Immunogenicity and protective efficacy of DnaJ (hsp40) of Streptococcus pneumoniae against lethal infection in mice.
The present study was carried out to evaluate the immunogenicity and protective efficacy of DnaJ (hsp40) of Streptococcus pneumoniae, by cloning the full-length DnaJ of S. pneumoniae and expressing in heterologous host E. coli BL-21 (DE3). PCR amplified DnaJ was ligated in pQE-30 expression vector and subsequently transformed in E. coli DH5alpha strain. Cloning of DnaJ was confirmed by double digestion and PCR, followed by DNA sequencing. The His-tag containing recombinant protein was purified by Ni-NTA affinity chromatography. To determine the immunogenicity of DnaJ, the mice (10 mice/group) were immunized by injecting 40 microg DnaJ protein/mouse i.p. There was a significant increase in IgG titres (2 x 10(5)) in mice immunized with DnaJ protein. Isotyping studies revealed that antibodies produced are predominantly IgG2a type indicating the predominance of Th1 response. A significant increase in lymphocyte proliferation was observed in mice immunized with DnaJ protein as compared to the control mice. Further, there was a significant increase in IL-2 and gamma-IFN levels in culture supernatants of splenocytes isolated from immunized mice. To determine the efficacy of DnaJ vaccination in eliciting protection, the mice were challenged with 1 x 10(5)cells of S. pneumoniae A66 type 3 capsular strain intra-nasally after 7 days of last immunization. All the control mice died within 2 days of post-infection, while 70% of animals immunized with DnaJ survived the lethal challenge by S. pneumoniae. The study reveals that immunization of mice with DnaJ elicits protective immunity against S. pneumoniae infection.
Khan
Mohd Nadeem
MN
Department of Experimental Biology, Defence Institute of Physiology and Allied Sciences, Lucknow Road, Timarpur, Delhi 110054, India.
Bansal
Anju
A
Shukla
Dhananjay
D
Paliwal
Piyush
P
Sarada
S K S
SK
Mustoori
Sai Ram
SR
Banerjee
Pratul Kumar
PK
D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology; D005260:Female; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D007074:Immunoglobulin G / Q000276:immunology; D007371:Interferon-gamma / Q000276:immunology; D007376:Interleukin-2 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011008:Pneumococcal Infections / Q000276:immunology / Q000517:prevention & control*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000494:pharmacology; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology
D050956:HSP40 Heat-Shock Proteins; D007074:Immunoglobulin G; D007376:Interleukin-2; D011994:Recombinant Proteins; D007371:Interferon-gamma