LDA Conference 2015 Marije Doppenberg-Oosting 1
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1 Host defence against Borrelia Dr. Marije Doppenberg-Oosting Lyme disease: after a tick bite The 14th LDA Conference September 12th 2015 Buckingham House Murray Edwards College University of Cambridge Marije.Doppenberg-Oosting@radboudumc.nl Borreliosis (Lyme disease) Life-cycle of ticks Borrelia sensu lato Ixodus species -Ricinus (EU) -Scapularis (USA) Erythema Migrans Lyme Disease Lyme Iceman's DNA reveals health risks and relations. Nature Communications Ötzi s genome hints at heart disease, bacterial infection and common ancestry with modern-day Sardinians. Ewen Callaway 28 February 2012 Ötzi s genome also hints at other health problems: Zink s team found almost two-thirds of the genome of Borrelia burgdorferi, a bacterium that causes Lyme disease. Doppenberg-Oosting 1
2 Borrelia infections in The Netherlands Infection of Humans with Borrelia Borrelia infections in The Netherlands 1 Early disease EM (75% of patients) 2 Early disseminated phase multiple EM, neurologic, and/or cardiac findings, malaise 3 Late (persistent) Lyme disease joint swelling and pain, neurologic manifestations, paralysis, chronic arthritis 1. Rash at bite site 2. Rashes at other sites 3. Fever, chills, sweats 4. Weight change (loss or gain) 5. Fatigue, tiredness 6. Unexplained hair loss 7. Swollen Glands 8. Sore Throat 9. Testicular pain or Pelvic pain 10. Unexplained menstrual irregularity 11. Irritable bladder or bladder dysfunction 12. Sexual dysfunction or loss of libido 13. Upset stomach 14. Change in bowel function, constipation 15. Chest pain or rib soreness 16. Shortness of breath, cough 17. Heart palpitations, pulse skips, heart block 18. Joint pain or swelling 19. Muscle pain or cramps 20. Twitching of the face or other muscles 21. Headache 22. Neck creaks and cracks, neck stiffness 23. Stiffness of the joints or back 24. Tingling, numbness, burning or a stabbing 25. Facial paralysis (mouth is crooked on one side when you talk in front of a mirror-bell's Palsy) 26. Eyes/vision: double, blurry, pain, increased floaters (specks in front of vision) 27. Ears/hearing: ringing, buzzing, ear pain 28. Dizziness, poor balance, increased motion sickness 29. Light-headedness, wooziness, difficulty walking, clumsy bumping into objects 30. Tremor 31. Confusion, difficulty in thinking, difficulty in concentrating, difficulty with sticking with task 32. Difficulty with concentration or reading 33. Decreased short term memory 34. Disorientation: driving past your turn, getting lost, going to wrong place 35. Difficulty with speech 36. Mood swings, irritability, depression 37. Disturbed sleep: too much, too little, early awakening- goes through cycles sometimes too much 38. Exaggerated symptoms or worse hangover from alcohol 39. Slowed pulse 40. Testicular enlargement 41. Nodules on earlobes 42. Stomach distension 43. Breathing with mouth open 44. Eye lesions 45. Nasal stuffiness /restriction 46. Shortness of breath 47. Hypersensitive olfactory organ (sensitive to the slightest odours / smells) 48. Feeling drunk without drinking 49. No desire to do anything- including favorite hobbies 50. Too slow a pulse when resting 51. Rapid pulse with slightest exertion 52. Perfuse sweating- soaking wet if you go snow skiing 53. Muscle pains, cramps or charley horse 54. Pains switching from one side of the body to the other 55. Enlarged lymph glands 56. Stiff aching neck 57. Changes in vision 58. Generalized acheyness 59. Limbs- especially arms feel heavier than. 60. Diminished or absent reflexes 61. Brain fog 62. Poor coordination/ataxia 63. Continual infections 64. Poor concentration 65. Decreased ability to spell correctly 66. GI distress/abdominal pain 67. Poor word retrieval/aphasia 68. Anxiety 69. Difficulty swallowing 70. Nausea/vomiting 71. Anorexia 72. Vasculitis 73. Loss of muscle tone 74. Changes in taste or smell 75. Obsessive-Compulsive symptoms 76. Panic attacks 77. Changes in cerebral blood flow/brain waves 78. Number reversal 79. Light Sensitivity 80. Trigeminal neuralgia (TMJ) 81. Dilated cardiomyopathy 82. Loss of temperature control 1 Early disease EM (75% of patients) 2 Early disseminated phase multiple EM, neurologic, and/or cardiac findings, malaise 3 Late (persistent) Lyme disease joint swelling and pain, neurologic manifestations, paralysis, chronic arthritis Doppenberg-Oosting 2
3 B.burgdorferi Arthritis How ticks get infected with Borrelia spirochetes B.afzelii Skin disorders; ACA B.garinii Neurological disorders; paralysis Borrelia belongs to the family of Spirochetes Causative agents: Borrelia burgdorferi sensu lato (B.burgdorferi, B.afzelii, B.garinii) B.burgdorferi B.afzelii B.garinii other Spirochetes of the Borrelia genus 8-30 µm Dissemination µm Corkscrew-shaped Gram negative 1. Tick-gut: Osps 2. Tick-human: Salp 3. Human-matrix:Adhesins 4. Human-endothelium:MMPs 5. Human-cell binding: p66 integrin NO LPS Protoplasmic cilinder Doppenberg-Oosting 3
4 Escape mechanisms Immune cell (macrophage) Recognition of Borrelia Change of shape? Change of coat Salivary glands Innate immune Pro- / anti- inflammatoiry IL-17 Adaptive immune IFN-γ IL-8 IL-6 IL-1β NFкB nucleus 20 Innate immunity Early defense mechanism; fast but unspecific Adaptive immunity Later defense mechanism; slower but specific Cells of immune system -Antibodies -Memory T B Y Th Focus of my research How does a macrophage know what to eat? RECOGNITION Surface of Borrelia exists out of building blocks Salivary glands Innate immune Pro- / anti- inflammatoiry IL-17 Adaptive immune IFN-γ IL-8 IL-6 IL-1β NFкB nucleus 23 Many types of building blocks Doppenberg-Oosting 4
5 How is Borrelia recognized? Borrelia bacteria 26 The host How is Borrelia burgdorferi recognized? (key-lock principle) PRR Pattern Recognition Receptor IL-1β Secreted PRRs TLRs TLR2 C-type lectins Macrophage T-helpercell NLRs 27 Recognition Cytokine PRR-focused research TLR2/2 TLR1/2 TLR6/2 TLR4 TLR5 MR? RICK NOD2 immune clinical outcome inhibition of immune modulates TLR1/2 s TLR7 TLR8 TLR9? blocking IL-6, and TNF clinical outcome spirochetal killing neutrophil influx Type I IFN DC Type I IFN Type I IFN Phagocytosis with TLR2 arthritis? silencing ko OspA CD14 PGN Doppenberg-Oosting 5
6 IL-1β Research using blocking antibodies Example: PBMC Recognition by PRRs blocking TLR2/6 TLR2/1 TLR10 NOD2 Conclusion Borrelia is recognized by TLR1 but not TLR6 32 wt ko G G A C G T T A C C C T G C A A T G Wild-type G G A C A T T A C C C T G T A A T G ko NEEDED: : Single Nucleotide polymorphism - functional data (PBMC) - DNA - MAF > 20% Do s in TLRx influence Borrelia-derived? Foresters (n=112) PBMC DNA Link s to clinical outcome IL-23R Wt IL-23 He IL-23R Wt IL-23 He n EM % 16% Stimulation 24h ELISA assay Clinical signs : % 36% -joint % 12% -back/pareshesia % 24% -heart 6 0 6% 0% -ACA 5 0 5% 0% TLRx Wt TLRx He TLRx Ho Similar for IL-6, IL-8, TNF-α no Lyme % 72% Lyme in past % 20% possible Lyme 9 1 9% 4% not known 3 1 3% 4% 35 Doppenberg-Oosting 6
7 What we demonstrated so far Future plans Study effect of microbiome and habits on immune s 1. N = Lyme patients -blood -swaps -urine -faeces -questionnaires -blood -swaps -urine -faeces functional data (PBMC, macrophages, transcriptomics) RNA sequencing Next generation sequencing ( analysis) metabolomics (plasma samples) microbiome analysis metabolomics microbiome and fungal-biome analysis What can we do with this knowledge? -In the future: inhibit or prevent infection with Borrelia -Screen patients on variants to predict clinical outcome -questionnaires demographic, education, illnesses, allergies, food, etc -Personalize treatment strategies Acknowledgements Radboudumc Department of Internal Medicine Marije Doppenberg-Oosting Sanne Smeekens Kathrin Buffen Hedwig Vrijmoeth Mariska Kerstholt Martin Jaeger Rahajeng Tunjungputri Jos van der Meer Leo Joosten Mihai Netea Microbiology Department C. Bartels Dept of Laboratory Medicine Hans Koenen Irma Joosten Esther Rijssen Marina van Welie Bram van Cranenbroek Esther Fasse Department of Genetics Joris Veltman Han Brunner Donders Institute Alejandro Arias-Vasquez Guillen Fernandez UMC Groningen Vinod Kumar Cisca Wijmenga Harvard/Broad Institute Dirk Gevers Aylwin Ng Curtis Huttenhower Ramnik Xavier Marije.Doppenberg-Oosting@radboudumc.nl Doppenberg-Oosting 7
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