rosettesep method  (STEMCELL Technologies Inc)

Journal: Frontiers in Immunology

Article Title: ERAP1 Controls the Interaction of the Inhibitory Receptor KIR3DL1 With HLA-B51:01 by Affecting Natural Killer Cell Function

doi: 10.3389/fimmu.2021.778103

Figure Lengend Snippet: Inhibition of ERAP1 enhances NK cell killing of 221-B*51:01 cells. (A) , Example of degranulation by human CD56 + NK cells from one healthy donor expressing KIR2DL1, KIR2DL3, and KIR3DL1 measured as CD107a cell surface expression in response to the indicated cells. The percentage of CD107a + cells within the indicated NK cell subsets is shown. (B) Summary of NK cell degranulation from donors after stimulation with 221 parental and transfectant cells in response to ERAP1 inhibition. P values, calculated by two-tailed paired Student’s t -test. ns, not significant.

Article Snippet: Human NK cells were isolated from peripheral blood mononuclear cells (PBMC) of healthy donors co-expressing KIR2DL1, KIR2DL2 and KIR3DL1 by using the RosetteSep NK cell enrichment mixture method (Stem-Cell Technologies).

Techniques: Inhibition, Expressing, Transfection, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: ERAP1 Controls the Interaction of the Inhibitory Receptor KIR3DL1 With HLA-B51:01 by Affecting Natural Killer Cell Function

doi: 10.3389/fimmu.2021.778103

Figure Lengend Snippet: Inhibition of ERAP1 enhanced NK cell killing of 221-B*51:01 cells by affecting the engagement of KIR3DL1. (A) , Example of degranulation by human CD56 + NK cells from one healthy donor co-expressing KIR2DL1, KIR2DL3, and KIR3DL1; CD107a cell surface expression was measured in specific KIR single positive (KIRsp) subsets (KIR3DL1sp for 221-B51 cells, KIR2DL1sp for 221-Cw4 cells and KIR2DL3 for 221-Cw3 and -Cw7 cells) in response to ERAP1-inhibited 221 transfectants and control cells. The percentage of CD107a + cells within the indicated NK cell subsets is shown. (B) , Summary of NK cell degranulation in the indicated KIRsp NK subsets from healthy donors after stimulation with 221 parental and transfectant cells in response to ERAP1 inhibition. P values, calculated by two-tailed paired Student’s t -test. ns, not significant.

Article Snippet: Human NK cells were isolated from peripheral blood mononuclear cells (PBMC) of healthy donors co-expressing KIR2DL1, KIR2DL2 and KIR3DL1 by using the RosetteSep NK cell enrichment mixture method (Stem-Cell Technologies).

Techniques: Inhibition, Expressing, Control, Transfection, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: ERAP1 Controls the Interaction of the Inhibitory Receptor KIR3DL1 With HLA-B51:01 by Affecting Natural Killer Cell Function

doi: 10.3389/fimmu.2021.778103

Figure Lengend Snippet: Inhibition of ERAP1 enhanced NK cell killing of 221-B*51:01 cells by affecting the engagement of NKG2A. (A) , Example of degranulation by human CD56 + NK cells from one healthy donor; CD107a cell surface expression was measured in NKG2A positive subset negative for KIR3DL1, KIR2DL1 and KIR2DL3 in response to ERAP1-inhibited 221 transfectants and control cells. The percentage of CD107a + cells within the indicated NK cell subset is shown. (B) , Summary of NK cell degranulation in the NKG2A positive KIR negative NK subset from healthy donors after stimulation with 221 parental and transfectant cells in response to ERAP1 inhibition. P values, calculated by two-tailed paired Student’s t -test. ns, not significant.

Article Snippet: Human NK cells were isolated from peripheral blood mononuclear cells (PBMC) of healthy donors co-expressing KIR2DL1, KIR2DL2 and KIR3DL1 by using the RosetteSep NK cell enrichment mixture method (Stem-Cell Technologies).

Techniques: Inhibition, Expressing, Control, Transfection, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: ERAP1 Controls the Interaction of the Inhibitory Receptor KIR3DL1 With HLA-B51:01 by Affecting Natural Killer Cell Function

doi: 10.3389/fimmu.2021.778103

Figure Lengend Snippet: Genetical and pharmacological inhibition of ERAP1 makes 221-B*51:01 cells more susceptible to lysis by NK cells expressing KIR3DL1*001. (A, B) 221 transfectant cells genetically (A) or pharmacologically (B) inhibited for ERAP1 were incubated with YTS or YTS-KIR3DL1*001 NK cells. A representative experiment of three independently performed is shown. Data represented mean ± SD of triplicate.

Article Snippet: Human NK cells were isolated from peripheral blood mononuclear cells (PBMC) of healthy donors co-expressing KIR2DL1, KIR2DL2 and KIR3DL1 by using the RosetteSep NK cell enrichment mixture method (Stem-Cell Technologies).

Techniques: Inhibition, Lysis, Expressing, Transfection, Incubation