Senior Member of the Staff
Head, Department of Cytokine Biology
Tuberculosis Video
email:
Medical School of Tringulo Mineiro, Uberaba, MG, Brazil, MD, 1968
Federal University of Rio de Janeiro, Brazil/Harvard Medical School, Ph.D., 1978, Immunology
University of Sao Paulo, Brazil, 1988, Professor of Microbiology and Immunology
Our laboratory focuses on research involving primarily global infectious diseases (tuberculosis and leishmaniasis). The World Health Organization estimates that tuberculosis claims approximately 2.8 million lives every year, and causes debilitating illness in many millions more and visceral leishmaniasis kills hundreds of thousands. These are the lives of infants, and children, and young adults in their most productive years. Unfortunately, there is still not an efficient vaccine for these diseases. Yet, vaccines have proven to be the most effective means of controlling many other life-threatening infectious diseases. Over the past ten years global efforts have been directed toward the development of vaccines against tuberculosis, malaria, and HIV/AIDS, leishmaniasis as well as to several other serious infectious diseases. As part of this effort, our laboratory has focused on antigen discovery for vaccine diagnostics development to tuberculosis and leishmaniasis. To achieve this goal we use modern molecular biology techniques combined with powerful immunological approaches. Thus far, several promising candidates are under testing for diagnosing and preventing tuberculosis and visceral leishmaniasis.
To reach these goals we screen for key pathogen antigens produced in vivo during infection. The candidate antigens are molecules identified in either bodily fluid of patients with tuberculosis or visceral leishmaniasis or are associated with MHC Class I molecules of adherent spleen cells of M. tuberculosis-infected mice.
Several proteins of both Mycobacterium tuberculosis and Leishmania infantum have thus far been identified in human secretions (urine) of patients. These antigens are apparently the only molecules reported to date that are derived directly from tuberculosis and visceral leishmaniasis patients from disease-endemic regions. This work is based on the central hypothesis that pathogen’s antigens that are actively produced during disease are interesting target molecules for vaccine and diagnostic development. We use Mass Spectroscopy to directly identify the pathogen’s antigens in the urine of the human patients. Following identification the antigens are selected for development of either vaccine (induction of CD4 mediated T cell responses) and/or for antigen detection assay for the diagnosis of tuberculosis or visceral leishmaniasis.
Because resistance to tuberculosis is also dependent CD8 T cell responses we have identified M. tuberculosis peptides that are associated with MHC Class I molecules during disease. We use Mass Spectroscopy to directly identify pathogen’s peptides stripped from purified MHC Class I molecules obtained from adherent spleen cells of M. tuberculosis-infected mice. This approach has allowed us the direct cloning of pathogen genes encoding proteins that potentially are involved in the stimulation of anti-microbial CD8+ T cell immunity.
Following the steps related to antigen discovery, gene cloning, and production of recombinant proteins prioritization of the antigens is based on their T cell recognition (CD4+ and CD8+) using peripheral blood mononuclear cells from healthy donors and by patients with active diseases as well as by T cells from lymphoid organs obtained from experimental models of tuberculosis and visceral leishmaniasis. Protection experiments are done in mice, hamsters, and guinea pigs, after conventional immunization with the antigens mixed with various adjuvants, or delivered in naked plasmid DNA format, or in a virus vectored platform of antigen delivery (e.g., Adenovirus).
Pollock NR, Kashino SS, Napolitano DR, Sloutsky A, Joshi S, Guillet J, Wong M, Nardell E, Campos-Neto A. (2009) Evaluation of the effect of treatment of latent tuberculosis infection on QuantiFERON-TB gold assay results. Infect. Control Hosp. Epidemiol. 30(4):392-395.
Kashino SS, Napolitano DR, Skobe Z, Campos-Neto A. (2008) Guinea pig model of Mycobacterium tuberculosis latent/dormant infection. Microbes Infect. 10(14-15):1469-1476.
Kashino SS, Pollack N, Napolitano DR, Rodrigues V Jr, Campos-Neto A. (2008) Indentification and characterization of Mycobacterium tuberculosis antigens in urine of patients with active pulmonary tuberculosis: an innovative and alterantive approach of antigen discovery of useful microbial molecules. Clin. Exp. Immunol. 153(1):56-62.
Leshem O, Kashino SS, Goncalves RB, Suzuki N, Onodera M, Fujimura A, Sasaki H, Stashenko P, Campos-Neto A. (2008) Th1 biased response to a novel Porphyromonas gingivalis protein aggravates bone resorption caused by this oral pathogen. Microbes Infect. 10(6):664-672.
Napolitano DR, Pollock N, Kashino SS, Rodrigues V Jr, Campos-Neto A. (2008) Identification of Mycobacterium tuberculosis ornithine carboamyltransferase in urine as a possible molecular marker of active pulmonary tuberculosis. Clin. Vaccine Immunol. 15(4):638-643.
Stashenko P, Goncalves RB, Lipkin B, Ficarelli A, Sasaki H, Campos-Neto A. (2007) Th1 immune response promotes severe bone resorption caused by Porphyromonas gingivalis. Am. J. Pathol. 170(1):203-213.
Kashino SS, Ovendale P, Izzo A, CamposNeto A. (2006). A unique model of dormant infection for vaccine development in tuberculosis. Clin. Vaccine Immunol.13(9):1014–1021.
Gonçalves RB, Lesham O, Bernards K, Webb JR, Stashenko PP, CamposNeto A. (2006) T-cell expression cloning of Porphyromonas gingivalis genes coding for T helper-biased immune responses during infection. Infect. Immun. 74(7) :3958–3966.
Mukherjee S, Kashino SS, Zhang Y, Daifalla N, Rodrigues V. Jr, Reed SG, Campos-Neto A. (2005) Cloning of the gene encoding a protective Mycobacterium tuberculosis secreted protein detected in vivo during the initial phases of the infectious process. J. Immunol.175(8):5298–5305.
Fujiwara RT, Vale AM, França Da Silva JC, Da Costa RT, Quetz J, Martins Filho OA, Reis AB, Corrêa Olivera R, Machado-Coelho GL, Bueno LL, Bethony JM, Frank G, Nascimento E, Genaro O, Mayrink W, Reed S, Campos-Neto A. (2005) Immunogenicity in dogs of three recombinant antigens (TSA, LeIF and LmSTI1) potential vaccine candidates for canine visceral leishmaniasis. Vet. Res. 36(5-6):827-836.
Campos-Neto A. (2005) What about Th1 /Th2 in cutaneous leishmaniasis vaccine discovery? Braz. J. Med. Biol. Res. 38(7):979–984.
Skeiky YAW, Alderson MR, Ovendale PJ, Lobet Y, Dalemans W, Orme IM, Reed SG, Campos-Neto A. (2005) Protection of mice and guinea pigs against tuberculosis induced by immunization with a single Mycobacterium tuberculosis recombinant antigen, MTB41. Vaccine 23(3):3937–3945.
Reece ST, Stride S, Ovendale P, Reed SG, Campos-Neto A. (2005) Skin test performed with highly purified Mycobacterium tuberculosis recombinant protein triggers tuberculin shock in infected guinea pigs. Infect. Immun. 73(6):3301–3306.
Assistant Research Investigator
Mark Cayabyab, Ph.D.
Staff Associate
Suely S. Kashino, Ph.D.
Postdoctoral Fellow
Kelly Kaufine
