Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1994 Sep;62(9):3696–3704. doi: 10.1128/iai.62.9.3696-3704.1994

Epitopes of the Onchocerca volvulus RAL1 antigen, a member of the calreticulin family of proteins, recognized by sera from patients with onchocerciasis.

L A Rokeach 1, P A Zimmerman 1, T R Unnasch 1
PMCID: PMC303020  PMID: 7520419

Abstract

RAL1 is an antigen (Ag) encoded by the filarial nematode Onchocerca volvulus, the parasite causing onchocerciasis (river blindness). RAL1 shares 64.4% identity with the autoantigen calreticulin. The striking similarity of the parasite Ag and the human autoantigen has led to the hypothesis that RAL1 may induce a cross-reactive immune response to calreticulin, which in turn may be involved in the pathogenesis of onchocerciasis. To test this hypothesis, we explored the immune response to RAL1 recombinant Ag (RAL1 rAg) and human calreticulin in patients with O. volvulus infection. A total of 86% of the O. volvulus-infected individuals produced antibodies recognizing RAL1 rAg. Antibody reactivity to RAL1 rAg in patient sera was confined primarily to the central and carboxyl-terminal parts of the molecule. No significant correlations were found to associate recognition of RAL1 rAg, or any particular portion thereof, with a particular disease state. Antibodies against RAL1 thus appear to be produced as a general immune reaction to O. volvulus infection and do not necessarily lead to a cross-reacting response with the host protein. In contrast, 33% of the patient sera tested bound recombinant human calreticulin. All of these sera also recognized a polypeptide encompassing the carboxyl-terminal portion of the RAL1 rAg. These results suggest that recognition of an epitope encoded in the carboxyl-terminal portion of RAL1 is at least in part responsible for inducing a cross-reacting immune response to the host protein.

Full text

PDF
3696

Images in this article

3700Image
on p.3700

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baksh S., Michalak M. Expression of calreticulin in Escherichia coli and identification of its Ca2+ binding domains. J Biol Chem. 1991 Nov 15;266(32):21458–21465. [PubMed] [Google Scholar]
  2. Bartlett A., Turk J., Ngu J., Mackenzie C. D., Fuglsang H. Variation in delayed hypersensitivity in onchocerciasis. Trans R Soc Trop Med Hyg. 1978;72(4):372–377. doi: 10.1016/0035-9203(78)90130-x. [DOI] [PubMed] [Google Scholar]
  3. Bockarie M. J., Davies J. B. The transmission of onchocerciasis at a forest village in Sierra Leone. II. Man-fly contact, human activity and exposure to transmission. Ann Trop Med Parasitol. 1990 Dec;84(6):599–605. doi: 10.1080/00034983.1990.11812515. [DOI] [PubMed] [Google Scholar]
  4. Burns K., Helgason C. D., Bleackley R. C., Michalak M. Calreticulin in T-lymphocytes. Identification of calreticulin in T-lymphocytes and demonstration that activation of T cells correlates with increased levels of calreticulin mRNA and protein. J Biol Chem. 1992 Sep 25;267(27):19039–19042. [PubMed] [Google Scholar]
  5. Chan C. C., Nussenblatt R. B., Kim M. K., Palestine A. G., Awadzi K., Ottesen E. A. Immunopathology of ocular onchocerciasis. 2. Anti-retinal autoantibodies in serum and ocular fluids. Ophthalmology. 1987 Apr;94(4):439–443. doi: 10.1016/s0161-6420(87)33452-9. [DOI] [PubMed] [Google Scholar]
  6. Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
  7. Dadzie K. Y., Remme J., Baker R. H., Rolland A., Thylefors B. Ocular onchocerciasis and intensity of infection in the community. III. West African rainforest foci of the vector Simulium sanctipauli. Trop Med Parasitol. 1990 Dec;41(4):376–382. [PubMed] [Google Scholar]
  8. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Donelson J. E., Duke B. O., Moser D., Zeng W. L., Erondu N. E., Lucius R., Renz A., Karam M., Flores G. Z. Construction of Onchocerca volvulus cDNA libraries and partial characterization of the cDNA for a major antigen. Mol Biochem Parasitol. 1988 Dec;31(3):241–250. doi: 10.1016/0166-6851(88)90154-5. [DOI] [PubMed] [Google Scholar]
  10. Donelson J. E., Zeng W. A comparison of trans-RNA splicing in trypanosomes and nematodes. Parasitol Today. 1990 Oct;6(10):327–334. doi: 10.1016/0169-4758(90)90177-6. [DOI] [PubMed] [Google Scholar]
  11. Donnelly J. J., Rockey J. H., Bianco A. E., Soulsby E. J. Ocular immunopathologic findings of experimental onchocerciasis. Arch Ophthalmol. 1984 Apr;102(4):628–634. doi: 10.1001/archopht.1984.01040030500036. [DOI] [PubMed] [Google Scholar]
  12. Donnelly J. J., Taylor H. R., Young E., Khatami M., Lok J. B., Rockey J. H. Experimental ocular onchocerciasis in cynomolgus monkeys. Invest Ophthalmol Vis Sci. 1986 Apr;27(4):492–499. [PubMed] [Google Scholar]
  13. Dorfman D. M., Zon L. I., Orkin S. H. Rapid amplification of lambda gt11 bacteriophage library inserts from plaques using the polymerase chain reaction (PCR). Biotechniques. 1989 Jun;7(6):568–570. [PubMed] [Google Scholar]
  14. Dupuis M., Schaerer E., Krause K. H., Tschopp J. The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T lymphocytes. J Exp Med. 1993 Jan 1;177(1):1–7. doi: 10.1084/jem.177.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Freedman D. O., Lujan-Trangay A., Steel C., Gonzalez-Peralta C., Nutman T. B. Immunoregulation in onchocerciasis. Functional and phenotypic abnormalities of lymphocyte subsets and changes with therapy. J Clin Invest. 1991 Jul;88(1):231–238. doi: 10.1172/JCI115282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gallin M. Y., Murray D., Lass J. H., Grossniklaus H. E., Greene B. M. Experimental interstitial keratitis induced by Onchocerca volvulus antigens. Arch Ophthalmol. 1988 Oct;106(10):1447–1452. doi: 10.1001/archopht.1988.01060140611033. [DOI] [PubMed] [Google Scholar]
  17. Gallin M. Y., Tan M., Kron M. A., Rechnitzer D., Greene B. M., Newland H. S., White A. T., Taylor H. R., Unnasch T. R. Onchocerca volvulus recombinant antigen: physical characterization and clinical correlates with serum reactivity. J Infect Dis. 1989 Sep;160(3):521–529. doi: 10.1093/infdis/160.3.521. [DOI] [PubMed] [Google Scholar]
  18. Gallin M., Edmonds K., Ellner J. J., Erttmann K. D., White A. T., Newland H. S., Taylor H. R., Greene B. M. Cell-mediated immune responses in human infection with Onchocerca volvulus. J Immunol. 1988 Mar 15;140(6):1999–2007. [PubMed] [Google Scholar]
  19. Garner A., Duke B. O. Fundus lesions in the rabbit eye following inoculation of onchocerca volvulus microfilariae into the posterior segment. II. Pathology. Tropenmed Parasitol. 1976 Mar;27(1):19–29. [PubMed] [Google Scholar]
  20. Greene B. M., Gbakima A. A., Albiez E. J., Taylor H. R. Humoral and cellular immune responses to Onchocerca volvulus infection in humans. Rev Infect Dis. 1985 Nov-Dec;7(6):789–795. doi: 10.1093/clinids/7.6.789. [DOI] [PubMed] [Google Scholar]
  21. Guldner H. H. Mapping of epitopes recognized by anti-(U1)RNP autoantibodies. Mol Biol Rep. 1992 Jun;16(3):155–164. doi: 10.1007/BF00464703. [DOI] [PubMed] [Google Scholar]
  22. Habets W. J., Hoet M. H., De Jong B. A., Van der Kemp A., Van Venrooij W. J. Mapping of B cell epitopes on small nuclear ribonucleoproteins that react with human autoantibodies as well as with experimentally-induced mouse monoclonal antibodies. J Immunol. 1989 Oct 15;143(8):2560–2566. [PubMed] [Google Scholar]
  23. Handford P. A., Mayhew M., Baron M., Winship P. R., Campbell I. D., Brownlee G. G. Key residues involved in calcium-binding motifs in EGF-like domains. Nature. 1991 May 9;351(6322):164–167. doi: 10.1038/351164a0. [DOI] [PubMed] [Google Scholar]
  24. Hawn T. R., Tom T. D., Strand M. Molecular cloning and expression of SmIrV1, a Schistosoma mansoni antigen with similarity to calnexin, calreticulin, and OvRal1. J Biol Chem. 1993 Apr 15;268(11):7692–7698. [PubMed] [Google Scholar]
  25. Horsfall A. C. Molecular mimicry and autoantigens in connective tissue diseases. Mol Biol Rep. 1992 Jun;16(3):139–147. doi: 10.1007/BF00464701. [DOI] [PubMed] [Google Scholar]
  26. Khalife J., Trottein F., Schacht A. M., Godin C., Pierce R. J., Capron A. Cloning of the gene encoding a Schistosoma mansoni antigen homologous to human Ro/SS-A autoantigen. Mol Biochem Parasitol. 1993 Feb;57(2):193–202. doi: 10.1016/0166-6851(93)90195-4. [DOI] [PubMed] [Google Scholar]
  27. Koerner T. J., Hill J. E., Myers A. M., Tzagoloff A. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods Enzymol. 1991;194:477–490. doi: 10.1016/0076-6879(91)94036-c. [DOI] [PubMed] [Google Scholar]
  28. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  29. Lieu T. S., Newkirk M. M., Capra J. D., Sontheimer R. D. Molecular characterization of human Ro/SS-A antigen. Amino terminal sequence of the protein moiety of human Ro/SS-A antigen and immunological activity of a corresponding synthetic peptide. J Clin Invest. 1988 Jul;82(1):96–101. doi: 10.1172/JCI113607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lux F. A., McCauliffe D. P., Büttner D. W., Lucius R., Capra J. D., Sontheimer R. D., Lieu T. S. Serological cross-reactivity between a human Ro/SS-A autoantigen (calreticulin) and the lambda Ral-1 antigen of Onchocerca volvulus. J Clin Invest. 1992 Jun;89(6):1945–1951. doi: 10.1172/JCI115801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mackenzie C. D., Williams J. F., Sisley B. M., Steward M. W., O'Day J. Variations in host responses and the pathogenesis of human onchocerciasis. Rev Infect Dis. 1985 Nov-Dec;7(6):802–808. doi: 10.1093/clinids/7.6.802. [DOI] [PubMed] [Google Scholar]
  32. McCauliffe D. P., Zappi E., Lieu T. S., Michalak M., Sontheimer R. D., Capra J. D. A human Ro/SS-A autoantigen is the homologue of calreticulin and is highly homologous with onchocercal RAL-1 antigen and an aplysia "memory molecule". J Clin Invest. 1990 Jul;86(1):332–335. doi: 10.1172/JCI114704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Meilof J. F., Van der Lelij A., Rokeach L. A., Hoch S. O., Smeenk R. J. Autoimmunity and filariasis. Autoantibodies against cytoplasmic cellular proteins in sera of patients with onchocerciasis. J Immunol. 1993 Nov 15;151(10):5800–5809. [PubMed] [Google Scholar]
  34. Michalak M., Milner R. E., Burns K., Opas M. Calreticulin. Biochem J. 1992 Aug 1;285(Pt 3):681–692. doi: 10.1042/bj2850681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Munro S., Pelham H. R. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. doi: 10.1016/0092-8674(87)90086-9. [DOI] [PubMed] [Google Scholar]
  36. Netter H. J., Guldner H. H., Szostecki C., Will H. Major autoantigenic sites of the (U1) small nuclear ribonucleoprotein-specific 68-kDa protein. Scand J Immunol. 1990 Aug;32(2):163–176. doi: 10.1111/j.1365-3083.1990.tb02906.x. [DOI] [PubMed] [Google Scholar]
  37. Newland H. S., White A. T., Greene B. M., D'Anna S. A., Keyvan-Larijani E., Aziz M. A., Williams P. N., Taylor H. R. Effect of single-dose ivermectin therapy on human Onchocerca volvulus infection with onchocercal ocular involvement. Br J Ophthalmol. 1988 Aug;72(8):561–569. doi: 10.1136/bjo.72.8.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ngu J. L. Immunological studies on onchocerciasis. Varying skin hypersensitivity and leucocyte migration inhibition in a clinical spectrum of the disease. Acta Trop. 1978 Sep;35(3):269–279. [PubMed] [Google Scholar]
  39. Oldstone M. B. Molecular mimicry and autoimmune disease. Cell. 1987 Sep 11;50(6):819–820. doi: 10.1016/0092-8674(87)90507-1. [DOI] [PubMed] [Google Scholar]
  40. Pearson C. A. A major cause of third world rheumatism largely overlooked--onchocerciasis. Br J Rheumatol. 1988 Oct;27(5):414–416. doi: 10.1093/rheumatology/27.5.414. [DOI] [PubMed] [Google Scholar]
  41. Rokeach L. A., Haselby J. A., Hoch S. O. High-level bacterial expression, purification and characterization of human calreticulin. Protein Eng. 1991 Dec;4(8):981–987. doi: 10.1093/protein/4.8.981. [DOI] [PubMed] [Google Scholar]
  42. Rokeach L. A., Haselby J. A., Meilof J. F., Smeenk R. J., Unnasch T. R., Greene B. M., Hoch S. O. Characterization of the autoantigen calreticulin. J Immunol. 1991 Nov 1;147(9):3031–3039. [PubMed] [Google Scholar]
  43. Rokeach L. A., Hoch S. O. B-cell epitopes of Sm autoantigens. Mol Biol Rep. 1992 Jun;16(3):165–174. doi: 10.1007/BF00464704. [DOI] [PubMed] [Google Scholar]
  44. Rokeach L. A., Jannatipour M., Haselby J. A., Hoch S. O. Mapping of the immunoreactive domains of a small nuclear ribonucleoprotein-associated Sm-D autoantigen. Clin Immunol Immunopathol. 1992 Dec;65(3):315–324. doi: 10.1016/0090-1229(92)90163-i. [DOI] [PubMed] [Google Scholar]
  45. Rokeach L. A., Jannatipour M., Hoch S. O. Heterologous expression and epitope mapping of a human small nuclear ribonucleoprotein-associated Sm-B'/B autoantigen. J Immunol. 1990 Feb 1;144(3):1015–1022. [PubMed] [Google Scholar]
  46. Rougemont A., Boisson-Pontal M. E., Pontal P. G., Gridel F., Sangare S. Tuberculin skin tests and B.C.G. vaccination in hyperendemic area of onchocerciasis. Lancet. 1977 Feb 5;1(8006):309–309. doi: 10.1016/s0140-6736(77)91857-8. [DOI] [PubMed] [Google Scholar]
  47. Routsias J. G., Tzioufas A. G., Sakarellos-Daitsiotis M., Sakarellos C., Moutsopoulos H. M. Calreticulin synthetic peptide analogues: anti-peptide antibodies in autoimmune rheumatic diseases. Clin Exp Immunol. 1993 Mar;91(3):437–441. doi: 10.1111/j.1365-2249.1993.tb05921.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  49. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Schulz-Key H. A simple technique to assess the total number of Onchocerca volvulus microfilariae in skin snips. Tropenmed Parasitol. 1978 Mar;29(1):51–54. [PubMed] [Google Scholar]
  51. Sekharudu Y. C., Sundaralingam M. A structure-function relationship for the calcium affinities of regulatory proteins containing 'EF-hand' pairs. Protein Eng. 1988 Jul;2(2):139–146. doi: 10.1093/protein/2.2.139. [DOI] [PubMed] [Google Scholar]
  52. Smith M. J. A C. elegans gene encodes a protein homologous to mammalian calreticulin. DNA Seq. 1992;2(4):235–240. doi: 10.3109/10425179209020808. [DOI] [PubMed] [Google Scholar]
  53. Tan E. M. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 1989;44:93–151. doi: 10.1016/s0065-2776(08)60641-0. [DOI] [PubMed] [Google Scholar]
  54. Unnasch T. R., Gallin M. Y., Soboslay P. T., Erttmann K. D., Greene B. M. Isolation and characterization of expression cDNA clones encoding antigens of Onchocerca volvulus infective larvae. J Clin Invest. 1988 Jul;82(1):262–269. doi: 10.1172/JCI113581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Wada I., Rindress D., Cameron P. H., Ou W. J., Doherty J. J., 2nd, Louvard D., Bell A. W., Dignard D., Thomas D. Y., Bergeron J. J. SSR alpha and associated calnexin are major calcium binding proteins of the endoplasmic reticulum membrane. J Biol Chem. 1991 Oct 15;266(29):19599–19610. [PubMed] [Google Scholar]
  56. Ward D. J., Nutman T. B., Zea-Flores G., Portocarrero C., Lujan A., Ottesen E. A. Onchocerciasis and immunity in humans: enhanced T cell responsiveness to parasite antigen in putatively immune individuals. J Infect Dis. 1988 Mar;157(3):536–543. doi: 10.1093/infdis/157.3.536. [DOI] [PubMed] [Google Scholar]
  57. Watson M. E. Compilation of published signal sequences. Nucleic Acids Res. 1984 Jul 11;12(13):5145–5164. doi: 10.1093/nar/12.13.5145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Zeng W. L., Alarcon C. M., Donelson J. E. Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans. Mol Cell Biol. 1990 Jun;10(6):2765–2773. doi: 10.1128/mcb.10.6.2765. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES