HIV An bodies Frequently Cross- React with Intes nal Microbiota An gens. Tony Moody Duke Human Vaccine Ins tute Duke University 8 April 2015

Transcription

1 HIV An bodies Frequently Cross- React with Intes nal Microbiota An gens Tony Moody Duke Human Vaccine Ins tute Duke University 8 April 2015

2 Commensal Bacteria and the Immune System The intes ne is home for about 100 trillion microbes. Coloniza on with gut microbiota is cri cal for the normal development of the immune system. B cells and T cells within the intes ne block the entry of bacterial into systemic circula on. 2

3 An body responses in Early Infec on and Vaccina on In acute HIV- 1 infec on (AHI), virus replica on is prominent in the gastrointes nal tract with early deple on of CD4+ T cells and destruc on of germinal centers. Brenchley et al., J Exp Med, 2004; Guadalupe et al., J. Virol, 2003; Mehandru et al., PLoS Med 2006; Pope and Haase, Nat Med, 2003; Veazey et al., Science 1998; Trends in Immunology 2001 The ini al B cell response to HIV is focused on non- neutralizing gp41 epitopes, with IgG and IgM gp41 an bodies arising at the same me. Tomaras et al., J. Virol, 2003; Liao et al. J Exp Med,

4 Hypothesis Pre- exis ng commensal bacteria or other environmental an gens that cross- react with HIV- 1 an gens may shape the B cell response to HIV- 1 infec on and vaccina on. 4

5 Environmental An gens in Shaping the Response to HIV Plasma cell repertoires in acute HIV- 1 infec on Plasma and memory B cells in terminal ileum during early and chronic HIV- 1 infec on Memory B cells in HIV- 1 Env vaccina on (Wilton Williams) Memory B cells in colostrum 5

6 Environmental An gens in Shaping the Response to HIV Plasma cell repertoires in acute HIV- 1 infec on Plasma and memory B cells in terminal ileum during early and chronic HIV- 1 infec on Memory B cells in HIV- 1 Env vaccina on (Wilton Williams) Memory B cells in colostrum 6

7 Non-Neutralizing gp41 Antibody Responses in Acute HIV-1 Infection Intes nal bacteria lysate (0.6%) Cardiolipin (0.3%) Influenza (1.2%) Hep- 2 (2.5%) HIV gp41 (6.2%) Cryptococcus (0.5%) Candida (0.2%) Tetanus toxoid (0.4%) Lipid A (0.8%) 977 mabs Polyreac ve (0.8%) Non- HIV reac ve (86.5%) Liao et al. J Exp Med,

8 High Mutation VH Frequencies of gp41 Antibodies from Acute HIV-1 Infection Days mean 5.0% ± 0.4% % mutation AHI (n=67) Liao et al. J Exp Med,

9 High Mutation VH Frequencies of gp41 Antibodies from Acute HIV-1 Infection Days mean 5.0% ± 0.4% % mutation AHI (n=67) Liao et al. J Exp Med,

10 High Mutation VH Frequencies of gp41 Antibodies from Acute HIV-1 Infection Days Days 182 Days % mutation mean 5.0% ± 0.4% % mutation mean 1.1% ± 0.5% mean 4.8% ± 0.5% AHI (n=67) 0 Primary Immuniza on, 2x (n=14) Secondary Immuniza on, 4x (n=21) Liao et al. J Exp Med, 2011 Moody et al. J Virol,

11 Two Possibilities of gp41 Antibody Clonal Origin HIV gp41 triggers a germline BCR on naïve B cell to expand. HIV gp41 triggers preexisting antibody clones in which the germline doesn t react with gp41 but an intermediate clone antibody acquires gp41 reactivity. 11

12 Clonal Lineage 558 from An Acute HIV-1 Infected Individual 8 H733-K535 6 Clone H726-K528 3 H622-K442 0 (3) H610-K433 8 H834-K619 4 gp H703-K505 gp41+ H828-K H789-K584 2 H657-K470 H723-K525 (24) H764-K567 1 H595-K423 1 H658-K H648-K462 (11) H752-K557 3 H624-K444 gp41+ H712-K513 12

13 Clonal Lineage 558 from An Acute HIV-1 Infected Individual 5 3 Clone UCA 6 H733-K535 H726-K528 3 H622-K442 0 (3) H610-K433 8 H834-K Produc on of Inferred Intermediate Clone An bodies H703-K505 0 H828-K616 1 gp H789-K584 2 H657-K470 H723-K525 (24) H764-K567 1 H595-K423 gp41+ H658-K471 1 H648-K462 1 (11) H752-K557 4 H624-K gp41+ Where did gp41 reac vity occur? 3 H712-K513 UCA H828- K616 13

14 HIV-1 EnvReactivity of Clonal Lineage 558 from a HIV-1 Acutely Infected Individual UCA )*+/ UCA 56789:8;<=78/ >!/ :8;<=78/ >"/ 56789:8;<=78/ >#/ H828- K616 2?@89A8;/=6B?C;D/ $%&%'("!"/ 30000! E!F#.&/ E&F"#./ &#FEE#/ IJKC98@L86L8/*6<7@/ ! 1000 G.#/!F.#"/!F0../ HFHEG/ &F&H"/ 500 &0#/ 0,-.!! "%.'"/,-!.0! gp gp140 CON-S gp '4/,-!.0!,-.!! "%.'"/,-!.0! gp gp140 CON-S gp '4/,-!.0!,-.!! "%.'"/,-!.0! gp gp140 CON-S gp '4/,-!.0!,-.!! "%.'"/,-!.0! gp gp140 CON-S gp '4/,-!.0!,-.!! "%.'"/,-!.0! gp gp140 CON-S gp '4/,-!.0! Liao et al. J Exp Med,

15 Clonal Lineage 558 from An Acute HIV-1 Infected Individual gp Clone UCA 6 H733-K535 H726-K528 3 H622-K442 0 (3) H610-K433 8 H834-K H703-K505 0 H828-K616 1 gp H789-K584 2 H657-K470 H723-K525 (24) H764-K567 1 H595-K423 gp41+ gp H658-K471 1 H648-K462 (11) H752-K557 gp H624-K444 3 H712-K513 UCA H828- K616 15

16 Clonal Lineage 558 from An Acute HIV-1 Infected Individual gp Clone UCA 6 H733-K535 H726-K528 3 H622-K442 0 (3) H610-K433 8 H834-K H703-K505 0 H828-K616 1 gp H789-K584 2 H657-K470 H723-K525 (24) H764-K567 1 H595-K423 gp41+ gp H658-K H648-K462 (11) H752-K557 3 H624-K444 gp41+ H712-K513 Addi onal 5 UCAs of gp41 clonal lineages tested and found none reacted with their autologous Env gp140 UCA H828- K616 16

17 Clonal Lineage 558 from An Acute HIV-1 Infected Individual gp41 - Suggests antibodies may be present before infection that crossreact with gp41 gp41 + Clone UCA H658-K H733-K535 H726-K528 3 H622-K442 0 (3) H610-K433 8 H834-K H648-K462 (11) H752-K557 3 H624-K444 H703-K505 0 H828-K616 1 gp H789-K584 2 H657-K470 H723-K525 (24) gp41+ H712-K513 H764-K567 1 H595-K423 gp41+ Addi onal 5 UCAs of gp41 clonal lineages tested and found none reacted with their autologous Env gp140 UCA H828- K616 17

18 What is the polyreactivity of this gp41 antibody clonal lineage with intestinal bacteria lysates and HEp-2 epithelial cells 18

19 Polyreactivity of Clonal Lineage 558 Inferred Germline and Intermediate and Observed gp41 Antibodies +"" +"" +"" gp41:"!"!"" +"" 5" 2" 228""""""""""""""""""""859" H648)K462" 167" +"" UCA RUA" 1" 6" Intestinal Gut"flora:"""172""""""""""""""""124"""""""""""""""""""391""""""""""""""""" : bacteria +"" +"" +"" H828)K616" 133" +"" 7" 9" 13" H789 K584" 158" 221""""""""""""""""""""160"""""""""""""""""""""330" +"" H834 K619" 47" Liao et al. J Exp Med,

20 I. Summary Of 5 HIV-1 acute HIV infection gp41 antibody clones analyzed, no UCA reacted with autologous Env. Only 6 of a total of 67 observed gp41 antibodies from 5 individuals were unmutated suggesting infrequent triggering of naïve B cells by gp41. HIV gp41 triggers a preexisting antibody clone in which the germline does not react with gp41 but an intermediate clone antibody acquires gp41 crossreactivity and then expands. HIV gp41 antibodies were reactive with intestinal bacterial antigens. 20

21 Environmental An gens in Shaping the Response to HIV Plasma cell repertoires in acute HIV- 1 infec on Plasma and memory B cells in terminal ileum during early and chronic HIV- 1 infec on Memory B cells in HIV- 1 Env vaccina on (Wilton Williams) Memory B cells in colostrum 21

22 Can HIV-1 reactive antibodies be isolated from the terminal ileum of individuals with early HIV-1 infection? 22

23 We studied the terminal ileum of 6 individuals with early HIV-1 infection (EHI), ~ days after transmission. Trama et al Cell Host Microbe. 16:215 23

24 Isolation of Plasma Cell and Memory B Cell VH and VL from Terminal Ileum of Early HIV-1-infected individuals 254 mab Screened Plasma Cells: 114 mab Memory B Cells: 140 mab 3 mab = HIV-1 gp41 reactive (1.2%) Thus, there is also a dearth of HIV-1-reactive antibodies in the terminal ileum as in the blood of acute HIV-1 infected individuals 24

25 Is there a similar scarcity of HIV-1-reactive plasma cells in the terminal ileum of chronically-infected individuals? 25

26 We studied the terminal ileum of 3 chronically-infected subjects (CHI), >200 days after transmission. 26

27 Chronic HIV-1 infected Individuals: HIV-1 Reactive Ab Isolated from Terminal Ileum Plasma Cells gp41 (5.7%) gp120 (0.6%) p24 (2.5%) non HIV- 1/ non- defined (92.1%) 9/158 mab = HIV- 1 Env gp41- reac ve (5.7%) Total: 14/158 mab = HIV- 1 reac ve (8.8%) Trama et al Cell Host Microbe. 16:215 27

28 Are gp41-reactive antibodies isolated from the terminal ileum cross-reactive with intestinal bacterial whole cell lysate? 28

29 HIV-1 gp41-reactive Terminal Ileum Antibodies Cross-React with intestinal Bacteria Whole Cell Lysate 17 gp41-reactive Abs from early and chronic HIV-1 infection: 15 (88.2%) are intestinal bacteria cross-reactive; 2 (11.8%) are not intestinal bacteria cross-reactive. 29

30 What are antigens in intestinal bacteria that crossreact with gp41-reactive antibodies? 30

31 Western Blot of Intestinal Bacteria Antigens on BN-PAGE with gp41 mab 276 KDa gp41- reac ve mab 276 binds to ~520kDa protein band LC- MS analysis iden fied the reac ve band as E coli RNA polymerase Blue Na ve Gel Western Blot with mab

32 Do antibodies isolated from the terminal ileum that react with both HIV-1 gp41 and intestinal bacteria whole cell lysate also react with recombinant purified E coli RNA polymerase? 32

33 HIV-1 gp41 and intestinal bacteria Cross- Reactive Antibodies Also React with E coli RNA Polymerase 15 gp41+ intestinal bacteria+ Abs from early and chronic HIV-1 infection: 4 (26.7%) are reactive with re coli RNA polymerase; 11 (73.3%) are not reactive with re coli RNA polymerase. 33

34 Can HIV gp41 commensal bacterial cross-reactive antibodies be isolated from the terminal ileum of uninfected individuals? 34

35 Uninfected Individual: HIV-1 gp41- intestinal bacteria cross-reactive Abs Isolated from Terminal Ileum Plasma Cells and Memory B Cells Trama et al Cell Host Microbe. 16:215 35

36 II. Summary Similar to blood plasma cells in AHI, there is a dearth of HIV-1 reactive plasma cells in the terminal ileum in early and chronic HIV-1 infection. 17 (77.3%) of the 22 HIV-1 reactive antibodies isolated from the terminal ileum were reactive to gp41. Of 17 gp41-reactive antibodies isolated from the terminal ileum, 15 (88.2%) cross-reacted with intestinal bacterial antigens. 4 (26.7%) of the 15 gp41-intestinal bacteria cross-reactive antibodies isolated from the terminal ileum also react with re coli RNA polymerase. Mutated, class-switched, gp41-intestinal bacteria cross-reactive antibodies could be isolated from HIV uninfected individuals. 36

37 Environmental An gens in Shaping the Response to HIV Plasma cell repertoires in acute HIV- 1 infec on Plasma and memory B cells in terminal ileum during early and chronic HIV- 1 infec on Memory B cells in HIV- 1 Env vaccina on (Wilton Williams) Memory B cells in colostrum 37

38 Environmental An gens in Shaping the Response to HIV Plasma cell repertoires in acute HIV- 1 infec on Plasma and memory B cells in terminal ileum during early and chronic HIV- 1 infec on Memory B cells in HIV- 1 Env vaccina on (Wilton Williams) Memory B cells in colostrum 38

39 Breast Milk Antibodies Myriad benefits of breastfeeding to the growing infant, including decreased risk of obesity. Breast milk is rich in IgG and IgA antibodies that contribute to protection against infection. Breastfeeding by HIV-infected mothers is encouraged in resource-limited areas because of the overall mortality benefits despite ongoing exposure. Despite ongoing exposure, many infants remain uninfected. 39

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43 Colostrum B Cells Isolation of memory B cells from colostrum yielded gp120-reactive antibodies with isotype restriction. B cell clonal lineage members can be found across compartments. 43

44 Colostrum B Cells Isolation of colostrum B cells by antigen-specific memory B cell sorting. 44

45 Unpublished Data 5 slides showing unpublished data. 45

46 III. Summary In contrast with intestinal B cells and the plasmablast response seen in early HIV-1 infection, HIV-1-specfic colostrum B cells are primarily gp120-reactive. Differential patterns of reactivity with intestinal flora antigens are observed. HIV-1 antibody cross-reactivity with intestinal microbiota antigens is not restricted to E coli RNA polymerase. 46

47 Conclusions Microbes or other environmental antigens with similar sequences or structures to HIV-1 Env can stimulate naïve B cells, which in turn differentiate into memory B cells. HIV-1 gp41 after HIV-1 infection can stimulate these memory B cells to proliferate and mature in affinity in response to HIV-1 gp41. Data from colostrum B cells cross-reactive between intestinal microbiota antigens and gp120 suggest that the phenomenon is not restricted to single microbial antigens or gp41 epitopes. 47

48 DHVI Barton Haynes Larry Liao Stephen Chen Ashely Trama Wilton Williams Munir Alam Kevin Wiehe David Montefiori Feng Gao John Whitesides Dawn Jones Marshall Guido Ferrari Justin Pollara Nathan Vandergrift Thomas Kepler Acknowledgments CHAVI Myron Cohen George Shaw Beatrice Hahn Stanford Andrew Fire Scott Boyd VRC Barney Graham John Mascola Richard Koup Gary Nabel UAB Christina Ochsenbauer DHVI Sallie Permar Giny Fouda Lee Jeffries Caitlin Sacha Erika Kunz Josh Eudailey Thad Gurley Lawrence Armand Tarra Von Holle Ashley Allen Lauren Stiegel

49 Acknowledgements Supported by: Na onal Ins tute of Allergy and Infec ous Diseases (NIAID) Na onal Ins tutes of Health (NIH) Division of AIDS (DAIDS) U.S. Department of Health and Human Services (HHS) Duke Center for HIV/AIDS Vaccine Immunology and Immunogen Discovery (CHAVI- ID) #UM1- AI