Public Health Research Institute Center
New Jersey Medical School - Rutgers, The State University of New Jersey
225 Warren Street
Newark, New Jersey 07103
Phone: (973) 854-3320
Most Mycobacterium tuberculosis (M. tb) infections are dormant; the human hosts are latently infected. Tuberculosis occurs when a failure of immunity, including that caused by HIV/AIDS, allows bacterial growth in macrophages. Thus, studying infected macrophages is central to improving control of the host-pathogen interaction. The long-term goal of the Pine laboratory is to understand regulation of gene expression in macrophages infected by M. tb well enough to open avenues for manipulating the immune response as a means to control infection and prevent or treat tuberculosis. The laboratory uses macrophages infected by M. tb in vitro as a model of active infection to investigate specific responses to immune modulators, and to uncover general effects on regulation of gene expression.
These studies started with a project to test the hypothesis that interferon (IFN)-regulated gene expression is altered in macrophages infected with M. tb. Original observations that cells infected by M. tb secrete IFNα/β (Weiden et al., 2000) were followed by studies of IFNα/β-stimulated signal transduction and gene expression in infected cells. Infection was found to limit response to IFNα/β (Prabhakar, et al., 2003), in part through negative feedback regulation caused by the secreted IFNα/β (Prabhakar, et al., 2005). The net effect is chronic, low-level response to IFNα/β. The next steps are to investigate the consequences of IFNα/β production during infection for macrophage apoptosis and antigen presentation; both processes are responsive to IFNα/β. The effects of the IFNα/β on interactions of infected macrophages with CD8+ cytotoxic T and Natural Killer (NK) cells are also of great interest, since IFNα/β profoundly affects the functions of both cell types.
Additional studies focused on the effects of infection and stimulation with IFNγ on expression of interferon regulatory factor 1 (IRF1). Both IFNγ and IRF1 are essential for host defense against M. tb. Both positive and negative regulation occurred (Qiao et al., 2002, 2004); this limit to perturbation of the host cell involves post-transcriptional regulation in the nucleus. Altered RNA stability failed to account for differences in the abundance of primary and processed IRF1 transcripts. Moreover, when macrophages are infected, a reprogrammed response to IFNγ affects mediators of post-initiation regulation. Therefore, the laboratory hypothesizes that post-initiation steps such as transcript elongation or maturation are regulated. Future studies will focus on the basis and role for such regulation in the host response to M. tb.
An ongoing study of co-infection by M. tuberculosis and HIV-1 uses functional genomics to test the hypotheses that infection by HIV-1 alters cellular response to subsequent infection by M. tb and vice versa, and that the final gene expression profile will depend on the order of infection. The nature of differences in gene expression will generate additional hypotheses to guide future studies. The data will also extend the hypothesis on post-initiation regulation as a response to M. tb to address the consequences of co-infection, which relates to the HIV life cycle through effects on the species of HIV transcripts that are produced.
These in vitro studies are complemented by clinical studies performed in collaboration with Drs. Rany Condos, Michael Weiden, and William Rom at New York University School of Medicine and Bellevue Hospital Center nearby in New York City. Functional-genomic and hypothesis-driven approaches are being employed to understand the molecular basis for clinical responses to adjunctive aerosolized IFNγ therapy for tuberculosis patients with and without AIDS (Condos et al., 2003; Raju et al., 2004, Weiden et al., 2007).
Vir P, Arrigucci R, Lakehal K, Davidow AL, Pine R, Tyagi S, Bushkin Y, Lardizabal A, Gennaro ML (2015) Single-Cell Cytokine Gene Expression in Peripheral Blood Cells Correlates with Latent Tuberculosis Status. PLoS One 10: e0144904. PMI: 26658491
Bushkin Y, Radford F, Pine R, Lardizabal A, Mangura BT, Gennaro ML, Tyagi S (2015) Profiling T cell activation using single-molecule fluorescence in situ hybridization and flow cytometry. J Immunol 194: 836-841. PMI: 25505292
Skerry C, Pinn ML, Bruiners N, Pine R, Gennaro ML, Karakousis PC (2014) Simvastatin increases the in vivo activity of the first-line tuberculosis regimen. J Antimicrob Chemother 69: 2453-2457. PMI: 24855121
Raju B, Hoshino Y, Belitskaya-Levy I, Dawson R, Ress S, Gold JA, Condos R, Pine R, Brown S, Nolan A, Rom WN, Weiden MD (2008) Gene expression profiles of bronchoalveolar cells in pulmonary TB. Tuberculosis (Edinb) 88: 39-51. PMI: 17921069
Fontan PA, Aris V, Alvarez ME, Ghanny S, Cheng J, Soteropoulos P, Trevani A, Pine R, Smith I (2008) Mycobacterium tuberculosis sigma factor E regulon modulates the host inflammatory response. J Infect Dis 198: 877-885. PMI: 18657035
Hoshino Y, Hoshino S, Gold JA, Raju B, Prabhakar S, Pine R, Rom WN, Nakata K, Weiden M (2007) Mechanisms of polymorphonuclear neutrophil-mediated induction of HIV-1 replication in macrophages during pulmonary tuberculosis. J Infect Dis 195: 1303-1310. PMI: 17396999
Singh A, Singh Y, Pine R, Shi L, Chandra R, Drlica K (2006) Protein kinase I of Mycobacterium tuberculosis: cellular localization and expression during infection of macrophage-like cells. Tuberculosis (Edinb) 86: 28-33. PMI: 16256441
Tanaka N, Hoshino Y, Gold J, Hoshino S, Martiniuk F, Kurata T, Pine R, Levy D, Rom WN, Weiden M (2005) Interleukin-10 induces inhibitory C/EBPbeta through STAT-3 and represses HIV-1 transcription in macrophages. Am J Respir Cell Mol Biol 33: 406-411. PMI: 16014896
Prabhakar S, Qiao Y, Canova A, Tse DB, Pine R (2005) IFN-alpha beta secreted during infection is necessary but not sufficient for negative feedback regulation of IFN-alpha beta signaling by Mycobacterium tuberculosis. J Immunol 174: 1003-1012. PMI: 15634924
Raju B, Hoshino Y, Kuwabara K, Belitskaya I, Prabhakar S, Canova A, Gold JA, Condos R, Pine RI, Brown S, Rom WN, Weiden MD (2004) Aerosolized gamma interferon (IFN-gamma) induces expression of the genes encoding the IFN-gamma-inducible 10-kilodalton protein but not inducible nitric oxide synthase in the lung during tuberculosis. Infect Immun 72: 1275-1283. PMI: 14977928
Qiao Y, Prabhakar S, Canova A, Hoshino Y, Weiden M, Pine R (2004) Posttranscriptional inhibition of gene expression by Mycobacterium tuberculosis offsets transcriptional synergism with IFN-gamma and posttranscriptional up-regulation by IFN-gamma. J Immunol 172: 2935-2943. PMI: 14978096
Hoshino Y, Tse DB, Rochford G, Prabhakar S, Hoshino S, Chitkara N, Kuwabara K, Ching E, Raju B, Gold JA, Borkowsky W, Rom WN, Pine R, Weiden M (2004) Mycobacterium tuberculosis-induced CXCR4 and chemokine expression leads to preferential X4 HIV-1 replication in human macrophages. J Immunol 172: 6251-6258. PMI: 15128813
Prabhakar S, Qiao Y, Hoshino Y, Weiden M, Canova A, Giacomini E, Coccia E, Pine R (2003) Inhibition of response to alpha interferon by Mycobacterium tuberculosis. Infect Immun 71: 2487-2497. PMI: 12704120
Condos R, Raju B, Canova A, Zhao BY, Weiden M, Rom WN, Pine R (2003) Recombinant gamma interferon stimulates signal transduction and gene expression in alveolar macrophages in vitro and in tuberculosis patients. Infect Immun 71: 2058-2064. PMI: 12654826
Chelbi-alix MK, Bobe P, Benoit G, Canova A, Pine R (2003) Arsenic enhances the activation of Stat1 by interferon gamma leading to synergistic expression of IRF-1. Oncogene 22: 9121-9130. PMI: 14668793
Qiao Y, Prabhakar S, Coccia EM, Weiden M, Canova A, Giacomini E, Pine R (2002) Host defense responses to infection by Mycobacterium tuberculosis. Induction of IRF-1 and a serine protease inhibitor. J Biol Chem 277: 22377-22385. PMI: 11948194
Pine R (2002) IRF and tuberculosis. J Interferon Cytokine Res 22: 15-25. PMI: 11846972
Chang TL, Mosoian A, Pine R, Klotman ME, Moore JP (2002) A soluble factor(s) secreted from CD8(+) T lymphocytes inhibits human immunodeficiency virus type 1 replication through STAT1 activation. J Virol 76: 569-581. PMI: 11752148
Weiden M, Tanaka N, Qiao Y, Zhao BY, Honda Y, Nakata K, Canova A, Levy DE, Rom WN, Pine R (2000) Differentiation of monocytes to macrophages switches the Mycobacterium tuberculosis effect on HIV-1 replication from stimulation to inhibition: modulation of interferon response and CCAAT/enhancer binding protein beta expression. J Immunol 165: 2028-2039. PMI: 10925286
Zhao BY, Pine R, Domagala J, Drlica K (1999) Fluoroquinolone action against clinical isolates of Mycobacterium tuberculosis: effects of a C-8 methoxyl group on survival in liquid media and in human macrophages. Antimicrob Agents Chemother 43: 661-666. PMI: 10049284
Uddin S, Majchrzak B, Woodson J, Arunkumar P, Alsayed Y, Pine R, Young PR, Fish EN, Platanias LC (1999) Activation of the p38 mitogen-activated protein kinase by type I interferons. J Biol Chem 274: 30127-30131. PMI: 10514501
Yan H, Piazza F, Krishnan K, Pine R, Krolewski JJ (1998) Definition of the interferon-alpha receptor-binding domain on the TYK2 kinase. J Biol Chem 273: 4046-4051. PMI: 9461596
Honda Y, Rogers L, Nakata K, Zhao BY, Pine R, Nakai Y, Kurosu K, Rom WN, Weiden M (1998) Type I interferon induces inhibitory 16-kD CCAAT/ enhancer binding protein (C/EBP)beta, repressing the HIV-1 long terminal repeat in macrophages: pulmonary tuberculosis alters C/EBP expression, enhancing HIV-1 replication. J Exp Med 188: 1255-1265. PMI: 9763605
Rayanade RJ, Patel K, Ndubuisi M, Sharma S, Omura S, Etlinger JD, Pine R, Sehgal PB (1997) Proteasome- and p53-dependent masking of signal transducer and activator of transcription (STAT) factors. J Biol Chem 272: 4659-4662. PMI: 9030516
Pine R (1997) Convergence of TNFalpha and IFNgamma signalling pathways through synergistic induction of IRF-1/ISGF-2 is mediated by a composite GAS/kappaB promoter element. Nucleic Acids Res 25: 4346-4354. PMI: 9336467
Krishnan K, Pine R, Krolewski JJ (1997) Kinase-deficient forms of Jak1 and Tyk2 inhibit interferon alpha signaling in a dominant manner. Eur J Biochem 247: 298-305. PMI: 9249040
Improta T, Pine R (1997) Susceptibility to virus infection is determined by a Stat-mediated response to the autocrine effect of virus-induced type I interferon. Cytokine 9: 383-393. PMI: 9199872