Cross Species Analysis: Disease Suscep3bility

Transcription

1 Cross Species Analysis: Disease Suscep3bility Gerard J. Nau, M.D., Ph.D. October 3, : Cross species analysis of genomics data

2 Cross- species comparisons: Overview To review innate immune recogni3on by TLR s. To review TLR5 func3on and disease suscep3bility. To review TLR11 and disease suscep3bility using cross- species comparisons.

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5 TLR Ligands Zymosan (yeast) peptidoglycan ssrna TLR8 CD14 most bacteria bacteria with flagella * * most bacteria most Gram neg * viruses Seminars in Immunology 16 (2004) 3 9

6 Motility and Bacterial Pathogenesis Mul3ple types of bacterial movement. Swimming Gliding Twitching Swimming mo3lity: Mediated by flagella. Likely helpful in low viscosity fluids. Direc3onal swimming through mucus (lung, intes3ne) or urinary tract. Vibrio cholerae, Listeria monocytogenes, Legionella pneumophila, UPEC

7 Flagella and Bacterial Pathogenesis h[p://en.wikipedia.org/wiki/file:empylori.jpg

8 Flagella and Bacterial Pathogenesis h6p://en.wikipedia.org/wiki/file:flagellum_base_diagram_en.svg

9 Flagella and Bacterial Pathogenesis Nature Immunology 4, (2003)

10 Flagella and Bacterial Pathogenesis Nature 410, (15 March 2001) h[p:// u.ac.jp/labs/namba/npn/index.html Nature Reviews Molecular Cell Biology 2, 232 (April 2001)

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12 Basic TLR5 Function Reporter construct: NFkB binding site Luciferase Nature Apr 26;410(6832):

13 Stimulation of TLR5 by Microbes Reporter construct: NFkB binding site Luciferase Microbial molecules Supernatants from bacterial cultures Nature Apr 26;410(6832):

14 Fraction of Supernatant that Stimulates Nature Apr 26;410(6832):

15 Flagellin Protein Identification Nature Apr 26;410(6832):

16 Flagellin Expression in Bacteria Nature Apr 26;410(6832):

17 Flagellin Stimulates Innate Immune Response in vivo Wild- type mice myd88 - /- mice Nature Apr 26;410(6832):

18 TLR5 Function: Conclusions and Questions A specific TLR detects a specific bacterial protein. Unclear if monomer (flagellin) or polymer (flagella) are detected (or both). Does TLR5 contribute to protect host against infec3on?

19 Legionella pneumophila Maria Ericsson (Harvard Medical School Electron Microscopy Facility)

20 TLR5 SNPs in 40 Healthy Controls

21 Legionella Flagellin Activates TLR5 CHO cells + TLR5 Bacteria strains LPS L. pneumophila Mouse strains

22 SNP Associated with Legionella Pneumonia

23 Signaling Defect in Cells from 392R* Donors A549 cells Purified LPS Purified flagellin Wild- type Legionella Flagellin nega3ve Legionella Calu- 3 cells Pa3ent cells Flagellin LPS

24 TLR5 Function: Conclusions and Questions - 2 SNPs in TLR5 are common. SNPs lead to biological effects on TLR5 func3on. Unfavorable haplotype increases risk for Legionnaire s disease by 2- fold. What other alleles predispose to infec3on? TLR5? Other TLRs?

25 Toll-like Receptors: Cross species comparison Human TLR: 1-10 Mouse TLR: 1-9, 11, 12 Chicken TLR: 1-5, 7, 15, 21 Drosophila TLR: 1-9 C. elegans TLR: 1 (?) D. dictyostelium TLR: 0 Molecular Immunology 40 (2004)

26 Urinary Tract Infections Among most common infec3ons in community and in the hospital. Diagnosed by symptoms and culture of urine. Most common e3ology is Escherichia coli. Upper tract Lower tract h[p://en.wikibooks.org/wiki/human_physiology/the_urinary_system

27 Uropathogenic Escherichia coli Uropathogenic = cause disease in urinary system. Rela3vely small subset of all E. coli strains. Have characteris3c features: o Especially notable for adhesins = structures to a[ach to cells of urinary system. Further subdivided by ability to cause upper or lower tract disease. Kidney InternaOonal (2007) 72, 19 25

28 Figure 1 Overview of uropathogen virulence factors Nielubowicz, G. R. & Mobley, H. L. T. (2010) Host pathogen interac3ons in urinary tract infec3on Nat. Rev. Urol. doi: /nrurol

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30 Toll-like Receptor Alignment Science Mar 5;303(5663):

31 mtlr11 Expression Science Mar 5;303(5663):

32 mtlr11 Signaling Science Mar 5;303(5663):

33 mtlr11 Function Science Mar 5;303(5663):

34 Making a Knockout Mouse h[p://en.wikipedia.org/wiki/knockout_mouse

35 Making a Knockout Mouse h[p://en.wikipedia.org/wiki/knockout_mouse

36 Interna3onal Knockout Mouse Consor3um h[p://

37 mtlr11 Knockout Mouse Science Mar 5;303(5663):

38 mtlr11 Knockout Mouse Science Mar 5;303(5663):

39 mtlr11 Knockout Mouse Science Mar 5;303(5663):

40 Toll-like Receptors Comparisons Allelic varia3on contributes to suscep3bility to infec3on within a species. Ortholog varia3on may contribute to suscep3bility to infec3on between species. Exploit differences to determine func3on. Challenge lies in the degree of difficulty execu3ng the biological studies.

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