RESULTS: A novel approach of utilizing an in-trans protein surface display system of Lactobacillus plantarum carrying and displaying combination of Mycobacterium tuberculosis subunit epitope antigens (Ag85B, CFP-10, ESAT-6, Rv0475 and Rv2031c) fused with LysM anchor motif designated as ACERL was constructed, cloned and expressed in Esherichia coli Rossetta expression host. Subsequently the binding capability of ACERL to the cell wall of L. plantarum was examined via the immunofluorescence microscopy and whole cell ELISA where successful attachment and consistent stability of cell wall binding up to 4 days was determined. The immunization of the developed vaccine of L. plantarum surface displaying ACERL (Lp ACERL) via the oral route was studied in mice for its immunogenicity effects. Lp ACERL immunization was able to invoke significant immune responses that favor the Th1 type cytokine response of IFN-γ, IL-12 and IL-2 as indicated by the outcome from the cytokine profiling of spleen, lung, gastrointestinal tract (GIT), and the re-stimulation of the splenocytes from the immunized mice. Co-administration of an adjuvant consisting of Lactococcus lactis secreting mouse IL-12 (LcIL-12) with Lp ACERL was also investigated. It was shown that the addition of LcIL-12 was able to further generate significant Th1 type cytokines immune responses, similar or better than that of Lp ACERL alone which can be observed from the cytokine profiling of the immunized mice's spleen, lung and GIT.
CONCLUSIONS: This study represents a proof of concept in the development of L. plantarum as a carrier for a non-genetically modified organism (GMO) tuberculosis vaccine, which may be the strategy in the future for tuberculosis vaccine development.
METHODOLOGY: This cross-sectional study was conducted using a validated self-administered questionnaire at 18 hospitals in six states of Malaysia, namely, Selangor, Kuala Lumpur, Penang, Kelantan, Sabah, and Sarawak in 2015. The study sample comprised 1600 TB patients who were randomly selected using data obtained from the Disease Control Division, Ministry of Health Malaysia. A total of 1368 of the completed questionnaires were considered usable and included in the statistical analysis.
RESULTS: Overall, the level of TB awareness was found to be high, and the respondents possessed positive attitudes towards TB and health-seeking behaviours. Self-preventive care among the TB patients was determined as being at a moderate level. With regard to contact with others, the patients were more comfortable around their families than their friends and neighbours.
CONCLUSIONS: More health education programmes are recommended to cultivate positive attitudes towards TB, to encourage communities to have a better understanding of TB, and to create awareness among patients of the proper ways to practice self-preventive care.