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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 May;85(10):3613–3617. doi: 10.1073/pnas.85.10.3613

Production of a monoclonal antibody against the Snow Mountain agent of gastroenteritis by in vitro immunization of murine spleen cells.

J Treanor 1, R Dolin 1, H P Madore 1
PMCID: PMC280264  PMID: 2835775

Abstract

The Snow Mountain agent (SMA) is a 27- to 32-nm noncultivatable virus that causes acute gastroenteritis in humans. SMA is morphologically similar to but immunologically distinct from the Norwalk agent. SMA has been partially purified from the stool of experimentally infected volunteers and contains a single structural protein of Mr 62,000 as well as one or more non-virion-associated soluble proteins. Further characterization of this important human pathogen and other Norwalk-like viruses has been hindered by the lack of reagents with which to study them. To further characterize SMA, we developed a monoclonal antibody to SMA using in vitro immunization--a technique that permitted use of small quantities of antigen for immunization. The monoclonal antibody, SM-4, was specific for SMA and did not react with the Norwalk or Hawaii agents. In addition, SM-4 reacted with purified virion but not with the soluble protein. SM-4 also blocked the ability of labeled postinfection human IgG to bind to purified virion. Finally, both SM-4 and human postinfection sera specifically recognized the Mr 62,000 virion-associated protein. Thus, SM-4 is directed against an epitope present on the SMA structural protein that is not shared by the Norwalk or Hawaii agents and that is not present on the soluble protein. The availability of a monoclonal antibody against SMA should facilitate further purification and characterization of this agent. The techniques utilized in these studies provide a method for the production of additional monoclonal antibodies to this group of viruses and also should be useful for the study of other occult viral agents.

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Selected References

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  1. Acute infectious nonbacterial gastroenteritis: etiology and pathogenesis. Ann Intern Med. 1972 Jun;76(6):993–1008. doi: 10.7326/0003-4819-76-6-993. [DOI] [PubMed] [Google Scholar]
  2. Brondum J., Spitalny K. C., Vogt R. L., Godlewski K., Madore H. P., Dolin R. Snow Mountain agent associated with an outbreak of gastroenteritis in Vermont. J Infect Dis. 1985 Oct;152(4):834–837. doi: 10.1093/infdis/152.4.834. [DOI] [PubMed] [Google Scholar]
  3. Dolin R., Blacklow N. R., DuPont H., Buscho R. F., Wyatt R. G., Kasel J. A., Hornick R., Chanock R. M. Biological properties of Norwalk agent of acute infectious nonbacterial gastroenteritis. Proc Soc Exp Biol Med. 1972 Jun;140(2):578–583. doi: 10.3181/00379727-140-36508. [DOI] [PubMed] [Google Scholar]
  4. Dolin R., Reichman R. C., Roessner K. D., Tralka T. S., Schooley R. T., Gary W., Morens D. Detection by immune electron microscopy of the Snow Mountain agent of acute viral gastroenteritis. J Infect Dis. 1982 Aug;146(2):184–189. doi: 10.1093/infdis/146.2.184. [DOI] [PubMed] [Google Scholar]
  5. Dolin R., Roessner K. D., Treanor J. J., Reichman R. C., Phillips M., Madore H. P. Radioimmunoassay for detection of the Snow Mountain Agent of viral gastroenteritis. J Med Virol. 1986 May;19(1):11–18. doi: 10.1002/jmv.1890190103. [DOI] [PubMed] [Google Scholar]
  6. Gary G. W., Jr, Kaplan J. E., Stine S. E., Anderson L. J. Detection of Norwalk virus antibodies and antigen with a biotin-avidin immunoassay. J Clin Microbiol. 1985 Aug;22(2):274–278. doi: 10.1128/jcm.22.2.274-278.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goodman M. G., Weigle W. O. Derivatized guanine nucleosides: a new class of adjuvant for in vitro antibody responses. J Immunol. 1983 Jun;130(6):2580–2585. [PubMed] [Google Scholar]
  8. Greenberg H. B., Valdesuso J. R., Kalica A. R., Wyatt R. G., McAuliffe V. J., Kapikian A. Z., Chanock R. M. Proteins of Norwalk virus. J Virol. 1981 Mar;37(3):994–999. doi: 10.1128/jvi.37.3.994-999.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greenberg H. B., Valdesuso J., Kapikian A. Z., Chanock R. M., Wyatt R. G., Szmuness W., Larrick J., Kaplan J., Gilman R. H., Sack D. A. Prevalence of antibody to the Norwalk virus in various countries. Infect Immun. 1979 Oct;26(1):270–273. doi: 10.1128/iai.26.1.270-273.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Greenberg H. B., Wyatt R. G., Valdesuso J., Kalica A. R., London W. T., Chanock R. M., Kapikian A. Z. Solid-phase microtiter radioimmunoassay for detection of the Norwalk strain of acute nonbacterial, epidemic gastroenteritis virus and its antibodies. J Med Virol. 1978;2(2):97–108. doi: 10.1002/jmv.1890020204. [DOI] [PubMed] [Google Scholar]
  11. Guest C., Spitalny K. C., Madore H. P., Pray K., Dolin R., Herrmann J. E., Blacklow N. R. Foodborne Snow Mountain agent gastroenteritis in a school cafeteria. Pediatrics. 1987 Apr;79(4):559–563. [PubMed] [Google Scholar]
  12. Herrmann J. E., Nowak N. A., Blacklow N. R. Detection of Norwalk virus in stools by enzyme immunoassay. J Med Virol. 1985 Oct;17(2):127–133. doi: 10.1002/jmv.1890170205. [DOI] [PubMed] [Google Scholar]
  13. Jackson S., Sogn J. A., Kindt T. J. Microdetermination of rabbit immunoglobulin allotypes by ELISA using specific antibodies conjugated with peroxidase or with biotin. J Immunol Methods. 1982;48(3):299–309. doi: 10.1016/0022-1759(82)90331-3. [DOI] [PubMed] [Google Scholar]
  14. Kapikian A. Z., Wyatt R. G., Dolin R., Thornhill T. S., Kalica A. R., Chanock R. M. Visualization by immune electron microscopy of a 27-nm particle associated with acute infectious nonbacterial gastroenteritis. J Virol. 1972 Nov;10(5):1075–1081. doi: 10.1128/jvi.10.5.1075-1081.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Luben R. A., Brazeau P., Böhlen P., Guillemin R. Monoclonal antibodies to hypothalamic growth hormone-releasing factor with picomoles of antigen. Science. 1982 Nov 26;218(4575):887–889. doi: 10.1126/science.6813967. [DOI] [PubMed] [Google Scholar]
  16. Luben R. A., Mohler M. A. In vitro immunization as an adjunct to the production of hybridomas producing antibodies against the lymphokine osteoclast activating factor. Mol Immunol. 1980 May;17(5):635–639. doi: 10.1016/0161-5890(80)90161-3. [DOI] [PubMed] [Google Scholar]
  17. Madore H. P., Treanor J. J., Dolin R. Characterization of the Snow Mountain agent of viral gastroenteritis. J Virol. 1986 May;58(2):487–492. doi: 10.1128/jvi.58.2.487-492.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Madore H. P., Treanor J. J., Pray K. A., Dolin R. Enzyme-linked immunosorbent assays for Snow Mountain and Norwalk agents of viral gastroenteritis. J Clin Microbiol. 1986 Sep;24(3):456–459. doi: 10.1128/jcm.24.3.456-459.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pardue R. L., Brady R. C., Perry G. W., Dedman J. R. Production of monoclonal antibodies against calmodulin by in vitro immunization of spleen cells. J Cell Biol. 1983 Apr;96(4):1149–1154. doi: 10.1083/jcb.96.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reading C. L. Theory and methods for immunization in culture and monoclonal antibody production. J Immunol Methods. 1982 Sep 30;53(3):261–291. doi: 10.1016/0022-1759(82)90175-2. [DOI] [PubMed] [Google Scholar]
  21. Schaffer F. L., Bachrach H. L., Brown F., Gillespie J. H., Burroughs J. N., Madin S. H., Madeley C. R., Povey R. C., Scott F., Smith A. W. Caliciviridae. Intervirology. 1980;14(1):1–6. doi: 10.1159/000149155. [DOI] [PubMed] [Google Scholar]
  22. Thornhill T. S., Kalica A. R., Wyatt R. G., Kapikian A. Z., Chanock R. M. Pattern of shedding of the Norwalk particle in stools during experimentally induced gastroenteritis in volunteers as determined by immune electron microscopy. J Infect Dis. 1975 Jul;132(1):28–34. doi: 10.1093/infdis/132.1.28. [DOI] [PubMed] [Google Scholar]
  23. Thornhill T. S., Wyatt R. G., Kalica A. R., Dolin R., Chanock R. M., Kapikian A. Z. Detection by immune electron microscopy of 26- to 27-nm viruslike particles associated with two family outbreaks of gastroenteritis. J Infect Dis. 1977 Jan;135(1):20–27. doi: 10.1093/infdis/135.1.20. [DOI] [PubMed] [Google Scholar]
  24. Truman B. I., Madore H. P., Menegus M. A., Nitzkin J. L., Dolin R. Snow Mountain agent gastroenteritis from clams. Am J Epidemiol. 1987 Sep;126(3):516–525. doi: 10.1093/oxfordjournals.aje.a114684. [DOI] [PubMed] [Google Scholar]
  25. Van Ness J., Laemmli U. K., Pettijohn D. E. Immunization in vitro and production of monoclonal antibodies specific to insoluble and weakly immunogenic proteins. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7897–7901. doi: 10.1073/pnas.81.24.7897. [DOI] [PMC free article] [PubMed] [Google Scholar]

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