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Microscopy-based Assays for High-throughput Screening of Host Factors Involved in Brucella Infection of Hela Cells
Published on: August 5, 2016
Comparative experimental study of Brucella melitensis and its lipopolysaccharide in mouse model infected via
Abdinasir Yusuf Osman1, Abdul Aziz Saharee2, Faez Firdaus Jesse2
1Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Pengkalan Chepa, 16100 Kota Bharu, Kelantan, Malaysia; Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Abstract:
Brucella melitensis is a major zoonotic pathogen in which lipopolysaccharide (LPS) is believed to play a major role in the diseases pathogenesis. To study the immunopathophysiological aspects, we established a mouse model experimentally infected with whole cell of B. melitensis and its lipopolysaccharide via subcutaneous route of exposure. Eighty four mice, BALB/c, both sexes with equal gender distribution and 6-8 weeks-old were randomly assigned into 3 groups. Group 1 (n = 36) were subcutaneoulsy inoculated with 0.4 mL 109 of B. melitensis while group 2 (n = 36) were subcutaneously challenged with 0.4 mL 109 of LPS. Group 3 (n = 12) was challenged subcuatneously with phosphate buffered saline and served as a control group. Animals were observed for clinical signs, haematological and histopathological analysis for a period of 24 days post-inoculation. Our results revealed that B. melitensis infected group demonstrated significant clinical signs and histopathological evidence than LPS infected group. However, both infected groups showed elevated levels of interleukins (IL-1β & IL6), antibody levels (IgM & IgG) as early as 3 days post-infection with predominance in LPS infected group. For hormone analysis, low levels of progesterone, estradiol and testosterone were observed in both B. melitensis and LPS challenged groups throughout the study period. Moreover, in B. melitensis infected groups, the organism was re-isolated from the organs and tissues of gastrointestinal, respiratory and reproductive systems; thereby confirming the possible transmission of the disease dynamics. Moreover, LPS stimulated significantly the innate and acquired immune system without significant systemic dysfunction suggesting the potentiality of the protective properties of this component as an alternative vaccine for brucellosis infection. This report is the first detailed investigation comparing the infection progression and host responses in relation to the immunopathophysiological aspects in mouse model after subcutaneous inoculation with B. melitensis and its lipopolysaccharide.
Insights
Brucella melitensis infection caused more severe disease than its lipopolysaccharide (LPS) in mice. However, LPS significantly boosted immune responses, suggesting its potential as a brucellosis vaccine candidate.
Area of Science:
- Immunology
- Pathogen Research
- Zoonotic Diseases
Background:
- Brucella melitensis is a significant zoonotic pathogen.
- Lipopolysaccharide (LPS) from B. melitensis is implicated in disease pathogenesis.
Purpose of the Study:
- To investigate the immunopathophysiological differences between B. melitensis and its LPS.
- To establish a mouse model for comparative analysis of infection progression and host response.
Main Methods:
- BALB/c mice were subcutaneously inoculated with B. melitensis, LPS, or PBS (control).
- Animals were monitored for clinical signs, hematology, and histopathology over 24 days.
- Interleukin, antibody, and hormone levels were analyzed; bacterial re-isolation was performed.
Main Results:
- B. melitensis infection induced more pronounced clinical and histopathological changes than LPS.
- Both groups showed elevated interleukins (IL-1β, IL-6) and antibodies (IgM, IgG), with LPS predominance.
- Hormonal imbalances (progesterone, estradiol, testosterone) were noted in both infected groups.
- B. melitensis was re-isolated from multiple organ systems, confirming systemic spread.
- LPS stimulated innate and adaptive immunity without significant systemic dysfunction.
Conclusions:
- B. melitensis causes more severe disease manifestations compared to its LPS.
- LPS significantly modulates immune responses, indicating potential as a protective antigen for brucellosis vaccines.
- This study provides a comprehensive comparison of B. melitensis and LPS immunopathophysiology in a murine model.
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