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Senevirathne, Amal,Hewawaduge, Chamith,Lee, John Hwa Elsevier 2019 Veterinary immunology and immunopathology Vol.209 No.-
<P><B>Abstract</B></P> <P>We demonstrated the use of attenuated <I>Salmonella</I> strains secreting <I>Brucella</I> antigens SodC, Omp19, BLS, and PrpA as live vaccine candidates against <I>Brucella abortus</I> infection and presented their cross-protection against <I>Salmonella</I> infections using a BALB/c mice model. Here, a single immunization with each individual strain was capable of establishing significantly high (p < 0.05) <I>Brucella</I>-specific systemic immunoglobulin (Ig)G and secretory IgA (sIgA) responses compared to control mice. Upon stimulation of the splenocytes harvested from immunized mice with the respective antigens SodC, Omp19, BLS, and PrpA, significant increases in splenocyte proliferative responses against all four antigens versus PBS and vector controls were observed (p < 0.05). Additionally, interferon-γ and interleukin-4 secretion clearly demonstrated an uplift of these cytokines in all four strains upon immunization compared to the control groups. However, a significantly high response was noted in the mice groups immunized with <I>Salmonella</I> secreting SodC and Omp19 only. Upon virulent <I>Brucella abortus</I> 544 challenge, all four antigens presented a significantly high protection index (PI) in the spleen, as follows: 0.85 for SodC; 0.96 for Omp19; 0.6 for BLS; and 0.66 for PrpA. In contrast, in the liver, the same antigens resulted in PI values of 1.37, 1.14, 1.12, and 1.81, respectively. Immunological profiling of immunized mice against <I>Salmonella</I>-specific immune responses also showed significant elicitation of both humoral and cell-mediated immune responses as measured by IgG, sIgA, splenocyte proliferation, and cytokine induction. In addition, full protection against virulent <I>Salmonella</I> challenge was shown with no mortality in immunized mice, whereas 100% (8/8) mortality was observed in control mice over a two-week post-<I>Salmonella</I> challenge. In conclusion, we show that the live attenuated <I>Salmonella</I> delivering <I>Brucella</I> protective antigens may efficiently confer dual protection against both brucellosis and salmonellosis in immunized mice.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Development of a safe and effective vaccine candidate against Brucellosis. </LI> <LI> <I>Salmonella</I> delivering <I>Brucella</I> antigens sodC, Omp19, BLS and PrpA were evaluated. </LI> <LI> Immunization with attenuated <I>Salmonella</I> conferred protection against virulent <I>Brucella</I> &<I>Salmonella</I> challenge. </LI> </UL> </P>
Senevirathne, Amal,Hewawaduge, Chamith,Hajam, Irshad A.,Lalsiamthara, Jonathan,Lee, John Hwa Elsevier 2019 Veterinary microbiology Vol.228 No.-
<P><B>Abstract</B></P> <P>The present study was aimed to develop a safe and effective anti-<I>Brucella</I> subunit vaccine for mucosal protection against the respiratory exposure of <I>Brucella</I> infection. A chitosan-based <I>Brucella</I> nasal vaccine (BNV) was formulated using well-known <I>Brucella</I> immunogens, sodC, omp19, BLS and PrpA and tested against nasal <I>Brucella</I> challenge in BALB/c mice. The mice were intra-nasally vaccinated with sterile phosphate buffer saline (PBS), BNV or BNV plus <I>Brucella</I> LPS, and humoral (systemic IgG and mucosal IgA) and cell-mediated immune responses were analyzed. Results showed that mice vaccinated with either BNV or BNV plus LPS elicited significantly (p < 0.05) high IgG and IgA responses compared to the PBS control. The IgG responses were significantly (p < 0.05) higher than IgA levels, which showed almost comparable levels observed in either intestines or in lungs. Furthermore, the IgG and IgA responses against each individual component of the BNV formulation indicated that omp19 induced highest levels of both IgG and IgA levels than the other constituents of BNV formulation. Upon re-stimulation of the splenocytes with <I>Brucella</I> whole cell lysate, significantly (p < 0.05) high IFN-γ levels, lymphocyte proliferation, and CD4<SUP>+</SUP> T cell responses were observed in mice vaccinated with BNV or BNV plus LPS. Upon sub-lethal nasal challenge with wild-type <I>Brucella strain</I>, vaccinated mice showed significant reduction of <I>Brucella</I> recovery in lungs and spleen compared to the PBS control. This study indicates that BNV formulation with or without <I>Brucella</I> LPS efficiently induced humoral and cell-mediated immune responses and conferred significant protection against the sub-lethal <I>Brucella</I> challenge.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Development of a safe and effective mucosal vaccine for <I>Brucellosis</I>. </LI> <LI> A chitosan-based vaccine was formulated using <I>Brucella</I> immunogens, sodC, omp19, BLS and PrpA. </LI> <LI> Highly conserved Brucella antigens was able to give significant protection against the nasal challenge. </LI> </UL> </P>
Kim, Tae-Hwan,Lee, Hyun-Cheol,Kim, Jae-Hoon,Hewawaduge, C. Y.,Chathuranga, Kiramage,Chathuranga, W. A. Gayan,Ekanayaka, Pathum,Wijerathne, H. M. S. M.,Kim, Chul-Joong,Kim, Eunhee,Lee, Jong-Soo Public Library of Science 2019 PLoS pathogens Vol.15 No.8
<P> Fas-associated factor 1 is a death-promoting protein that induces apoptosis by interacting with the Fas receptor. Until now, FAF1 was reported to interact potentially with diverse proteins and to function as a negative and/or positive regulator of several cellular possesses. However, the role of FAF1 in defense against bacterial infection remains unclear. Here, we show that FAF1 plays a pivotal role in activating NADPH oxidase in macrophages during <I>Listeria monocytogenes</I> infection. Upon infection by <I>L. monocytogenes,</I> FAF1 interacts with p67phox (an activator of the NADPH oxidase complex), thereby facilitating its stabilization and increasing the activity of NADPH oxidase. Consequently, knockdown or ectopic expression of FAF1 had a marked effect on production of ROS, proinflammatory cytokines, and antibacterial activity, in macrophages upon stimulation of TLR2 or after infection with <I>L. monocytogenes.</I> Consistent with this, FAF1<SUP>gt/gt</SUP> mice, which are knocked down in FAF1, showed weaker inflammatory responses than wild-type mice; these weaker responses led to increased replication of <I>L. monocytogenes.</I> Collectively, these findings suggest that FAF1 positively regulates NADPH oxidase-mediated ROS production and antibacterial defenses. </P>
Chowdhury, Mohammed Y.E.,Kim, Tae-Hwan,Uddin, Md Bashir,Kim, Jae-Hoon,Hewawaduge, C.Y.,Ferdowshi, Zannatul,Sung, Moon-Hee,Kim, Chul-Joong,Lee, Jong-Soo Elsevier 2017 Veterinary microbiology Vol.201 No.-
<P><B>Abstract</B></P> <P>To develop a safe and effective mucosal vaccine that broad cross protection against seasonal or emerging influenza A viruses, we generated a mucosal influenza vaccine system combining the highly conserved matrix protein-2 (sM2), fusion peptide of hemagglutinin (HA<SUB>2</SUB>), the well-known mucosal adjuvant cholera toxin subunit A1 (CTA1) and poly-γ-glutamic acid (γ-PGA)-chitosan nanoparticles (PC NPs), which are safe, natural materials that are able to target the mucosal membrane as a mucosal adjuvant. The mucosal administration of sM2HA2CTA1/PC NPs could induce a high degree of systemic immunity (IgG and IgA) at the site of inoculation as well as at remote locations and also significantly increase the levels of sM2- or HA2-specific cell-mediated immune response. In challenge tests in BALB/c mice with 10 MLD<SUB>50</SUB> of A/EM/Korea/W149/06(H5N1), A/Puerto Rico/8/34(H1N1), A/Aquatic bird/Korea/W81/2005(H5N2), A/Aquatic bird/Korea/W44/2005 (H7N3) or A/Chicken/Korea/116/2004(H9N2) viruses, the recombinant sM2HA2CTA1/PC NPs provided cross protection against divergent lethal influenza subtypes and also the protection was maintained up to six months after vaccination. Thus, sM2HA2CTA1/PC NPs could be a promising strategy for a universal influenza vaccine.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Construction of poly-γ-glutamic acid (γ-PGA)-chitosan nanoparticles (PC NPs) containing conserved antigens of Influenza virus and mucosal adjuvant cholera toxin subunit A1 (CTA1) as a safe and effective mucosal vaccine against emerging influenza A viruses. </LI> <LI> The mucosal administration of sM2HA2CTA1/PC NPs could induce a high degree of systemic, mucosal and cell-mediated immune response. </LI> <LI> Induced immune responses could protect mice against 10MLD<SUB>50</SUB> of divergent influenza subtypes. </LI> <LI> The immune response and efficacy of protection was maintained up to six months after final inoculation. </LI> </UL> </P>