Mycobacterium tuberculosisESAT-6 antigen immunogenicity in Owl Monkeys

Rosalba Alfonso, Paola Barato, Martha Calderon, Diana Giraldo, Martha Pinto, Carlos Parra-López, Manuel A. Patarroyo.

Resumen


La única vacuna disponible contra la tuberculosis es la cepa Mycobacterium bovis BCG, que ofrece una eficacia protectiva variable (0%-80%), siendo urgente un nuevo agente profiláctico. Se han evaluado diversos candidatos a vacuna contra este patógeno, en los modelos animales de experimentación convencionales (murino, cobayo, conejo), obteniéndose información básica sobre el efecto de la vacuna en la carga bacterial frente a un reto infeccioso, así como también la reducción o prevención de la patología en los pulmones u otros órganos blanco; además de los aspectos relacionados con la respuesta inmune hacia el Mycobacterium tuberculosis. Los primates no humanos tienen ventajas sobre los modelos convencionales en la evaluación de vacunas, de hecho se ha verificado el comportamiento de agentes terapéuticos en humanos después de haber sido medida la capacidad protectiva de éstos en monos con tuberculosis inducida. Los primates más estudiados en la infección por micobacterias son el cynomulgus, y el rhesus, observándose que estos animales mantienen la infección en un estado subclínico, muy similar a la tuberculosis humana donde el 90% de la población infectada mantiene la infección en un estado latente.

Dado que el modelo animal debe semejar el comportamiento de las proteínas estudiadas en el ser humano, el mono Aotus puede representar ventajas en investigación de tuberculosis por ser un primate con aproximadamente un 90% de similitud al humano en cuanto a las moléculas del sistema inmune estudiadas hasta hoy. La proteína ESAT-6 de (early secretory antigenic target 6 kD) de Mycobacterium tuberculosis es un componente minoritario del filtrado de cultivo de corto tiempo (CFP), ha sido genética y químicamente caracterizada e induce una potente respuesta inmunogénica del tipo TH1. Este antígeno es secretado durante la fase inicial de crecimiento siendo fuertemente reconocido por animales y humanos infectados por Mycobacterium tuberculosis, este hecho hace que su inclusión en una futura vacuna anti-tuberculosis por subunidades y en pruebas de inmunodiagnóstico.

En este trabajo se estudió la respuesta inmune del mono Aotus frente al antígeno micobacteriano ESAT-6 en forma recombinante (rESAT-6). El antígeno fue inmunizado junto con el adyuvante Montanide 720 produciendo una fuerte respuesta humoral y celular, no solo hacia rESAT-6 sino también hacia la proteína nativa presente en el filtrado de medio de cultivo de Mycobacterium tuberculosis. La respuesta celular fue medida por la incorporación de [3H]timidina en ensayos de linfoproliferación. La respuesta humoral se analizó por ensayos de ELISA e inmunoblot, obteniéndose altos títulos de anticuerpos (hasta de 1:12,800) dirigidos rESAT- 6. Se demostró la naturaleza multiepitope de este antígeno ya que en general péptidos que mapean la proteína fueron específicamente reconocidos tanto a nivel celular como humoral. Se observó además variación en la respuesta a la vacunación entre animal y animal, siendo una desventaja para un modelo de experimentación en cuanto a la predicción de la respuesta. Este estudio se constituye en un primer paso de evaluación del mono Aotus como modelo animal en tuberculosis.


Palabras clave


Owl monkey; animal model; Mycobacterium tuberculosis; ESAT-6; Inmune response.

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Referencias


Colditz G, Brewer T, Berkey C, Wilson M, Burdick E, Fineberg H, Mosteller F: Efficacy of BCG vaccine in the prevention of tuberculosis. Metaanalysis of the published literature. Jama 271:698-702, 1994

Fine PE: Variation in protection by BCG: implications of and for heterologous immunity. Lancet 346:1339 1345, 1995

Ginsberg AM: What.s new in tuberculosis vaccines? Bull World Health Organ 80:483-488, 2002

. Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D, Gordon SV, Eiglmeier K, Gas S, Barry CE, 3rd, Tekaia F, Badcock K, Basham D, Brown D, Chillingworth T, Connor R, Davies R, Devlin K, Feltwell T, Gentles S, Hamlin N, Holroyd S, Hornsby T, Jagels K, Barrell BG, et al.: Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393:537-544, 1998

. Tekaia F, Gordon SV, Garnier T, Brosch R, Barrell BG, Cole ST: Analysis of the proteome of Mycobacterium tuberculosis in silico. Tuber Lung Dis 79:329-342, 1999

. Jungblut PR, Schaible UE, Mollenkopf HJ, Zimny-Arndt U, Raupach B, Mattow J, Halada P, Lamer S, Hagens K, Kaufmann SH: Comparative proteome analysis of Mycobacterium tuberculosis and Mycobacterium bovis BCG strains: towards functional genomics of microbial pathogens. Mol Microbiol 33:1103-1117, 1999

. Domenech P, Barry CE, 3rd, Cole ST: Mycobacterium tuberculosis in the post-genomic age. Curr Opin Microbiol 4:28-34, 2001

Covert BA, Spencer JS, Orme IM, Belisle JT: The application of proteomics in defining the T cell antigens of Mycobacterium tuberculosis. Proteomics 1:574-586, 2001

Brennan PJ: Structure, function, and biogenesis of the cell wall of Mycobacterium tuberculosis. Tuberculosis (Edinb) 83:91-97, 2003

Brennan PJ, Besra GS: Structure, function and biogenesis of the mycobacterial cell wall. Biochem Soc Trans 25:188-194, 1997

Orme IM, Cooper AM: Cytokine/chemokine cascades in immunity to tuberculosis. Immunol Today 20:307-312, 1999

Flynn JL, Chan J: Immunology of tuberculosis. Annu Rev Immunol 19:93-129, 2001

Pym AS, Brodin P, Majlessi L, Brosch R, Demangel C, Williams A, Griffiths KE, Marchal G, Leclerc C, Cole ST: Recombinant BCG exporting ESAT-6 confers enhanced protection against tuberculosis. Nat Med 9:533-539, 2003

Bao L, Chen W, Zhang H, Wang X: Virulence, immunogenicity, and protective efficacy of two recombinant Mycobacterium bovis bacillus Calmette-Guerin strains expressing the antigen ESAT-6 from Mycobacterium tuberculosis. Infect Immun 71:1656-1661, 2003

Jackson M, Phalen SW, Lagranderie M, Ensergueix D, Chavarot P, Marchal G, McMurray DN, Gicquel B, Guilhot C: Persistence and protective efficacy of a Mycobacterium tuberculosis auxotroph vaccine. Infect Immun 67:2867-2873, 1999

Kamath AT, Feng CG, Macdonald M, Briscoe H, Britton WJ: Differential protective efficacy of DNA vaccines expressing secreted proteins of Mycobacterium tuberculosis. Infect Immun 67:1702-1707, 1999

Li Z, Howard A, Kelley C, Delogu G, Collins F, Morris S: Immunogenicity of DNA vaccines expressing tuberculosis proteins fused to tissue plasminogen activator signal sequences. Infect Immun 67:4780-4786, 1999

Huygen K: On the use of DNA vaccines for the prophylaxis of mycobacterial diseases. Infect Immun 71:1613-1621, 2003

Nor NM, Musa M: Approaches towards the development of a vaccine against tuberculosis: recombinant BCG and DNA vaccine. Tuberculosis (Edinb) 84:102-109, 2004

Orme IM, McMurray DN, Belisle JT: Tuberculosis vaccine development: recent progress. Trends Microbiol 9:115-118, 2001

Dannenberg AM Jr, Collins FM: Progressive pulmonary tuberculosis is not due to increasing numbers of viable bacilli in rabbits, mice and guinea pigs, but is due to a continuous host response to mycobacterial products. Tuberculosis (Edinb);81(3):229-42, 2001

Baldwin SL, D.Souza C, Roberts AD, Kelly BP, Frank AA, Lui MA, Ulmer JB, Huygen K, McMurray DM, Orme IM: Evaluation of new vaccines in the mouse and guinea pig model of tuberculosis. Infect Immun 66:2951-2959, 1998

Scanga CA, Mohan VP, Yu K, Joseph H, Tanaka K, Chan J, Flynn JL: Depletion of CD4(+) T cells causes reactivation of murine persistent tuberculosis despite continued expression of interferon gamma and nitric oxide synthase 2. J Exp Med 192:347-358, 2000

Mogues T, Goodrich ME, Ryan L, LaCourse R, North RJ: The relative importance of T cell subsets in immunity and immunopathology of airborne Mycobacterium tuberculosis infection in mice. J Exp Med 193:271-280, 2001

McMurray DN: Recent progress in the development and testing of vaccines against human tuberculosis. Int J Parasitol 33:547-554, 2003

McMurray DN: A nonhuman primate model for preclinical testing of new tuberculosis vaccines. Clin Infect Dis 30 Suppl 3:S210-212, 2000

Good RC: Diseases in Nonhuman Primates. In The Mycobacteria. A Sourcebook. Kubica G, Wayne L (eds). New York, Marcel Dekker. Inc., 1984, pp 903-924

VandeBerg JL, Williams-Blangero S: Advantages and limitations of nonhuman primates as animal models in genetic research on complex diseases. J Med Primatol 26:113-119, 1997

Walsh GP, Tan EV, dela Cruz EC, Abalos RM, Villahermosa LG, Young LJ, Cellona RV, Nazareno JB, Horwitz MA: The Philippine cynomolgus monkey (Macaca fasicularis) provides a new nonhuman primate model of tuberculosis that resembles human disease. Nat Med 2:430-436, 1996

Capuano SV, III, Croix DA, Pawar S, Zinovik A, Myers A, Lin PL, Bissel S, Fuhrman C, Klein E, Flynn JL: Experimental Mycobacterium tuberculosis infection of cynomolgus macaques closely resembles the various manifestations of human M. tuberculosis infection. Infect Immun 71:5831-5844, 2003

Langermans JA, Andersen P, van Soolingen D, Vervenne RA, Frost PA, van der Laan T, van Pinxteren LA, van den Hombergh J, Kroon S, Peekel I, Florquin S, Thomas AW: Divergent effect of bacillus Calmette Guerin (BCG) vaccination on Mycobacterium tuberculosis infection in highly related macaque species: implications for primate models in tuberculosis vaccine research. Proc Natl Acad Sci U S A 98:11497 11502, 2001

Baer JF: Husbandry and medical management of the owl monkey. In Aotus: The owl monkey Baer JF, Weller RE, Kakoma I (eds). New York, Academic Press Inc., 1994, pp 133-164

Attanasio R, Pehler K, McClure HM: Immunogenicity and safety of Mycobacterium tuberculosis culture filtrate proteins in nonhuman primates. Clin Exp Immunol 119:84-91, 2000

Pehler K, Brasky KM, Butler TM, Attanasio R: Mycobacterium tuberculosis-secreted protein antigens: immunogenicity in baboons. J Clin Immunol 20:306-316, 2000

Berthet FX, Rasmussen PB, Rosenkrands I, Andersen P, Gicquel B: A Mycobacterium tuberculosis operon encoding ESAT-6 and a novel low-molecular-mass culture filtrate protein (CFP-10). Microbiology 144 ( Pt 11):3195-3203, 1998

Gey Van Pittius NC, Gamieldien J, Hide W, Brown GD, Siezen RJ, Beyers AD: The ESAT-6 gene cluster of Mycobacterium tuberculosis and other high G+C Gram-positive bacteria. Genome Biol 2:RESEARCH0044, 2001

Sorensen AL, Nagai S, Houen G, Andersen P, Andersen AB: Purification and characterization of a low molecular-mass T-cell antigen secreted by Mycobacterium tuberculosis. Infect Immun 63:1710-1717, 1995

Harboe M, Oettinger T, Wiker HG, Rosenkrands I, Andersen P: Evidence for occurrence of the ESAT-6 protein in Mycobacterium tuberculosis and virulent Mycobacterium bovis and for its absence in Mycobacterium bovis BCG. Infect Immun 64:16-22, 1996

Renshaw PS, Panagiotidou P, Whelan A, Gordon SV, Hewinson RG, Williamson RA, Carr MD: Conclusive evidence that the major Tcell antigens of the Mycobacterium tuberculosis complex ESAT-6 and CFP-10 form a tight, 1:1 complex and characterization of the structural properties of ESAT-6, CFP-10, and the ESAT-6*CFP-10 complex. Implications for pathogenesis and virulence. J Biol Chem 277:21598-21603, 2002

Skjot RL, Oettinger T, Rosenkrands I, Ravn P, Brock I, Jacobsen S, Andersen P: Comparative evaluation of low-molecular-mass proteins from Mycobacterium tuberculosis identifies members of the ESAT-6 family as immunodominant T-cell antigens. Infect Immun 68:214-220, 2000

Mustafa AS, Oftung F, Amoudy HA, Madi NM, Abal AT, Shaban F, Rosen Krands I, Andersen P: Multiple epitopes from the Mycobacterium tuberculosis ESAT-6 antigen are recognized by antigen-specific human T cell lines. Clin Infect Dis 30 Suppl 3:S201-205, 2000

Brandt L, Elhay M, Rosenkrands I, Lindblad EB, Andersen P: ESAT-6 subunit vaccination against Mycobacterium tuberculosis. Infect Immun 68:791-795, 2000

Olsen AW, Hansen PR, Holm A, Andersen P: Efficient protection against Mycobacterium tuberculosis by vaccination with a single subdominant epitope from the ESAT-6 antigen. Eur J Immunol 30:1724-1732, 2000

Mori T, Sakatani M, Yamagishi F, Takashima T, Kawabe Y, Nagao K, Shigeto E, Harada N, Mitarai S, Okada M, Suzuki K, Inoue Y, Tsuyuguchi K, Sasaki Y, Mazurek GH, Tsuyuguchi I: Specific Detection of Tuberculosis Infection with an Interferongamma Based Assay Using New Antigens. Am J Respir Crit Care Med, 2004

Goodman M, Porter CA, Czelusniak J, Page SL, Schneider H, Shoshani J, Gunnell G, Groves CP: Toward a phylogenetic classification of Primates based on DNA evidence complemented by fossil evidence. Mol Phylogenet Evol 9:585-598, 1998

Nino-Vasquez JJ, Vogel D, Rodriguez R, Moreno A, Patarroyo ME, Pluschke G, Daubenberger CA: Sequence and diversity of DRB genes of Aotus nancymaae, a primate model for human malaria parasites. Immunogenetics 51:219-230, 2000

Daubenberger CA, Salomon M, Vecino W, Hubner B, Troll H, Rodriques R, Patarroyo ME, Pluschke G: Functional and structural similarity of V gamma 9V delta 2 T cells in humans and Aotus monkeys, a primate infection model for Plasmodium falciparum malaria. J Immunol 167:6421-6430, 2001

Montoya GE, Vernot JP, Patarroyo ME: Partial characterization of the CD45 phosphatase cDNA in the owl monkey (Aotus vociferans). Am J Primatol 57:1-11, 2002

Guerrero JE, Pacheco DP, Suarez CF, Martinez P, Aristizabal F, Moncada CA, Patarroyo ME, Patarroyo MA: Characterizing Tcell receptor gamma-variable gene in Aotus nancymaae owl monkey peripheral blood. Tissue Antigens 62:472-482, 2003

Webster D, Hill AV: Progress with new malaria vaccines. Bull World Health Organ 81:902-909, 2003

Espejo F, Bermudez A, Torres E, Urquiza M, Rodriguez R, Lopez Y, Patarroyo ME: Shortening and modifying the 1513 MSP-1 peptide.s alpha-helical region induces protection against malaria. Biochem Biophys Res Commun 315:418-427, 2004

Rodriguez R, Moreno A, Guzman F, Calvo M, Patarroyo ME: Studies in owl monkeys leading to the development of a synthetic vaccine against the asexual blood stages of Plasmodium falciparum. Am J Trop Med Hyg 43:339-354, 1990

Houghten RA: General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigenantibody interaction at the level of individual amino acids. Proc Natl Acad Sci U S A 82:5131 5135, 1985

Alfonso R, Romero RE, Diaz A, Calderon MN, Urdaneta G, Arce J, Patarroyo ME, Patarroyo MA: Isolation and identification of mycobacteria in New World primates maintained in captivity. Vet Microbiol 98:285-295, 2004

Palomino JC, Portaels F: Effects of decontamination methods and culture conditions on viability of Mycobacterium ulcerans in the BACTEC system. J Clin Microbiol 36:402-408, 1998

Espinosa AM, Sierra AY, Barrero CA, Cepeda LA, Cantor EM, Lombo TB, Guzman F, Avila SJ, Patarroyo MA: Expression, polymorphism analysis, reticulocyte binding and serological reactivity of two Plasmodium vivax MSP-1 protein recombinant fragments. Vaccine 21:1033-1043, 2003

Bollag DM, Edelstein SJ: Immunoblotting. In Protein Methods. (ed). New York, John Wiley and sons, Inc, 1993, pp 181-211

Andersen P, Andersen AB, Sorensen AL, Nagai S: Recall of longlived immunity to Mycobacterium tuberculosis infection in mice. J Immunol 154:3359-3372, 1995

Ulrichs T, Munk ME, Mollenkopf H, Behr-Perst S, Colangeli R, Gennaro ML, Kaufmann SH: Differential T cell responses to Mycobacterium tuberculosis ESAT6 in tuberculosis patients and healthy donors. Eur J Immunol 28:3949-3958, 1998

Brock I, Weldingh K, Lillebaek T, Follmann F, Andersen P: Comparison of a New Specific Blood Test and the Skin Test in Tuberculosis Contacts. Am J Respir Crit Care Med, 2004

Brandt L, Oettinger T, Holm A, Andersen AB, Andersen P: Key epitopes on the ESAT-6 antigen recognized in mice during the recall of protective immunity to Mycobacterium tuberculosis. J Immunol 157:3527-3533, 1996

Pollock JM, Andersen P: Predominant recognition of the ESAT-6 protein in the first phase of interferon with Mycobacterium bovis in cattle. Infect Immun 65:2587-2592, 1997

Kanaujia GV, Motzel S, Garcia MA, Andersen P, Gennaro ML: Recognition of ESAT-6 sequences by antibodies in sera of tuberculous nonhuman primates. Clin Diagn Lab Immunol 11:222-226, 2004

Morris S, Kelley C, Howard A, Li Z, Collins F: The immunogenicity of single and combination DNA vaccines against tuberculosis. Vaccine 18(20):2155-63, 2000

Minion FC, Menon SA, Mahairas GG, Wannemuehler MJ: Enhanced murine antigen-specific gamma interferon and immunoglobulin G2a responses by using mycobacterial ESAT-6 sequences in DNA vaccines. Infect Immun 71:2239-2243, 2003

Ravn P, Demissie A, Eguale T, Wondwosson H, Lein D, Amoudy HA, Mustafa AS, Jensen AK, Holm A, Rosenkrands I, Oftung F, Olobo J, von Reyn F, Andersen P: Human T cell responses to the ESAT-6 antigen from Mycobacterium tuberculosis. J Infect Dis 179:637-645, 1999

Vincenti D, Carrara S, De Mori P, Pucillo LP, Petrosillo N, Palmieri F, Armignacco O, Ippolito G, Girardi E, Amicosante M, Goletti D: Identification of early secretory antigen target-6 epitopes for the immunodiagnosis of active tuberculosis. Mol Med 9:105-111, 2003

Mustafa AS, Shaban FA, Al-Attiyah R, Abal AT, El-Shamy AM, Andersen P, Oftung F: Human Th1 cell lines recognize the Mycobacterium tuberculosis ESAT-6 antigen and its peptides in association with frequently expressed HLA class II molecules. Scand J Immunol 57:125-134, 2003

Patarroyo ME, Romero P, Torres ML, Clavijo P, Moreno A, Martinez A, Rodriguez R, Guzman F, Cabezas E: Induction of protective immunity against experimental infection with malaria using synthetic peptides. Nature 328:629-632, 1987

Collins WE, Galland GG, Sullivan JS, Morris CL: Selection of different strains of Plasmodium falciparum for testing bloodstage vaccines in Aotus nancymai monkeys. Am J Trop Med Hyg 51:224-232, 1994

Diaz D, Naegeli M, Rodriguez R, Nino-Vasquez JJ, Moreno A, Patarroyo ME, Pluschke G, Daubenberger CA: Sequence and diversity of MHC DQA and DQB genes of the owl monkey Aotus nancymaae. Immunogenetics 51:528-537, 2000

Pinzon-Charry A, Vernot JP, Rodriguez R, Patarroyo ME: Proliferative response of peripheral blood lymphocytes to mitogens in the owl monkey Aotus nancymae. J Med Primatol 32:31-38, 2003

Chaparro PE, García I, Guerrero MI, León CI: Situación de la tuberculosis en Colombia. Biomédica 2004;24(Supl.):102-14, 2002




DOI: https://doi.org/10.22490/24629448.344

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