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able of Contents Acknowledgements ......................................................................................................................... iv List of Figures ................................................................................................................................ vi List of Tables ................................................................................................................................. vii List of Abbreviations .................................................................................................................... viii CHAPTER 1 .................................................................................................................................... 1 Literature Review ............................................................................................................................ 1 Lyme Disease Epidemiology ........................................................................................................... 1 The Enzootic Cycle of Borrelia burgdorferi ................................................................................... 2 Borrelia burgdorferi Physiology ..................................................................................................... 3 Lyme Disease Pathogenesis ............................................................................................................ 5 The innate immune response to Borrelia burgdorferi .................................................................... 6 The adaptive immune response to Borrelia burgdorferi ............................................................... 10 Borrelia burgdorferi evasion and dissemination .......................................................................... 11 References ..................................................................................................................................... 14 CHAPTER II ................................................................................................................................. 22 Insights in the Characterization of the Immune Response to B. burgdorferi Peptidoglycan ....... 22 Introduction ................................................................................................................................... 22 Methodology ................................................................................................................................. 23 Results ........................................................................................................................................... 27 Discussion ..................................................................................................................................... 32 References ..................................................................................................................................... 67 CHAPTER III ................................................................................................................................ 71 NapA is a Peptidoglycan Associating Protein in B. burgdorferi .................................................. 71 Introduction ................................................................................................................................... 71 Methodology ................................................................................................................................. 72 Results ........................................................................................................................................... 80 Discussion ..................................................................................................................................... 88 References ................................................................................................................................... 108 CHAPTER IV .............................................................................................................................. 114 Conclusions and Future Directions ............................................................................................ 114 References ................................................................................................................................... 118 M -562*-+74)5 @>LI< *!JKIL:KLI<8E;;@==<I<EK@8K@FE @>LI<FOGI<JJJ@FEE8CPJ@J @>LI<!)EI@:?D<EKE8CPJ@J,<JLCKJ=FIK?<*'J-K@DLC8K<;N@K?8:K<I@8C*!J=FI 8E;"FLIJ @>LI<!<E<OGI<JJ@FEE8CPJ@J@E*'J-K@DLC8K<;N@K?8:K<I@8C*!J=FI 8E; "FLIJ @>LI<%!!EI@:?D<EKE8CPJ@J,<JLCKJ=FIK?<*'J-K@DLC8K<;N@K?8:K<I@8C*!J=FI 8E;"FLIJ @>LI<CLJK<IE8CPJ@JF=@==<I<EK@8CCPOGI<JJ<;!<E<J @>LI<!)EI@:?D<EKE8CPJ@J,<JLCKJ=FIK?<*'J-K@DLC8K<;N@K?*!9FI&@M< 8K :<CCJ D&=FI 8E;?FLIJ @>LI<%!!EI@:?D<EKE8CPJ@J,<JLCKJ=FIK?<*'JJK@DLC8K<;N@K?*!9FI&@M< 8K :<CCJ D&=FI 8E;"FLIJ @>LI<!<E<OGI<JJ@FEE8CPJ@J@E*'J-K@DLC8K<;N@K?8:K<I@8C*!J=FI 8E; "FLIJ @>LI< !<E<OGI<JJ@FEE8CPJ@J@E*'J-K@DLC8K<;N@K?&@M<8K :<CCJ D&=FI 8E;"FLIJ @>LI< #;<EK@=@:8K@FEF=G<GK@;F>CP:8E 8JJF:@8K<;GIFK<@EJ**J@E @>LI< (8G@J8** @>LI< &PJFQPD<8E;(8CJKI<JJK<JKJ @>LI< <CC<EM<CFG<JKI<JJ8E;;<=<:KJF=(8G;<=@:@<EK98:K<I@8 @>LI< (8G *!@JI<C<8J<;@EFLK<I D<D9I8E<M<J@:C<J8E;KF><K?<I8:K8J8E<LKIFG?@C :?<DF8KKI8:K8EK @>LI< *?PCF><E<K@:E8CPJ@JF=(8G M@ -562*"%&/)5 .89C< ,(-<HL<E:@E>-8DGC<(8D<J .89C< @==<I<EK@8CCPOGI<JJ<;!<E<JF=#EK<I<JK@E*'J-K@DLC8K<;N@K?*!9=FI "FLIJ .89C< @==<I<EK@8CCPOGI<JJ<;!<E<JF=#EK<I<JK@E*'J-K@DLC8K<;N@K?*!9=FI ?FLIJ .89C< @==<I<EK@8CCPOGI<JJ<;!<E<JF=#EK<I<JK@E*'J-K@DLC8K<;N@K?&@M< =FI "FLIJ .89C< @==<I<EK@8CCPOGI<JJ<;!<E<JF=#EK<I<JK@E*'J-K@DLC8K<;N@K?&@M< =FI"FLIJ .89C< ,<8:KFD<E8CPJ@J*FJK "FLI*'-K@DLC8K@FEN@K?*!-D .89C< ,<8:KFD<E8CPJ@J*FJK "FLI*'-K@DLC8K@FEN@K?*!-8 .89C< ,<8:KFD<E8CPJ@J*FJK "FLI*'-K@DLC8K@FEN@K?*!: .89C< ,<8:KFD<E8CPJ@J*FJK "FLI*'-K@DLC8K@FEN@K?*!J .89C< ,<8:KFD<E8CPJ@J*FJK "FLI*'-K@DLC8K@FEN@K?*!9 .89C< !)E8CPJ@J!I8ELCF:PK<:K@M8K@FE*8K?N8P .89C< !)E8CPJ@J!I8ELCF:PK<:K@M8K@FE*8K?N8P .89C< ,<8:KFD<E8CPJ@J*FJK "FLI*'-K@DLC8K@FEN@K?&@M< .89C< ,<8:KFD<E8CPJ@J*FJK"FLI*'-K@DLC8K@FEN@K?&@M< .89C< )EI@:?D<EKE8CPJ@J .89C< -LDD8IP& '-I<JLCKJ=IFD**J:I<<E .89C< *8I<EK8C:CFE< DLK8K@FEJI<C8K@M<KF