TWI355940B - Vaccine adjuvant and vaccine composition - Google Patents

Description

135.5940 IX. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to a vaccine adjuvant' and particularly relates to a vaccine oily adjuvant containing acid hydrolyzed dextran.

[Prior Art J Adjuvant refers to any substance that increases the immune response to antigenic humor and/or cells. Traditional vaccines are prepared by the use of killed pathogenic microorganisms, and impurities associated with pathological microbial culture can be used as adjuvants to enhance the immune response. However, when a pathological microorganism or a purified protein-unit is used as an antigen, 'because of the immunogenicity caused by this antigen, it is necessary to add an additional foreign substance as an adjuvant. In addition, since both synthetic and subunit vaccines are very expensive to produce, antigen doses can be reduced by adjuvants to save vaccine production costs. The mechanism of action of the adjuvant is extremely complicated and is not fully understood at present. The main # thinks that (1) the adjuvant is mixed with the aqueous solution to form water-in-oil or oil-in-water (〇il-in-water). Milky particles to increase the contact area of antigen with macrophages; (2) adjuvant can prolong the retention time of antigen in vivo; (3) adjuvant can induce inflammation of the antigen injection site and its local lymph nodes, Use to stimulate the proliferation of immune cells. At present, adjuvants can be divided into aluminum glue and oil adjuvant, and oil adjuvants can be divided into W/0 emulsion type, 0/W emulsion type and W/0/W emulsion type. W/0 emulsion-type adjuvants, for example, Freund's complete and incomplete adjuvants, are a common adjuvant that prolongs the residence time of antigens within tissues and can persist

Client’s Docket No.: TT's Docket No:0973-A41273-TW/fmal/Kai/7/5/2007 1.355940 Inflammatory response, but its viscosity is too high, prone to granuloma or abscess. 0/W emulsion type has low viscosity, so there is no serious local reaction at the injection site, and it can induce a short-term immune reaction with high safety. However, since the antigen is in the continuous aqueous phase, it is rapidly spread at the injection site after the injection, and the retention time is short, and it is difficult to exert the effect of long-term immunity. Although the 'W/0/W emulsion type has the advantages of both W/O emulsion type and 0/W emulsion type, its price is too high, and the antibody yield is still insufficient. Therefore, the development of vaccine adjuvants that increase protection, protection, and safety is an important issue at present. Generally, oily adjuvants often need to be combined with an expensive and cytotoxic immunopotentiator to provide two adjuvant effects, such as muramyl dipeptide, CpG and bacterial nucleic acids, cytokines, and exotoxins. Endotoxin and the like. Β-glucan is a natural polysaccharide that is widely present in the cell wall of plants and is easy to obtain. Β-glucan is a high molecular weight polysaccharide. It is generally extracted and purified from fungi or plants. After drying and grinding into powder, it is hardly soluble in cold water. After water coating, it is easy to produce agglomeration. The difficulty in using β-glucan also reduces the immune effect. The present invention utilizes acid hydrolysis to degrade the β-glucan molecule to reduce the molecular weight. The β-glucan is dissolved in the aqueous phase of the adjuvant to increase and enhance the immunopotency. SUMMARY OF THE INVENTION A vaccine adjuvant is provided, including - acid hydrolyzed sugar ((4) ns), which is dispersed in an aqueous phase solution, the aqueous phase solution _ emulsifier and oil Equilibrium emulsification (4) 彡成诚料剂, 1 The weight average molecular weight of the ruthenium-free ruthenium

Client's Docket No.: ΤΤ^ Docket No:0973-A41273-TW/final/Kai/7/5/2007 1355940 〇_, and the acid-hydrolyzed μ screaming degree is on ^_. The present invention further provides a vaccine composition comprising at least, - comprising - a biologically acceptable effective antigen and - lyophile W; and - a fat, wherein the aqueous phase solution is homogenized with the fat = to form the vaccine The composition, the weight of the acid hydrolysis __; = the amount between about 丨 丨 to 丨 __, and the concentration of the glucosamine in the tank is above 0.1 mg / ml. The above and other objects, features, and advantages of the present invention are set forth in the <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; It provides a vaccine adjuvant, including an acid-hydrolyzed compartment, __cans), which can increase the protective value of the vaccine and prolong the vaccine's time-keeping. The "hydrolyzed (four) poly_♦_) according to the present invention refers to a molecular weight obtained by acid hydrolysis of ruthenium (IV), which has a weight average molecular weight of from about 1 〇〇〇〇 to face (four). Between 2 and 5 _ _ Μ Μ 是 Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ Μ. The /3-glucan of the present invention includes /Μ, 3-glucanase, 州···················································· , small ^, rye, buckwheat, corn, rice or millet, etc.), category (Lingzhi, Qizhizhi, Yunzhi, Cordyceps sinensis, Sang Brazil

Client's Docket No.: TT's Docket No:0973-A41273-TW/fmal/Kai/7/5/2〇〇7 1355940, willow mushroom, coral mushroom or shiitake mushroom, etc. or yeast, preferably barley]8- Glucan. In the present invention, the granules are first ground into a powder, and the granule powder is treated with ethanol, and then an appropriate concentration of amylase, proteolytic enzyme or the like is added to remove starch and protein in the barley. Next, the non-glucan was precipitated by using a high concentration of ethanol, and the precipitated cold-dextran precipitate was washed with ethanol. The concentration of ethanol may range from 80 ° / ❶ to 95% (ν / ν). The cleaning step can be repeated 2-3 times. Finally, the stone-dextran was vacuum dried. For the purification procedure of non-glucan, please refer to Xu Rongcheng (20〇4) “The effect of different molecular weight barley dextran on the physicochemical properties of rice starch”, Master's thesis “National Taiwan University and other literature. The /5-glucan is dissolved in water and added to the acidic solution for several tens to several hundred minutes, for example, 20 to 150 minutes. Next, /?-glucan was precipitated with a high concentration of ethanol and the precipitated sputum-dextran precipitate was washed. The concentration of ethanol can range from 80% to 95% (v/v). The cleaning step can be repeated 2-3 times. Finally, it is vacuum dried by acid hydrolysis/5-glucan. For the acid hydrolysis procedure for dextran-free dextran, see Doubleier, J. L. and Wood Wood, P. J. (1995)

Cereal Chem. 72:355·34〇 and other literature. Those skilled in the art can appropriately change the steps and conditions of purification and hydrolysis according to the source and hydrazine of 鲁 々 葡. The acid hydrolyzed dextran is dispersed in the aqueous phase. The aqueous phase solution may be a general food or pharmaceutical sigma component such as an amino acid, a salt, a sugar or a sugar alcohol. For example, the amino acid or a salt thereof may be a glycinate, an alanine: a arginine, a histidine, a phenylalanine, an aspartate, a sodium citrate, (10) Na salt, Lin potassium salt, and its hydrate. The sugars and sugar alcohols may be selected from the group consisting of trehalose, xylitol, mannitol, maltose pure, and lactitol.

Client’s Docket No.: TT*s Docket No: 0973-A41273-TW/fmal/Kai/7/5/2007 1355940 and so on. Other ingredients include various phosphates of the buffer solution. In addition, the aqueous phase solution (or antibody containing) can be a solution or suspension. The aqueous phase solution described above is emulsified and emulsified with an emulsifier and a fat to form an oil adjuvant. The fats and oils may be light liquid paraffin (mineral oil), edible oil, cod liver oil, squalene, squalane, monoglyceride, diglyceride, triglyceride, and the like. The oil phase emulsifier may be Alarcel P-135, Alarcel 83, Alarcel A, Span 80, Span 83, Span 85, and its HLB value is preferably less than 10. The aqueous phase emulsifier may be Tween 20, Tween 80, Tween 85, etc., and its HLB value is preferably greater than 1 Torr. The adjuvant of the present invention may be in various emulsified forms such as 'water in 〇丨1, w/O type, oil in water (0/W) type or double emulsified (d〇) There are no particular restrictions on the type of uble einuisi〇n, w/O/W). For the emulsification step, refer to Carstensen JT. Disperse systems. In. In: Theory of pharmaceutical systems II. Heterogeneous systems· New York: Academic Press; 1973. Those skilled in the art can vary the steps and conditions of emulsification according to the type of dispersant and the type of emulsification. In another embodiment, the grease of the present invention further comprises a nonionic block copolymer, for example, an ethylene oxide-propylene oxide copolymer (E〇_p〇copolymer), ethylene oxide-styrene. Copolymer, propylene oxide-styrene copolymer, styrene-acrylonitrile copolymer, styrene-butadiene copolymer. The nonionic block copolymer further promotes the immune response to increase the protective cost. The vaccine adjuvant of the invention can promote the immune response of the animal, so as to increase the protective value of the vaccine and prolong the protection time, without adversely affecting the animal.

Client's Docket No.: TT's Docket No:0973-A41273-TW/final/Kai/7/5/2007 9 1355940 The present invention further provides a vaccine composition: an effective antigen and a And (2) and oil, wherein the aqueous phase material is homogeneously emulsified with the oil to form the vaccine composition, and the weight of the anaerobic hydrazine (4) is between (4) and 1__, preferably 5 sides. Between 3 〇〇 _, and the acid hydrolysis of /5-glucan _ is more than 〇 爪 咖, preferably between 0.3 mg / ml and 3 mg / ml.

The vaccine composition of the present invention is an animal vaccine which is preferably a non-human animal vaccine, for example, a livestock or poultry vaccine. Livestock or poultry is an animal that is domesticated by humans and can be artificially controlled for its use in eating, labor, fur, dragon, experiment, etc. 'It includes' but not limited to pigs, sheep, cattle, deer, dogs , rats, mice, rabbits 'horses, donkeys, mushrooms, turkeys, ducks, geese, turkeys, pigeons or ostriches. The antigen of the present invention may be an attenuated antigen, an antigen or a recombinant antigen, and the antigen may be derived from various livestock or poultry pathogens, for example, Hog cholera virus, Hepesvirus suis, bovine cancer virus. (Rinderpest virus), Marek's disease virus, paramyxovirus, adenoviruses, infectious bursal disease virus, Feline Parvovirus , Canine Parvovirus, Canine distemper vims, Lyssaviruses, bovine parainfluenza virus, Foot-and-mouth disease virus (FMDVchitis) Virus), virus

Client's Docket No.: TT's Docket No: 0973-A41273-TW/fmal/Kai/7/5/2007 10 1355940 parvovirus) or pseudorabies virus (pseud〇rabies virus). The vaccine composition of the present invention can be immunized to animals by intramuscular injection, subcutaneous injection or intraperitoneal injection. When the vaccine composition of the present invention has a good protective value and protection, the protective strength of the &apos;H # a month vaccine composition is 〇 $ to 1.5 times that of a general commercial vaccine. [Examples] 1. /3-Glucan Firstly, the barley grain was ground into a barley flour capable of passing through a 1 mm aperture sieve. 400 g of barley flour was placed in 80% (v/v) ethanol. Minutes, the treated barley flour was obtained by centrifugation at 8000 g for 1 minute, and the treated barley flour was spread on an iron pan and dried overnight at 40X. Take 500 g of treated barley flour and 2000 mi of deionized water in 4 (after 30 minutes of reaction at TC), centrifuge at 8000 g for 2 minutes to obtain supernatant. Take 5 mla_amylase (Temamyl® 12〇, Cat. NO). .A3403, Sigma) was added to the above supernatant to react at 95 ° C for 30 minutes. After the reaction, the water bath was cooled to 5 (TC, and 1 ml of proteolytic enzyme (Fiav〇urZymeTM 500L, Cat. No. P6110, Sigma) was reacted at 50 ° C for 60 minutes, and then boiled in a boiling water bath for 30 minutes to activate the enzyme activity. After centrifugation at 3000 g for 1 minute, an equal volume of 95% (v / v) ethanol was slowly added under vigorous stirring. 4. (: Let stand for 180 minutes to obtain /3-glucan precipitation. Concentrate the dextran precipitate with 2000 ml of deionized water in a homogenizer (17,000 rpm, PT-3000, Polytron, Switzerland) After 2 minutes, the precipitate was uniformly dispersed, and then continuously stirred at 7 ° C for 30 minutes to completely dissolve. The temperature bath was cooled to

Client's Docket No.: TT*s Docket No:0973-A41273-TW/final/Kai/7/5/2007 1355940 3〇C 'After adding an equal volume of 95% (v/v) ethanol slowly under vigorous stirring, The mixture was allowed to stand at 4 C for 180 minutes, and centrifuged at 3 〇〇〇g for 1 minute to obtain a precipitate. The precipitate was homogenized with 8 〇〇mi 95% (v/v) ethanol in a homogenizer for 1-2 minutes to uniformly disperse the precipitate, and then filtered (4 〇〇mesh) and vacuum dried to obtain Barley dextran. 2. Acid hydrolysis / 3-glucan pickle 30 g of barley, - dextran at 7 〇 t: dissolved in 1500 ml of deionized water 'Add 70 ° C equal volume of 〇. 4M HC1, at 70 分别 respectively The reaction was carried out for 30, 60, and 120 minutes. The temperature was quickly reduced to about 3 〇 ac and the pH was adjusted to 6.5-7.0 with 1 M NaOH. An equal volume of 95% (v/v) ethanol was slowly added under vigorous stirring, and allowed to stand at 4 ° C for 180 minutes to obtain a stone-dextran precipitate, and the sediment was collected by centrifugation at 3000 g for 1 minute. Next, 47.5% (v/v) of ethanol was added to the precipitate, and after washing with a homogenizer, the precipitate was collected by centrifugation. Finally, add 1200 ml of 95% (v/v) ethanol to the precipitate, homogenize for 1-2 minutes in a homogenizer, and uniformly disperse the precipitate. After filtration (400 mesh) and vacuum drying, acid can be obtained. Hydrolyzed barley/5-glucan. For the related steps of acid hydrolysis, please refer to Doubleier, JL and Wood Wood, PJ (1995) Cereal Chem. 72:355-340. Reference 0 Referring to Figure 1, as the time of acid hydrolysis increases, barley iS-glucan The longer the residence time, the smaller the molecular weight of the barley-glucan. The molecular weight, purity and recovery of barley/3-glucan are shown in Table 1. Table 1. Molecular weight, purity and recovery of barley f-glucan. Acid hydrolysis. Acid hydrolysis for 60 minutes. Acid hydrolysis for 120 minutes.

Client's Docket No.: TT's Docket No:0973-A41273-TW/fmal/Kai/7/5/2007 1355940 Molecular Weight 280 ± 25 KD 130 ± 15 KD 70 ± 33 KD Purity 78.4 Soil 3.4% 87.5 ± 5.6% 79.5 Soil 6.2 % Recovery rate 0.46% 0.35% 0.21% 3. Wj/O/W, preparation of sputum

Wi/O production: take 30% to 60% phosphate buffer solution 'utilize HLB value below 10 to know emulsifier such as Span 80 (HLB: 4.3, concentration between 2% and 10% dispersed between 70% to 40% Between the mineral oil and heating to 5 CTC. High-speed homogenizer (Polytron PT-MR 3000, Kinematica AG, Littaau, Switzerland, 6000 rpm) first homogenized for 5 min, then with micro-jet high pressure homogenizer (Haskel hochdruck System GmbH ] \4〇(1611^〇:1^1600?101^, 10,000?3丨) is made into a 1/0 type emulsion, which is homogenized for 1, 3, and 5 cycles and sampled for analysis. Production of WVO/W2: Utilization Tween 80 (HLB: 12.1, concentration between 0.1% and 10%) was vacuum-high-speed homogenizer (ΙΚΑ T25, Germany) secondary emulsification (20,000 rpm; 5 min) of the above Wi/O emulsion to form Wi/O/ V^ type composite emulsion. The above emulsification process is carried out at room temperature, if necessary, using cold water to cool down, and each component is sterilized according to standard sterilization method. 4. Mouse immunization Π) Take 48 Balb/c mice, divided into 8 Group, each group of 6. The experimental conditions of each group are shown in Table 2. The W/0/W emulsions shown in Table 2 were exempted every two weeks by subcutaneous injection. A mouse, the second co-immunoprecipitation, each dose of 0.3ml. Two weeks after each immunization to enzyme-linked immunosorbent assay (ELISA)

Client's Docket No.: TTss Docket No: 0973-A41273-TW/final/Kai/7/5/2007 13 1355940 The antibody titer of the mouse serum was examined. Referring to Fig. 2, neither Ctll nor Ctl2 could promote the production of antibodies in mice, while the glucan group (Lb, Hb, Lb60, Hb60) can significantly increase the antibody titer, and the hydrolyzed glucan enhances the antibody titer. The ability is greater than the unhydrolyzed / 3 - glucan. In addition, the /3-glucan of the present invention is more capable of promoting antibody production than the commercially available 206 adjuvant (Seppic, France), MVP Emulsigen adjuvant (MVP, USA). Table 2, Experimental condition group of Example 4 Ctll Bovine serum albumin (antigen) Ctl2 Bovine serum albumin + L121 nonionic block copolymer MVP Bovine serum albumin + MVP Commercial adjuvant (MVP, USA) 206 Bovine blood · Qing Protein +206 commercially available adjuvant (Seppic, France) Lb bovine jk albumin + unacidified 0.3 mg/ml /3-glucan Hb bovine jk albumin + unacidified 3 mg/ml /3- Glucan group Lb60 Bovine serum albumin + acid hydrolysis for 60 minutes 0.3 mg/ml /3-glucan group Hb60 Bovine serum albumin + acid hydrolysis for 60 minutes 3 mg/ml /3-glucan group 5. Mice Exemption (2) The procedure of this example is the same as that of Example 4, and only the experimental conditions of each group are changed, as shown in Table 3. Referring to Figure 3, the Ctll group failed to promote antibody production in mice. The barley/?-glucan group (Lb60, Hb60) and yeast/3-glucan all significantly increased the antibody titer. Table 3, Experimental conditions of Example 5

Client's Docket No.: TT^ Docket No:0973-A41273-TW/final/Kai/7/5/2007 14 1355940 Group cm Cow jk albumin ion incorporation copolymer Jkr Bovine serum albumin + unacidified 〇3 Mg/ml f glucan group _ _ bovine serum albumin + unacidified 3 mg/ml glucan group Lb60 bovine serum albumin + acid hydrolysis for 60 minutes 0.3 mg/ml /3-glucan group Hb60 cattle Serum protein + acid hydrolysis for 60 minutes 3 mg / ml / 3 - glucan group ^ ~ ~ sales. $ 8 specific pathogens (spf, specific pathogen free), divided into 3 groups, respectively Negative control group, positive control group and & dextran group. The negative control group was not immunized, and the positive control group was immunized with the MVP Emulsigen adjuvant mixed with the swine fever non-activated virus antigen. The non-glucan/w adjuvant group was mixed with the swine fever non-activated virus antigen. The (3 mg/ml) acid hydrolyzed heart glucan 0/W adjuvant of the present invention was immunized. Each pig was immunized once every two weeks by subcutaneous injection of Lu, and was immunized three times with a dose of 2 ml each. The antibody titer of the pig serum was measured by enzyme-linked immunoassay (ELISA) two weeks after each immunization. Referring to Table 4, the antibodies produced by the glucan group of the present invention have the ability to protect pigs, and their antibody titers are greater than those of the other two groups. Table 4, antibody strength of each group of pigs, pig numbering group P0 P1 P2 P3 1 negative control group &lt; 4 &lt; 4 &lt; 4 &lt; 4 2 negative control group &lt; 4 &lt; 4 &lt; 4 &lt ;4

Client's Docket No.: TT5s Docket No:0973-A41273-TW/fmal/Kai/7/5/2007 15 1355940 3 Positive control group &lt;4 &lt;4 8 32 4 Positive control group &lt;4 &lt;4 6 8 5 positive control group &lt;4 &lt;4 23 362 6 /3-glucan group &lt;4 &lt;4 45 128 7 /3-glucan group &lt;4 &lt;4 11 256 8 /3-glucan Sugar group &lt;4 &lt;4 23 &gt; 724 P0: pre-immune; P1: first immunization; P2: second immunization; P3: third immunization 7. chicken sales immunity 36 non-specific diseases The chickens were divided into three groups: negative control group, positive control group and iS-glucan 0/W type adjuvant group. The experimental conditions and results of each group are shown in Table 5. Two to three weeks after immunization of the chicken, blood was collected to detect the HI titer of the erythrocyte agglutination inhibitor (Newcastle disease virus, NDV) in the blood of the chicken. As can be seen from Table 5, both the commercially available adjuvant and the/5-glucan adjuvant of the present invention can promote the antibody titer of the chicken, and the antibody of the iS-glucan adjuvant of the present invention has a higher antibody price than the commercially available one. Adjuvant. Table 5. Immunization conditions and antibody valence of each group of chickens were negative control group or control group / 3-glucan group number 10 16 16 Immune content acid buffer Buffer commercially available w/o Fu adjuvant adjuvant + NDV Antigen 1 | S-glucan adjuvant + NDV antigen 2 pre-immune antibody 22-23 22-23 22-23

Client’s Docket No.: TT's Docket No:0973-A41273-TW/final/Kai/7/5/2007 Antibody price 19 days after immunization

29 (8), 28 (4), 27 (2) 21〇 (2), 29 (6), 28 (2), 26 (4), 2. Ben:: adjuvant + Xincheng chicken plague virus antigen, dosage: ° · 8 (6) dose · _2ιηΓ / W Feng Xincheng virus antigen, the ratio is! · 限定 限定 = = = = = = = = = = = = = = = = = = = = = = ======================================================================================= The order is subject to the definition of Patent Park. [Simple description of the diagram] Figure 1 shows the molecular weight of barley (four) poly (four), by the first! It can be seen that the increase in barley (four) poly_ molecular weight with the increase in hydrolysis (4). Fig. 2 shows the effect of various adjuvants on the immunity of mice. It can be seen from Fig. 2 that the acid-hydrolyzed barley (tetra) polyacetate can increase the antibody titer of mice. Figure 3 shows the effect of various adjuvants on the immunity of mice. As can be seen from Figure 3, the acid-hydrolyzed barley intercropping can increase the antibody cost of small gas. [Main component symbol description]

Client's Docket No.: &lt; S &gt; TT's Docket No:0973-A41273-TW/fmal/Kai/7/5/2007 17

Claims (1)

1355940 J, %. The original case number 096124457 September 15, 2014 Revision This patent application scope 1 · A vaccine adjuvant, including -β-glucan (β-glucans), which is dispersed in the aqueous phase solution 2 ^ The aqueous phase solution is homogeneously emulsified with an emulsifier and a grease to form a vaccine adjuvant, wherein the ρ_glucan is extracted from the steroid, and the weight average molecular weight of the dextran is between about 50,000 and 3 〇. Between mash, and the β-glucan content is above 01 mg/ml. 2. The vaccine adjuvant according to claim 1, wherein the β-glucan is present in an amount of between mgmg/ml and 1〇mg/ml. 3. The vaccine adjuvant according to claim 2, wherein the β-glucan comprises P_U_Glucan, &quot;, cardiac glucan or dextran. The vaccine adjuvant according to claim 1, wherein the mites include barley, oats, wheat, rye, buckwheat, corn, rice or millet. _ 5. The vaccine adjuvant according to the scope of claim 2, wherein the oil component comprises light liquid paraffin (mineral oil), edible oil, cod liver oil, squalene, squalane, monoglyceride, double Acid glyceride or triglyceride. 6. The vaccine adjuvant of claim 1, wherein the emulsifier comprises an oil phase emulsifier or an aqueous phase emulsifier. 7. The vaccine adjuvant of claim 6, wherein the oil phase emulsifier has an HLB value of less than 1 〇. 8. The vaccine adjuvant of claim 6, wherein the oil phase emulsifier comprises Alarcel P-135, Alarce 83, Alarcel A, Span 80, Span 83 or Span 85. 0973A-A41273TWF1 9. The vaccine adjuvant of claim 6, wherein the aqueous phase emulsifier has an HLB value greater than 1 〇. '10. The vaccine adjuvant according to claim 6, wherein the aqueous phase emulsifier comprises Tween 20, Tween 80 or Tween 85 ° 11. The vaccine adjuvant according to claim 1 of the patent application, Wherein the adjuvant is an oily adjuvant. 12. The vaccine adjuvant according to claim 1, wherein the adjuvant comprises water-in-water (w/0) type, oil-in-water (ml ln water, 〇/w) type or Double emulsified bie e signal ^ 丨〇 w / o / w) type. 13. A vaccine composition, at least one aqueous phase solution comprising a beta-glucan, and comprising: a biologically acceptable effective antigen and a fat, wherein the aqueous phase solution is homogeneously emulsified with the oil The composition of the epidemic field, the β-glucan is derived from a moss, and the weight average molecular weight of the glucan is between about lion (9) to 3_(9), and the concentration of the glucan is 0.1 mg. Above /ml, wherein the vaccine composition is a livestock or poultry vaccine. The vaccine composition according to claim 13, wherein the concentration of the poly(4) is from 0.1 mg/rnl to l〇mg/ml. 15. The vaccine composition as described in claim 13 The glycerol comprises (M,3-glucan, β-Μ-glucan or β-M-glucan. HJ ^ 16 · The vaccine composition as described in claim 13 of the patent scope 0973A-A41273TWF1 19, wherein the mites include barley, oats, wheat, rye, buckwheat, corn, rice, or millet. The vaccine composition of claim 13, wherein the fat component comprises light liquid paraffin ( Mineral oil), edible oil, cod liver oil, squalene, squalane, monoglyceride, diglyceride or triglyceride. 18. The vaccine composition as described in the scope of claim 1 ' Livestock includes pigs, sheep, cattle, deer, dogs, cockroaches, mice, rabbits, horses, donkeys or cockroaches. 19. The vaccine composition of claim 13 wherein the poultry includes chickens and ducks , goose, turkey, pigeon or ostrich. 0973A-A41273TWF1 20