# US 9,040,059 B2 · Recombinant bacterium capable of eliciting a protective immune response against C. perfringens

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- Number: US 9,040,059 B2
- Document ID: US9040059B2
- Status: Granted
- Kind: B2
- Application number: 12/681,711
- Filed: 2008-10-06
- Published: 2011-10-20
- Granted: 2015-05-26
- Earliest claimed: 2007-10-05
- Assignees: Arizona Board of Regents for and on behalf of Arizona State University
- Applicants: Roy Curtiss, III; Bereket Zekarias; Kenneth Roland
- Inventors: Roy Curtiss, III; Bereket Zekarias; Kenneth Roland
- Web page: https://patentgrep.com/patent/US9040059B2

## CPC

- A61K 39/0275: {Salmonella}
- A61K 39/025: {Enterobacteriales, e.g. Enterobacter}
- A61P 37/04: Immunostimulants
- C12N 15/74: Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- A61K 2039/522: {avirulent or attenuated}
- A61K 2039/523: {expressing foreign proteins}
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

The present invention encompasses recombinant bacteria and immunogenic compositions comprising the bacteria. The immunogenic composition may be used to induce an immune response against C. perfringens.

## Claims

### Claim 1

A recombinant attenuated Salmonella enterica bacterium, wherein the bacterium is capable of expression of one nucleic acid encoding at least two protein antigens of Clostridium perfringens, or wherein the bacterium is capable of expression of more than one nucleic acid, each encoding a Clostridium perfringens protein antigen, wherein the C. perfringens protein antigen(s) are selected from PlcC (C-terminal domain of C. perfringens alpha toxin), NetB, GDP (glyceraldehyde 3-phosphate dehydrogenase), PFOR (pyruvate:ferredoxin oxidoreductase), FBA (fructose 1,6-biphosphate aldolase), HP (a C. perfringens hypothetical protein), and a fragment thereof that is capable of eliciting an immune response, wherein the bacterium upon administration to a mammal or a bird, elicits an immune response against C. perfringens in the mammal or the bird.

### Claim 2 (depends on 1)

The recombinant attenuated Salmonella enterica bacterium of claim 1, wherein the bacterium is capable of colonizing the gut of the mammal or the bird upon oral administration.

### Claim 3 (depends on 1)

The recombinant attenuated Salmonella enterica bacterium of claim 1, wherein the bacterium further comprises the □pmi-2426 mutation and is capable of eliciting an immune response against at least two Salmonella enterica serotypes.

### Claim 4 (depends on 1)

The recombinant attenuated Salmonella enterica bacterium of claim 1, wherein the codons of the nucleic acid encoding the antigen(s) have been modified so as to optimize the expression level of the antigen(s).

### Claim 5 (depends on 1)

A vaccine composition comprising the recombinant attenuated Salmonella enterica bacterium of claim 1.

### Claim 6 (depends on 5)

A method of inducing an immune response against Clostridium perfringens, the method comprising administering the vaccine composition of claim 5 to a mammal or a bird.

### Claim 7 (depends on 6)

The method of claim 6, wherein the recombinant attenuated Salmonella enterica bacterium is Salmonella enterica serovar Typhimurium and the bird is a commercial poultry bird.

### Claim 8

A recombinant attenuated Salmonella enterica bacterium, wherein the bacterium is capable of expression of one nucleic acid encoding at least three protein antigens of Clostridium perfringens, or wherein the bacterium is capable of expression of more than two nucleic acids, each encoding a Clostridium perfringens protein antigen, wherein the C. perfringens protein antigen(s) are selected from PlcC (C-terminal domain of C. perfringens alpha toxin), NetB, GDP (glyceraldehyde 3-phosphate dehydrogenase), PFOR (pyruvate:ferredoxin oxidoreductase), FBA (fructose 1,6-biphosphate aldolase), HP (a C. perfringens hypothetical protein), and a fragment thereof that is capable of eliciting an immune response, wherein the bacterium upon administration to a mammal or a bird, elicits an immune response against C. perfringens in the mammal or the bird.

### Claim 9

A recombinant attenuated Salmonella enterica bacterium, wherein the bacterium is capable of expression of one or more than one nucleic acid, which encodes at least three protein antigens of Clostridium perfringens, wherein the protein antigens are PlcC (C-terminal domain of C. perfringens alpha toxin), NetB, and FBA (fructose 1,6-biphosphate aldolase) and wherein the bacterium upon administration to a mammal or a bird, elicits an immune response against C. perfringens in the mammal or the bird.