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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2016, Vol. 10 Issue (4) : 490-498     DOI: 10.1007/s11684-016-0470-y
RESEARCH ARTICLE |
Immunogenicity and protective immunity against otitis media caused by pneumococcus in mice of Hib conjugate vaccine with PsaA protein carrier
Zeyu Chen1,2(),Rong Guo3,Jianghong Xu1,2,Chuangjun Qiu4
1. Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai 200031, China
2. Shanghai Clinical Medical Center of Hearing Medicine, Shanghai 200031, China
3. The Laboratory of Bacterial Vaccine, Wuhan Institute of Biological Products, Wuhan 430207, China
4. Dingtai-Haigui Biotechnology Co. Ltd., Gu’an 065500, China
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Abstract  

This study evaluated the immunogenicity and protective immunity of a Hemophilus influenzae b (Hib) polysaccharide conjugate vaccine with the pneumococcal surface adhesin A (PsaA) protein carrier in young mice. The Hib polysaccharide was conjugated with the rPsaA protein carrier, which was produced using recombinant DNA technology. A total of 15 young mice aged 3 weeks to 5 weeks were immunized with the conjugate vaccine, and another 15 young mice of the same age were immunized with the licensed Hib-tetanus toxoid (TT) vaccine. Furthermore, the third group of 15 young mice was inoculated with phosphate buffer saline as control. The immunized mice were inoculated with pneumococcus in the middle ear. Results showed that IgG antibody responses against both the PsaA protein and Hib polysaccharide were observed in the Hib-PsaA group. However, no statistical difference was observed in the titer of IgG against the Hib polysaccharide between Hib-PsaA and Hib-TT groups. The elimination rate of pneumococcus and the inflammation of the middle ear showed the effectiveness of protective immunity against otitis media caused by pneumococcus. Our results suggest that the Hib polysaccharide can be successfully conjugated with rPsaA via amide condensation. This new Hib-PsaA conjugate vaccine can induce both anti-PsaA and anti-Hib immune responses in young mice and elicit effective protection against acute otitis media caused by pneumococcus.

Keywords conjugate vaccine      pneumococcal surface adhesin A      Hemophilus influenzae b      immunogenicity      otitis media     
Corresponding Authors: Zeyu Chen   
Just Accepted Date: 19 August 2016   Online First Date: 20 September 2016    Issue Date: 01 December 2016
URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-016-0470-y     OR     http://academic.hep.com.cn/fmd/EN/Y2016/V10/I4/490
Fig.1  PCR amplification of the PsaA gene. Line M, marker; Line PsaA, PsaA gene.
Fig.2  Identification of the purity of rPsaA protein with SDS-PAGE. Line 1, the first 1 ml eluent eluted by 20 mmol/L imidazole; Line 2, the second 1 ml eluent eluted by 20 mmol/L imidazole; Line 3, the third 1 ml eluent eluted by 20 mmol/L imidazole; Line 4, the fourth 1 ml eluent eluted by 20 mmol/L imidazole; Line 5: the fifth 1 ml eluent eluted by 20 mmol/L imidazole; Line 6, the first 1 ml eluent eluted by 200 mmol/L imidazole; Line 7, the second 1 ml eluent eluted by 200 mmol/L imidazole; Line 8, the third 1 ml eluent eluted by 200 mmol/L imidazole; Line 9, the fourth 1 ml eluent eluted by 200 mmol/L imidazole; Line M, protein marker.
Fig.3  Chromatography of rPsaA obtained by using a Sepharose 4FF column.
Fig.4  Chromatography of the rPsaA-Hib polysaccharide conjugate obtained by using a Sepharose 4FF column.
GMT of anti-PsaA IgG
(GMT±S)
GMT of anti-Hib IgG
(GMT±S)
Hib-PsaA group 68.42±15.55 51.64±16.91
Hib-TT group
Control group
N/A
8.71±2.09
49.29±15.93
8.52±2.10
Tab.1  Serum antibody titers of IgG against PsaA and IgG against Hib polysaccharide for each group of mice after immunization
Fig.5  GMT of anti-PsaA IgG and anti-Hib polysaccharide IgG. 1, GMT of anti-PsaA IgG; 2, GMT of anti-Hib polysaccharide IgG.
Fig.6  Histopathological changes of the middle ear of mice three and seven days after the challenge with pneumococcus (HE staining, 50 ×). (A) Three days after challenge, Hib-PsaA group. (B) Three days after challenge, PBS group. (C) Seven days after challenge, Hib-PsaA group. (D) Seven days after challenge, PBS group.
Fig.7  Histopathological changes of the drum membrane of mice three and seven days after challenge with pneumococcus (HE staining, 50×; the arrow indicates the drum membrane). (A) Three days after challenge, Hib-PsaA group. (B) Three days after challenge, PBS group. (C) Seven days after challenge, Hib-PsaA group. (D) Seven days after challenge, PBS group.
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