Oligonucleotide primers.

<p>Oligonucleotide primers.</p

Immunological study and analysis of <i>E</i>. <i>faecium</i> L3 Bac +protective properties.

<p>General setup.</p

The bacterial count at different stages of GBS infection in mice after vaginal and oral vaccination.

<p>Vaginally vaccinated immune mice (n = 60, 10 mice per each point) were infected intravaginally with GBS (H36 Iac) (A). At the same time, orally vaccinated immune mice (n = 40, 10 mice per each point) were infected intraperitoneally with the same strain (B). After infection, the bacterial burden in CFU within the vaginal cavity (A) and the spleen (B) was calculated and expressed as log10. (A) Means with letter a and b differ significantly (p<0,05).(B) Means with letter c and d differ significantly (p<0,05).</p

The bacterial count at different stages of GBS infection in mice after nasal vaccination.

<p>Nasally vaccinated immune mice (n = 160, 10 mice per each point) were infected with GBS (H36 Iac) by intranasal (A), intraperitoneal (B) and vaginal (C) routes. After infection, the bacterial burden in CFU within lungs (A), spleens (B), and vaginal lavages (C) was calculated and expressed as log10. (A) Means with letter a and b differ significantly (p<0,05) (B) Means with letter c and d differ significantly (p<0,05).</p

Integration of the plasmid pT7ermB with the <i>ent-bac</i> into the chromosome of the strain <i>E</i>. <i>faecium</i> L3.

<p>P-promoter of the gene <i>d2</i>; <i>d2</i>-1-a region of the <i>d2</i> gene encoding for N- terminal part of D2 protein; <i>bac</i>- a fragment of the <i>bac</i> gene, encoding for IgA binding; D2-end–end of the <i>d2</i> gene encoding for the C terminus of D2 protein; pT7 ErmB—integrative plasmid. Arrows correspond to the open reading frames in the integrated element. The entire integrated element <i>ent-bac</i> with plasmid pT7ErmB is shown in brackets.</p

Development of experimental GBS vaccine for mucosal immunization

<div><p><i>Streptococcus agalactiae</i>, or group B streptococcus (GBS), is an important pathogen as it is the leading cause of neonatal deaths due to sepsis, meningitis or bacterial pneumonia. Although the development of an effective and safe GBS vaccine is on the agenda of many research labs, there is no GBS vaccine on the market yet. In the present study we attempted to engineer a live vaccine strain based on Bac, a surface protein of GBS, incorporated into a surface fimbrial protein of probiotic Enterococcus. The resulting strain induced specific systemic and local immune responses in mice and provided protection against GBS when administered via the intranasal, oral or intravaginal immunization routes.</p></div

Confocal microscopy of the original enterococcal strain <i>E</i>. <i>faecium</i> L3 and its derivative with the fragment of streptococcal <i>bac</i> gene using an HRP labeled IgA.

<p>A- <i>E</i>. <i>faecium L3</i>, <i>B- E</i>. <i>faecium L3</i> Bac +.</p

The immune response in mice after vaccination with live vaccine.

<p>Mice were vaccinated vaginally (A), per os (B) nasally, (C) with <i>E</i>. <i>faecium</i> L3Bac +. The Bac-specific secretory IgA and/or serum IgG levels were measured after vaccination. Each point on the chart represents the average value from 10 measurements. In controls, the concentration of specific mucosal IgA and serum IgG did not exceed 3 mg/L. Means with different letter differ significantly (p<0,05). Symbols a-b are valid for IgA, c-d for IgG.</p