Cryptosporidiosis vaccine
The invention is based on the finding that incubating a Cryptosporidium gp40 protein with an aziridine, significantly increases its immunogenicity. When used as a vaccine, this allows a reduction of the dose, which improves economic feasibility and safety. Consequently the aziridine-treated gp40 can now be used as a safe and effective subunit-vaccine for humans or non-human-animals against Cryptosporidiosis. Specifically for new-born ruminants a vaccination by way of colostral transfer was found to be very effective in reducing clinical signs of Cryptosporidiosis, especially diarrhoea.
1. A Cryptosporidium gp40 protein, wherein that the gp40 protein comprise one or more amino acids alkylated by an aziridine.
2. The gp40 protein of claim 1 , wherein the alkylated amino acid is alkylated with an alkyl group of Formula (2):
wherein R1 is selected from the group consisting of: H, alkyl, alkylsulfonyl, mesyl, tosyl, nosyl, brosyl, alkenyl, alkynyl, alkylaryl, arylalkyl, and cycloalkyl, wherein each of the alkyl, alkenyl, alkynyl, alkylaryl, arylalkyl, and cycloalkyl is optionally substituted with a substituent selected from the group consisting of: carbonyl, hydroxyl, alkyl, and haloalkyl;
wherein R2′ and R2″ are each independently selected from H and alkyl; and
wherein R3′ and R3″ are each independently selected from the group consisting of H and alkyl.
3. The gp40 protein of claim 1 , wherein the alkyl group of Formula (2), has one of the combinations of substituents selected from the group consisting of:
R1 is C(═O)CH3, R2′ is H, R2″ is H, R3′ is H, and R3″ is H;
R1 is H, R2′ is CH2CH3, R2″ is H, R3′ is H, and R3″ is H;
R1 is H, R2′ is CH3, R2″ is H, R3′ is H, and R3″ is H;
R1 is CH2CH2OH, R2′ is H, R2″ is H, R3′ is H, and R3″ is H; and
R1 is H, R2′ is C(CH3)3, R2″ is H, R3′ is H, and R3″ is H.
4. The gp40 protein of claim 1 , wherein the alkylated amino acid is one or more selected from the group consisting of: cysteine, methionine, serine, threonine, tyrosine, lysine, arginine, valine, glutamic acid, and aspartic acid.
5. The gp40 protein of claim 4 , wherein the alkylated amino acid is one or more selected from the group consisting of:
valine, glutamic acid, and aspartic acid.
6. The Cryptosporidium gp40 protein of claim 1 , wherein one or more or all of the features are applied, selected from the group consisting of:
the aziridine is ethylenimine or is binary ethylenimine,
the Cryptosporidium gp40 is from Cryptosporidium parvum,
the composition comprising the Cryptosporidium gp40 protein is the supernatant or filtrate from a baculovirus-insect cell expression system culture, and
said supernatant or filtrate is purified by column chromatography.
7. A vaccine for a human-or non-human animal target against Cryptosporidiosis, said vaccine comprising the Cryptosporidium gp40 protein of claim 1 , and a pharmaceutically acceptable carrier.
8. The vaccine of claim 7 , wherein the vaccine further comprises an adjuvant.