Dr. William R. Jacobs, Jr., is the Principal Investigator for an NIH-funded P01 Program Project Grant ( AI063537) in collaboration with the laboratories of Dr. Steven A. Porcelli, and Dr. John Chan. This five-year project funded in 2005 is focused on the molecular analysis of Tuberculosis immunology. The P01 Program Project Grant has the following projects and cores:
Project 1 - Genetic Approches for Improving Tuberculosis Vaccines (William R. Jacobs, Jr, Project Leader)
Project 2 - Determinants of T cell Immunity to Tuberculosis Vaccines (Steven Porcelli, Project Leader)
Project 3 - The Tuberculous Granuloma (John Chan, Project Leader)
Core A - Adminstrative (William R. Jacobs, Jr, Core Leader)
Core B - Animals and Infection (John Chan, Core Leader)
Core C - Flow Cytometry (Steven Porcelli, Core Leader)
During its first four years, the P01-funded program has been extremely successful both in catalyzing the interaction of tuberculosis investigators at Einstein, and in generating major scientific advances that are pointing the way to more effective prevention and treatment of tuberculosis. The following is a summary of the major scientific achievements of the P01-funded program to date that are directly relevant to the primary goal of improved TB vaccine development:
1. The first discovery of anti-apoptosis genes in Mtb. Earlier work had shown that the ability of Mtb to block apoptosis of infected macrophages is an important virulence trait. Our work identified for the first time several specific genes of the mycobacterium that are required for this immune evasion strategy.
2. The demonstration that pro-apoptotic mutants of Mtb dramatically increase the priming of CD8+ T cells against mycobacterial antigens. While a few earlier studies had suggested a link between apoptosis and priming of CD8+ T cell responses, we have created the first specific mutant strains of Mtb that engage this mechanism. This has major implications for understanding adaptive immunity to tuberculosis and for the development of better live attenuated mycobacterial vaccine strains.
3. The demonstration that immunization with pro‑apoptotic mutant strains of Mtb can give protective immunity superior to BCG. Prior to our studies, there were few if any examples of experimental tuberculosis vaccines that reproducibly gave better protection against Mtb challenge in terms of tissue bacterial burden and survival when compared head to head with BCG in animal models. This important finding was replicated in both mouse and guinea pig models of TB. In guinea pigs, the protective effect following vaccination with a pro-apoptotic mutant strain of Mtb was associated with an extraordinary reduction of infection in the draining pulmonary lymph nodes, which was almost two logs greater in animals vaccinated with a pro-apoptotic mutant strain of Mtb compared to BCG vaccinated animals.
4. The development of a high throughput genome-wide screen for Mtb genes that regulate cytokine production by infected host cells. This included the development of a macrophage cell line expressing a reporter system for IL-12 production, which was used to identify Mtb genes that attenuate the production of IL-12 by infected macrophages. More recently, this approach was also extended to a genome-wide screen for Mtb genes that regulate production of TNF by macrophages.
5. The first identification of a specific mycobacterial gene involved in suppression of IL-12 production by Mtb infected macrophages. It has been known for some time that Mtb modulates the cytokine responses of infected cells and tissues. However, prior to our studies virtually nothing was known of the mycobacterial genes that controlled these alterations in cytokine responses. Our genome-wide screen for Mtb mutants generating enhanced IL-12 responses identified mmaA4 (Rv0642c), which encodes a modifier of cell wall mycolic acids, as a key gene involved in suppression of the IL-12 response by Mtb.
6. The generation of a large collection of over 50 strains of recombinant BCG that give enhanced host cell apoptosis and cytokine responses. We extended our search for mutants with enhanced immunogenicity to BCG, and have been richly rewarded by the identification of large numbers clones with transposon insertion mutations in genes that interfere with host cell apoptosis and cytokine production. These represent potentially safer and more effective forms of BCG, one or more of which could potentially enter into clinical trials of vaccine efficacy in the near future. We have fully established the methods required to identify the mutagenized genes, and to precisely delete these genes using the specialized transduction system developed in our laboratories.
7. Similarly, we have developed strains of Mtb in which deletion of immune-evasion genes have been combined with deletion of other genes required for virulence to produce safe attenuated vaccine strains. These also have the potential to enter into clinical testing in the near future.
8. As a first step toward assessing the feasibility of clinical trials, a non-human primate study was carried out by collaborators evaluate the safety of attenuated Mtb vaccines. A follow up study in non-human primates using pro-apoptotic Mtb mutants generated through support of this P01 has just finished and a manuscript is in preparation.
9. In addition to the above accomplishments that have direct relevance to the development of improved vaccines, major scientific progress was also achieved in several related areas of immunopathogenesis and microbiology that directly addressed specific aims of the original P01 proposal. For example:
10. It was known prior to our studies that significant protective immunity against TB can be induced by vaccination of CD4-/- knockout mice, which have been considered as a possible model for vaccination in the setting of HIV-associated T cell depletion. However, the mechanism of vaccine-induced protection in these CD4 deficient mice was unknown. We completed and published studies that determined the mechanisms of vaccine-induced protective immunity in this model of T cell-deficient mice.
11. The role of B cells and antibodies has been a contentious and largely neglected area of research on tuberculosis immunity. We demonstrated in well-controlled mouse models that B cells are required for optimal protection against Mtb, and can enhance Th1 response and increased activation of antigen presenting cells through effects mediated by Fcγ receptors. These findings could greatly alter the current thinking about the role of B cells and antibodies in protective immunity to Mtb, and will be pursued in detail because of their potential importance in the context of vaccine design
12. Acid-fast staining has long been used as a criterion for identifying mycobacteria in tissues and clinical specimens, but the precise mechanism that accounts for this staining characteristic remained obscure. We identified the first specific mutation in mycobacteria that eliminates their acid-fast staining property, which involves a gene that carries out a step in the synthesis and modification of mycolic acids. These studies have led to an improved understanding of what it means to be "acid-fast bacilli" and may help in the development of better diagnostic techniques
Publications from P01 Investigators, 2005 to 2009 (Items in bold are papers that report work that was entirely or predominately supported by the P01 grant).
1. Ndonye RM, Izmirian DP, Dunn MF, Yu KO, Porcelli SA, Khurana A, Kronenberg M, Richardson SK, Howell AR. Synthesis and evaluation of sphinganine analogues of KRN7000 and OCH. J Org Chem 2005;70(25):10260-70.
2. Rao V, Fujiwara N, Porcelli SA, Glickman MS. Mycobacterium tuberculosis controls host innate immune activation through cyclopropane modification of a glycolipid effector molecule. J Exp Med 2005;201(4):535-43.
3. Sambandamurthy VK, Derrick SC, Jalapathy KV, Chen B, Russell RG, Morris SL, Jacobs WR, Jr. Long-term protection against tuberculosis following vaccination with a severely attenuated double lysine and pantothenate auxotroph of Mycobacterium tuberculosis. Infect Immun 2005;73(2):1196-203.
4. Sambandamurthy VK, Jacobs WR, Jr. Live attenuated mutants of Mycobacterium tuberculosis as candidate vaccines against tuberculosis. Microbes Infect 2005;7(5-6):955-61.
5. Sieling PA, Torrelles JB, Stenger S, Chung W, Burdick AE, Rea TH, Brennan PJ, Belisle JT, Porcelli SA, Modlin RL. The human CD1-restricted T cell repertoire is limited to cross-reactive antigens: implications for host responses against immunologically related pathogens. J Immunol 2005;174(5):2637-44.
6. Yu KO, Im JS, Molano A, Dutronc Y, Illarionov PA, Forestier C, Fujiwara N, Arias I, Miyake S, Yamamura T, Chang YT, Besra GS, Porcelli SA. Modulation of CD1d-restricted NKT cell responses by using N-acyl variants of alpha-galactosylceramides. Proc Natl Acad Sci U S A 2005;102(9):3383-8.
7. Cayabyab MJ, Hovav AH, Hsu T, Krivulka GR, Lifton MA, Gorgone DA, Fennelly GJ, Haynes BF, Jacobs WR, Jr., Letvin NL. Generation of CD8+ T-cell responses by a recombinant nonpathogenic Mycobacterium smegmatis vaccine vector expressing human immunodeficiency virus type 1 Env. J Virol 2006;80(4):1645-52.
8. Junqueira-Kipnis AP, Basaraba RJ, Gruppo V, Palanisamy G, Turner OC, Hsu T, Jacobs WR, Jr., Fulton SA, Reba SM, Boom WH, Orme IM. Mycobacteria lacking the RD1 region do not induce necrosis in the lungs of mice lacking interferon-gamma. Immunology 2006;119(2):224-31.
9. Lee S, Jeon BY, Bardarov S, Chen M, Morris SL, Jacobs WR, Jr. Protection elicited by two glutamine auxotrophs of Mycobacterium tuberculosis and in vivo growth phenotypes of the four unique glutamine synthetase mutants in a murine model. Infect Immun 2006;74(11):6491-5.
10. Rao V, Gao F, Chen B, Jacobs WR, Jr., Glickman MS. Trans-cyclopropanation of mycolic acids on trehalose dimycolate suppresses Mycobacterium tuberculosis -induced inflammation and virulence. J Clin Invest 2006;116(6):1660-7.
11. Roy E, De Silva AD, Sambandamurthy VK, Clark SO, Stavropoulos E, Jacobs WR, Jr., Brennan J, Chan J, Williams A, Colston MJ, Tascon RE. Induction of high levels of protective immunity in mice after vaccination using dendritic cells infected with auxotrophic mutants of Mycobacterium tuberculosis. Immunol Lett 2006;103(2):196-9.
12. Sambandamurthy VK, Derrick SC, Hsu T, Chen B, Larsen MH, Jalapathy KV, Chen M, Kim J, Porcelli SA, Chan J, Morris SL, Jacobs WR, Jr. Mycobacterium tuberculosis DeltaRD1 DeltapanCD: a safe and limited replicating mutant strain that protects immunocompetent and immunocompromised mice against experimental tuberculosis. Vaccine 2006;24(37-39):6309-20.
13. Tsai MC, Chakravarty S, Zhu G, Xu J, Tanaka K, Koch C, Tufariello J, Flynn J, Chan J. Characterization of the tuberculous granuloma in murine and human lungs: cellular composition and relative tissue oxygen tension. Cell Microbiol 2006;8(2):218-32.
14. Tufariello JM, Mi K, Xu J, Manabe YC, Kesavan AK, Drumm J, Tanaka K, Jacobs WR, Jr., Chan J. Deletion of the Mycobacterium tuberculosis resuscitation-promoting factor Rv1009 gene results in delayed reactivation from chronic tuberculosis. Infect Immun 2006;74(5):2985-95.
15. Vilcheze C, Wang F, Arai M, Hazbon MH, Colangeli R, Kremer L, Weisbrod TR, Alland D, Sacchettini JC, Jacobs WR, Jr. Transfer of a point mutation in Mycobacterium tuberculosis inhA resolves the target of isoniazid. Nat Med 2006;12(9):1027-9.
16. Yu JS, Peacock JW, Vanleeuwen S, Hsu T, Jacobs WR, Jr., Cayabyab MJ, Letvin NL, Frothingham R, Staats HF, Liao HX, Haynes BF. Generation of mucosal anti-human immunodeficiency virus type 1 T-cell responses by recombinant Mycobacterium smegmatis. Clin Vaccine Immunol 2006;13(11):1204-11.
17. Bhatt A, Fujiwara N, Bhatt K, Gurcha SS, Kremer L, Chen B, Chan J, Porcelli SA, Kobayashi K, Besra GS, Jacobs WR, Jr. Deletion of kasB in Mycobacterium tuberculosis causes loss of acid-fastness and subclinical latent tuberculosis in immunocompetent mice. Proc Natl Acad Sci U S A 2007;104(12):5157-62.
18. Bhatt K, Gurcha SS, Bhatt A, Besra GS, Jacobs WR, Jr. Two polyketide-synthase-associated acyltransferases are required for sulfolipid biosynthesis in Mycobacterium tuberculosis. Microbiology 2007;153(Pt 2):513-20.
19. Chakravarty SD, Xu J, Lu B, Gerard C, Flynn J, Chan J. The chemokine receptor CXCR3 attenuates the control of chronic Mycobacterium tuberculosis infection in BALB/c mice. J Immunol 2007;178(3):1723-35.
20. Derrick SC, Evering TH, Sambandamurthy VK, Jalapathy KV, Hsu T, Chen B, Chen M, Russell RG, Junqueira-Kipnis AP, Orme IM, Porcelli SA, Jacobs WR, Jr., Morris SL. Characterization of the protective T-cell response generated in CD4-deficient mice by a live attenuated Mycobacterium tuberculosis vaccine. Immunology 2007;120(2):192-206.
21. Hinchey J, Lee S, Jeon BY, Basaraba RJ, Venkataswamy MM, Chen B, Chan J, Braunstein M, Orme IM, Derrick SC, Morris SL, Jacobs WR, Jr., Porcelli SA. Enhanced priming of adaptive immunity by a proapoptotic mutant of Mycobacterium tuberculosis. J Clin Invest 2007;117(8):2279-88.
22. Hovav AH, Cayabyab MJ, Panas MW, Santra S, Greenland J, Geiben R, Haynes BF, Jacobs WR, Jr., Letvin NL. Rapid memory CD8+ T-lymphocyte induction through priming with recombinant Mycobacterium smegmatis. J Virol 2007;81(1):74-83.
23. Larsen MH, Biermann K, Jacobs WR, Jr. Analyses of Mycobacterium tuberculosis proteins. Curr Protoc Microbiol 2007;Chapter 10:Unit 10A 4.
24. Larsen MH, Biermann K, Jacobs WR, Jr. Laboratory maintenance of Mycobacterium tuberculosis. Curr Protoc Microbiol 2007;Chapter 10:Unit 10A 1.
25. Larsen MH, Biermann K, Tandberg S, Hsu T, Jacobs WR, Jr. Genetic Manipulation of Mycobacterium tuberculosis. Curr Protoc Microbiol 2007;Chapter 10:Unit 10A 2.
26. Li Q, Ndonye RM, Illarionov PA, Yu KO, Jerud ES, Diaz K, Bricard G, Porcelli SA, Besra GS, Chang YT, Howell AR. Rapid identification of immunostimulatory alpha-galactosylceramides using synthetic combinatorial libraries. J Comb Chem 2007;9(6):1084-93.
27. Maglione PJ, Xu J, Chan J. B cells moderate inflammatory progression and enhance bacterial containment upon pulmonary challenge with Mycobacterium tuberculosis. J Immunol 2007;178(11):7222-34.
28. Mo Y, Quanquin NM, Vecino WH, Ranganathan UD, Tesfa L, Bourn W, Derbyshire KM, Letvin NL, Jacobs WR, Jr., Fennelly GJ. Genetic alteration of Mycobacterium smegmatis to improve mycobacterium-mediated transfer of plasmid DNA into mammalian cells and DNA immunization. Infect Immun 2007;75(10):4804-16.
29. Velmurugan K, Chen B, Miller JL, Azogue S, Gurses S, Hsu T, Glickman M, Jacobs WR, Jr., Porcelli SA, Briken V. Mycobacterium tuberculosis nuoG is a virulence gene that inhibits apoptosis of infected host cells. PLoS Pathog 2007;3(7):e110.
30. Vilcheze C, Jacobs WR. Isolation and analysis of Mycobacterium tuberculosis mycolic acids. Curr Protoc Microbiol 2007;Chapter 10:Unit 10A 3.
31. Waters WR, Palmer MV, Nonnecke BJ, Thacker TC, Scherer CF, Estes DM, Jacobs WR, Jr., Glatman-Freedman A, Larsen MH. Failure of a Mycobacterium tuberculosis DeltaRD1 DeltapanCD double deletion mutant in a neonatal calf aerosol M. bovis challenge model: comparisons to responses elicited by M. bovis bacille Calmette Guerin. Vaccine 2007;25(45):7832-40.
32. Yu JS, Peacock JW, Jacobs WR, Jr., Frothingham R, Letvin NL, Liao HX, Haynes BF. Recombinant Mycobacterium bovis bacillus Calmette-Guerin elicits human immunodeficiency virus type 1 envelope-specific T lymphocytes at mucosal sites. Clin Vaccine Immunol 2007;14(7):886-93.
33. Yu KO, Im JS, Illarionov PA, Ndonye RM, Howell AR, Besra GS, Porcelli SA. Production and characterization of monoclonal antibodies against complexes of the NKT cell ligand alpha-galactosylceramide bound to mouse CD1d. J Immunol Methods 2007;323(1):11-23.
34. Chakravarty SD, Zhu G, Tsai MC, Mohan VP, Marino S, Kirschner DE, Huang L, Flynn J, Chan J. Tumor necrosis factor blockade in chronic murine tuberculosis enhances granulomatous inflammation and disorganizes granulomas in the lungs. Infect Immun 2008;76(3):916-26.
35. Dao DN, Sweeney K, Hsu T, Gurcha SS, Nascimento IP, Roshevsky D, Besra GS, Chan J, Porcelli SA, Jacobs WR. Mycolic acid modification by the mmaA4 gene of Mtb modulates IL-12 production. PLoS Pathog 2008;4(6):e1000081.
36. Huang D, Shen Y, Qiu L, Chen CY, Shen L, Estep J, Hunt R, Vasconcelos D, Du G, Aye P, Lackner AA, Larsen MH, Jacobs WR, Jr., Haynes BF, Letvin NL, Chen ZW. Immune distribution and localization of phosphoantigen-specific Vgamma2Vdelta2 T cells in lymphoid and nonlymphoid tissues in Mycobacterium tuberculosis infection. Infect Immun 2008;76(1):426-36.
37. Im JS, Kang TJ, Lee SB, Kim CH, Lee SH, Venkataswamy MM, Serfass ER, Chen B, Illarionov PA, Besra GS, Jacobs WR, Jr., Chae GT, Porcelli SA. Alteration of the relative levels of iNKT cell subsets is associated with chronic mycobacterial infections. Clin Immunol 2008;127(2):214-24.
38. Maglione PJ, Xu J, Casadevall A, Chan J. Fc gamma receptors regulate immune activation and susceptibility during Mycobacterium tuberculosis infection. J Immunol 2008;180(5):3329-38.
39. Ojha AK, Baughn AD, Sambandan D, Hsu T, Trivelli X, Guerardel Y, Alahari A, Kremer L, Jacobs WR, Jr., Hatfull GF. Growth of Mycobacterium tuberculosis biofilms containing free mycolic acids and harbouring drug-tolerant bacteria. Mol Microbiol 2008;69(1):164-74.
40. Park JJ, Lee JH, Ghosh SC, Bricard G, Venkataswamy MM, Porcelli SA, Chung SK. Synthesis of all stereoisomers of KRN7000, the CD1d-binding NKT cell ligand. Bioorg Med Chem Lett 2008;18(14):3906-9.
41. Porcelli SA, Jacobs WR, Jr. Tuberculosis: unsealing the apoptotic envelope. Nat Immunol 2008;9(10):1101-2.
42. Chen CY, Huang D, Wang RC, Shen L, Zeng G, Yao S, Shen Y, Halliday L, Fortman J, McAllister M, Estep J, Hunt R, Vasconcelos D, Du G, Porcelli SA, Larsen MH, Jacobs WR, Jr., Haynes BF, Letvin NL, Chen ZW. A critical role for CD8 T cells in a nonhuman primate model of tuberculosis. PLoS Pathog 2009;5(4):e1000392.
43. Lim J, Derrick SC, Kolibab K, Yang AL, Porcelli S, Jacobs WR, Morris SL. Early pulmonary cytokine and chemokine responses in mice immunized with three different vaccines against Mycobacterium tuberculosis determined by PCR array. Clin Vaccine Immunol 2009;16(1):122-6.
44. Maglione PJ, Chan J. How B cells shape the immune response against Mycobacterium tuberculosis. Eur J Immunol 2009;39(3):676-86.
45. Piuri M, Jacobs WR, Jr., Hatfull GF. Fluoromycobacteriophages for rapid, specific, and sensitive antibiotic susceptibility testing of Mycobacterium tuberculosis. PLoS ONE 2009;4(3):e4870.
46. Waters WR, Palmer MV, Nonnecke BJ, Thacker TC, Scherer CF, Estes DM, Hewinson RG, Vordermeier HM, Barnes SW, Federe GC, Walker JR, Glynne RJ, Hsu T, Weinrick B, Biermann K, Larsen MH, Jacobs WR, Jr. Efficacy and immunogenicity of Mycobacterium bovis DeltaRD1 against aerosol M. bovis infection in neonatal calves. Vaccine 2009;27(8):1201-9.
47. Zimmerman DM, Waters WR, Lyashchenko KP, Nonnecke BJ, Armstrong DL, Jacobs WR, Jr., Larsen MH, Egan E, Dean GA. Safety and immunogenicity of the Mycobacterium tuberculosis DeltalysA DeltapanCD vaccine in domestic cats infected with feline immunodeficiency virus. Clin Vaccine Immunol 2009;16(3):427-9.
48. Venkataswamy MM, Baena A, Goldberg MF, Bricard G, Im JS, Besra GS, Chan J, Jacobs WR, Jr., Porcelli SA. NKT cell activating glycolipid adjuvants enhance protective efficacy of M. bovis BCG vaccine against tuberculosis. J Immunol. 2009 Aug 1;183(3):1644-56.PMID: 19620317.
49. Im JS, Arora P, Bricard G, Molano A, Venkataswamy MM, Baine I, Jerud ES, Goldberg MF, Yu KO, Ndonye RM, Howell AR, Chang YT, Illarionov PA, Besra GS, Porcelli SA. Kinetics and cellular site of glycolipid loading control the outcome of Natural Killer T cell acitivation. Immunity. 2009 Jun 19;30(6):888-98.PMID: 19538930
50. Larsen MH, Biermann K, Chen B, Hsu T, Samdandamurthy VK, Lackner AA, Aye PP, Didier P, Huang D, Shao L, Wei H, Letvin NL, Frothingham R, Haynes BH, Chen ZW, Jacobs WR, Jr. Efficacy and safety of live attenuated persistent and rapidly cleared Mycobacterium tuberculosis vaccine candidates in non-human primates. Vaccine. 2009 Jul 23;27(34):4709-17. Epub 2009 Jun 2.PMID: 19500524
51. Ranganathan U, Larsen MH, Kim J, Porcelli SA, Jacobs WR Jr., Fennelly GJ. Recombinant proapoptotic M. tuberculosis generates CD8+ T-cell responses against human immunodeficiency virus type 1 Env and M. tuberculosis in neonatal mice. Vaccine 28:152-61. PMID: 19808028 (2009).
52. Park JJ, Lee JH, Seo KC, Bricard G, Venkataswamy MM, Porcelli SA, Chung SK. Syntheses and biological activities of KRN7000 analogues having aromatic residues in the acyl and backbone chains with varying stereochemistry. Bioorg Med Chem Lett. 2010 Jan 4. [Epub ahead of print] PMID: 20061147
53. Yam, K.C., I. D'Angelo, R. Kalscheuer, H. Zhu, J.X. Wang, V. Snieckus, L.H. Ly, P.J. Converse, W.R. Jacobs, Jr., N. Strynadka, and L.D. Eltis. 2009. Studies of a ring-cleaving dioxygenase illuminate the role of cholesterol metabolism in the pathogenesis of Mycobacterium tuberculosis. PLoS Pathog 5:e1000344.
54. Waters, W.R., M.V. Palmer, B.J. Nonnecke, T.C. Thacker, D.M. Estes, M.H. Larsen, W.R. Jacobs, Jr., P. Andersen, J. McNair, F.C. Minion, K.P. Lyashchenko, R.G. Hewinson, H.M. Vordermeier, and R.E. Sacco. 2009. Signal regulatory protein alpha (SIRPalpha) cells in the adaptive response to ESAT-6/CFP-10 protein of tuberculous mycobacteria. PLoS One 4:e6414.
55. Parra, M., A.L. Yang, J. Lim, K. Kolibab, S. Derrick, N. Cadieux, L.P. Perera, W.R. Jacobs, M. Brennan, and S.L. Morris. 2009. Development of a murine mycobacterial growth inhibition assay for evaluating vaccines against Mycobacterium tuberculosis. Clin Vaccine Immunol 16:1025-1032.
56. Nascimento, I.P., W.O. Dias, W. Quintilio, T. Hsu, W.R. Jacobs, Jr., and L.C. Leite. 2009. Construction of an unmarked recombinant BCG expressing a pertussis antigen by auxotrophic complementation: protection against Bordetella pertussis challenge in neonates. Vaccine 27:7346-7351.
57. Lee, W.L., B. Gold, C. Darby, N. Brot, X. Jiang, L.P. de Carvalho, D. Wellner, G. St John, W.R. Jacobs, Jr., and C. Nathan. 2009. Mycobacterium tuberculosis expresses methionine sulphoxide reductases A and B that protect from killing by nitrite and hypochlorite. Mol Microbiol 71:583-593.
58. Ioerger, T.R., S. Koo, E.G. No, X. Chen, M.H. Larsen, W.R. Jacobs, Jr., M. Pillay, A.W. Sturm, and J.C. Sacchettini. 2009. Genome analysis of multi- and extensively-drug-resistant tuberculosis from KwaZulu-Natal, South Africa. PLoS One 4:e7778.
59. Endsley, J.J., W.R. Waters, M.V. Palmer, B.J. Nonnecke, T.C. Thacker, W.R. Jacobs, Jr., M.H. Larsen, A. Hogg, E. Shell, M. McAlauy, C.F. Scherer, T. Coffey, C.J. Howard, B. Villareal-Ramos, and D.M. Estes. 2009. The calf model of immunity for development of a vaccine against tuberculosis. Vet Immunol Immunopathol 128:199-204.
60. Cirillo, S.L., S. Subbian, B. Chen, T.R. Weisbrod, W.R. Jacobs, Jr., and J.D. Cirillo. 2009. Protection of Mycobacterium tuberculosis from reactive oxygen species conferred by the mel2 locus impacts persistence and dissemination. Infect Immun 77:2557-2567.
61. Chen, J., J. Kriakov, A. Singh, W.R. Jacobs, Jr., G.S. Besra, and A. Bhatt. 2009. Defects in glycopeptidolipid biosynthesis confer phage I3 resistance in Mycobacterium smegmatis. Microbiology 155:4050-4057.
62. Cayabyab, M.J., B. Korioth-Schmitz, Y. Sun, A. Carville, H. Balachandran, A. Miura, K.R. Carlson, A.P. Buzby, B.F. Haynes, W.R. Jacobs, and N.L. Letvin. 2009. Recombinant Mycobacterium bovis BCG prime-recombinant adenovirus boost vaccination in rhesus monkeys elicits robust polyfunctional simian immunodeficiency virus-specific T-cell responses. J Virol 83:5505-5513.
63. Baughn, A.D., S.J. Garforth, C. Vilcheze, and W.R. Jacobs, Jr. 2009. An anaerobic-type alpha-ketoglutarate ferredoxin oxidoreductase completes the oxidative tricarboxylic acid cycle of Mycobacterium tuberculosis. PLoS Pathog 5:e1000662.
64. Banaei, N., E.Z. Kincaid, S.Y. Lin, E. Desmond, W.R. Jacobs, Jr., and J.D. Ernst. 2009. Lipoprotein processing is essential for resistance of Mycobacterium tuberculosis to malachite green. Antimicrob Agents Chemother 53:3799-3802.