dickinson FACScan Flow Cytometer (BD Biosciences Immunocytometry Systems, San Jose, CA).
Cells stimulated by medium and cells stimulated by YCP (400 nM) were collected at various times (0 h, 6 h, 12 h, 24 h, 48 h, and 96 h) after washing by PBS and stained with anti-mouse CD3e PE-Cyanine5, anti-mouse CD4 FITC, and anti-mouse CD8a PE monoclonal antibodies (eBioscience, San Diego, CA, USA).
RT-PCR primers for pluripotency (Oct3/4, Nanog, Sox2), MSC surface markers (CD4, CD8A, CD25, CD33, CD34, CD44, CD54, CD61, CD80, CD105, CD117, Flk-1 and HMGA2) and GAPDH internal control were shown in
Fluorochrome- or biotinlabeled human monoclonal antibodies specific for the following antigens were purchased from BD Biosciences (San Jose, CA, USA), eBioscience (San Diego, CA, USA), and BioLegend (San Diego, CA, USA): CD8a (SK1), CD4 (L200), CD3 (SP342), CD28 (CD28.2), CD95 (DX2), CD1b (SN13), CD8[beta] (SIDI8BEE), HLA-DR (G46-6), CXCR5 (MU5UBEE), and PD-1 (EH12.2H7).