IL-1β以及IL-18都是介白素家族的成員之一，IL-1β的產生是由巨噬細胞所活化的，但是為了獲得與IL-1受體結合的能力，IL-1β前體先要由胱天蛋白酶-1剪切。
IL-1β是炎症反應的重要介質並且參與各種細胞活動，包括細胞增殖，分化與細胞凋亡。IL-18是先天和後天免疫反應的重要調節蛋白，最初被發現是用於結合活化T細胞以及自然殺手細胞使其誘導產生IFN-γ，IL-18在增強對外來病原體的免疫反應是有效的。以前的文獻表示，IL-18是IFN-γ誘導因子，文獻中以IL-18刺激小鼠的脾臟細胞誘導產生IFN-γ。
新城疫病毒（NDV）已被認為是對家禽業的最嚴重的威脅之一。在這項研究中，我們構建了一系列重組的IL-1β作為疫苗佐劑。
實驗中指出我們的蛋白可以有效的加強IBD和NDV的保護能力。除了先天免疫受IgG和IgA的影響而增加，後天免疫受IFN-gamma的影響所增加。從實驗中可以看出IL-1β 突變體做為疫苗佐劑的部分是有優勢的。
另外，雞的IL-18及其受體的結構目前尚未解出，基於從我們合作夥伴共同討論指示，在雞的IL-18的一些胺基酸上可和受體結合和訊號傳導中發揮重要作用，參考初步的IL-18/IL-18R複合物的結構。我們建立分析平台，採用初級的脾臟細胞利用IL-18及其突變體偵測IFN-γ。

Interleukin-1β and interleukin-18 (IL-18) are members of the IL-1 cytokine superfamily. IL-1β is produced by activated macrophages as a pro-protein, which is proteolytically processed to its active form by caspase 1. IL-1β is an important mediator of the inflammatory response, and it also involved in a variety of cellular activities, including cell proliferation, differentiation, and apoptosis. IL-18 is an important regulator of innate and acquired immune responses. IL-18 was originally found to bind the receptors on activated T cells, natural killer cells and it has the ability to induce IFN-γ. IL-18 was potent in enhancing cell-mediated immune responses against foreign pathogens. Previous literature has shown that IL-18 is an interferon-gamma (IFN-γ) inducing factor as well as an endotoxin-induced serum factor that stimulates IFN-γ production in murine splenocytes.
Newcastle disease virus (NDV) has been considered as one of the most severe threats to poultry industry. In this study, we constructed a series of recombinant IL-1βs as vaccine adjuvants.
Our experiments showed that proteins can effectively enhance the protection of IBD and NDV. The increasing of IFN-gamma can also promote the acquired immunity. ChIL-1β mutant 1 has higher amount of IgG and IgA than any other groups. It is an advantage for ChIL-1β mutants as vaccine adjuvants.
In addition, the complex structure of chicken IL-18 and its receptor has not been determined. Based on the preliminary IL-18/IL-18R complex structure from our collaborator showing that some residues in chicken IL-18 may play important roles in receptor binding and signaling. We are working on the establishment of the analyzed platform to compare the biological activities between IL-18 and its mutants using IFN-production measurement in primary splenocyte. The biological implication in this study will elucidate the roles of IL-18 hot spot residues in the biological processes.

口試委員會審定書 #
ABSTRACT i
中文摘要 iii
CONTENTS iv
LIST OF FIGURES vi
LIST OF TABLES vii
Chapter 1 Introduction 1
1.1 The IL-1 family 1
1.2 IL 1β 1
1.3 IL-1β and IL-18 2
1.4 Newcastle disease viruses (NDV) 2
1.5 Cytokine as adjuvant 3
1.6 Chickens vaccine in Taiwan 3
1.7 Aim of the study 4
Chapter 2 Materials and methods 5
2.1 Materials 5
2.2 Cloning and overexpression of IL-1β mutants 7
2.3 Proteins expression and purification 8
2.4 Circular dichroism spectroscopy 9
2.5 Sample collection and processing 10
2.6 Cortisol experiment 11
2.7 HI titers experiment 12
2.8 Plasma of IgA and IgG by enzymelinked immunosorbent assay ( ELISA ) 13
2.9 Cell mediated of IFN-γ assay 14
Chapter 3 Results and discussion 15
3.1 Circular dichroism and secondary structure 15
3.2 Plasma cortisol levels in vivo in response to chicken IL-1β and mutants 15
3.3 Assay of serum hemagglutination inhibition (HI) antibody 16
3.4 Assay of IgG and IgA antibody 17
3.5 Assay of IFN-gamma 18
3.6 IBD titers and H&E stains 19
3.7 Biological activity of IL-18 19
Conclusion 20
Figures 22
Tables 40
Reference 44

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