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

Phone: (973) 854-3210
e-mail: gennarma@umdnj.edu


Research Summary

The Gennaro laboratory studies infection with the disease-causing bacterium Mycobacterium tuberculosis. About one-third of the world population is infected with M. tuberculosis. Most infected persons remain symptom-free for their entire life because their immune system keeps the infection in check. However, conditions that impair immune responses greatly increase the risk of progression from latent infection to active tuberculosis. The disease, which usually affects the lung, is life threatening when untreated. Diseased persons transmit infection to their close contacts, usually by coughing. Proper drug treatment not only cures the diseased persons but also drastically decreases the time they are contagious. Thus, early and accurate diagnosis of disease leading to timely treatment is a key element of tuberculosis control.

One area of research in the laboratory is identifying immune biomarkers of M. tuberculosis infection and disease, primarily for diagnostic purposes. Bacterial proteins characterized in our laboratory over the years have become part of multi-protein, diagnostic tests for latent infection and active disease that are commercially available or are being developed. To expand our current knowledge of biomarkers of disease, our laboratory has engaged in genome-scale interrogation of the antibody response to M. tuberculosis infection and disease. This project is being conducted by probing all 4,000 proteins of M. tuberculosis -- arrayed on a platform developed by ImmPORT Therapeutics Inc. -- with hundreds of sera collected from tuberculosis patients, infected and non-infected individuals worldwide.

A second area of research is the characterization of M. tuberculosis during latent infection. It is known that M. tuberculosis responds to unfavorable environmental conditions by entering a state characterized by viable but non-growing bacteria. This non-replicative state can be investigated by use of in vitro and in vivo models. We have shown that arrest of M. tuberculosis growth induced by expression of lung immunity in mice is characterized by transcriptional profiles indicative of upregulation of a “dormancy” regulon, changes in respiratory metabolism and in carbon flux distribution, and remodeling of the mycobacterial cell wall. We have also seen that growth and persistence of M. tuberculosis in an animal model are associated with different profiles of bacterial antigens, a finding that may have profound implications on new vaccine design. We are currently moving toward genome-scale explorations of the dormant tubercle bacillus to obtain an integrated knowledge of the transcriptional and metabolic changes associated with M. tuberculosis dormancy. Understanding the changing physiology of M. tuberculosis with growth state is key to the development of new drugs and new vaccines.

Useful links:

Secreted Proteins of Mycobacterium tuberculosis
The Gennaro laboratory has created a Web site that publishes the output of a bioinformatic search for secreted proteins of Mycobacterium tuberculosis. Go to http://tbsp.phri.org for methodology and tables that identify the secreted proteins.

Protocols for Measuring Bacterial mRNA
To help overcome technical difficulties that may be encountered in measuring bacterial mRNA in infected tissues, the Gennaro laboratory provides access to a PDF document describing detailed protocols for lung RNA extraction, reverse transcription, real time RT-PCR, and data analysis. To access this document, click Research Protocol.




Recent Articles

Bothamley G. and Gennaro M.L. (2008).
The antibody response to infection with Mycobacterium tuberculosis
in TB Handbook, S.H.E. Kaufmann and W. Britton, eds., Wiley-VCH pp. 227-244.

Shi L., Sohaskey C.D., North R.J., and Gennaro M.L. (2008).
Transcriptional characterization of the antioxidant response of Mycobacterium tuberculosis in vivo and during adaptation to hypoxia in vitro.
Tuberculosis (Edinb.) 88:1-6.

Khan I.H., Ravindran R., Yee J., Ziman M., Lewinsohn D.M., Gennaro M.L., Flynn J.L., Goulding C.W., DeRiemer K., Lerche N.W., Luciw P.A. (2008).
Profiling antibodies to Mycobacterium tuberculosis by multiplex microbead suspension arrays for serodiagnosis of tuberculosis.
Clin Vaccine Immunol. 15:433-438.

Gennaro ML, Affouf M, Kanaujia GV, Brusasca PN, Mangura B, and Reichman L. (2007).
Antibody markers of incident tuberculosis among HIV-infected adults in the USA: a historical prospective study.
Int J Tuberc Lung Dis. 2007 Jun;11(6):624-31.

Azzurri A., Kanaujia G.V., Sow O.Y., Bah B., Diallo A., Del Prete G., and Gennaro M.L. (2006).
Serological markers of pulmonary tuberculosis and of response to anti-tuberculosis treatment in a patient population in Guinea.
Int J Immunopathol Pharmacol. 19:199-208.

Shi L., Sohaskey C.D., Kana B.D., Dawes S., North R.J., Mizrahi V. and Gennaro M.L. (2005).
Changes in energy metabolism in Mycobacterium tuberculosis in mouse lung and under in vitro conditions affecting aerobic respiration.
Proc. Natl. Acad. Sci. USA 102:15629-34.

Davidow, A., Kanaujia G.V., Shi L., Kaviar, J, Guo, X.D., Sung N., Kaplan G., Menzies D. and
Gennaro M.L. (2005).
Antibody profiles characteristic of Mycobacterium tuberculosis infection state.
Infect. Immun. 73:6846-6851.

Kanaujia G.V., Lam P.K., Perry S., Brusasca P.N., Catanzaro A., and Gennaro M.L. (2005).
Integration of microscopy and serodiagnostic tests to screen for active tuberculosis.
Int. J. Tuberc. Lung Dis. 9:1120-1126.

Spencer JS, Kim HJ, Marques AM, Gonzalez-Juarerro M, Lima MC, Vissa VD, Truman RW,
Gennaro ML, Cho SN, Cole ST, and Brennan PJ. (2004).
Comparative analysis of B- and T-cell epitopes of Mycobacterium leprae and Mycobacterium tuberculosis culture filtrate protein 10.
Infect Immun. 2004 Jun;72(6):3161-70
PMID: 15155617
Abstract  |  Full Article

Lewinsohn DA, Gennaro ML, Scholvinck L, Lewinsohn DM. (2004).
Tuberculosis immunology in children: diagnostic and therapeutic challenges and opportunities.
Int J Tuberc Lung Dis. 2004 May;8(5):658-74
PMID: 15137550
Abstract

Goulding CW, Apostol MI, Gleiter S, Parseghian A, Bardwell J, Gennaro M, Eisenberg D. (2004).
Gram-positive DsbE proteins function differently from Gram-negative DsbE homologs. A structure to function analysis of DsbE from Mycobacterium tuberculosis.
J Biol Chem. 2004 Jan 30;279(5):3516-24. Epub 2003 Nov 03
PMID: 14597624
Abstract  |  Full Article

Shi L., North R, and Gennaro M.L. (2004).
Effect of growth state on transcription levels of genes encoding major secreted antigens of Mycobacterium tuberculosis in the mouse lung.
Infect Immun. 2004 Apr;72(4):2420-4
PMID: 15039373
Abstract  |  Full Article

Kanaujia GV, Motzel S, Garcia MA, Andersen P, Gennaro ML. (2004).
Recognition of ESAT-6 sequences by antibodies in sera of tuberculous nonhuman primates.
Clin Diagn Lab Immunol. 2004 Jan;11(1):222-6
PMID: 14715573
Abstract  |  Full Article

Kanaujia GV, Garcia MA, Bouley DM, Peters R, Gennaro ML. (2003).
Detection of early secretory antigenic target-6 antibody for diagnosis of tuberculosis in non-human primates.
Comp Med. 2003 Dec;53(6):602-6
PMID: 14727807
Abstract

Silva VM, Kanaujia G, Gennaro ML, Menzies D. (2003).
Factors associated with humoral response to ESAT-6, 38 kDa and 14 kDa in patients with a spectrum of tuberculosis.
Int J Tuberc Lung Dis 2003 May;7(5):478-84
PMID: 12757050
Abstract

Rolinck-Werninghaus C, Magdorf K, Stark K, Lyashchenko K, Gennaro ML, Colangeli R, Doherty TM, Andersen P, Plum G, Herz U, Renz H, Wahn U. (2003).
The potential of recombinant antigens ESAT-6, MPT63 and mig for specific discrimination of Mycobacterium tuberculosis and M. avium infection.
Eur J Pediatr. 2003 Jul;162(7-8):534-6. Epub 2003 Apr 25.
PMID: 12715165

Brusasca PN, Peters RL, Motzel SL, Klein HJ, Gennaro ML. (2003).
Antigen recognition by serum antibodies in non-human primates experimentally infected with Mycobacterium tuberculosis.
Comp Med. 2003 Apr;53(2):165-72
PMID: 12784850
Abstract

Brown RM, Cruz O, Brennan M, Gennaro ML, Schlesinger L, Skeiky YA, Hoft DF. (2003).
Lipoarabinomannan-reactive human secretory immunoglobulin A responses induced by mucosal bacille Calmette-Guerin vaccination.
J Infect Dis 2003 Feb 1;187(3):513-7
PMID: 12552438
Abstract  |  Full Article

Shi L, Jung YJ, Tyagi S, Gennaro ML, North RJ. (2003).
Expression of Th1-mediated immunity in mouse lungs induces a Mycobacterium tuberculosis transcription pattern characteristic of nonreplicating persistence.
Proc Natl Acad Sci USA 2003 Jan 7;100(1):241-6
PMID: 12506197
Abstract  |  Full Article

Goulding CW, Parseghian A, Sawaya MR, Cascio D, Apostol MI, Gennaro ML, Eisenberg D. (2002).
Crystal structure of a major secreted protein of Mycobacterium tuberculosis-MPT63 at 1.5-A resolution.
Protein Sci 2002 Dec;11(12):2887-93
PMID: 12441386
Abstract

Chandramuki A, Lyashchenko K, Kumari HB, Khanna N, Brusasca P, Gourie-Devi M, Satishchandra P, Shankar SK, Ravi V, Alcabes P, Kanaujia GV, Gennaro ML. (2002).
Detection of antibody to Mycobacterium tuberculosis protein antigens in the cerebrospinal fluid of patients with tuberculous meningitis.
J Infect Dis 2002 Sep 1;186(5):678-83
PMID: 12195355
Abstract  |  Full Article

Amadori M, Lyashchenko KP, Gennaro ML, Pollock JM, Zerbini I (2002).
Use of recombinant proteins in antibody tests for bovine tuberculosis.
Vet Microbiol 2002 Apr 2;85(4):379-89
PMID: 11856587
Abstract  |  Full Article

Brusasca PN, Colangeli R, Lyashchenko KP, Zhao X, Vogelstein M, Spencer JS, McMurray DN, Gennaro ML. (2001).
Immunological characterization of antigens encoded by the RD1 region of the Mycobacterium tuberculosis genome.
Scand J Immunol 2001 Nov;54(5):448-52
PMID: 11696195
Abstract

Johnson S, Brusasca P, Lyashchenko K, Spencer JS, Wiker HG, Bifani P, Shashkina E, Kreiswirth B, Harboe M, Schluger N, Gomez M, Gennaro ML (2001).
Characterization of the secreted MPT53 antigen of Mycobacterium tuberculosis.
Infect Immun 2001 Sep;69(9):5936-9
PMID: 11500477
Abstract  |  Full Article

Landowski CP, Godfrey HP, Bentley-Hibbert SI, Liu X, Huang Z, Sepulveda R, Huygen K, Gennaro ML, Moy FH, Lesley SA, Haak-Frendscho M.J. (2001).
Combinatorial use of antibodies to secreted mycobacterial proteins in a host immune system-independent test for tuberculosis.
J Clin Microbiol 2001 Jul;39(7):2418-24
PMID: 11427548
Abstract  |  Full Article



Lyashchenko KP, Singh M, Colangeli R, Gennaro ML. (2000).
A multi-antigen print immunoassay for the development of serological diagnosis of infectious diseases.
J Immunol Methods. 2000 Aug 28;242(1-2):91-100

Gennaro ML. (2000).
Immunologic diagnosis of tuberculosis.
Clin Infect Dis. 2000 Jun;30 Suppl 3:S243-6

Gomez M, Johnson S, Gennaro ML. (2000).
Identification of secreted proteins of Mycobacterium tuberculosis by a bioinformatic approach.
Infect Immun. 2000 Apr;68(4):2323-7

Colangeli R, Spencer JS, Bifani P, Williams A, Lyashchenko K, Keen MA, Hill PJ, Belisle J, Gennaro ML. (2000).
MTSA-10, the product of the Rv3874 gene of Mycobacterium tuberculosis, elicits tuberculosis-specific, delayed-type hypersensitivity in guinea pigs.
Infect Immun. 2000 Feb;68(2):990-3

Colangeli R, Antinori A, Cingolani A, Ortona L, Lyashchenko K, Fadda G, Gennaro ML. (1999).
Humoral immune responses to multiple antigens of Mycobacterium tuberculosis in tuberculosis patients co-infected with the human immunodeficiency virus.
Int J Tuberc Lung Dis. 1999 Dec;3(12):1127-31

Lyashchenko KP, Pollock JM, Colangeli R, Gennaro ML. (1998).
Diversity of antigen recognition by serum antibodies in experimental bovine tuberculosis.
Infect Immun. 1998 Nov;66(11):5344-9

Colangeli R, Heijbel A, Williams AM, Manca C, Chan J, Lyashchenko K, Gennaro ML. (1998).
Three-step purification of lipopolysaccharide-free, polyhistidine-tagged recombinant antigens of Mycobacterium tuberculosis.
J Chromatogr B Biomed Sci Appl. 1998 Sep 4;714(2):223-35

Lyashchenko K, Colangeli R, Houde M, Al Jahdali H, Menzies D, Gennaro ML. (1998).
Heterogeneous antibody responses in tuberculosis.
Infect Immun. 1998 Aug;66(8):3936-40

Lyashchenko K, Manca C, Colangeli R, Heijbel A, Williams A, Gennaro ML. (1998).
Use of Mycobacterium tuberculosis complex-specific antigen cocktails for a skin test specific for tuberculosis.
Infect Immun. 1998 Aug;66(8):3606-10

Manca C, Lyashchenko K, Colangeli R, Gennaro ML. (1997).
MTC28, a novel 28-kilodalton proline-rich secreted antigen specific for the Mycobacterium tuberculosis complex.
Infect Immun. 1997 Dec;65(12):4951-7

Zhang Q, Soares de Oliveira S, Colangeli R, Gennaro ML. (1997).
Binding of a novel host factor to the pT181 plasmid replication enhancer.
J Bacteriol. 1997 Feb;179(3):684-8

Colombo D, Iordanescu S, Gennaro ML. (1995).
Replication enhancer requirement for recognition of heterologous replication origin by an initiator protein.
Plasmid. 1995 May;33(3):232-4

Gennaro ML. (1993).
Genetic evidence for replication enhancement from a distance.
Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5529-33

Gennaro ML, Novick RP. (1988).
An enhancer of DNA replication.
J Bacteriol. 1988 Dec;170(12):5709-17

PubMed Lisitings>




C.V.

M.D. University of Palermo, Italy, M.D., 1977; London School of Hygiene & Tropical Medicine, M.S., 1981; Istituto Superiore di Sanita, Rome, 1978-1986; PHRI, 1984-present.