Immunogenic Activity of Lipopolysaccharide Isolated from Salmonella enterica serovar Typhimurium and Conjugated with Liposome in Albino Male Mice.

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12/5/2014 11:42 AM

Immunogenic Activity of Lipopolysaccharide Isolated from Salmonella enterica serovar Typhimurium and Conjugated with Liposome in Albino Male Mice.


Ayad M. ALAubaidy(Department of Biotechnology/College of Science,Al-Nahrain University.)
Farah T. O. Al-Jumaili(Department of Biotechnology/College of Science,Al-Nahrain University.)
Ali H. Ad'hiah(Tropical-Biological Research Unit/ College of Science,University of Baghdad.)


Abstract

This study was designed to evaluate the role of Salmonella enterica serovar Typhimurium as a causative pathogen in hospitalized diarrhoeal patients younger than five years old, extract and purify endotoxin (lipopolysaccharide; LPS) from isolated and identified S. Typhimurium, and determine the immunogenic activity (humoral and cellular immune responses) of LPS-liposome conjugate in albino male mice. The patients (95 cases) were admitted to the Central Pediatric Hospital and Al- Kadhiymiah Pediatric Hospital in Baghdad during the period 24/10/2010 - 30/11/2010, because of severe diarrhoea and fever.
Bacterial evaluation of stool samples revealed the identification of two (2.1%) S. Typhimurium isolates (S1 and S2). Antibiotic sensitivity test demonstrated that S1 isolate was more resistance than S2 isolate; therefore it was considered more virulent and subjected for extraction and purification of LPS. Chemical characterization of the extracted LPS revealed that the carbohydrate content was 2.34 mg/ml, while the protein concentration was very low (0.52 μg/ml). Partial purification of extracted LPS by using gel-filtration chromatography (Sephacryl S200) showed three peaks, and after determination of protein and carbohydrate concentrations for each peak, peak two was observed to have the highest carbohydrate content (25%) and the lowest contaminated protein (0.001%). The LPS of this peak was immunologically evaluated in mice at a concentration of 100 μg/ml, alone or in conjugation with a commercially available liposome (LIP). These evaluations revealed that LPS-LIP conjugate was able to modulate the humoral and cellular immune responses against live S. Typhimurium.
Keywords: Diarrhea, Salmonella, Lipopolysaccharide, Liposome.



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