Expression of Mycobacterium leprae HSP65 in tobacco and its effectiveness as an oral treatment in adjuvant-induced arthr
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ORIGINAL PAPER
Expression of Mycobacterium leprae HSP65 in tobacco and its effectiveness as an oral treatment in adjuvant-induced arthritis Ce´sar Rodrı´guez-Narciso • Mayra Pe´rez-Tapia • Rosa Marı´a Rangel-Cano Celio L. Silva • Mariana Meckes-Fisher • Rafael Salgado-Garciglia • Sergio Estrada-Parra • Rodolfo Lo´pez-Go´mez • Iris Estrada-Garcı´a
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Received: 25 November 2009 / Accepted: 13 May 2010 / Published online: 6 June 2010 Ó Springer Science+Business Media B.V. 2010
Abstract Transgenic plants are able to express molecules with antigenic properties. In recent years, this has led the pharmaceutical industry to use plants as alternative systems for the production of recombinant proteins. Plant-produced recominant proteins can have important applications in therapeutics, such as in the treatment of rheumatoid arthritis (RA). In this study, the mycobacterial HSP65 protein expressed in tobacco plants was found to be effective as a treatment for adjuvant-induced arthritis (AIA). We cloned the hsp65 gene from Mycobacterium leprae into plasmid pCAMBIA 2301 under the control of the double 35S promoter from cauliflower mosaic virus. Agrobacterium
tumefaciens bearing the pChsp65 plasmid was used to transform tobacco plants. Incorporation of the hsp65 gene was confirmed by PCR, reverse transcriptionPCR, histochemistry, and western blot analyses in several transgenic lines of tobacco plants. Oral treatment of AIA rats with the HSP65 protein allowed them to recover body weight and joint inflammation was reduced. Our results suggest a synergistic effect between the HSP65 expressed protein and metabolites presents in tobacco plants. Keywords Transgenic plants HSP65 protein Tobacco Adjuvant induced arthritis Oral treatment Rats
Electronic supplementary material The online version of this article (doi:10.1007/s11248-010-9404-7) contains supplementary material, which is available to authorized users. C. Rodrı´guez-Narciso R. Salgado-Garciglia R. Lo´pez-Go´mez (&) Instituto de Investigaciones Quı´mico-Biolo´gicas, Universidad Michoacana de San Nicola´s de Hidalgo, Edificio B1, Francisco J Mu´jica S/N Col. Felicitas del Rio, CP 58060 Morelia, Michoaca´n, Mexico e-mail: [email protected] M. Pe´rez-Tapia S. Estrada-Parra I. Estrada-Garcı´a Departamento de Inmunologı´a, Escuela Nacional de Ciencias Biolo´gicas IPN, Prol. Carpio y Plan de Ayala s/n, Col. Santo Tomas., CP 11340 Me´xico, D.F., Mexico
C. L. Silva Department of Biochemistry and Immunology, Medicine School of Ribeira˜o Preto, The Centre for Tuberculosis Research , University of Sa˜o Paulo, Sa˜o Paulo, SP CEP 14049-900, Brazil M. Meckes-Fisher Farmacologı´a de Productos Naturales, Unidad Pedia´trica, Centro Me´dico Nacional Siglo XXI, IMSS. Av. Cuauhtemoc 330, Col Doctores, CP 06725 Me´xico, D.F., Mexico
R. M. Rangel-Cano Deparatamento de Ingenierı´a Gene´tica, CINVESTAV Campus Guanajuato IPN, Km 9,5 Libramiento Norte Carr. Irapuato-Leo´n., Irapuato, Guanajuato, Mexico
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Abbreviations AIA Adjuvant-induced arthritis RA Rheumatoid arthritis
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