A mutated Bordetella strain comprising at least a mutated ptx gene, a deleted or mutated dnt gene and a heterologous ampG gene is provided. The attenuated mutated Bordetella strain can be used in an immunogenic composition or a vaccine for the treatment or prevention of a Bordetella infection. Use of the attenuated Bordetella strain for the manufacture of a vaccine or immunogenic composition, as well as methods for protecting mammals against infection by Bordetella are also provided.

A mutated Bordetella strain comprising at least a mutated ptx gene, a deleted or mutated dnt gene and a heterologous ampG gene is provided. The attenuated mutated Bordetella strain can be used in an immunogenic composition or a vaccine for the treatment or prevention of a Bordetella infection. Use of the attenuated Bordetella strain for the manufacture of a vaccine or immunogenic composition, as well as methods for protecting mammals against infection by Bordetella are also provided.

A mutated Bordetella strain comprising at least a mutated ptx gene, a deleted or mutated dnt gene and a heterologous ampG gene is provided. The attenuated mutated Bordetella strain can be used in an immunogenic composition or a vaccine for the treatment or prevention of a Bordetella infection. Use of the attenuated Bordetella strain for the manufacture of a vaccine or immunogenic composition, as well as methods for protecting mammals against infection by Bordetella are also provided.

The present invention describes a second-generation tetanus toxoid vaccine and a process for the preparation thereof, comprising the steps of: inducing an E. coli culture OD.sub.600=0.5 by adding 0.2 mM IPTG; growing the culture at 14-16 C. for 14 to 20 hours; suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride; adding 1% of triton-X-100 to the phosphate buffer, and adding the buffer to the culture; sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4 C. on cold beads; centrifuging the culture for 60 to 90 minutes; collecting and purifying a supernatant using Ni-NTA affinity column with an eluant; and combining the supernatant into a pool with contaminated bands and concentrating using Centriprep-30 centrifuge filters (30 kDa pores).

The present invention describes a second-generation tetanus toxoid vaccine and a process for the preparation thereof, comprising the steps of: inducing an E. coli culture OD.sub.600=0.5 by adding 0.2 mM IPTG; growing the culture at 14-16 C. for 14 to 20 hours; suspending the culture in 25 mM phosphate buffer containing 200 mM sodium chloride; adding 1% of triton-X-100 to the phosphate buffer, and adding the buffer to the culture; sonicating the culture for a period of 3 minutes (at 5 sec on/off pulse) at 4 C. on cold beads; centrifuging the culture for 60 to 90 minutes; collecting and purifying a supernatant using Ni-NTA affinity column with an eluant; and combining the supernatant into a pool with contaminated bands and concentrating using Centriprep-30 centrifuge filters (30 kDa pores).

Provided are a recombinant expression vector for whooping cough antigens, an engineered bacterium and use thereof. The recombinant expression vector for whooping cough antigens comprises a protein expression cassette for pertactin, a gene for resistance screening, and an upstream recombinant nucleic acid fragment and a downstream recombinant nucleic acid fragment of filamentous haemagglutinin gene, wherein the protein expression cassette for pertactin is located between the upstream recombinant nucleic acid fragment and the downstream recombinant nucleic acid fragment of the filamentous haemagglutinin gene, and the upstream recombinant nucleic acid fragment and the downstream recombinant nucleic acid fragment of the filamentous haemagglutinin gene can undergo homologous recombination with the upstream and downstream of the filamentous haemagglutinin gene, respectively. The gene of FHA protein in the recombinant expression vector is knocked out and the copies PRN protein gene are increased, so that the Bordetella pertussis genetically engineered strain can efficiently overexpress PRN protein antigen and PT protein, and the expressed PRN protein antigen and PT protein exist in the cells and the supernatant respectively, thereby facilitating subsequent separation and purification of the antigens.

Provided are a recombinant expression vector for whooping cough antigens, an engineered bacterium and use thereof. The recombinant expression vector for whooping cough antigens comprises a protein expression cassette for pertactin, a gene for resistance screening, and an upstream recombinant nucleic acid fragment and a downstream recombinant nucleic acid fragment of filamentous haemagglutinin gene, wherein the protein expression cassette for pertactin is located between the upstream recombinant nucleic acid fragment and the downstream recombinant nucleic acid fragment of the filamentous haemagglutinin gene, and the upstream recombinant nucleic acid fragment and the downstream recombinant nucleic acid fragment of the filamentous haemagglutinin gene can undergo homologous recombination with the upstream and downstream of the filamentous haemagglutinin gene, respectively. The gene of FHA protein in the recombinant expression vector is knocked out and the copies PRN protein gene are increased, so that the Bordetella pertussis genetically engineered strain can efficiently overexpress PRN protein antigen and PT protein, and the expressed PRN protein antigen and PT protein exist in the cells and the supernatant respectively, thereby facilitating subsequent separation and purification of the antigens.

The invention relates to a chimeric protein comprising or consisting of, from N-terminal to C-terminal, (a) a N-terminal part of a Bordetella CyaA protein (b) a heterologous polypeptide, and (c) a C-terminal part of a Bordetella CyaA protein. The invention also relates to a polynucleotide encoding a deleted version of a Bordetella CyaA, as well as a polynucleotide encoding this chimeric protein. A composition comprising at least one chimeric protein(s) of the invention and the prophylactic and/or therapeutic uses of said composition are also part of the invention.

The invention relates to a chimeric protein comprising or consisting of, from N-terminal to C-terminal, (a) a N-terminal part of a Bordetella CyaA protein (b) a heterologous polypeptide, and (c) a C-terminal part of a Bordetella CyaA protein. The invention also relates to a polynucleotide encoding a deleted version of a Bordetella CyaA, as well as a polynucleotide encoding this chimeric protein. A composition comprising at least one chimeric protein(s) of the invention and the prophylactic and/or therapeutic uses of said composition are also part of the invention.

A massive clonal expansion of activated CD8.sup.+ T-cells with increased frequency of HPV 16-specific CD8.sup.+ T-cells was discovered to be a characteristic of oral lichen planus (OLP), indicating a causal link between HPV infection and the dysimmune process. The invention relates to compositions and methods for the diagnosis and treatment of OLP patients.