Ascaris suum: partial fractionation of metabolic antigens from "in vitro" cultured larvae

Guerrero, Jorge; Silverman, Paul H.

doi:10.1590/S0037-86821973000500005

Abstracts

Ascaris suum metabolic antigens were obtained frorn second and early third stage larvae cultured in vitro in supplemented Eagle's minimum essential medium. Metabolic antigens harvested after 12 and 16 days from in vitro cultures were eluted through Bio-Gel Al.5. Three main elution peaks were identified, dialysed, lyophilized and injected into mice with 4% sodium alginate. Peak 11 from elution of two preparations of metabolic antigens protected mice against a chállenge infection of 10,000 A. suum embryonated eggs.

Os antígenos metabólicos de Ascaris suum foram obtidos de larvas de segundo e de terceiro estágio (precoce) cultivadas in vitro em meio mínimo essencial de Eeagle com suplemento proteico. Os antígenos metabólicos coletados depois de 12 a 16 dias das culturas in vitro foram eluidos em Bio-Gel Al 5. Três picos de eluição foram identificados, dializados, liofilizados e injetados em camundongos, com alginato de sódio a 4%. O pico II da eluição de duas preparações dos antígenos metabólicos protegeram os camundongos da infecção com 10.000 ovos infectivos de A. suum

Ascaris suum: partial fractionation of metabolic antigens from "in vitro" cultured larvae

Jorge Guerrero^I; Paul H. Silverman^II

^ICentro de Pesquisas em Fisiologia e Imunologia de Helmintos, Departamento de Microbiologia e Parasitologia, Escola Paulista de Medicina. São Paulo. Brasil

^IIDepartment of Biology, University of New Mexico, Albuquerque, New Mexico. U.S.A.

ABSTRACT

Ascaris suum metabolic antigens were obtained frorn second and early third stage larvae cultured in vitro in supplemented Eagle's minimum essential medium. Metabolic antigens harvested after 12 and 16 days from in vitro cultures were eluted through Bio-Gel Al.5.

Three main elution peaks were identified, dialysed, lyophilized and injected into mice with 4% sodium alginate. Peak 11 from elution of two preparations of metabolic antigens protected mice against a chállenge infection of 10,000 A. suum embryonated eggs.

RESUMO

Os antígenos metabólicos de Ascaris suum foram obtidos de larvas de segundo e de terceiro estágio (precoce) cultivadas in vitro em meio mínimo essencial de Eeagle com suplemento proteico. Os antígenos metabólicos coletados depois de 12 a 16 dias das culturas in vitro foram eluidos em Bio-Gel Al 5.

Três picos de eluição foram identificados, dializados, liofilizados e injetados em camundongos, com alginato de sódio a 4%. O pico II da eluição de duas preparações dos antígenos metabólicos protegeram os camundongos da infecção com 10.000 ovos infectivos de A. suum.

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Submitted to publication on 6.25.73.

This study was performed at the Department of Zoology, Uníversity of Illinois, Urbana, Illinois, U.S.A., under partial support of grant AI05910 from the National Institute of Allergy and Infectious Diseases.

References

1. ADLER, F. L. and MOLLER, G. - Antigenic competition. In: Progress in Immunology. Ed. B. Amos. Academic Press. 1511-1514, 1971.
2. BJÖRNBOE, M. - Serum protein und Antistoffprotein bei Immunisierung mit mehreren Antigen. Z. Immunitatsf. Exper. therapie, 99: 245- 256, 1941.
3. Bio-Rad Laboratoires. Gel chromatography. Bio-Rad Laboratoires. Richmond, Califórnia. 89 pp. 1971.
4. BOYD, W. C. - Fundamentls of immunology . Interscience Publishers. New York. 74 pp. 1966.
5. BROWN, H. W., and CHAN, K. F. - The effect of piperazine HC1 on migrating larvae of Ascaris suum Goeze 1782. Am. J. Vet. Res., 16: 613-615, 1955.
6. DETERMAN, H. - Gel chromatography. Springer-Verlag. New York. 195 pp. 1968.
7. DOUVRES, F. W. and TROMBA, F. G. - Influence of pH, serum and cell cultures on development of Ascaris suum to fourth stage in vitro Journal Parasit., 56: 238-248, 1970.
8. DUNCAN, D. B. - A Bayesian approach to multiple comparison. Technometrics, 7: 171- 222.
9. FLODIN, P. - Methodogical aspects of gel filtration with special reference to desalting opera tion. J. Chrom., 5: 103-115, 1961.
10. GELOTTE, B. - Fractionation of proteins peptides and amino acids by gel filtration. In: A. T. James and L. D. Morris (eds). New Biochemical Separations. D. Van Nostrand Col. Ltd., p. 93-109, 1964.
11. GLENNY, A. T. - Diptheria antitoxin in the blood of normal horses. J. Path. Bact. 28: 241-250, 1925.
12. GLENNY, A. T. - The relation between dosage and death time. J. Path. Bact., 28: 251-260, 1925.
13. GUERRERO, J. - Studies on the immunogenicity of Ascaris suum larval antigens. Pt. D. Thesis, University of Illinois, Urbana, Illinois, 1971.
14. GUERRERO, J., and SILVERMAN, P. H. - Ascaris suum: Immune reactions in mice. I. Larval metabolic and somatic antigens. Exp. Parasit., 26: 272-281, 1969.
15. GUERRERO, J., and SILVERMAN, P. H. - Ascaris suum: Immune reactions in mice. II. Metabolic and somatic antigens from in vitro cultured larvae. Exp. Parasit., 29: 110-115, 1971.
16. HASKINS, W. T., and WEINSTEIN, P.O. - The amine constitue.nts from the excretory products of Ascaris lumbricoides and Trichinella spiralis larvae. J. Parasit., 43: 28-32, 1957.
17. HJERTÉN, S. - Chromatography separation according to size of macromolecules and cell particles on columns of agarose suspensions. Arch. Biochem. Bioph., 99: 466-475, 1962.
18. HJERTÉN, S. - Chromatography on agarose spheres. In: Methods in Immunology and Immunochemistry. Vol. II. Academic Press. New York. 149- 154, 1966.
19. HOGARTH-SCOTT, R. S. - The molecular weight range of nematode allergens. Immunology, 13: 535-537, 1967.
20. JASKOSKI, J. B. and COLUCCI, A. V. - In vitro hatching of Ascaris suum eggs. Trans. Am. Micros. Soc., 83: 294-300, 1964.
21. KABAT, E. A., and MAYER, M. M. - Experimental Immunochemistry. Charles C. Thomas. 2nd ed. Springfield. 905 pp. 1964.
22. LEHNERT, J. P. - Studies on the biology and immunology of Ascaris suum in mice. Ph. D. thesis, University of Illinois, 1967.
23. LEVINE, H. S., and SILVERMAN, P. H. - Cultivation of Ascaris suum larvae in supplemented chemically defined media. J. Parasit., 55: 17-21, 1969.
24. LOWRY, O. H., ROSEBROUGH, N. J.( FARR, A. L. and RANDALL, R. J. - Protein measurements with the phenol reagent. J. Biol. Ohem., 193: 265-274, 1951.
25. MITCHISON, N. A. - Immunological paralysis as a dosage phenomenon. In: Regulation of the Antibody Response. Cinader, B. Ed. Charles C. Tbnmas. Springfield, Illinois, 54-65, 1968.
26. OLIVER-GONZALES, J. and GADEA, D. - Difference in serological properties of macroglobulins from câncer and normal sera (31608). Proc. Soc. Exp. Biol. Med., 123: 802-805, 1968.
27. OLIVER-GONZALES, J., HURLBRINK, P., CONDE, C. and KAGAN, I. G. - Serological activity of antigen isolated from the body flu d of Ascaris suum J. Imm., 102: 15-19, 1969.
28. OZEROL, N. H., and SILVERMAN, P. H. - Further characterization of active metabolites from histotropic larvae of Haemonchus contortus cultured in vitro. J. Parasit., 56: 1199- 1205, 1970.
29. SELA, M. - Antigenicity: Some molecular aspects. Science, 166: 1365- 1374, 1969.

Publication Dates

Publication in this collection
21 June 2013
Date of issue
Oct 1973

History

Accepted
25 June 1973
Received
25 June 1973

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. ADLER, F. L. and MOLLER, G. - Antigenic competition. In: Progress in Immunology. Ed. B. Amos. Academic Press. 1511-1514, 1971.

[2] 2. BJÖRNBOE, M. - Serum protein und Antistoffprotein bei Immunisierung mit mehreren Antigen. Z. Immunitatsf. Exper. therapie, 99: 245- 256, 1941.

[3] 3. Bio-Rad Laboratoires. Gel chromatography. Bio-Rad Laboratoires. Richmond, Califórnia. 89 pp. 1971.

[4] 4. BOYD, W. C. - Fundamentls of immunology . Interscience Publishers. New York. 74 pp. 1966.

[5] 5. BROWN, H. W., and CHAN, K. F. - The effect of piperazine HC1 on migrating larvae of Ascaris suum Goeze 1782. Am. J. Vet. Res., 16: 613-615, 1955.

[6] 6. DETERMAN, H. - Gel chromatography. Springer-Verlag. New York. 195 pp. 1968.

[7] 7. DOUVRES, F. W. and TROMBA, F. G. - Influence of pH, serum and cell cultures on development of Ascaris suum to fourth stage in vitro Journal Parasit., 56: 238-248, 1970.

[8] 8. DUNCAN, D. B. - A Bayesian approach to multiple comparison. Technometrics, 7: 171- 222.

[9] 9. FLODIN, P. - Methodogical aspects of gel filtration with special reference to desalting opera tion. J. Chrom., 5: 103-115, 1961.

[10] 10. GELOTTE, B. - Fractionation of proteins peptides and amino acids by gel filtration. In: A. T. James and L. D. Morris (eds). New Biochemical Separations. D. Van Nostrand Col. Ltd., p. 93-109, 1964.

[11] 11. GLENNY, A. T. - Diptheria antitoxin in the blood of normal horses. J. Path. Bact. 28: 241-250, 1925.

[12] 12. GLENNY, A. T. - The relation between dosage and death time. J. Path. Bact., 28: 251-260, 1925.

[13] 13. GUERRERO, J. - Studies on the immunogenicity of Ascaris suum larval antigens. Pt. D. Thesis, University of Illinois, Urbana, Illinois, 1971.

[14] 14. GUERRERO, J., and SILVERMAN, P. H. - Ascaris suum: Immune reactions in mice. I. Larval metabolic and somatic antigens. Exp. Parasit., 26: 272-281, 1969.

[15] 15. GUERRERO, J., and SILVERMAN, P. H. - Ascaris suum: Immune reactions in mice. II. Metabolic and somatic antigens from in vitro cultured larvae. Exp. Parasit., 29: 110-115, 1971.

[16] 16. HASKINS, W. T., and WEINSTEIN, P.O. - The amine constitue.nts from the excretory products of Ascaris lumbricoides and Trichinella spiralis larvae. J. Parasit., 43: 28-32, 1957.

[17] 17. HJERTÉN, S. - Chromatography separation according to size of macromolecules and cell particles on columns of agarose suspensions. Arch. Biochem. Bioph., 99: 466-475, 1962.

[18] 18. HJERTÉN, S. - Chromatography on agarose spheres. In: Methods in Immunology and Immunochemistry. Vol. II. Academic Press. New York. 149- 154, 1966.

[19] 19. HOGARTH-SCOTT, R. S. - The molecular weight range of nematode allergens. Immunology, 13: 535-537, 1967.

[20] 20. JASKOSKI, J. B. and COLUCCI, A. V. - In vitro hatching of Ascaris suum eggs. Trans. Am. Micros. Soc., 83: 294-300, 1964.

[21] 21. KABAT, E. A., and MAYER, M. M. - Experimental Immunochemistry. Charles C. Thomas. 2nd ed. Springfield. 905 pp. 1964.

[22] 22. LEHNERT, J. P. - Studies on the biology and immunology of Ascaris suum in mice. Ph. D. thesis, University of Illinois, 1967.

[23] 23. LEVINE, H. S., and SILVERMAN, P. H. - Cultivation of Ascaris suum larvae in supplemented chemically defined media. J. Parasit., 55: 17-21, 1969.

[24] 24. LOWRY, O. H., ROSEBROUGH, N. J.( FARR, A. L. and RANDALL, R. J. - Protein measurements with the phenol reagent. J. Biol. Ohem., 193: 265-274, 1951.

[25] 25. MITCHISON, N. A. - Immunological paralysis as a dosage phenomenon. In: Regulation of the Antibody Response. Cinader, B. Ed. Charles C. Tbnmas. Springfield, Illinois, 54-65, 1968.

[26] 26. OLIVER-GONZALES, J. and GADEA, D. - Difference in serological properties of macroglobulins from câncer and normal sera (31608). Proc. Soc. Exp. Biol. Med., 123: 802-805, 1968.

[27] 27. OLIVER-GONZALES, J., HURLBRINK, P., CONDE, C. and KAGAN, I. G. - Serological activity of antigen isolated from the body flu d of Ascaris suum J. Imm., 102: 15-19, 1969.

[28] 28. OZEROL, N. H., and SILVERMAN, P. H. - Further characterization of active metabolites from histotropic larvae of Haemonchus contortus cultured in vitro. J. Parasit., 56: 1199- 1205, 1970.

[29] 29. SELA, M. - Antigenicity: Some molecular aspects. Science, 166: 1365- 1374, 1969.