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. 1995 Nov;102(2):262–267. doi: 10.1111/j.1365-2249.1995.tb03775.x

Dengue virus-induced human cytotoxic factor: production by peripheral blood leucocytes in vitro.

R Mukerjee 1, U C Chaturvedi 1, R Dhawan 1
PMCID: PMC1553409  PMID: 7586676

Abstract

During dengue type 2 virus (DV) infection of mice a unique cytokine, the cytotoxic factor (CF), is produced which reproduces the pathological lesions seen in patients of dengue haemorrhagic fever (DHF). Recently we have observed a CF-like protein in the sera of DHF cases. The present study was undertaken to investigate whether DV can stimulate human peripheral blood mononuclear cells (PBMC) in vitro to produce human CF (hCF). Cultures prepared from PBMC or its enriched subpopulations were inoculated with 1000 LD50 of DV and controls with normal mouse brain suspension (NMB). Aliquots of cultures were harvested daily from 24 h to 96 h and their supernatant (CS) and cells were separated. CS were screened for viral titres and for the presence of hCF by cytotoxicity assay, inhibition ELISA, dot blot and Western blot tests using anti-mouse-CF antibodies. The RNA from the cells was screened in Northern blot and dot blot tests for the presence of mRNA for CF. It was observed that hCF appeared in the CS of DV-infected culture of PBMC and T-enriched cells at 48 h and was present until 96 h; no CF was detected in CS of B cells or monocyte cultures. The production of hCF was abrogated by pretreatment of the T cells with anti-CD4 antibodies but not with anti-CD8 antibodies, indicating that hCF was produced by CD4+ T cells. The Northern blot analysis using oligonucleotide probes prepared on the basis of amino-terminal sequence of mouse CF, showed presence of mRNA for hCF in PBMC and T cell cultures. DV replicated in PBMC cultures with peak titres at 48 h. The findings of the present study demonstrate that DV-induced hCF is produced by human T cells.

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Selected References

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

  1. Andrews B. S., Theofilopoulos A. N., Peters C. J., Loskutoff D. J., Brandt W. E., Dixon F. J. Replication of dengue and junin viruses in cultured rabbit and human endothelial cells. Infect Immun. 1978 Jun;20(3):776–781. doi: 10.1128/iai.20.3.776-781.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chaturvedi U. C., Bhargava A., Mathur A. Production of cytotoxic factor in the spleen of dengue virus-infected mice. Immunology. 1980 Aug;40(4):665–671. [PMC free article] [PubMed] [Google Scholar]
  3. Chaturvedi U. C., Dhawan R., Khanna M., Mathur A. Breakdown of the blood-brain barrier during dengue virus infection of mice. J Gen Virol. 1991 Apr;72(Pt 4):859–866. doi: 10.1099/0022-1317-72-4-859. [DOI] [PubMed] [Google Scholar]
  4. Chaturvedi U. C., Mathur A., Tandon P., Natu S. M., Rajvanshi S., Tandon H. O. Variable effect on peripheral blood leucocytes during JE virus infection of man. Clin Exp Immunol. 1979 Dec;38(3):492–498. [PMC free article] [PubMed] [Google Scholar]
  5. Chaturvedi U. C., Mukerjee R., Dhawan R. Active immunization by a dengue virus-induced cytokine. Clin Exp Immunol. 1994 May;96(2):202–207. doi: 10.1111/j.1365-2249.1994.tb06542.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chaturvedi U. C., Nagar R., Mathur A. Effect of dengue virus infection on Fc-receptor functions of mouse macrophages. J Gen Virol. 1983 Nov;64(Pt 11):2399–2407. doi: 10.1099/0022-1317-64-11-2399. [DOI] [PubMed] [Google Scholar]
  7. Chaturvedi U. C., Tandon P., Mathur A. Effect of immunosuppression on dengue virus infection in mice. J Gen Virol. 1977 Sep;36(3):449–458. doi: 10.1099/0022-1317-36-3-449. [DOI] [PubMed] [Google Scholar]
  8. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  9. Khanna M., Chaturvedi U. C. Purification and amino-terminal sequence of the dengue virus-induced cytotoxic factor. Int J Exp Pathol. 1992 Feb;73(1):43–49. [PMC free article] [PubMed] [Google Scholar]
  10. Khanna M., Chaturvedi U. C., Sharma M. C., Pandey V. C., Mathur A. Increased capillary permeability mediated by a dengue virus-induced lymphokine. Immunology. 1990 Mar;69(3):449–453. [PMC free article] [PubMed] [Google Scholar]
  11. Liew F. Y., Simpson E. Delayed-type hypersensitivity responses to H-Y: characterization and mapping of Ir genes. Immunogenetics. 1980;11(3):255–266. doi: 10.1007/BF01567792. [DOI] [PubMed] [Google Scholar]
  12. Navikas V., Link J., Wahren B., Persson C., Link H. Increased levels of interferon-gamma (IFN-gamma), IL-4 and transforming growth factor-beta (TGF-beta) mRNA expressing blood mononuclear cells in human HIV infection. Clin Exp Immunol. 1994 Apr;96(1):59–63. doi: 10.1111/j.1365-2249.1994.tb06230.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rizvi N., Chaturvedi U. C., Mathur A. Inhibition of the presentation of dengue virus antigen by macrophages to B cells by serine-protease inhibitors. Int J Exp Pathol. 1991 Feb;72(1):23–29. [PMC free article] [PubMed] [Google Scholar]
  14. Rizvi N., Chaturvedi U. C., Mathur A. Obligatory role of macrophages in dengue virus antigen presentation to B lymphocytes. Immunology. 1989 May;67(1):38–43. [PMC free article] [PubMed] [Google Scholar]
  15. Shukla M. I., Dalakoti H., Chaturvedi U. C. Ly phenotype of T lymphocytes producing dengue virus-induced immunosuppressive factors. Indian J Exp Biol. 1982 Jul;20(7):525–528. [PubMed] [Google Scholar]
  16. Trizio D., Cudkowicz G. Separation of T and B lymphocytes by nylon wool columns: evaluation of efficacy by functional assays in vivo. J Immunol. 1974 Oct;113(4):1093–1097. [PubMed] [Google Scholar]
  17. Voller A., Bidwell D. E., Bartlett A. Enzyme immunoassays in diagnostic medicine. Theory and practice. Bull World Health Organ. 1976;53(1):55–65. [PMC free article] [PubMed] [Google Scholar]

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