Public Health Research Institute Center
UMDNJ - New Jersey Medical School
225 Warren Street
Newark, New Jersey 07103

Phone: (973) 854-3320
e-mail: pineri@umdnj.edu


Research Summary

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).




References

Condos, R., Raju, B., Canova, A., Zhao, B. Y., Weiden, M., Rom, W. N., and 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.

Prabhakar, S., Qiao, Y., Canova, A., Tse, D. B., and Pine, R. (2005). IFNab secreted during infection is necessary but not sufficient for down-regulation of IFNab signaling by M. tuberculosis. J Immunol 173, 1003-1012.

Prabhakar, S., Qiao, Y., Hoshino, Y., Weiden, M., Canova, A., Giacomini, E., Coccia, E., and Pine, R. (2003). Inhibition of Response to Alpha Interferon by Mycobacterium tuberculosis. Infect Immun 71, 2487-2497.

Qiao, Y., Prabhakar, S., Canova, A., Hoshino, Y., Weiden, M., and Pine, R. (2004). Posttranscriptional Inhibition of Gene Expression by Mycobacterium tuberculosis Offsets Transcriptional Synergism with IFN-g and Posttranscriptional Up-Regulation by IFN-g. J Immunol 172, 2935-2943.

Qiao, Y., Prabhakar, S., Coccia, E. M., Weiden, M., Canova, A., Giacomini, E., and Pine, R. (2002). "Host defense responses to infection by Mycobacterium tuberculosis: induction of IRF-1 and a serine protease inibitor." J. Biol. Chem. 277, 22377-22385.

Raju, B., Hoshino, Y., Kuwabara, K., Belitskaya, I., Prabhakar, S., Canova, A., Gold, J. A., Condos, R., Pine, R. I., Brown, S., et al. (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.

Weiden, M., Tanaka, N., Qiao, Y., Zhao, B. Y., Honda, Y., Nakata, K., Canova, A., Levy, D. E., Rom, W. N., and 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 b Expression. J Immunol 165, 2028-2039.

Raju, B., Hoshino, Y., Belitskaya-Levy, I., Dawson, R., Ress, R., Gold, J., Condos, R., Pine, R., Brown, S., Nolan, A. Rom, W., Weiden, M. "Gene expression profiles of bronchoalveolar cells in Pulmonary TB." Tuberculosis (in press)






Additional Recent Articles

Hoshino, Y., Hoshino, S., Gold, J., Raju, B., Prabhakar, S., Pine, R., Rom, W., Nakata, K., and Weiden, M. (2007) "Mechanisms of PMN-mediated induction of HIV-1 replication in macrophages during pulmonary tuberculosis." J. Inf. Dis. 195, 1303-1310. *

Singh, A., Singh, Y. Pine, R., Shi, L., Chandra, R. and Drlica, K. (2006) "Protein kinase I of Mycobacterium tuberculosis: cellular localization and expression during infection of macrophage-like cells." Tuberculosis (Edinb). 86, 28-33. *

Tanaka, N., Hoshino, Y., Gold, J., Hoshino, S., Martiniuk, F., Kurata, T., Pine, R., Levy, D., Rom, W.N., and Weiden, M. (2005) "IL-10 induces inhibitory C/EBP{beta} through STAT-3 and represses HIV-1 transcription in macrophages" Am J Respir Cell Mol Biol. 33, 406-411. *

Hoshino, Y., Tse, D., Rochford, G., Prabhakar, S., Hoshino, S., Kuwabara, K., Ching, E., Raju, B., Gold, J., Borkowsky, W., Rom, W., Pine, R., and Weiden, M. (2004). "Tuberculosis induces CXCR4 and beta-chemokine expression enhancing HIV-1-X4 replication in Alveolar Macrophages." J. Immunol. 172, 6251-6258. *

Chelbi-Alix, M., Bobe, Pl, Benoit,, G., Canova, A., Pine, R. (2003). "Arsenic enhances the activation of Stat1 by interferon g leading to synergistic expression of IRF-1." Oncogene 22, 9121-9130.

Pine, R. (2002). "Interferon Regulatory Factors and Tuberculosis." J. Interferon and Cytokine Res. 22, 15-25 (invited review). *

Chang, T., Mosoian, A., Pine, R., Klotman, M. E., and Moore, J. P. (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. *

Uddin, S., Majchrzak, B., Woodson, J. Arunkumar, P., Alsayed, Y., Pine, R., Young, P. R., Fish, E. N., and Platanias, L. C. (1999). "Activation of the p38 mitogen-activated protein kinase by type I interferons." J. Biol. Chem. 274, 30127-30131.

Zhao, B.-Y., Pine, R., Domagala, J., and Drlica, K. (1999). "Fluoroquinolone action against clinical isolates of Mycobacterium tuberculosis: effects of a C8-methoxyl group on survival in liquid media and in human macrophages." Antimicrobial Agents and Chemotherapy 43, 661-666. *

Honda, Y., Nakata, K. Zhao, B-Y., Pine, R., Rogers, L. Nakai, Y. Rom, W. N., and Weiden, M. (1998). "Type I interferon induces inhibitory 16 kDa C/EBPb repressing the HIV-1 LTR in macrophages: Pulmonary tuberculosis alters C/EBP expression enhancing HIV-1 replication." J. Exp. Med. 188, 1255-1265. *

Yan, H., Piazza, F., Krishnan, K., Pine, R., and Krolewski, J. J. (1998). "Definition of the interferon-a receptor binding domain on the tyk2 kinase." J. Biol. Chem. 273, 4046-4051.

Pine, R. (1997) "Convergence of TNFa and IFNγ signaling pathways through synergistic induction of IRF-1/ISGF-2 is mediated by a composite GAS/kB promoter element." Nucleic Acids Res. 25, 4346-4354.

Krishnan, K., Pine, R., and Krolewski, J. J. (1997). "Kinase-deficient forms of Jak1 and Tyk2 inhibit interferon a signaling in a dominant manner." Eur. J. Biochem. 247, 298-305.

Rayanade, R. J., Patel, K. Ndubuisi, M., Sharma, S., Omura, S., Etlinger, J. D., Pine, R., and Sehgal, P. B. (1997). "Proteasome-and p53-dependent masking of STAT transcription factors." J. Biol. Chem. 272, 4659-4662.

Improta, T. and 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.

*M. tb- and HIV-related

PubMed Lisitings>