Current Issues of Pharmacy and Medical Sciences

Liposomal-lipopolysaccharide vaccine extracted from P.mirabilis induces moderate TLR4 and CD14 production

Curr Issues Pharm Med Sci., Vol. 32, No. 2, 81-86

Ruaa SH.Nile, Mayyada F. Darweesh*, Mohauman M. Al-Rufaie

Biology dep. Faculty of science, Kufa University, Najaf, Iraq


https://doi.org/10.2478/cipms-2019-0016

Abstract

Proteus mirabilis is a common cause of recurrent urinary tract infections in individuals with functional or structural abnormalities. It also forms bladder and kidney stones. Lipopolysaccharide (LPS) is a potential Proteus virulence factor that plays a key role in pathogenesis, as well as in stimulating innate immune response. Therefore, this study aimed to extract LPS from a highly resistant isolate and incorporate it in a delivery system (liposome) to stimulate an immune response against virulent pathogens. In the work, 50 isolates of P. mirabilis were taken from 200 urine specimens obtained from recurrent-urinary tract infections (UTI) of patients of AL-Sadar Hospital. Specimens were cultured on specific media, and then bacterial isolates were identified via morphological, biochemical and Vitek-2 systems. The results showed that P. mirabilis was expressed in 11 (22%), 30 (60%) and 9 (18%) recurrent UTI, kidney stone and catheter samples, respectively. All isolates were assessed through antibiogram testing, with the results revealing that most isolates were multidrug resistant to more than 3 classes of antibiotics. Herein, P. mirabilis NO 50 revealed particularly high resistance, so it was chosen for LPS extraction. Lethal dose 50 (LD50) observations indicated that a live suspension of P. mirabilis was at 4.5×107 CFU/ml, while LPS was at 270 μg/ml. LPS was used as an immunogenic to stimulate the immune system through injecting Rats intraperitoneally (I.P.) with 1 ml of LD50%. Subsequently, the efficiency of immunogenes in stimulating the immune response was evaluated by determining the Toll-like receptor and CD14 levels. The results indicate that LPS incorporated in the Liposome released moderate levels of Toll-like receptors-4 (TLR4) that enabled the immune system to clear pathogens. The LPS+ complete Freund’s adjuvant (CFA) and LPS vaccinated groups recorded hyper production for TLR4 (52.2 and 40.9 pg/ml, respectively), this was followed by liposome (LIP) and bacterial suspension (11 and 20.5 pg/ml, respectively) in ranking effectiveness. This study reveals a mean of CD14 that was higher in both LPS and LPS+CFA
and moderate in LPS+LIP, in comparison with control and liposome groups.
In conclusion, LPS-Liposomes are a promising nanomedicine for modulating the hyper response of LPS. This may lead to tissue inflammation but appeared beneficial in stimulating the immune response at moderate levels so as to eradicate infection without tissue damage.

Keywords

Lipopolysaccharide, Liposome (LIP), P. mirabilis, immunomodulater activity, cytokine.

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