Crosley CRSH268MW2 Bedienungsanleitung PDF herunterladen (Seite 4)

Contents

3.2.9 Subtractive proteome analysis (SUPRA) . . . . . . . . . . . . . . . . 27

3.2.10 Cloning, expression and puriﬁcation of vaccine candidates . . . . . . 27

3.2.11 Enrichment of speciﬁc IgGs from IVIG . . . . . . . . . . . . . . . . . 28

3.2.12 Surface localisation of vaccine candidates . . . . . . . . . . . . . . . 29

3.2.13 In vitro opsonophagocytosis of S. aureus by human neutrophils . . . 29

3.2.14 In vitro opsonophagocytic killing of S. aureus by human neutrophils . 30

3.2.15 Immunisation of mice . . . . . . . . . . . . . . . . . . . . . . . . . . 30

3.2.16 Murine model of sepsis . . . . . . . . . . . . . . . . . . . . . . . . . 30

3.2.17 Statistical analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

4 Results 32

4.1 Characterisation of the bacteriostatic effect mediated by S. aureus speciﬁc

IgGs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

4.1.1 Human serum inhibits in vitro growth of S. aureus . . . . . . . . . . . 32

4.1.2 Intravenous immunoglobulin preparation (IVIG) speciﬁcally inhibits in

vitro growth of S. aureus . . . . . . . . . . . . . . . . . . . . . . . . 32

4.1.3 Bacteriostatic effect on S. aureus is mediated by S. aureus speciﬁc

IgGs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

4.1.4 Gene expression proﬁling of S. aureus over the course of bacteriostasis 37

4.2 Identiﬁcation and characterisation of potential vaccine candidates against

S. aureus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

4.2.1 Subtractive proteome analysis (SUPRA) of anchorless cell wall (ACW)

proteins enables identiﬁcation of potential vaccine candidates against

S. aureus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

4.2.2 BT1, BT2 and BT3 are localised on the surface of S. aureus . . . . . 49

4.2.3 Afﬁnity puriﬁed IgGs speciﬁc for BT1, BT2 and BT3 trigger opsonophago-

cytosis of S. aureus by human neutrophils in vitro . . . . . . . . . . 49

4.2.4 Afﬁnity puriﬁed IgGs speciﬁc for BT1 and BT2 mediate killing of S. aureus

by human neutrophils in vitro . . . . . . . . . . . . . . . . . . . . . . 51

4.2.5 Immunisation with recombinant BT1 and BT3 protects mice from death

upon lethal challenge with S. aureus . . . . . . . . . . . . . . . . . . 52

5 Discussion 56

5.1 Characterisation of the bacteriostatic effect mediated by S. aureus speciﬁc

IgGs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56

5.2 Identiﬁcation and characterisation of potential vaccine candidates against

Staphylococcus aureus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59

6 Abstract 64

1 2 3 4 5 6 7 8 9 ... 87 88

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