WO2015013739A1 - Vaccin atténué contre babesia - Google Patents

Vaccin atténué contre babesia Download PDF

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WO2015013739A1
WO2015013739A1 PCT/AU2014/000750 AU2014000750W WO2015013739A1 WO 2015013739 A1 WO2015013739 A1 WO 2015013739A1 AU 2014000750 W AU2014000750 W AU 2014000750W WO 2015013739 A1 WO2015013739 A1 WO 2015013739A1
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babesia
immunogenic composition
stage
blood
parasites
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Michael Good
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Griffith University
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Griffith University
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    • 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
    • A61K39/002—Protozoa antigens
    • A61K39/015—Hemosporidia antigens, e.g. Plasmodium antigens
    • A61K39/018—Babesia antigens, e.g. Theileria antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/52—Bacterial cells; Fungal cells; Protozoal cells
    • A61K2039/522—Bacterial cells; Fungal cells; Protozoal cells avirulent or attenuated

Definitions

  • TECHNICAL FIELD relates to babesiosis. More particularly, this invention relates to the use of chemically-attenuated babesia parasites as a vaccine against babesia.
  • Babesiosis is a group of diseases and conditions of mammals caused by, or otherwise associated, protozoan protists of the genus Babesia. Babesiosis may occur in mammals such as cattle, buffalo, pigs, deer, dogs, sheep, donkeys, horses and humans, although without limitation thereto. Babesiosis is typically associated with severe haemolytic anaemia and a positive erythrocyte-in-saline-agglutination test indicating an immune-mediated component to haemolysis.
  • Babesiosis Human babesiosis is uncommon although some cases occur due to tick-borne Babesia divergens or Babesia microti, a parasite of small mammals. A few human cases have been fatal- Generally, in non-human mammals the main pathogeic species of Babesia include Babesia bovis, Babesia canis, Babesia bigemena, Babesia divergetis, Babesia ovis, Babesia av ta, Babesia occultans, Babesia eaballi and Babesia motasi.
  • bovine mammals The main parasites of bovine mammals are Babesia bovis, Babesia bigemena and Babesi divergent, which cause hemolytic anemia, such that an infected animal will also show pale to yellow mucous membranes due to the failure of the liver to metabolise excess bilirubin.
  • babesiosis In catde, babesiosis is also known as "tick fever” or "cattle fever” and is economically the most important arthropod-borne disease of cattle worldwide, with cattle populations in most areas of Australia, Africa, South and Central America and the United States at risk.
  • Outbreaks of babesiosis can have considerable economic impact not only from mortality, but also calf abortions, loss of milk and/or meat production, the cost of control measures (such as acaricide treatments, vaccines and therapeutics) and negative impacts on cattle trade.
  • B. bige ia are Rhipicephalus microplus and R. atm latus, while R. de ohratus, R. 2 geigyi, and R. everisi can also transmit this species.
  • the major vectors for B. bovis are R. microplw and R. ammiaim, hut R. geigyi can also be a vector.
  • B. divergent is transmitted mainly by Ixodes ricm ' rn.
  • B jakimovi may also be transmitted by an Ixodes species.
  • Haemaphysalis punctata transmits B. major, Haemaphysalis longieornis transmits B. ovata, and Hya mma marginatum transmits B. o cultans.
  • the typical Babesia infection cycle begins when an infected tick sends sporozoites into a mammalian host while taking a blood meal. The sporozoites then invade red blood cells, where some transition from a trophozoite to a merozoite stage. Merozoites then asexually reproduce by binary fission, producing male and female gametes which may be ingested by a tick, where they undergo sporogony, producing sporozoites which are then transferred to a new mammalian host when the infected tick takes a blood meal.
  • This repHcative blood-stage is approximately of about 10 hrs duration, compared to a much longer 24-48 hrs in other protozoan diseases such as malaria. It is also noted that there is no liver stage in babesiosis, unlike the case with malaria.
  • Babesia are maintained in cattle populations by asymptomatic carriers that have recovered from acute disease.
  • B. bovis persists in cattle for years, and B bigemina survives for a few months. Recrudescence of parasitemia can occur at irregular intervals.
  • the invention is therefore broadly directed to an iraunogenic composition comprising chemically attenuated whole, blood-stage Babesia parasites, preferably in the absence of adjuvant.
  • a preferred approach includes the administration of attenuated blood-stage Babesia parasites, either as parasite-infected red blood ceils or as purified blood- stage parasites in the absence of adjuvant Even more preferably, a single dose of attenuated blood-stage Babesia parasites or infected red blood cells is sufficient to immunize against subsequent Babesia infection.
  • the parasite-infected red blood cells are intact cells.
  • the invention provides a method of producing an immunogenic composition including the step of treating blood-stage Babesia parasites or red blood cells infected with said blood-stage Babesia parasites with tafuramycin A or an analog or derivative thereof to thereby produce said immunogenic composition.
  • the method excludes the step of including an adjuvant.
  • the invention provides an immunogenic composition
  • an immunogenic composition comprising blood-stage Babesia parasites or red blood cells infected with said Wood- stage Babesia parasites that have been treated with tafuramycin A or an analog or derivative thereof; and an immunologically acceptable carrier, diluent or excipient.
  • the immunogenic composition does not include an adjuvant.
  • the invention provides an immunogenic composition for use in treating or preventing babesiosis, said immunogenic composition comprising blood-stage Babesia parasites or red blood cells infected with said blood- stage Babesia parasites that have been treated with tafuramycin A or an analog or derivative thereof; and an immunologically acceptable carrier, diluent or excipient.
  • the immunogenic composition does not include an adjuvant.
  • the invention provides a method of treating or preventing babesiosis, said method including the step of administering the immunogenic composition of the aforementioned aspect to an animal to thereby prevent or inhibit Babesia infection or treat an existing Babesia infection in said animal. 4
  • the method includes administering a single dose of said immunogenic composition to thereby prevent or inhibit Babesia infection or treat an existing Babesia infection in said animal.
  • the method does not include administration of an adjuvant.
  • the blood-stage Babesia parasites include ⁇ or are, merozoites, sc izonts, rings or trophozoites, although without limitation thereto.
  • the animal of the aforementioned aspects is preferably a mammal, inclusive of humans, bovines, deer, pigs, horses, sheep, goats, donkeys and dogs, although without limitation thereto.
  • FIG. 1 Rectal temperatures post vaccination with 1 x 10 9 iRBCs untreated (D1578); treated with 2 ⁇ tafuramycin A (TfA; D1574); or 20 ⁇ TfA (01577). Calf 1599 received 10* uninfected RBC
  • FIG. 2 PC V post vaccination. The treatment of these calves was as described in Fig. 1.
  • Figure 3 Positive control parasitaemia percentage post vaccination.
  • Figure 5 Packed cell volume of animals as in Fig. I post challenge with 10 7 iRBCs.
  • Figure d Parasitaemia count (xlO* /mL) from jugular vein blood smear post challenge with lO 7 iRBCs . Animals are as described in Fig. 1.
  • the present invention is predicated at least partly on the discovery that a single dose of tafuramycin A-treated blood-stage Babesia parasite-infected red blood cells of cattle can effectively immunize against subsequent Babesia infection. 5
  • Tafuramycin A is a rationally designed, DNA binding and alkylating agent that binds A-T rich regions of DMA
  • the efficacy of taftiramycm A in chemically attenuating Babesia parasites is unexpected, as compared to other protozoans such as Plasmodium, Babesia parasites typically have a relatively low proportion of A-T residues (about 58% compared to about 81% in Plasmadmm).
  • the invention provides a method of producing an immunogenic composition including the step of treating isolated or purified blood- stage Babesia parasites or red blood cells infected with said blood-stage Babesia parasites with tafuramycin A or an analog or derivative thereof, to thereby produce said immunogenic compositioa
  • the invention provides an immunogenic composition comprising isolated or purified blood-stage Babesia parasites or red blood cells infected with said blood-stage Babesia parasites that have been treated with tafuramycin A or an analog or derivative thereof.
  • Bobesia parasites are any pathogenic protists of the genus
  • Bobesia The genus “Babesia”.
  • the genus “Babesia” includes pathogenic species such as Babesia bovis, Babesia canis, Babesia bigemena, Babesia divergens and Babesia microti, although without limitation thereto.
  • the blood-stage Babesia parasites are, or include, any form of the parasite that exists within red blood cells, including but not limited to merozoites.
  • one embodiment of the invention relates to in vitro treatment of isolated or purified blood-stage Babesia parasites or red blood ceils infected with blood-stage parasites, with tafuramycin A or an analog or derivative of tafuramycin A.
  • This treatment is effective to chemically attenuate the blood-stage Babesia parasites without killing the parasite, such as by inhibiting parasite replication.
  • the attenuated blood-stage Babesia parasites are not capable of proliferation, or are capable of only limited proliferation, following attenuation by treatment with tafuramycin A.
  • Tafuramycin A is a rationally designed, DNA binding and alkylating agent based on duocarmycins that comprises a stereocenter.
  • tafuramycin A analogs or derivatives any molecule structurally related to tafuramycin A which exhibits binding to AT-containing nucleotide sequences to thereby induce DNA damage.
  • Tafuramycin, tafuramycin A, analogs or derivafrves inclusive of non-chiral, cbiral and racemic analogs and derivatives of duocarmycin and CC-1065, non-chiral, chiral and racemic isomers, salts or solvates thereof are also described in International Publications WO2002/030894, WO2008/050140, WO2009/064908 and WO2012/162731, Chavda. et almony 2010, Bioorganic ⁇ Med, Chem. 18:5016-5024, Howard, et or/., 2002, Bioorganic & Med. Chem. 10:2941-2952, Pornell et at., 2006, Med Chem.
  • Achiral se O-hydroxy-aza-CBI- ⁇ , a s ⁇ o-cyclopropy1pyrido[e]radolone (CPyl) compound is an example of an analog of centanamcyin as described in Chavda et l ⁇ 2010, supra. Racemic and chiral 5-methylfuran analogs of tafuramycin A are described in Purnell et al., 2006, supra.
  • tafuramycin A or the analog or derivative is a compound of formula I:
  • Ri is either MHR or OR, where R is selected from ⁇ , benzyl, benzyloxycarbonyl, 4-nitroberizyloxycarbonyl and T-n ethylpiperazmyl-N-carDonyl; 7
  • R 2 is selected from H and Ci-e alkyl
  • Ra is selected from H and alkyJ
  • fused ring selected from the group consisting of a benzene ring, a pyrrole ring, a pyridine ring, a furan ring and a 5-methylfuran ring, which fused ring may be optionally substituted with Cj ⁇ alkyl, CF3 or Cw, alkyloxycarbcmyl;
  • Y is an alkylene radical selected from - €3 ⁇ 4-, -C3 ⁇ 4CH 2 - or -CH 2 C3 ⁇ 4CH 2 -; or Y may be -CH- to form a five-merabered ring with R4;
  • X is an electrophilic leaving group
  • R4 is selected from H, Ci ⁇ $ alkyl, a -C3 ⁇ 4- group bonded to Y to form a five- membered ring or a -CH 2 - group bonded to the -CH 2 - to which X is attached to form a six membered ring;
  • R is selected from the group consisting of:
  • Yi and Y 2 are independently selected from O and NH;
  • Ri is OH.
  • R 2 and R together form a furan ring.
  • Y is -CH- and forms a five-membered ring with R .
  • X is selected from chloro, bromo, iodo, mesylate, tosylate, thio, ammonium, Ci-3 ⁇ 4 alkylsulfonyi and d-6 alkylsulfoxyl.
  • X is selected from chloro, bromo, iodo, mesylate and tosylate.
  • X is chloro
  • R4 is a -CH2- group bonded to Y to form a five membered ring structure.
  • R 5 is:
  • a preferred form of this embodiment is a compound of Formula ⁇ (mfuTamyci A):
  • Ri is NH 2
  • R 2 and Rj together form a benzene ring.
  • Y is -CH-.
  • X is selected from chloro, bromo, iodo, mesylate, tosylate, thio, ammonium,, alkylsulf nyl and alkylsulfoxyL
  • X is selected from chloro, bromo, iodo, mesylate and tosylate.
  • X is chloro
  • R* is H or C 1-6 alkyl.
  • a preferred form of this embodiment is a compound of Formula 111 (centanatrrycin):
  • Ri is OH.
  • Rj and R3 together form a pyridine ring optionally substituted with Ci ⁇ alkyl.
  • Y is -CH-.
  • X is selected from chloro, bromo, iodo, mesylate, tosylate, thio, ammonium,, alkylsulfonyl and Ci- ⁇ alkylsulfoxyl.
  • X is selected from chloro, bromo, iodo, mesylate and tosylate.
  • X is chloro
  • R is B or Ci ⁇ alkyl.
  • R* is
  • a preferred form of this embodiment is a compound of Formula IV (AS-VII Q4):
  • Typical, although non-limiting concentrations of tafuramycin A or analogs or derivatives for treatment of blood-stage Babesia parasites, or red blood cells infected with Babesia parasites are in the range of about 0.1 to 100 ⁇ .
  • the concentration is in the range of about 0.5-50 ⁇ , more preferably i tbe range about 1-40 ⁇ or even more preferably i the range of about 2-20 ⁇ .
  • Particular concentrations include about 0.5 ⁇ , i.O ⁇ , 1.5 ⁇ , 2.0 ⁇ , 3 ⁇ , 4 ⁇ , 5.0 ⁇ , 6.0 ⁇ , 7.0 ⁇ , 8.0 ⁇ , 9.0 ⁇ , 10 ⁇ , 1 J ⁇ , 12 ⁇ , 13 ⁇ , 14 ⁇ , 15 ⁇ , 16 ⁇ , 17 ⁇ , 18 ⁇ , 19 ⁇ , 20 ⁇ , 25 ⁇ and 30 ⁇ , or in any range therebetween.
  • Treatment duration may be in the range 1 minute to 12 hours, preferably 10 minutes to 4 hours or more preferably about 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5 or 2 hours, or any range therebetween.
  • the tafuramycin A-treated blood-stage Babesia parasites may be used to prepare the immunogenic composition as an isolated, enriched, purified or partially purified parasite preparation, or the tafuramycin A-treated blood-stage Babesia parasites may be used to infect red blood cells, which are then used to prepare the immunogenic composition. Alternatively, red blood cells infected with blood-stage Babesia parasites may be treated with tafuramycin A and then used to prepare the immunogenic composition.
  • p BC may be administered as intact cells or as a lysate.
  • the pRBC are in intact form.
  • the pRBC may be obtained from blood of a parasite-infected animal prior to tafuramycin A treatment.
  • non-infected red blood cells may be obtained from an animal and then infected in vitro with blood- stage Babesia parasites pre-treated with tafuramycin A, or with untreated blood-stage Babesia parasites so that the pRBC are thereafter treated with tafuramychi A.
  • the dose of tafuramycin A-treated blood-stage Babesia parasites or red blood cells infected wi th said Babesi parasites is capable of eliciting an immune response to subsequent infection by Babesia parasites.
  • the immune response is characterised by inducing a T cell response and, optionally, inducing B cells to produce detectable levels, or only low levels, of antibodies.
  • a typical dose of pRBC suitable for administration to cattle is no more than 10 u pRBC such as including 10 TO , W 9 , 10 8 , 10 7 , IQ 6 , lO 5 , 10 4 , 10 3 , 10* or lO RBC, or any range between any of these values.
  • the dose may be in the form of purified blood-stage Babesia parasites.
  • a typical dose suitable for adrninistrarion to cattle is no more than 10* 1 pRBC such as including 10 1Q , 10 9 , 10 s , 10 7 , 10 6 , 10 s , 10 4 , 10 3 , 10 2 or 10 pRBC, or any range between any of these values.
  • the invention provides an immunogenic composition for preventing or treating babesiosis, said immunogenic composition comprising isolated or purified blood-stage Babesia parasites or red blood cells infected with said blood-stage Babesia parasites that have been treated with tafuramycin A or an analog or derivative thereof, said composition preferably excluding adjuvant.
  • the invention provides a method of treating or preventing babesiosis, said method including the step of administering the immunogenic composition disclosed herein to an animal to thereby prevent or inhibit Babesia infectio or treat an existing Babesia infection in said animal, said method preferably excluding administration of adjuvant.
  • babesiosis includes all forms of the disease caused by protozoan protists of the genus Babesia.
  • the genus "Babesia” includes pathogenic species such as include Babesia bovis, Babesia canis, Babesia bigemena, Babesia divergens, Babesia ovis, Babesia ovata, Babesia occultans, Babesia caballi and Babesia moiasi, although without limitation thereto.
  • the causative Babesia species are typically esia bovis, Babesia bigemena and Babesia divergens.
  • the immunogenic composition and method of prophylactic or therapeutic treatment of babesiosis comprises tafuramycin A-treated parasites of one or more strains or isolates of Babesia parasites such as Babesia bovis, Babesia bigemena and/or Babesia divergens, or pRBC infected with said Babesia parasites,
  • Babesiosis can include in non-specific flu-like symptoms, such as fever, chills, swears, headache, body aches, loss of appetite, nausea, or fatigue and other symptoms such as thrombocytopenia, low or unstable blood pressure and hemolytic anemia which can lead to jaundice and darkened urine, although without limitation tthereto.
  • non-specific flu-like symptoms such as fever, chills, swears, headache, body aches, loss of appetite, nausea, or fatigue and other symptoms such as thrombocytopenia, low or unstable blood pressure and hemolytic anemia which can lead to jaundice and darkened urine, although without limitation tthereto.
  • treating refers to a therapeutic intervention, course of action or protocol that at least ameliorates a symptom of the disease, disorder or condition after symptoms have at least started to develop.
  • ⁇ preventing, "preve “ or “prevention” refers to therapeutic intervention, course of action or protocol initiated prior to the onset of a symptom of the disease, disorder or condition so as to prevent, inhibit or delay or development or progression of the disease, disorder or condition or the symptom.
  • preventative therapies may be referred to as "prophylaxis" or a “prophylactic” therapy.
  • immunization or vaccination is a form of preventative or prophylactic therapy.
  • tafuramycin A-treated blood-stage Babesia parasites of a particular isolate, strain or species may, upon administration to an animal, immunize or vaccinate against infectio by heterologous Babesia isolates, strains and/or species.
  • etero gatis pathogens means related pathogens that may be different strains or variants of a same or related species.
  • th immunogenic composition obviates the need for an adjuvant, whether as a component of the immunogenic composition or when administered together with the immunogenic composition.
  • adjuvant is meant an agent which assists, augments or otherwise facilitates the elicitation of an immune response by an imraunogen.
  • excluded adjuvants include, Freund's adjuvant, aluminium hydroxide (ahim), aluminium phosphate, squalene, IL-12, CpG-oligonucleotide, Montanide ISA720, imiquimod, SBAS2, SBAS4, MF59, PL, Quil A, QS21 and iSCO s.
  • adjuvant is absent from the immunogenic composition
  • other components such as immunologically acceptable carriers, diluents and/or excipients may be included.
  • immunologically acceptable carriers include solid or liquid fillers, diluents or encapsulating substances that may be safety used in systemic administration.
  • carriers, diluents and/or excipients may be selected from a group including sugars, starches, cellulose and its derivatives, malt, gelatine, talc, calcium sulfate, vegetable oils, synthetic oils, polyois, alginic acid, isotonic saline, pyrogen- free water, wetting or emulsifying agents, bulking agents, coatings, binders, fillers, disintegrants, lubricants and pH buffering agents (e.g. phosphate buffers) although without limitation thereto.
  • a group including sugars, starches, cellulose and its derivatives, malt, gelatine, talc, calcium sulfate, vegetable oils, synthetic oils, polyois, alginic acid, isotonic saline, pyrogen- free water, wetting or emulsifying agents, bulking agents, coatings, binders, fillers, disintegrants, lubricants and pH buffering agents (e.g. phosphate
  • the immunogenic composition and method of prophylactic or therapeutic treatment of babesiosis may be administered to an animal in any one or more dosage forms that include tablets, dispersions, suspensions, injectable solutions, syrups, troches, capsules, suppositories, aerosols, transdermal patches and the like.
  • the immunogenic composition and/or method of prevention or treatment of babesiosis are effective against blood-stage Babesiosis.
  • Blood-stage babesiosis is a stage where the Babesia parasite (e.g. merozoite) enters erythrocytes. In the blood stage, the parasite divides several times to produce new merozoites, which leave the red blood cells and travel within the bloodstream to invade new red blood cells.
  • blood-stage babesiosis is typically characterized by successive waves of fever arising from simultaneous waves of merozoites escaping and infecting red blood cells.
  • the immunogenic composition and method of prevention or treatment of babesiosis may elicit an immune response that is characterized as a CD4 ' T cell- mediated response (including solely CD4 r T celi-mediated responses and mixed CD4 + and CD8 + T cell-mediated responses), possibly with little or no antibody response.
  • the immunogenic composition and/or method immunize the animal to prevent, inhibit or otherwise protect the animal against subsequent Babesia infection,
  • a single dose of the immunogenic composition prevents, inhibits or otherwise protects the animal against subsequent Babesia infection.
  • animal refers to any animal capable of infection by a Babesia parasite, particularly mammals inclusive of humans, bovines, pigs, deer, horses, donkeys, sheep and goats.
  • havines are members of the mammalian sub-family Bovinae and include cattle, buffalo, bison and yaks.
  • Cattle include all breeds and sub-species of the genus Bos, including Bos indica and Bos taunts and hybrids thereof
  • the aim of this trial was to assess infectivity, virulence and immunity induced by Dixie strain B. hovis treated with Tafuramycin A (TFA) compared to the current live attenuated vaccine Dixie strain B. b&w to gauge the plausibility of conducting a larger scale research trial involving susceptible intact yearling or adult cattle.
  • TFA Tafuramycin A
  • Tafuramycin A (TFA98) was prepared fresh from 2 mM stock solution.
  • Treated parasitized red blood cells pRBC
  • untreated parasitized red blood cells RBC
  • normal red blood cells nRBC
  • pRBC parasitized red blood cells
  • nRBC normal red blood cells
  • the parasitized red blood ceils were transferred into separate 50mL centrifuge tubes and the tubes filled up to 50mL widi RPMI-1640. They were centrifuged at 1500 rpm for 5 minutes. Supernatants were removed and the tubes filled up to 50mL with RPMI- 1640. Tubes were incubated at 37°C in a C0 2 incubator for 20 minutes.
  • tubes were centrifuged at 1500 rpm for 5 minutes. The supernatants were removed and the tubes filled up to 50 mL with PBS. Following a final centrifugation at 1500 rpm for 5 minutes, supernatants were removed and pRBC pellet was resuspended in original volume. After an accurate cell count m a hemacytometer the required amount of pRBC for the appropriate immunizing dose was removed and diluted m 10 mL PBS. Tubes were centrifuged at ISOOrpra for 5 minutes before dilution to the appropriate immunizing dose of I x 10 9 pRBC in 0.9% sodium chloride (6 mL). Trial method
  • EDTA blood samples collected from the jugular vein.
  • the EDTA samples were also stored frozen for later B> bovis PC - this continued twice weekly from Day 15 to 52.
  • Serum samples were collected on day 0 and day 28 to determine seroconversion by B. hovis ELISA.
  • a blood sample collected into lithium heparin was also collected on day 0 and leucocytes extracted and stored for later cell mediated immunity (CM I) evaluation in conjunction with post-challenge samples.
  • CM I cell mediated immunity
  • the calves were again monitored daily for 23 days (then every second day until Day 29) by rectal temperature and microscopic examination of tail-tip (and jugular as appropriate) smears for parasttaemia.
  • Packed cell volume (PCV) measurements were taken from blood collected from jugular vein EDTA samples until Day 14; then directly from the coccygeal artery/vein from Da 15 onwards until there was evidence of PCV recovery.
  • EDTA blood samples were collected daily until Day 14 and stored frozen for later PCR analysis. Lithium heparin samples were also collected at day 0, 3, 7 and 14 post challenge inoculation for later CM1 evaluation.
  • the negative control (D1599) developed the greatest maximum PCV depression (35%) and maximum parashaemia count (43.0 xl 0 6 /mL) compared to the other calves post challenge inoculation (Table 3; Figures 5 and 6). These findings correlated with pyrexia at 40.3°C on Day 8 post challenge ( Figure 4). This calf spontaneously recovered without treatment within four days from Day 10 when a rapid decline in parasitaemia occurred ( Figure 6). The PCV subsequently increased and plateaued from Day 15 at the lower end of the normal range, still 1 % below the pre-inocnlation value. The rectal temperature also returned to within normal limits from day 12, with occasional fluctuation.
  • the TFA 20 ⁇ ⁇ ⁇ calf ( 1577) also experienced pyrexia at 40.6°C on Day 12 post challenge. Instead of a sudden decline in PCV (as in the negative control calf)' * gradual reduction in PCV was seen in this calf to a minimum of 24% on Day 20-21 , representing a PCV depression of 25%. This was associated with a persistent low level parasitaemia until Day 21 (with microscopic evidence of B. bovte on thick tail-ti smear only). The PCV remained stable between 24-25% from Day 21 onwards and began to slowly rise on Day 29.
  • the TFA 2 pmol/L calf (D1574) developed a greater and more persistent parasitaemia than the TFA 20 ⁇ /L calf, in which B. bevts iRBCs were evident in the jugular vein blood smear once only (on Day 7) ( Figure 6). Moreover, both were sustantially leas than the negative control (Table 3). In addition, the TFA 2 ⁇ /L calf s PCV and temperature parameters remained within normal limits throughout the monitoring period ( Figures 4 and 5), although PCV did reduce by 18% compared with pre-inoculatiort values. Interestingly, both TFA calves showed a similar slow decline in PCV from Day 9-25 which is most likely associated with the persistent parasitaemias.

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Abstract

La présente invention concerne une composition immunogène et un procédé de production d'une composition immunogène comprenant des parasites Babesia au stade sanguin isolés ou purifiés, ou des érythrocytes infectés avec des parasites Babesia au stade sanguin, traités avec la tafuramycine A ou un analogue ou dérivé de celle-ci. Typiquement, les parasites Babesia au stade sanguin sont, ou comprennent, une forme quelconque du parasite qui existe dans les érythrocytes ce qui inclut, sous toutefois s'y limiter, des mérozoïtes. La composition immunogène est utilisée pour le traitement ou la prophylaxie de la babésiose chez des mammifères tels que des bovins.
PCT/AU2014/000750 2013-07-29 2014-07-25 Vaccin atténué contre babesia Ceased WO2015013739A1 (fr)

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AU2013902802A AU2013902802A0 (en) 2013-07-29 Attenuated babesia vaccine
AU2013902802 2013-07-29

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018049459A1 (fr) * 2016-09-16 2018-03-22 Griffith University Vaccin comprenant un médicament et administration de parasite
WO2022236370A1 (fr) * 2021-05-11 2022-11-17 Griffith University Composition immunogène

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008050140A2 (fr) * 2006-10-27 2008-05-02 Spirogen Limited Composés pour le traitement d'une infection parasitaire
WO2012162731A1 (fr) * 2011-05-27 2012-12-06 Griffith University Vaccin contre le paludisme au stade sanguin

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008050140A2 (fr) * 2006-10-27 2008-05-02 Spirogen Limited Composés pour le traitement d'une infection parasitaire
WO2012162731A1 (fr) * 2011-05-27 2012-12-06 Griffith University Vaccin contre le paludisme au stade sanguin

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018049459A1 (fr) * 2016-09-16 2018-03-22 Griffith University Vaccin comprenant un médicament et administration de parasite
US11406694B2 (en) 2016-09-16 2022-08-09 Griffith University Vaccine comprising drug and parasite administration
WO2022236370A1 (fr) * 2021-05-11 2022-11-17 Griffith University Composition immunogène

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