Title Page
Contents
ABSTRACT 8
1. INTRODUCTION 11
1.1. Trained immunity and its characteristics 11
1.2. Effects of Trained Immunity in Health and Diseases 12
1.3. Mechanism of Trained Immunity 13
1.4. Nuclear Lamin-A 14
1.5. Objectives 16
2. MATERIALS AND METHODS 16
2.1. Chemicals and reagents 16
2.2. Bone marrow-derived-macrophage isolation 17
2.3. In-vitro trained protocol 17
2.4. In-vivo trained protocol 18
2.5. Enzyme-linked immunosorbent assay (ELISA) 19
2.6. Isolation of peritoneal macrophages (PMO) and flow cytometry analysis 19
2.7. RNA Isolation and qRT-PCR 20
2.8. Seahorse assay 20
2.9. Statistical analysis 22
3. RESULTS 23
3.1. Macrophage lamin-A suppresses immune response in β-glucan-induced trained immunity 23
3.2. Macrophage lamin-A alters immune response in β-glucan-induced trained immunity in a sex-independent manner 26
3.3. Macrophage lamin-A decreased pro-inflammatory cytokine production from β-glucan-induced trained immunity mice 28
3.4. Reduction of β-glucan-induced trained immunity associated with reduction of monocyte-derived macrophages infiltration 30
3.5. Deficiency of lamin-A decreased β-glucan-induced hematopoietic stem cell proliferation 33
3.6. Macrophage lamin-A alters the glycolytic capacity in the β-glucan-induced trained immunity 36
4. DISCUSSION 39
5. CONCLUSION 45
REFERENCES 48
ABBREVIATION 56
Abstract 60
Table 1. Primers used for qRT-PCR 46
Table 2. Antibodies used in this study 46
Figure 1. Deficiency of lamin-A suppresses the β-glucan-induced trained immunity in BMDM in male mice. (A) Schematic diagram of the in-vitro training model.... 24
Figure 2. Deficiency of lamin-A lowers trained immunity in a sex-independent manner. Pro-inflammatory TNF-α and IL-6 cytokines released in the culture... 27
Figure 3. Deficiency of lamin-A in macrophages decreases the pro-inflammatory response in β-glucan-induced trained immunity in mice. Schematic diagram of in... 29
Figure 4. Lamin-A deficient mice display a reduction in monocyte-derived macrophage infiltration in the β-glucan experienced conditions. Peritoneal... 32
Figure 5. Reduction of trained immunity in lamin-A deficient mice altered hematopoietic stem cell proliferation. Bone marrow from trained mice was... 34
Figure 6. Deficiency of lamin-A decreases the glycolytic capacity in macrophages during trained immunity. BMDMs were trained using β-glucan (100ug/mL) as the... 38