GB2152510A - Method for the production of helminths vaccines - Google Patents
Method for the production of helminths vaccines Download PDFInfo
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Abstract
An easily cultured worm is genetically modified such that its surface antigens on its modified form are immunologically identical to the surface antigens of a parasitic worm. Specifically, C elegans is genetically modified to produce antigens corresponding to the antigens of D. immitis which causes heartworm infection. The antigens derived from the easily cultured worm are used to form a vaccine and/or diagnostic.
Description
SPECIFICATION
Method for the commercial production of Helminths antigens
This invention relates to the development and commercial production of helminths (parasitic worm) antigens.
The use of antigens for diagnostics and vaccines to detect and prevent infectious dieases such as polio, smallpox, diphtheria, tetanus and hoof and mouth diseases has been clearly shown in both humans and animals. Most antigens used as diagnostics and vaccines are derived from cultured infectious organisms.
The infectious organisms are grown in vitro or in vivo in animals or tissue cultures. An example is polio antigens which are derived from viruses grown in vitro in monkey kidney cells. The antigens are used as a diagnostic and to vaccinate humans.
If a small amount of attenuated viruses is innoculated in a host, the innoculated viruses which carry specific antigens will induce antibody formation in the host which in the future will recognize and destroy future invading polio viruses. The same polio virus antigens can also be used for the diagnosis of a polio infection. If a host is infected by polio, some specific antibodies will be elicited in the body and the presence of these antibodies will be indicative of an infection. The detection of antibodies can be assayed for their binding to polio antigens with the standard immunofluoroscent, radio-immune, and enzyme-linked, immunoelectrophoresis, hemagglutinin and immunodiffusion assays.
It is not possible, however, to commercially produce antigens for all infectious diseases. One limitation of production has been the inability to cultivate or produce a large amount of infectious organisms in vivo or in vitro from which the antigens are derived. This is especially true for vaccines in which the infectious organisms have complicated life cycles and/or have more than one host. Most parasitic worm diseases that infect dogs, cats, sheep, pigs, horses and humans fall into this category.
An example is the helminths disease of the heartworm which can infect a wide variety of organisms from dogs, cats, seals to humans (infrequently). The parasite, however, generally resides in the dog as its host with the mosquito as its intermediate host. It would be difficult to derive antigens necessary in the manufacture of vaccines or diagnostics since the parasitic worm undergoes several stages of larval development and only one larval stage contains the appropriate antigens. In the heartworm this particular stage, the infectious larvae, resides in the mosquito. Under these conditions, the capture of the intermediate hosts (mosquitos) and the dissection for the infectious larvae is necessary to generate required antigens for the manufacture of a heartworm vaccine.Wong, M.M., Guest, M.F., and Laviopierre, M.J. (1974) Dirofilaria immitis; Fate and Immunogenicity of Irradiated Infective Stage Larvae in Beagles Experimental Parasitology 35, 65-74. Similarly, antigens which are necessary to diagnose heartworm antibodies for infection must be derived from adult heartworms harboured in the heart of an infected dog. Desowitz, R.S. and una, S.R.
(1976), The Detection of Antibodies in Human and Animal Filariasis by Counter-immunoelectrophoresis with
Dirofiliaria immitis Antigen. Journal ofHelminthology, 50, 53-57, Grieve, R.B., Mika-Johnson, M., Jacobson,
R.H., and Raymond, C.H., (1981) Enzyme-Linked Immunosorbent Assay for Measurement of Antibody
Response to Dirofilaria immitis in Experimentally Infected Dogs. American-Journal of Veterinary Research 42, 66-69. These sources and methods of generating heartworm antigens for diagnostics (from the heart of a dog) and vaccines (from mosquitos) are neither commercially feasible nor may they be socially acceptable.
I have devised a method of producing helminths (parasitic worm) antigens wherein a related species of the infectious worm that can be easily cultured is genetically modified. The method involves identifying the surface antigens of the difficult to culture parasitic worm and then creating these same antigens in the easily cultured species.
The easily cultured species is genetically altered through mutations until said species hax e some of the same immunological antigens of the difficult to culture parasitic worm. Through these genetic manipulations, the types and amounts of antigens of interest in the easily cultured species are altered. Antigens derived from the genetically altered species are then used for the commercial production of antigens necessary for the manufacture of diagnostics and vaccines.
The principle of my invention is to alter the antigen genes of the easily cultured species to produce preferably in situ antigens of the hard to culture parasitic species. The expression of these altered genes to produce these antigens can also occur in bacteria or yeast with the cloning of the antigen genes, and thus, provide a potentially more efficient production of these same antigens.
According to the present invention therefore, there is provided a method for the production of parasitic worm antigens which comprises:
identifying the antigens of a parasitic worm;
modifying genetically an organism related to the parasitic worm until a mutant is formed, which mutant is characterised by antigens having immunological identity to the antigens of the parasitic worm;
cultivating the mutant; and
harvesting the mutant whereby the antigens may be isolated for the use as a diagnostic or vaccine for detection and prevention of the invention caused by said parasitic worm.
In the preferred embodiments, the antigens produced using this invention against the heartworm infection is by Dirofilaria immitis, and the substituted related species is the free-living hermaphroditic nematode
Caenorhabditis elegans.
Description of the preferred embodiment
Generation of antibodies against heartworm antigens of Dirofilaria immitis.
Antibodies must be first generated against parasitic antigens of interest. These antibodies which recognise and bind specifically to the hard to culture parasitic antigens can then be used as a probe to isolate mutants in the easily cultured species which are capable of producing the corresponding parasitic antigens. In the examples set forth below, I have generated antibodies against adult heartworm antigens and isolated mutants in C. elegans with the corresponding heartworm antigens. Although described in reference to C.
elegans, other species which are believed suitable are Panagrellas redivious, Turbatric acetic and C.
briggsae.
Adult heartworms were isolated from a dog infected with heartworm. The adult heartworms were excised from the heart and suspended in 0.15 M phosphate buffered saline pH 7.4 (PBS). The heartworms were homogenised and the homogenate were adjusted to a final concentration of 500 micrograms of protein (wet weight) per ml of PBS. This homogenate was then mixed with equal volume of Freund's complete adjuvant.
A 2 ml sample of this mix was used to immunize a rabbit for heartworm antigens. Before actual immunization, a preimmune serum was drawn from the rabbit. The rabbit was innoculated at days 25 and 35 after the first immunization with 2 ml of a mixture of equal volume of homogenate and Freund's incomplete adjuvant. After a positive test bleed at 45 days, the serum was collected and used to assay for antibody formation against heartworm antigens used for the immunization.
Assay ofserum containing antiheartworn: antibodies
The serum of the rabbit immunized with heartworm antigens was assayed for antibody activities against heartworm antigens and C. elegans antigens. My preferred method is to be use a immunofluorescent assay of conjugated fluoroscein or rhodamine goat anti-rabbit IgG. Other methods such as enzyme-linked, radio-immune, hemagglutinin and immundiffusion assay can also be used. Sections of heartworm tissues from dogs (containing the antigens) were washed three times with PBS and then incubated with the rabbit anti-heartworm antibodies at room temperature for 30 minutes, the tissues were then washed 3 times with
PBS to remove the rabbit antiheartworm antibodies. Following this, the tissues were incubated with fluoroscein goat anti-rabbit IgG antibodies.After incubation, the freegoat anti-rabbit IgG antibodies were removed by washing the heartworm tissues 3 times with PBS. The presence of rabbit antibody activities binding to specific heartworm antigens was indicated by the binding of the fluoroscent goat anti rabbit IgG antibodies which were then observed in a fluoroscent microscope, red (rhodamine) or green (fluoroscein) in colour.
C. elegans whole animals and tissues were also used to assay the anti-heartworm antibodies generated by the immunized rabbit serum. The results of the fluoroscent assays for anti-heartworm activities are summarized in Table I.
The control preimmune serum had no activity against either heartworm tissues or C. elegans tissues. In contrast, there was evidence of antibody activities in the immunized rabbit serum against heartworm tissues but not C. elegans. Thus, the generation of antibodies against heartworm antigens is only specific for heartworm but not C. elegans. This serum which appears specific was used as a probe to isolate mutants of
C. elegans which will, in accordance with my invention, carry antigens corresponding to heartworm antigens.
TABLE I
Antibody activities of serum from a rabbit which has been immunized with heartworm antigens
Titter Immune Serum Preimmune Serum
Heartworm C. elegans Heartworm C. elegans
tissue tissue tissue tissue
1:5 1:20 - - - 1:40 - - - 1:80 - - 1:160 - - = = fluorescent activity - = no activity
TABLE II
Antibody binding activity of six independently isolated mutants of C. ele gans with 1::50 titer of rabbit
anti-heartworm serum
Immune Serum reimmune Serum
Wild type -
(C.elegans)
Heartworm +
Mutant Strain
VGI-1 t
VGI-2 + VGI-3 +
VGI-4 +
VGI-5 +
VGI-6 +
TABLE Ill
Antibody binding activity of one mutant VGI-6 with rabbit anti-heartworm serum
Wild Type VGl-6 Heartworm (C.elegans) Titer tissue tissue tissue 1:5 + t 1:20 + + 1:40 t 1:80 < + 1:160 +
Mutagenization of C. elegans to generate mutants which have altered antigens that correspond to heartworm antigens.
The free-living soil nematode C. elegans is preferred since it can be readily cultured in large quantities and homozygous recessive mutations can be generated. Brenners, S. (1974), the Genetics of Caenorhabditis elegans Genetics 77,71-94. This is due to the hermaphroditic nature of the animal, that is, it is self4ertilizing.
A mutation introduced in the animal upon self-fertilization for two generations will produce a homozygous mutation.
Young larvae of C. elegans at the first and second larval stage of development were exposed to the mutagen, ethyl methyl sulfonate (any other mutagens can also be used, such as methyl methane sulfonate, acridine orange, nitrosoguanidine, hydroxynitrosamine; Drake, J.W. and R.H. Baltz (1976), The Biochemistry of Mutagenesis, Annual Review ofBiochemistry 45, 11) and allowed to self-fertilize and lay eggs. The F1 generation of this mutagenesis was allowed to reproduce and the F2 generation was then screened for animals which would bind to antibodies made against specific antigens of the heartworm. Because wild type
C. elegans do not bind to antibodies made against heartworm antigens generated in our rabbit heartworm antiserum (see Table I), a visual screen was made for mutants that had antigens that bound to the heartworm antibodies using the above fluorescent assay.Mutants which bound to the heartworm antibodies unlike the wild type (with no fluorescence) had a fluorescent colour of red (rhodamine) or green (fluorescein). These mutant animals were removed and cloned.
Animals which breed true for this characteristic are considered to be mutants which have an altered gene that would produce an antigen(s) corresponding to those of the heartworm.
Using the above methods, I have demonstrated with my invention, in Table II, that mutants of C. elegans with altered antigens corresponding to those of heartworm antigens can be generated. The antibody binding activities with anti-heartworm serum of six (6) independently isolated mutants is compared with heartworm tissues and wild type tissues in Table II.
Table Ill represents a further characterisation of one of the representative mutants among the six (6) in
Table II in which the binding of antibody activity behave very closely to heartworm tissue.
Furthermore, when one of these C elegans mutants was immunized to a rabbit, it was able to elicit an immunogenic response in which the antibodies produced in the serum had binding activities to a species of heartworm antigens.
Thus, the mutants isolated with my invention not only have an antigenic response of binding to heartworm antibodies, but also a immunogenic response in producing antibodies against heartworm antigens.
Using the above-described method, I also isolated a mutant of C. elegans which was altered in at least some of its surface antigens to be immunologically identical to the antigens of another parasitic worm,
Ancylostoma caninum, a hookworm which infests in the intestine of the dog.
Any worm species which are readily cultured and in which mutations can be introduced to cause an alteration in its antigens to a corresponding difficult to culture parasitic worm antigen can be used with my method.
Conceptually, using C. elegans and the heartworm, D. immitis as an example, once the antigens of the desired parasite are generated with the above method, they can be used to develop a range of diagnostics for the presence of antibodies induced upon a parasitic worm infection. Such an approach with heartworm antigens has been shown to be useful in the diagnosis of several types of helminths infections. Desowitz and
Una, supra; Hedge and Ridley (1977). Immunofluorescent Reactions with Microfilariae. 1 Diagnostic
Evaluation; Transaction of the Royal Society of Tropical Medicine and Hygiene 71, 304-307. In the case of the heartworm antigens which I have generated, I have found that one of the mutants isolated in C. elegans,
VGI-2, is useful in the diagnosis of heartworm infection.When serum samples of dogs diagnosed as infected and uninfected with heartworms were assayed with the VGI-2 mutant, the results were remarkedly accurate in that 6 dogs diagnosed with the standard filter assay as positively infected based on the presence of microfilarie were also found to be positive in this mutant with the immunofluorescent assay. In contrast, 5 samples of serum from clinically diagnosed uninfected dogs were also found negative in the mutant.
Besides the use of immunofluorescent assay to detect parasitic worm infection with mutant antigens, enzyme-linked, radio-immune, immunodiffusion or hemagglutinin assay can also be applied. D.Stites (1976),
Laboratory Methods for Detection of Antigens and Antibodies, Basic and Clinical Immunology, Lange
Medical Publications, pp. 281-315.
Antigens generated with the described method can also be used for the manufacture of parasitic worm vaccines. In the example of heartworm infection, it is known that the antigens derived from infectious larvae can be used as a vaccine for dogs against heartworm infection; Wond, M.M., Guest, M.F., and Lavoipierre,
M.J. (1974) Dirofilaria immitis; Fate and Immunogenicity of Irradiated Infective Stage Larvae in Beagles
ExperimentalParasitology35, 65-79. As long as antibodies can be specifically generated against these particular infectious larval antigens and mutants of C elegans with corresponding antigens are isolated, these mutants can be used to derive the immunogenic antigens necessary for the production and manufacture of heartworm vaccine.The vaccine generated can then be suspended for example in a buffered saline and then used for injection subcutaneously or intramuscu!arly in an animal for providing immunity against heartworm infection. W.J. Herbert (1978), Laboratory Animal Techniques for Immunology in "Handbook of Experimental Immunology", Vol. 1 ImmunoChemistry, 3rd Edition, Editor, D.M. Weir, Blackwell Scientific Publishers, Oxford, London.
The basic principle of my method is to alter the gene(s) of a closely related worm species that is easy to culture to generate antigens of a difficult to culture parasitic worm. In the described embodiment, the concept of the generation of antigens in situ in the species of the easily cultured worm. This concept can be extended further in the expression of antigens in another organism such as bacteria, yeast or other microorganisms which may provide a more efficient and productive yield of the desired antigens. The isolation of the antigen gene(s) and their insertion into bacteria with genetic vectors can be applied.The genes for example in C. elegans can be isolated by initially isolating the nucleic acid of the animal with the proteinase K-SDS method described by Emmons, S., Klass, M.R., and Hirsh, D. (1979); Anaysis of the
Constancy of DNA Sequences During Development and Evolution of the Nematode Caenorhabditis elegans
Proceeding of the National Academy Sciences 76, 1333-1337.
The poly A (RNA) is separated by ethanol precipitation and by binding to oligo(dT) cellulose according to
Aviv and Leder (1972), Purification of Biologically Active Globin Messanger, RNA by Chroatography on Oligothymidylic Acid Cellulose, Proceedings ofthe NationalAcademy ofScience 69, 1408-1412. The material binding to oligo(dT) cellulose are that used to purify poly A(RNA) which codes for the antigen gene product.
Since the poly A (RNA) will only hybridize to the DNA which codes for the antigen gene, this poly A (RNA) can be used to select recombinant poly A (DNA) molecules of C. elegans coding for the antigen gene of interest. These recombinant DNA molecules can then be inserted into bacteria or yeast with various genetic vector such as plasmids or bacteriophage to express the antigen gene and product of interest using the basic techniques described by David, R.W., Botstein, D., and Roth, J.R. (1980). A manual for genetic engineering
Advanced Bacterial Genetics, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York).
There is a possibility that antigens produced by the natural parasites may be more potent or specific than those originating from C. elegans through genetic mutations. Thus, the cloning of the naturally hard to culture parasitic genes for their antigens in an microorganism may be advantageous. If the cloning approach described above is used, it would be extremely difficult if not impossible to accomplish. This is due to the limitation of the amount of parasitic tissues available to isolate the poly A (RNA) of interest. The poly A (RNA) is only present in tissues which are expressing the antigen. In the case of poly A (RNA) corresponding to the antigens of the infectious larvae of the heartworm, the tissues must be derived from larval animals dissected from captured mosquitos which harbour the parasites. The availability of infectious larval tissues necessary to isolate enough poly A (RNA) corresponding to their antigens is limited therefore, making the task of poly A (RNA) isolation extremely difficult, if not impossible. However, if we use the poly A (RNA) isolated from C.
elegans mutant that correspond to these antigens of interest, we can through hybridization of the heartworm
DNA isolate the recombinant DNA genes corresponding to the heartworm infectious larvae antigens. The availability of heartworm DNA is not limited since all tissues have DNA containing all genes and a large quantity of tissues can be acquired from adult heartworms.
Claims (18)
1. A method for the production of parasitic worm antigens which comprises:
identifying the antigens of a parasitic worm;
modifying genetically an organism related to the parasitic worm until a mutant is formed, which mutant is characterised by antigens having immunological identity to the antigens of the parasitic worm;
cultivating the mutant; and
harvesting the mutant whereby the antigens may be isolated for the use as a diagnostic or vaccine for detection and prevention of the infection caused by said parasitic worm.
2. A method according to claim 1 wherein the parasitic worm is a helminths.
3. A method according to claim 1, wherein the parasitic worm is a trematode, cestode or nematode.
4. A method according to anyone of claims 1 to 3 wherein the related organism is a helminths.
5. A method according to claim 4 wherein the related organism is a trematode, cestode or nematode.
6. A method according to claim 5 wherein the related organism is a nematode.
7. A method according to claim 1 wherein the related organism is C. elegans, p. redivious, T. acetic or C.
brig gsa e.
8. A method according to claim 1 to 7 wherein the parasitic worm is D. immitis.
9. A method according to claim 1 or 7, wherein the parasitic worm isA. caninum.
10. A method according to any one of the preceding claims which includes identifying the mutants which have the immunological identity with the antigens of the parasitic worm by selecting those mutants which bind to antibodies generated by a host immunized with parasitic worm antigens.
11. A method according to claim 10 which includes removing the mutants so identified; and cloning the mutants so removed.
12. A method according to claim 1 wherein the genetic modification of the closely related organism includes:
isolating the nucleic acid of the related organism;
separating the poly A (RNA);
selecting subsequently the recombinant poly A (DNA) molecules of the closely related organism;
coding for the antigen gene of interest; and
inserting the recombinant DNA molecules into a support culture to express the antigen gene; and
harvesting subsequently the same.
13. A method according to claim 1 wherein the genetic modification of the closely related organism includes:
isolating the nucleic acid of the related organism;
separating the poly A (RNA) of the related organism;
selecting subsequently the recombinant poly A (DNA) molecules of the parasitic worm;
coding for the antigen gene of interest; and
inserting the recombinant DNA molecules into a support culture to express the antigen gene; and
harvesting subsequently the same.
14. A method for immunizing a host subject to infection by a parasitic worm which comprises:
administering to the host subject to an immune response caused by the administration of an antigen, a composition which comprises a biologically acceptable carrier; and the antigen produced from a mutant of a related organism of the parasitic worm, the antigens immunologically identical to the antigens of the parasitic worm.
15. A method of determining the presence of antibodies induced upon a parasitic worm infection which includes:
assaying a serum of an infected host with a composition comprising a biologically acceptable carrier and antigens produced from a mutant of a related organism of a parasitic worm, the antigens immunologically identical to the antigens of the parasitic worm.
16. The composition which comprises a biologically acceptable carrier and an effective amount of the mutant obtained by the method claimed in any one of claims 1 to 13.
17. The composition of claim 16 wherein the composition is a diagnostic.
18. The composition of claim 16 wherein the composition is a vaccine.
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB08400529A GB2152510B (en) | 1984-01-10 | 1984-01-10 | Method for the commercial production of helminths antigens |
| JP59004933A JPS60152423A (en) | 1984-01-10 | 1984-01-17 | Industrial manufacture of parasitic antigen |
| DE19843402492 DE3402492A1 (en) | 1984-01-10 | 1984-01-25 | ANTIGENT PARASITICAL WORMS AND THEIR PRODUCTION AND USE |
| AU34974/84A AU572908B2 (en) | 1984-01-10 | 1984-11-02 | Method for the commercial production of helminths antigens |
| FR848418373A FR2573983B1 (en) | 1984-01-10 | 1984-12-03 | PROCESS FOR THE COMMERCIAL PRODUCTION OF HELMINTHESA ANTIGENS |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB08400529A GB2152510B (en) | 1984-01-10 | 1984-01-10 | Method for the commercial production of helminths antigens |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| GB8400529D0 GB8400529D0 (en) | 1984-02-15 |
| GB2152510A true GB2152510A (en) | 1985-08-07 |
| GB2152510B GB2152510B (en) | 1988-06-15 |
Family
ID=10554758
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| GB08400529A Expired GB2152510B (en) | 1984-01-10 | 1984-01-10 | Method for the commercial production of helminths antigens |
Country Status (5)
| Country | Link |
|---|---|
| JP (1) | JPS60152423A (en) |
| AU (1) | AU572908B2 (en) |
| DE (1) | DE3402492A1 (en) |
| FR (1) | FR2573983B1 (en) |
| GB (1) | GB2152510B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2573983A1 (en) * | 1984-01-10 | 1986-06-06 | Lew Kenneth | PROCESS FOR THE COMMERCIAL PRODUCTION OF HELMINTHES ANTIGENS |
| WO1990011086A1 (en) * | 1989-03-17 | 1990-10-04 | Edward Albert Munn | Production and use of anthelmintic agents and protective immunogens |
| WO1994010201A1 (en) * | 1992-10-28 | 1994-05-11 | Mcgill University | Peptides and vaccines derived from nematode tubulin |
| WO1997029766A1 (en) * | 1996-02-15 | 1997-08-21 | Heska Corporation | Parasitic helminth larval thiol specific antioxidant proteins, nucleic acid molecules, and uses thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1988001277A1 (en) * | 1986-08-18 | 1988-02-25 | The Australian National University | Helminth parasite vaccine |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4396600A (en) * | 1980-12-18 | 1983-08-02 | Gus Gallucci | Adult schistosome worm-derived antigenic substance and method of obtaining same |
| US4345026A (en) * | 1981-01-12 | 1982-08-17 | The Children's Hospital Medical Center | Mutagenicity assay |
| GB2152510B (en) * | 1984-01-10 | 1988-06-15 | Kenneth K Lew | Method for the commercial production of helminths antigens |
| ES8705231A1 (en) * | 1984-11-15 | 1987-05-01 | Biotech Australia Pty Ltd | Anthelmintic vaccines comprising a nematode species. |
-
1984
- 1984-01-10 GB GB08400529A patent/GB2152510B/en not_active Expired
- 1984-01-17 JP JP59004933A patent/JPS60152423A/en active Pending
- 1984-01-25 DE DE19843402492 patent/DE3402492A1/en not_active Withdrawn
- 1984-11-02 AU AU34974/84A patent/AU572908B2/en not_active Ceased
- 1984-12-03 FR FR848418373A patent/FR2573983B1/en not_active Expired - Lifetime
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2573983A1 (en) * | 1984-01-10 | 1986-06-06 | Lew Kenneth | PROCESS FOR THE COMMERCIAL PRODUCTION OF HELMINTHES ANTIGENS |
| WO1990011086A1 (en) * | 1989-03-17 | 1990-10-04 | Edward Albert Munn | Production and use of anthelmintic agents and protective immunogens |
| EP0846760A1 (en) * | 1989-03-17 | 1998-06-10 | The Babraham Institute | Production and use of anthelmintic agents and protective immunogens |
| WO1994010201A1 (en) * | 1992-10-28 | 1994-05-11 | Mcgill University | Peptides and vaccines derived from nematode tubulin |
| WO1997029766A1 (en) * | 1996-02-15 | 1997-08-21 | Heska Corporation | Parasitic helminth larval thiol specific antioxidant proteins, nucleic acid molecules, and uses thereof |
| US5744593A (en) * | 1996-02-15 | 1998-04-28 | Heska Corporation | Parasitic helminth larval thiol specific antioxidant proteins and nucleic acid molecules |
| US6031077A (en) * | 1996-02-15 | 2000-02-29 | Heska Corporation | Parasitic helminth larval thiol specific antioxidant proteins, nucleic acid molecules and uses thereof |
| US6489448B1 (en) | 1996-02-15 | 2002-12-03 | Heska Corporation | Antiparasitic helminth larval thiol specific antioxidant-antibodies, and uses thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| AU572908B2 (en) | 1988-05-19 |
| GB2152510B (en) | 1988-06-15 |
| FR2573983A1 (en) | 1986-06-06 |
| AU3497484A (en) | 1986-05-08 |
| JPS60152423A (en) | 1985-08-10 |
| DE3402492A1 (en) | 1985-10-24 |
| GB8400529D0 (en) | 1984-02-15 |
| FR2573983B1 (en) | 1990-03-09 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PCNP | Patent ceased through non-payment of renewal fee |