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WO2011079627A1 - Adjuvant à composants multiples d'un vaccin par immersion pour poissons, son application et sa méthode d'utilisation - Google Patents

Adjuvant à composants multiples d'un vaccin par immersion pour poissons, son application et sa méthode d'utilisation Download PDF

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Publication number
WO2011079627A1
WO2011079627A1 PCT/CN2010/076978 CN2010076978W WO2011079627A1 WO 2011079627 A1 WO2011079627 A1 WO 2011079627A1 CN 2010076978 W CN2010076978 W CN 2010076978W WO 2011079627 A1 WO2011079627 A1 WO 2011079627A1
Authority
WO
WIPO (PCT)
Prior art keywords
fish
vaccine
adjuvant
saponin
triton
Prior art date
Application number
PCT/CN2010/076978
Other languages
English (en)
Chinese (zh)
Inventor
王秀华
黄倢
王玉娟
赵伟伟
Original Assignee
中国水产科学研究院黄海水产研究所
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 中国水产科学研究院黄海水产研究所 filed Critical 中国水产科学研究院黄海水产研究所
Publication of WO2011079627A1 publication Critical patent/WO2011079627A1/fr

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
    • A61K2039/552—Veterinary vaccine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55577—Saponins; Quil A; QS21; ISCOMS

Definitions

  • the invention relates to a marine biological disease immunodeficiency control technology, in particular to a multi-combination adjuvant for immunizing fish with a immersion vaccine, and an application and method thereof.
  • the object of the present invention is to provide a highly effective multi-group adjuvant for fish immersion vaccines.
  • N-acetylcysteine (NAC) and Triton X-100 (TRTTONX-100) have the ability to dissolve mucus on fish, and saponin has improved mucus immunity.
  • NAC N-acetylcysteine
  • TRTTONX-100 Triton X-100
  • the effect of cellular immunity, while racemic anisodamine (also known as 654-2) can relax muscles and facilitate the penetration of vaccine antigens into the body.
  • the four adjuvants listened to and combined with the 3 ⁇ 41 ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ , , , , , , il 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654 654
  • the multi-combination adjuvant for fish immersion vaccine of the present invention includes NAC, TRITON X-100 saponin and 654-2, wherein the range of NAC, TRITON X-100 saponin and 654-2 is 0.1-10 Mg L, 0.1-10 mg L, 0.1-50 mg L, 1-60
  • the present invention has a concentration of 0.1-10 mg/L, 0.1-10 mg L, 0.1-50 mg L, 1-60 mg L of NAC, TRITON X-100, saponin and 654-2 immunomulti-adjuvant adjuvant. Synergistic effects in the immersion vaccine of seawater-stained water fish to achieve high efficiency immunity -3 ⁇ 4 ⁇ 4.
  • the shed method of the present invention is as follows:
  • the fish to be immunized are transferred into a fine aqueous solution of NAC and TRITON X-100 in a concentration range of 0.1-10 mg/l, pre-soaked for 1-20 min, and then added with a concentration of 0.1-50 saponins and used.
  • Vaccine soak for 1 to 20 minutes, and then into the vaccine soaking solution to force the concentration of 1-60 654-2 to l-10min to complete the j3 ⁇ 43 ⁇ 4 epidemic.
  • the positive of the present invention lies in: using NAC, TRITON X-100, saponin, 654-2 multiple agents as a vaccine combination adjuvant can significantly improve the immunity of the vaccine - 3 ⁇ 43 ⁇ 4 is significantly better than a single adjuvant; Simple, n, safe for the shed, has no effect on the survival rate of the fish and the raw meal. > And the process and the process are not necessary, and it is also harmless to the ringworm.
  • the multi-combination adjuvant of the fine immersion vaccine of the present invention is characterized in that the multi-component adjuvant as a fish immersion vaccine includes NAC, TRITON X-100, saponin and 654-2, wherein NAC, TRITON X-100, saponin
  • the range of glucosides of glucosides and 654-2 is 0.1-10 mg L, 0.1-10 mg/L, 0.1-50 mg L, and l-60 mg/L, respectively.
  • the present invention has a concentration of 0.1-10 mg/L, 0.1-10 mg L, 0.1-50 mg L, 1-60 mg L of NAC, TRITON X-100, saponin and 654-2 immunomulti-adjuvant adjuvant. Synergistic effects in the immersion vaccine of seawater-stained water fish to achieve high efficiency immunity -3 ⁇ 4 ⁇ 4.
  • the shed method of the present invention is as follows:
  • the fish to be immunized are transferred into a fine aqueous solution of NAC and TRITON X-100 in a concentration range of 0.1-10 mg/l, pre-soaked for 1-20 min, and then added with a concentration of 0.1-50 saponins and used.
  • Vaccine immersion immunization for 1-20 min, then add 654-2 with a concentration of 1-60 to the vaccine soaking solution to maintain l-10 min.
  • dissolved oxygen in the water when i3 ⁇ 4 ⁇ 7m or more, ⁇ female $1 ⁇ 2 ⁇ 2 °C. After the game is immune, it is difficult to move the fish into the dance.
  • NAC and TRTTON-100 are used in a concentration range of 0.1-10 mg/L, and the soaking time is 3-50 min; the concentration of saponin is 0.1-50 mg L, the soaking time is 2-30 min; The concentration range is l-60mg L, and the soaking time is l-10min.
  • the ship is used for marine fish, 3 ⁇ 4feK fish and semi-fish.
  • Vaccine preparation Vanguillarum was inoculated into 2216E liquid medium, cultured at 28 ° C for 24 h; centrifuged at 4000 ip for 20 min at 4 ° C, and the cells were collected; the collected cells were placed in sterile PBS, pH 7.2, Made of 1.0 ⁇ 108 cells/mL of Vibrio suspension, 0.5% After being inactivated at 28 °C for 48 hours, it was used as a vaccine for i. It was stored at 4 °C. The inactivated cells were inoculated on TSB solid plate, and cultured at 28 °C for 24 hours and 48 hours. .
  • Immersion immune adjuvant NAC, TRITON X-100 saponin, 654-2, shed concentration of 0.1mg L, 0.1mg L, 50mg L, lmg L.
  • Group i Inch immunization group, no adjuvant vaccine soaking immunization group, Zuoqi vaccine immersion sensitivity, injection blank control group, soaking blank control group, each group was parallel, each group 3x30 fiber.
  • Table 1 uses NAC, TRITON X-100, saponin, 654-2 as the combination of Zuoqi warm bubble to avoid immune immunization relative to ⁇ 3 ⁇ 4 ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ ⁇ 0 death rate 0 / 0 RPS% average RPS%
  • Example 2 Application of the present invention to a concentration of 5 m, 5 m, lm, 30 mg L of NAC, TRITON X-100 saponin, 654-2 multi-adjuvant adjuvant and inactivated P. aeruginosa vaccine to Luo Fei Fish soaking immunity 1 material:
  • Aeromonas punctatus was inoculated into LB liquid medium and cultured at 28 ° C for 24 h ; The bacteria were collected by centrifugation at 4000 ipm for 20 min at C; the collected cells were placed in a sterile sterilized solution, pH 7.2, and made into a suspension of 1.0 X 109 cells/mL, and 0.5% formaldehyde was inactivated at 28 ° C. After 48 hours, the test vaccine was stored at 4 ° C. At the same time, the inactivated cells were inoculated on LB solid plates, and cultured at 28 ° C for 24 h and 48 h, and it was confirmed that no cells grew.
  • Immersion immunoadjuvant NAC, TRITON X-100 saponin, 654-2 combination adjuvant, 5m, 5mg L lmg/L 30mg ⁇ L.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Farming Of Fish And Shellfish (AREA)

Abstract

La présente invention concerne un adjuvant à composants multiples d'un vaccin par immersion pour poissons, comprenant de la N-acétylcystéine, du Triton X-100, de la saponine et de la raceanisodamine. Les concentrations de travail desdits composants sont, respectivement, de 0,1 à 10 mg/l, de 0,1 à 10 mg/l, de 0,1 à 50 mg/l et de 1 à 60 mg/l. L'invention concerne également l'application dudit adjuvant dans une immunisation par immersion pour poissons et sa méthode d'utilisation.
PCT/CN2010/076978 2009-12-31 2010-09-16 Adjuvant à composants multiples d'un vaccin par immersion pour poissons, son application et sa méthode d'utilisation WO2011079627A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN200910255826.1 2009-12-31
CN2009102558261A CN101791401B (zh) 2009-12-31 2009-12-31 鱼用浸泡疫苗多组合佐剂及其应用和使用方法

Publications (1)

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WO2011079627A1 true WO2011079627A1 (fr) 2011-07-07

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WO (1) WO2011079627A1 (fr)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101791401B (zh) * 2009-12-31 2012-04-18 中国水产科学研究院黄海水产研究所 鱼用浸泡疫苗多组合佐剂及其应用和使用方法
CN105816874A (zh) * 2016-03-21 2016-08-03 沈阳工学院 一种鱼用浸泡免疫复合疫苗佐剂及其应用方法
CN105833266A (zh) * 2016-04-26 2016-08-10 肇庆大华农生物药品有限公司 一种提高渔用浸泡疫苗免疫效果的佐剂及鱼类浸泡免疫方法
CN109568575A (zh) * 2018-12-02 2019-04-05 河南师范大学 一种增强嗜水气单胞菌OmpA疫苗接种效果的小分子代谢物佐剂及其应用

Citations (2)

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WO2007068907A2 (fr) * 2005-12-13 2007-06-21 Glaxosmithkline Biologicals Sa Nouvelle composition
CN101791401A (zh) * 2009-12-31 2010-08-04 中国水产科学研究院黄海水产研究所 鱼用浸泡疫苗多组合佐剂及其应用和使用方法

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CN1568678A (zh) * 2004-05-12 2005-01-26 中国水产科学研究院黄海水产研究所 一种鱼类疫苗的浸泡免疫接种方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007068907A2 (fr) * 2005-12-13 2007-06-21 Glaxosmithkline Biologicals Sa Nouvelle composition
CN101791401A (zh) * 2009-12-31 2010-08-04 中国水产科学研究院黄海水产研究所 鱼用浸泡疫苗多组合佐剂及其应用和使用方法

Non-Patent Citations (2)

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Title
LIU ZHENXING ET AL: "Adjuvants used in aquaculture", RESERVOIR FISHERIES, vol. 27, no. 2, 2007, pages 3 - 6 *
ZHANG NIANCI ET AL: "Supplementary effect of henbane on the bathing immunization of summer fingerlings of grass carp (Ctenopharyngodon idellus)", JOURNAL OF ZHEJIANG COLLEGE OF FISHERIES, vol. 9, no. 2, November 1990 (1990-11-01), pages 79 - 84 *

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CN101791401B (zh) 2012-04-18

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