Search

KR-102963606-B1 - Immunogenic composition

KR102963606B1KR 102963606 B1KR102963606 B1KR 102963606B1KR-102963606-B1

Abstract

The present invention relates particularly to bacterial strains for use in the field of vaccines, and to the field of prevention or treatment of infections caused by bacteria of the genus Bordetella.

Inventors

  • 데보스, 나탈리, 이사벨
  • 시몬스, 스티븐, 클레멘트

Assignees

  • 글락소스미스클라인 바이오로지칼즈 에스.에이.

Dates

Publication Date
20260513
Application Date
20200930
Priority Date
20191001

Claims (20)

  1. Sequence Identification Number: Genome LpxA gene encoding the LpxA protein containing the amino acid sequence of 2 As recombinant Bordetella pertussis bacteria including, In the above amino acid sequence, The amino acid residue corresponding to position 170 of sequence identification number 1 is a serine residue, and The amino acid residue corresponding to position 229 of sequence identification number 1 is an alanine residue Recombinant Bordetella pertussis bacteria.
  2. In paragraph 1, Sequence Identification Number: Genomic insertion of a heterologous LpxD gene encoding the LpxD protein containing the amino acid sequence of 4 Recombinant Bordetella pertussis bacteria containing additional
  3. In paragraph 1, a recombinant Bordetella pertussis bacterium in which the LpxA gene is inactivated.
  4. In paragraph 2, recombinant Bordetella pertussis bacteria in which the LpxD gene is inactivated.
  5. In claim 1, recombinant Bordetella pertussis bacteria producing lipid A characterized by one or more of the following: (i) At least 30% of the C3' acyl chain is reduced from length C14 to length C10 to C12 , (ii) at least 30% of the C2' acyl chain is reduced from length C14 to length C10 to C12 , and (iii) At least 30% of the C2 acyl chain is reduced from length C14 to length C10 to C12 .
  6. In claim 1, a recombinant Bordetella pertussis bacterium that produces lipid A having reduced toxicity activity compared to the toxicity activity of lipid A produced by the parent strain.
  7. Recombinant Bordetella pertussis bacteria according to claim 1, wherein the growth curve of the bacterial population of the bacteria differs from the growth curve of the parent strain population by less than 20%, less than 15%, less than 10%, or less than 5%.
  8. As an isolated outer membrane vesicle (OMV) derived from a recombinant Bordetella pertussis bacterium of any one of claims 1 to 7, comprising modified lipid A incorporated into the membrane, (i) All C3' acyl chains of lipid A have a length of C10 , or (ii) all C2 and C2' acyl chains of lipid A have a length of C12 , or (iii) both of the above (i) and the above (ii) Isolated outer membrane vesicle.
  9. In paragraph 8, an isolated outer membrane vesicle free from cutaneous necrotic toxins.
  10. In paragraph 8, an isolated outer membrane vesicle containing an endogenous, genetically decoded pertussis toxoid.
  11. A vaccine composition comprising isolated OMV according to claim 8 and pharmaceutically acceptable excipients for use in the prevention or treatment of an infection caused by bacteria of the genus Bordetella or Bordetella pertussis.
  12. In Paragraph 11, (1) Pertussis toxoid (PT), (2) FHA, (3) Pertactin (PRN), (4) FIM2/FIM3, (5) Adenylate cyclase, (6) Diphtheria toxoid (DT), (7) Tetanus toxoid (TT), (8) Inactivated polio virus (IPV), (9) Hepatitis B surface antigen and (10) Hib PRP A vaccine composition further comprising an additional antigen selected from the group consisting of
  13. In paragraph 11, a vaccine composition for use in mammals or humans.
  14. A method for modulating the reactogenicity of lipid A of Bordetella pertussis bacteria, comprising stably incorporating into the genome of said bacteria: (i) As an LpxA gene encoding an LpxA protein containing the amino acid sequence of sequence identification number 2, If the above protein is numbered according to sequence identification number: 2, Serine (S170) at position 170 and Alanine (A229) at location 229 The LpxA gene containing, or (ii) the LpxA gene of (i) above and Sequence identification number: LpxD gene encoding the LpxD protein containing the amino acid sequence of 4.
  15. delete
  16. delete
  17. delete
  18. delete
  19. delete
  20. delete

Description

Immunogenic composition The present invention relates to the field of vaccines, particularly to the prevention or treatment of infections caused by bacteria of the genus Bordetella, particularly Bordetella pertussis . Pertussis, caused by Bordetella pertussis, is a highly contagious respiratory infection. Acute infection can lead to severe disease characterized by respiratory failure, pulmonary hypertension, leukocytosis, and death. Pertussis is a vaccine-preventable disease, with acellular pertussis (aP) or whole-cell pertussis (wP) vaccines used worldwide. However, the disease has persisted in vaccinated populations, and epidemiological data have reported a global increase in pertussis incidence in recent years. Several assumptions have been made, including the evolution of pertussis strains and the reduced duration of protection from the aP vaccine compared to the wP vaccine. One area of research aimed at controlling the re-emergence of pertussis concerns the development of new vaccines. Vesicles derived from pathogens have been used in the development of immunogenic compositions, such as vaccines [1]. The use of outer membrane vesicles (OMVs) or their derivatives could potentially deliver a wide range of antigens in their natural forms. OMV comprises bacterial lipopolysaccharides (LPS) composed of a highly variable O antigen, a less variable core oligosaccharide, and a highly conserved lipid moiety designated as lipid A. Bacterial lipooligosaccharides (LOS) share a similar lipid A structure with many of the same functional activities as LPS but lack the O-antigen unit. In the relevant technical field, the term "LPS" is often used to refer to both LPS and LOS bacterial saccharides. However, the presence of LPS/LOS within OMV can present issues from clinical and regulatory perspectives. For example, when LPS/LOS is broken down in the human body, it can induce the production of excessive pro-inflammatory cytokines. Through their interaction with human TLR4, they can potentially cause numerous adverse effects (reactives), such as fever, chills, shock, and various other symptoms, depending on the specific organism and patient's condition. The endotoxic activity of LPS/LOS is mainly determined by the composition of its lipid A moiety, which consists of glucosamine disaccharides substituted with one or two phosphate groups and varying numbers of acyl chains [2]. WO2018/167061 [3] describes the use of LpxA variants derived from Pseudomonas aeruginosa (LpxA Pa ) or Neisseria meningitidis (LpxA Nm ) to reduce the length of the C3' acyl chain in Bordetella pertussis. The exogenous LpxA gene was expressed from a plasmid, while the genomic endogenous LpxA gene was knocked out. Episomal expression of LpxA Pa had the effect of reducing the length of some acyl chains, and the authors noted that TLR4 stimulation was reduced. However, the strain exhibited severe growth defects. Additionally, the expression of LpxA Nm was lethal. When an LpxD variant derived from Pseudomonas aeruginosa was expressed episomally in B. pertussis, similar severe growth defects were observed. The results confirm that reactogenicity can be modified by manipulation of lipid A, whereas growth defects in the strain are a major obstacle to the use of these strains in vaccine manufacturing. Therefore, improvements suitable for the manufacture of immunogenic compositions, such as vaccines, are still necessary. Summary of the Invention The applicant has developed a novel recombinant Bordetella pertussis bacterial strain that overcomes problems observed in the relevant art and is particularly suitable for the production of whole-cell antigens and/or outer membrane vesicle components. These components can be used in immunogenic compositions, such as vaccines. In a first aspect, the present invention relates to recombinant Bordetella pertussis bacteria. In particular, the present invention relates to recombinant Bordetella pertussis bacteria comprising: at least one genomic LpxA gene encoding an LpxA protein, wherein the LpxA protein comprises a mutation at position 170 and/or a mutation at position 229 compared to SEQ ID NO: 1; and/or at least one genomic insertion of a heterologous LpxD gene. In this first aspect, the present invention preferably relates to a recombinant Bordetella pertussis bacterium comprising at least one genomic LpxA gene encoding an LpxA protein, wherein the LpxA protein comprises a mutation at position 170 and/or a mutation at position 229 compared to sequence identification number: 1. The amino acid sequence of the wild-type LpxA protein from Bordetella pertussis (LpxA Bpe ) is provided as sequence identification number: 1. The amino acid sequence of the LpxA protein from Bordetella parapertussis (LpxA Bpa ) is provided as sequence identification number: 2. The present invention may relate to recombinant Bordetella pertussis bacteria comprising at least one genomic LpxA gene encoding an LpxA protein, wherein the LpxA gene is der