Expression of Mycobacterium leprae HSP65 in tobacco and its effectiveness as an oral treatment in adjuvant-induced arthritis

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 pCAM-BIA 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 transcription-PCR, 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 Á HSP65 protein Á Tobacco Á Adjuvant induced arthritis Á Oral treatment Á Rats Electronic supplementary material The online version of this article (

pVAXhsp65 Vaccination Primes for High IL-10 Production and Decreases Experimental Encephalomyelitis Severity

“…Again, the empty plasmid prompted a similar but less pronounced effect. This protective effect of hsp65 in distinct inflammatory conditions is highly supported by findings of many other authors [58–61]. …”

Hsp65-Producing Lactococcocus lactis Prevents Antigen-Induced Arthritis in Mice

“…To confirm our results, we also tested an acute model of arthritis induced by methylated bovine serum albumin (mBSA) (35). Although there are several pieces of evidence on the tolerogenic role of Hsp65 in different arthritis models, including ingestion of recombinant Mycobacterium Hsp65 (36,37), the use of recombinant L. lactis has advantages: it has anti-inflammatory properties of its own, it survives through the digestive tract and delivers large amounts of Hsp65 directly into the mucosa, boosting its tolerogenic effect (30). In addition, L. lactis is a gram-positive lactic bacterium that produces an endotoxin free version of HSP65.…”

Immunotherapeutic Activities of a DNA Plasmid Carrying the Mycobacterial hsp65 Gene (DNAhsp65)

“…Likewise, the DNAhsp65 vaccine has also been evaluated for its ability to induce other autoimmune diseases, such as arthritis and encephalomyelitis to determine whether it has therapeutic properties in combating these diseases. Previously, the work by Rodríguez-Narciso et al (68) showed a recovery of body weight and reduced inflammation in the joints after oral administration of recombinant HSP65 protein produced in tobacco in rat adjuvant-induced arthritis. Corroborating this data, other works found that the administration of the DNAhsp65 vaccine did not induce arthritis (69) or encephalomyelitis in animal models, even when using the BCG/DNAhsp65 prime-boost approach (70).…”