Active immunization using a siderophore receptor protein

Inactive Publication Date: 2008-11-27
EPITOPIX LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0009]The vaccine contains one or more siderophore receptor proteins extracted from the outer membrane of a single strain or species, or two or more different strains or species of gram-negative bacteria. The amount and type of siderophore receptor protein included in the vaccine is effective to stimulate production of antibodies reactive with a siderophore receptor protein of one, preferably two or more strains, species or genera of gram-negative bacteria. A preferred vaccine is composed of an amount and profile of siderophore receptor-proteins to e

Problems solved by technology

While vaccines provide adequate immunity for those pathogens against which a flock has been immunized, there are few vaccines which can provide broad-based cross-protection against unanticipated diseases or against those diseases for which an animal has not been specifically vaccinated.
While many vaccines are available for immunization against individual species and serotypes, non

Method used

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  • Active immunization using a siderophore receptor protein
  • Active immunization using a siderophore receptor protein
  • Active immunization using a siderophore receptor protein

Examples

Experimental program
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Effect test

example 1

Production and Purification of Siderophore Receptor Proteins

[0070]Escherichia coli serotype 078 (turkey isolate; serotyped by Pennsylvania State University, deposited with the American Type Culture Collection (ATCC), Bethesda, Md., U.S.A., as ATCC #55652, on Jan. 3, 1995) (700 ml at 108 colonies / ml) was inoculated into a Virtis bench-top fermenter (Virtis, Inc., Gardiner, NY), charged with 20-L of brain-heart infusion (BHI, Difco Laboratories, Detroit, Mich.) containing 50 μgrams / ml of dipyridyl (Sigma Chemical Co., St. Louis, Mo.) at 41° C. This isolate has been shown to produce four siderophore receptor proteins for (MW 89 kDa, 84 kDa, 78 kDa, 72 kDa) under iron-restrictive conditions. The pH was held constant at 7.4 by automatic titration with 5N NaOH. The fermenter was stirred at 400 rpm. The culture was grown continuously for 18 hours after which the bacteria were removed by continuous-flow centrifugation at 20,000×g at 4° C. using a Beckman (Model J2-21M) centrifuge (Beckman I...

example 2

Preparation of Vaccine with Siderophore Receptor Proteins

[0074]The precipitate from Example 1, hereinabove, containing siderophore receptor proteins of E. coli serotype 078, were resuspended in physiological saline (0.85%) containing 0.1% formalin as a preservative. The protein concentration was 300 μg / ml. The aqueous protein suspension (1,000 ml) was emulsified in a water-in-mineral oil adjuvant containing 972 ml Drakeol 6 mineral oil and 28 ml of Anlacel A as an emulsifier. The mixture was emulsified using an Ultra-Turnax T50 emulsifier (KIKA Works, Inc., Cincinnati, Ohio) at 4° C. The water-in-oil emulsion was stored at 4° C.

example 3

Vaccination of Poultry with Siderophore Receptor Protein Vaccine

[0075]Seventy-two turkey poults were raised in isolation from one day of age. At three weeks of age, the birds were divided into two equal groups. Group 1 was vaccinated subcutaneously with the vaccine from Example 2 above, at a dosage level of 150 μg of siderophore receptor protein per bird. Group 2 remained as non-vaccinated controls. Group 1 was given a booster vaccination with the vaccine at a dosage level of 250 μg siderophore receptor protein per bird at 18 days after the first vaccination.

[0076]The vaccinated and non-vaccinated birds were equally divided among four isolation rooms. Rooms A and B contained the vaccinated birds, and Rooms C and D contained the non-vaccinated controls. At seven weeks of age, birds in Groups A and C were challenged subcutaneously with Salmonella agona at 1.0×108 cfu / bird. At 24, 48, 72, 96 and 120 hours post-challenge, two controls and two vaccinated birds were killed. The spleens we...

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Abstract

The invention provides a vaccine for immunizing poultry and other animals against infection by a gram-negative bacteria, and a method of immunizing an animal using the vaccine. The vaccine may contain purified siderophore receptor proteins derived from a single strain or species of gram-negative bacteria or other organism, which are cross-reactive with siderophores produced by two or more strains, species or genera of gram-negative bacteria. The invention further provides a process for isolating and purifying the siderophore receptor proteins, and for preparing a vaccine containing the proteins. Also provided is a method for diagnosing gram-negative sepsis.

Description

BACKGROUND OF INVENTION[0001]The economic impact of infectious diseases in the poultry industry is well-appreciated. Immunization of birds has helped reduce the cost of production by decreasing the incidence of gastrointestinal, respiratory and systemic diseases. While vaccines provide adequate immunity for those pathogens against which a flock has been immunized, there are few vaccines which can provide broad-based cross-protection against unanticipated diseases or against those diseases for which an animal has not been specifically vaccinated.[0002]A number of important diseases of domestic poultry are caused by bacteria able to invade host tissues, such as Salmonella spp., Escherichia spp. and Pasteurella spp. While many vaccines are available for immunization against individual species and serotypes, none provide cross-protection or stimulate broad-based immunity against multiple serotypes, species or genera.[0003]One essential factor required for a bacteria to induce clinical d...

Claims

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Application Information

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IPC IPC(8): G01N33/569A61K39/102
CPCA61K39/0258A61K39/0275A61K39/102G01N33/56911Y10S424/823Y10S424/824Y10S424/826A61P31/04A61P37/04Y02A50/30
Inventor EMERY, DARYLL A.STRAUB, DARREN E.
Owner EPITOPIX LLC
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