A novel DNA vaccine expressing the Ag85A-HA2 fusion protein provides protection against influenza A virus and Staphylococcus aureus

Virology Journal

Mingyuan Li¹  Hong Li³  Zhonghua Jiang²  Sha Li²  Jingjing Shao²  Bin Zuo²  Kang Cao²  Laifeng Ren²  Yu Kuang²  Baoning Wang²  Decui Pei²  Jun Dai² 

[1]State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, Sichuan 610041, PR China

[2]Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041, PR China

[3]Key Laboratory of Obstetric & Pediatic Disease and Birth Defects of Ministry of Education, West China Second University Hospital, Sichuan University, Chengdu, Sichuan, 610041, China

关键词: DNA vaccine;    Mycobacterium tuberculosis secreted antigen Ag85A;    Influenza a viruse;    Staphylococcus aureus;   

Others  :  1151993
DOI  :  10.1186/1743-422X-10-40

 received in 2012-01-19, accepted in 2013-01-14,  发布年份 2013

【 摘 要 】

Secondary pneumonia due to Staphylococcus aureus (S. aureus) causes significant morbidity and mortality. The aim of the research was designed a novel DNA vaccine encoding the Mycobacterium tuberculosis secreted antigen Ag85A fused with the influenza A virus (IAV) HA2 protein to provide protection against both influenza and secondary infection with S. aureus. The DNA vaccine vector efficiently expressed the encoded antigen in mammalian cells, as determined by RT-PCR, Western blotting and immunofluorescence analysis. Mice were immunized with the vaccine by intramuscular injection before challenge with IAV and S. aureus. The pulmonary and the splenocyte culture IFN-γ levels were significant higher in immunized mice than their respective controls. Although the antibody titer in the HI test was low, the sera of mice immunized with the novel vaccine vector were effective in neutralisation assay in vitro. The vaccine could reduce the loss of body weight in mice during IAV challenge. Both Western blotting and RT-PCR showed that the vaccine markedly enhanced toll like receptor 2 (TLR2) expression in splenocytes after the secondary infection with S. aureus. The survival rate of mice with high TLR2 expression (pEGFP/Ag85A-HA2 or iPR) was significantly increased compared with mice immunized with pEGFP/HA2 after challenge with S. aureus. However, the pulmonary IL-10 concentration and S. aureus titer were significantly decreased in immunized mice, and expression of TLR2 was increased after challenge with S. aureus. These results demonstrated that Ag85A could strengthen the immune response to IAV and S. aureus, and TLR2 was involved in the host response to S. aureus.

【 授权许可】

   
2013 Dai et al; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 8.

【 参考文献 】

• [1]Tsigrelis C, Mohammad M, Fraimow HS, Dellinger RP, Marchesani D, Reboli AC: Secondary bacterial pneumonia due to Staphylococcus aureus complicating 2009 influenza A (H1N1) viral infection. Infection 2010, 38(3):237-239.
• [2]Murray RJ, Robinson JO, White JN, Hughes F, Coombs GW, Pearson JC, Tan HL, Chidlow G, Williams S, Christiansen KJ, Smith DW: Community-acquired pneumonia due to pandemic A(H1N1)2009 influenzavirus and methicillin resistant Staphylococcus aureus co-infection. PLoS One 2010, 5(1):e8705.
• [3]Chandran SS, Verhoeven D, Teijaro JR, Fenton MJ, Farber DL: TLR2 engagement on dendritic cells promotes high frequency effector and memory CD4 T cell responses. J Immunol 2009, 183(12):7832-7841.
• [4]Kim JH, Skountzou I, Compans R, Jacob J: Original antigenic sin responses to influenza viruses. J Immunol 2009, 183(5):3294-3301.
• [5]Sui J, Hwang WC, Perez S, et al.: Structural and functional bases for broad-spectrum neutralization of avian and human influenza a viruses. Nat Struct Mol Biol 2009, 16(3):265-273.
• [6]Steel J, Lowen AC, Wang TT, Yondola M, Gao Q, Haye K, García-Sastre A, Palesea P: Influenza virus vaccine based on the conserved hemagglutinin stalk domain. Mbio 2010, 1(1):e00018-10.
• [7]Moulton RG, Gerner GD: Antibacterial activity of BCG-induced, interferon-containing sera. Can J Microbiol 1986, 32(5):442-445.
• [8]Romano M, Roupie V, Hamard M, Huygen K: Evaluation of the immunogenicity of pBudCE4.1 plasmids encoding mycolyl-transferase Ag85A and phosphate transport receptor PstS-3 from Mycobacterium tuberculosis. Vaccine 2006, 24(21):4640-4643.
• [9]Holley MM, Zhang Y, Lehrmann E, Wood WH, Becker KG, Kielian T: Toll-like receptor 2 (TLR2)-TLR9 crosstalk dictates IL-12 family cytokine production in microglia. Glia 2011,  .
• [10]Jann NJ, Schmaler M, Ferracin F, Landmann R: TLR2 enhances NADPH oxidase activity and killing of Staphylococcus aureus by PMN. Immunol Lett 2011, 135(1–2):17-23.
• [11]Müller-Anstett MA, Müller P, Albrecht T, Nega M, Wagener J, Gao Q, Kaesler S, Schaller M, Biedermann T, Götz F: Staphylococcal peptidoglycan co-localizes with Nod2 and TLR2 and activates innate immune response via both receptors in primary murine keratinocytes. PLoS One 2010, 5(10):e13153.
• [12]Dessing MC, van der Sluijs KF, Florquin S, Akira S, van der Poll T: Toll-like receptor 2 does not contribute to host response during postinfluenza pneumococcal pneumonia. Am J Respir Cell Mol Biol 2007, 36(5):609-614.
• [13]Tan CK, Kao CL, Shih JY, Lee LN, Hung CC, Lai CC, Huang YT, Hsueh PR: Coinfection with Mycobacterium tuberculosis and pandemic H1N1 influenza A virus in a patient with lung cancer. J Microbiol Immunol Infect 2011, 44(4):316-318.
• [14]Sharp JK, Hereth J, Fasanello J: Bronchoscopic findings in a child with pandemic novel H1N1 influenza A and methicillin-resistant Staphylococcus aureus. Pediatr Pulmonol 2011, 46(1):92-95.
• [15]Passariello C, Nencioni L, Sgarbanti R, Ranieri D, Torrisi MR, Ripa S, Garaci E, Palamara AT: Viral hemagglutinin is involved in promoting the internalisation of Staphylococcus aureus into human pneumocytes during influenza A H1N1 virus infection. Int J Med Microbiol 2011, 301(2):97-104.
• [16]Kim MR, Hong SW, Choi EB, Lee WH, Kim YS, Jeon SG, Jang MH, Gho YS, Kim YK: Staphylococcus aureus-derived extracellular vesicles induce neutrophilic pulmonary inflammation via both Th1 and Th17 cell responses. Allergy 2012, 67(10):1271-1281.
• [17]Iverson AR, Boyd KL, McAuley JL, Plano LR, Hart ME, McCullers JA: Influenza virus primes mice for pneumonia from Staphylococcus aureus. J Infect Dis 2011, 203(6):880-888.
• [18]Tisoncik JR, Billharz R, Burmakina S, Belisle SE, Proll SC, Korth MJ, Garcíia-Sastre A, Katze MG: The NS1 protein of influenza A virus suppresses interferon-regulated activation of antigen-presentation and immune-proteasome pathways. J Gen Virol 2011, 92(9):2093-2104.
• [19]Antunes I, Kassiotis G: Suppression of innate immune pathology by regulatory T cells during Influenza A virus infection of immunodeficient mice. J Virol 2010, 84(24):12564-12575.
• [20]Zivkovic A, Sharif O, Stich K, Doninger B, Biaggio M, Colinge J, Bilban M, Mesteri I, Hazemi P, Lemmens-Gruber R, Knapp S: TLR 2 and CD14 mediate innate immunity and lung inflammation to staphylococcal Panton-Valentine leukocidin in vivo. J Immunol 2011, 186(3):1608-1617.
• [21]Vu AT, Baba T, Chen X, Le TA, Kinoshita H, Xie Y, Kamijo S, Hiramatsu K, Ikeda S, Ogawa H, Okumura K, Takai T: Staphylococcus aureus membrane and diacylated lipopeptide induce thymic stromal lymphopoietin in keratinocytes through the Toll-like receptor 2-Toll-like receptor 6 pathway. J Allergy Clin Immunol 2010, 126(5):985-993.
• [22]Beran O, Potměšil R, Holub M: Differences in Toll-like receptor expression and cytokine production after stimulation with heat-killed Gram-positive and Gram-negative bacteria. Folia Microbiol (Praha). 2011, 56(3):283-287.
• [23]van der Sluijs KF, van der SluijsKoenraad F, van Elden Leontine JR, Monique N, Rob S, Pater JM, Sandrine F, Michel G, Jansen HM, René L, van der Poll T: IL-10 is an important mediator of the enhanced susceptibility to pneumococcal pneumonia after influenza infection. J Immunol 2004, 172:7603-7609.
• [24]Olivier D, Audery T, Kamiel P, Fabienne J, Van Den Thierry-p B, Albert V, Josette O, Eric S, Ulmer JB, Jean C, Kris H: Vaccination with plasmid DNA encoding Mycobacterial antigen 85A stimulates a CD41 and CD81 T-cell epitopic repertoire broader than that stimulated by Mycobacterium tuberculosis H37Rv infection. Infect Immun 1998, 66(4):1527-1533.
• [25]Borremans M, De Wit L, Volckaert G, Ooms J, De Bruyn J, Huygen K, Van Vooren J-p, Stelander M, Verhofstadt R, Content J: Cloning, sequence determination, and expression of a 32-kilodalton-protein gene of Mycobacterium tuberculosis. Infect Immun 1989, 57(10):3123-3130.
• [26]Zhang W, Li W, Li Y, Li H, Wang B, Wang F, Zhu Y, Jiang Z, Li LZM: Immune effects against influenza A virus and a novel DNA vaccine with co-expression of haemagglutinin- and neuraminidase-encoding genes. J Med Microbiol 2009, 58:845-854.
• [27]Smith SM, Roger B, Klein M’l R, Malin AS, Lukey PT, King AS, Ogg GS, Hill AVS, Dockrell HM: Human CD81 CTL Specific for the Mycobacterial Major Secreted Antigen 85A. J Immunol 2000, 165:7088-7095.
• [28]Harris K, Ream R, Gao J, Eichelberger MC: Intramuscular immunization of mice with live influenza virus is more immunogenic and offers greater protection than immunization with inactivated virus. Virol J 2011, 8(1):251-261. BioMed Central Full Text
• [29]Tamura SI, Asanuma H, Ito Y, Hirabayashi Y, Suzuki Y, Nagamine T, Aizawa C, Kurata T, Oya A: Superior cross-protective effect of nasal vaccination to subcutaneous inoculation with influenza hemagglutinin vaccine. Eur J Immunol 1992, 22(2):477-481.