Method to prevent fertilization in mammals by administering a single dose of zona pellucida derived antigens, liposome and adjuvant

a technology of zona pellucida and antigens, applied in the field of vaccine compositions for immunocontraception of mammals, can solve the problems of permanent sterility, surgical sterilization, culling, etc., and achieve the effect of prolonging the period of antibody production and prolonging the period of contraception

Inactive Publication Date: 2001-06-12
IMMUNOVACCINE TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention relates to an immunocontraceptive vaccine preparation which comprises zona pellucida antigens incorporated into a liposome delivery system. The liposome system effects the slow release of antigen resulting in an extended period of antibody production and thereby an extended period of contraception. Therefore, the present invention provides a method to achieve immunocontraception of mammals using a single injection of zona pellucida glycoproteins.
In one embodiment of the present invention, the liposome vaccine composition is freeze-dried and incorporated into a BALLISTIVET biobullet. Such an embodiment makes the vaccine easier to deliver to the animal.

Problems solved by technology

Methods such as surgical sterilization, or more drastically, culling are generally not acceptable or even allowable in most countries.
Unfortunately, the increasing population of seals is eating away at the diminishing fish stocks which poses serious problems for the fishing industry.
Secondly, the infertility caused by anti-ZP antibodies is reversible, although, hyper- immunization may cause permanent sterility.
Multiple injections are clearly not practical for wild populations as it entails recapturing the same wild animal each time an injection is necessary.
Multiple injections are also cumbersome in any situation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

experiment 1

Production of Anti-ZP Antibodies in Rabbits

Protocol.

Rabbits were injected i.m. (two rabbits for each treatment, details of each treatment are given in Table 1) as follows (a) ZP in three injections at monthly intervals, (b) ZP with Freund's complete adjuvant plus two boosters with ZP in Freund's incomplete adjuvant at 4 and 8 weeks and (c) ZP with muramyl dipeptide adjuvant (Sigma chemicals). In a second series, rabbits were immunized as above but ZP, plus adjuvant where applicable, were encapsulated in liposomes and in all cases in the second series only a single injection was administered. Preimmunization serum samples were taken from all rabbits. Titers of antibodies were measured by Elisa assay using both SIZP and ZP3 as antigen.

Results

The results of this study are illustrated in Table 1. Immunization of rabbits with ZP (no adjuvant) raised a low antibody titer after the third injection. A single injection of ZP in liposomes with no adjuvant raised no detectable antibodies. Use ...

experiment 2

Immunization of Captive Seals

Protocol.

Captive female grey seals (13) were divided into six groups of two animals each, except for group d which comprised three animals. The groups were immunized with a single injection i.m. as follows:

(a) ZP (15 .mu.g) in liposomes with Freund's complete adjuvant (FCA)

(b) ZP (90 .mu.g) in liposomes with FCA

(c) ZP (90 .mu.g) in FCA

(d) ZP (90 .mu.g) in liposomes with TITERMAX*

(e) ZP (90 .mu.g) in TITERMAX

(f) ZP (90 .mu.g) in liposomes with BCG adjuvant in water

All animals received in injection having a volume of 1.0 ml. Bacillus Calmette-Guerin or BCG is one component of FCA. Liposomes were formulated as previously described. TITERMAX (0.5ml) mixed with saline (0.5 ml) containing SIZP or liposomes (0.5 ml) containing SIZP as described by the manufacturer. TITERMAX (Cytrx Corporation, Norcross, Ga.) is a new adjuvant system which claims to induce higher titers of antibodies than FCA with a single injection while being less toxic to animals.

Results

The r...

experiment 3

Immunization of Harbour Seals

Twelve harbour seals, divided into three groups of four, were immunized with ZP in a single injection (Table 3). The seals were immunized immediately after giving birth. Samples were taken in the two or three week period until the seals left the pupping area. The results are shown in Table 3. Immunization (with 30 and 90 .mu.g ZP) of harbour seals that had just given birth resulted in anti-ZP titers of only 9% (range 6 to 12%) of immunocontraceptive levels after 3 weeks. As discussed previously, anti-ZP antibodies can act

TABLE 2 Anti-ZP Antibody Production by Captive Seals. Anti-ZP titer (% of standard serum)* Time (months) Antigen 1 2 3 4 5 6 7 8 9 10 11 12 13 16 18 20 22 (a) ZP (15 .mu.g) 4 70 60 155 146 141 136 94 34 20 -- -- -- -- -- -- -- FCA / liposomes 3 92 166 216 192 182 224 185 150 117 135 95 87 23 35 20 18 (b) ZP (90 .mu.g) 1 66 206 186 74 169 125 103 64 67 59 68 20 10 3 5 5 FCA / liposomes 18 78 230 227 230 247 223 164 127 122 115 74 31 14 10 7 4...

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Abstract

A vaccine for the immunocontraception of mammals is described. The vaccine consists of zona pellucida antigens and an adjuvant encapsulated in a liposome delivery system. The liposome delivery system allows for the slow release of antigen resulting in a prolonged immune response. In particular, after a single injection of the vaccine, levels of anti-zona pellucida antibodies were detected for up to 22 months in seals. Thus, the vaccine according to the present invention is effective after a single dose and is therefore very useful in immunocontraceptive protocols.

Description

FIELD OF THE INVENTIONThe present invention relates to a vaccine composition for the immunocontraception of mammals.BACKGROUND OF THE INVENTIONThere is a real need for population control in several species of domestic and wild animals. Methods such as surgical sterilization, or more drastically, culling are generally not acceptable or even allowable in most countries. For example, in Canada the culling or harvesting of seals was prohibited in the early 1980's, resulting in an increased seal population from 10,000 in 1978 to approximately 45,000 at present. Increases in harp seal populations have been much greater. Unfortunately, the increasing population of seals is eating away at the diminishing fish stocks which poses serious problems for the fishing industry. The seals also contain parasites such as seal worms that they pass on to the fish. In 1986 it was estimated that the cost to the fishing industry of removing seal worms from fish by hand was upwards of $30 million a year. Th...

Claims

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Application Information

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Patent Type & Authority Patents(United States)
IPC IPC(8): A61K39/00A61K9/127A61P15/18
CPCA61K9/127A61K39/0002A61K39/0006A61P15/18
Inventor BROWN, ROBERTMEZEI, MICHAELMEZEI, CATHERINEPOHAJDAK, BILLKIMMINS, WARWICK CHARLES
Owner IMMUNOVACCINE TECH INC
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