Title Page
Contents
Abstract 10
I. INTRODUCTION 12
II. MATERIALS AND METHODS 15
1. Cell lines, PBMCs and NK cells 15
2. Flow cytometry 17
3. PBMC-mediated cytotoxicity 18
4. ELISA 19
5. Microarray 20
6. qRT-PCR 21
7. CD161 blocking assay 22
8. Statistical analysis 23
9. Data sharing statement 24
III. RESULTS 25
1. Lenalidomide treatment further enhances the proliferation and cytotoxicity of NK92 cells induced by IL-2 25
2. Lenalidomide further increases the cell number and cytotoxicity of peripheral blood NK cells induced by long-term treatment of IL-2. 30
3. Phenotypic analysis of activated peripheral blood NK cells in the presence of lenalidomide, IL-2 or IL-15 42
4. Lenalidomide increases secretion of IFN-γ by PBMCs in the presence of IL-2 or IL-15 47
5. Transcriptomal analysis of NK cells activated by lenalidomide and IL-2 50
6. Validation of downregulation of mRNA and surface expression of CD161 55
7. Blocking interaction between CD161 on PBMC and LLT1 on K562 by anti-CD161 antibody enhances PBMC-mediated lysis of K562 60
IV. DISCUSSION 64
V. CONCLUSION 68
REFERENCES 69
ABSTRACT IN KOREAN 80
Table 1. List of isotype control, primary and secondary antibody 29
Table 2. Modulation of cell populations within PBMCs stimulated by lenalidomide. 41
Figure 1. Lenalidomide increases the proliferation and cytotoxicity... 28
Figure 2. In the presence of IL-2 signaling, lenalidomide at... 35
Figure 3. Lenalidomide alters expression of specific NK cell receptors. 44
Figure 4. Lenalidomide increases IFN-γ secretion on PBMCs in... 48
Figure 5. Whole transcriptome analysis of NK cells activated by lenalidomide. 52
Figure 6. Lenalidomide downregulates expression of CD161 mRNA and protein. 57
Figure 7. Blocking CD161 on PBMCs increases PBMC-mediated cytotoxicity. 62
Supplementary Figure 1. Lenalidomide increases the proliferation of... 33
Supplementary Figure 2. Individual proliferative effects of NK cells... 36
Supplementary Figure 3. FACS gating strategies for cytotoxicity... 38
Supplementary Figure 4. Lenalidomide has no effects on NK cell... 40
Supplementary Figure 5. In the arm of IL-15, lenalidomide... 46
Supplementary Figure 6. Lenalidomide increases IFN-γ secretion... 49
Supplementary Figure 7. Transcriptomic modulation of NK cells... 53
Supplementary Figure 8. Lenalidomide upregulates expression of... 58
Supplementary Figure 9. Expression of CD25 is negatively... 59
Supplementary Figure 10. PBMCs stimulated with lenalidomide... 63