Skip to main content

Advertisement

Log in

Effect of the calcium phosphate-mediated RNA uptake on the transfer of cellular immunity of a synthetic peptide of HIV-1 to human lymphocytes by exogenous RNA

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

It is known that exogenous RNA molecules can be taken up by eukaryotic cells and can exert a variety of biological effects both in vitro and in vivo. The modulation of human lymphocytes by exogenous RNAs has medical implications. The exogenous RNA used in this study was obtained from lymphoid organs of animals immunized with the synthetic peptide p12 of HIV-1 and was referred to as p12-RNA. Human lymphocytes were transfected with the p12-RNA and the transfer of immunoreactivity of p12 was assessed by the lymphocyte proliferation and the leukocyte adherence inhibition assays. Our results indicate that the transfer of cellular immune response to the p12 occurred in 9 donors (60%) who were named responsive individuals whereas 6 donors (40%) were non-responsives. We also found that the calcium phosphate-mediated RNA uptake method is effective in converting non-responsive into responsive donors. The calcium phosphate-mediated RNA uptake may also be used to increase the efficiency of RNA transfection in other models with medical implications and to contribute to a better understanding of the molecular events involved in the uptake of RNA. Our findings give support for the use of exogenous RNAs obtained from lymphoid organs of immunized animals with synthetic peptides of HIV-1 in the immune reconstitution of individuals infected with HIV-1.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Bhargava PM, Shanmugam G: Uptake of nonviral nucleic acids by mammalian cells. In: J.N. Davidson, W.E. Cohn (eds). Progress in Nucleic Acid Research and Molecular Biology. Academic Press, New York, 1971, vol 11, pp 103–192

    Google Scholar 

  2. Solo S, Christian JL, Moon RT, Melon DA: Injected NWT RNA induces a complete body axis in Xenopus embryos. Cell 67: 741–752, 1991

    Google Scholar 

  3. Boczkowski D, Nair SK, Snyder D, Gilboa E: Dendritic cells pulsed with RNA are potent antigen-presenting cells in vitro and in vivo. J Exp Med 184: 465–472, 1996

    Google Scholar 

  4. Timmerman JM, Levy R: Dendritic cell vaccines for cancer immunotherapy. Annu Rev Med 50: 507–529, 1999

    Google Scholar 

  5. Kleinschmidt AM, Pederson T: RNA Processing and ribonucleoprotein assembly studied in vivo by RNA transfection. Proc Natl Acad Sci USA 87: 1283–1287, 1990

    Google Scholar 

  6. Wang JJG, Steel S, Wisniewolski R, Wang CY: Detection of antibodies to human T-lymphotropic virus type III by using a synthetic peptide of 21 amino acid residues corresponding to a highly antigenic segment of gp 41 envelope protein. Proc Natl Acad Sci USA 83: 6159–6183, 1986

    Google Scholar 

  7. Merrifield RB: Solid phase peptide synthesis I. The synthesis of a tetrapeptide. J Am Chem Soc 85: 2149–2154, 1963

    Google Scholar 

  8. Sales VSF: Doctorate Thesis. School of Medicine of Ribeirão Preto University of São Paulo, 1998

  9. White BN, De Lucca FL: Preparation and analysis of RNA. In: R.B. Turner (ed). Analytical Biochemistry of Insects. Elsevier Scientific Publishing, Amsterdam, 1977, pp 85–130

    Google Scholar 

  10. Watanabe MAE, Almeida DO, Serrano SV, Sales VSF, Soares FA, De Lucca FL: Immunotherapy of metastases with lymphocytes treated with exogenous RNA in mice bearing B16 melanoma. Cell Mol Biol 43: 393–398, 1997

    Google Scholar 

  11. Artigas C, Thomson DM, Dusko M, Sutherland M, Scanzano R, Shenouda G, Dubois AEJ: Identification of a Mr 40,000 polypeptide from colorectal cancer with express organ-specific cancer neoantigen activity as determined by leukocyte adherence inhibition. Cancer Res 46: 1847–1881, 1986

    Google Scholar 

  12. De Lucca FL, Bertolini MC, Zini MN: In vitro transfer of reactivity to Trypanosoma cruzi antigens from rat cells to human cells with immune RNA. J Infect Dis 145: 148–151, 1982

    Google Scholar 

  13. Dray S, Braun DP: Some perspectives on the transfer of cell-mediated immunity by immune RNA. Mol Cell Biochem 25: 15–30, 1979

    Google Scholar 

  14. Mroczkowski B, Dym HP, Siegel EJ, Heywood SM: Uptake and utilization of RNA by myogenic cells in culture. J Cell Biol 87: 65–71, 1980

    Google Scholar 

  15. Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A, Felgner PL: Directed gene transfer into mouse muscle in vivo. Science 247: 1465–1468, 1990

    Google Scholar 

  16. Ribeiro RA, De Lucca FL, Flores CA, Cunha FQ, Ferreira SH: RNA from LPS-stimulated macrophages induces de release of tumor necrosis factor-á and interleukin-1 by resident macrophages. Med Inflam 2: 435–442, 1993

    Google Scholar 

  17. Ribeiro RA, Flores CA, Cunha FQ, Ferreira SH, De Lucca FL: Partial characterization of the RNA from LPS-stimulated macrophages that induces the release of chemotactic cytokines by resident macrophages. Mol Cell Biochem 148: 105–113, 1995

    Google Scholar 

  18. Watanabe MAE, De Lucca FL: Anti-tumor effect of splenocytes treated with RNA from animals immunized with bovine mielin basic protein. Cell Mol Biol 42: 249–255, 1996

    Google Scholar 

  19. Passos GAS Jr, De Lucca FL: RNA-mediated transfer of cellular immunity to a synthetic env antigen of the human immunodeficiency virus (HIV-1). Mol Cell Biol 108: 1–8, 1991

    Google Scholar 

  20. Sales VSF, Sawan FM, Watanabe MAE, Silva JS, Voltarelli JC, De Lucca FL: In vitro transfer of cellular immunity to synthetic peptides of HIV-1 to human lymphocytes with exogenous RNA. Mem Inst Oswaldo Cruz 91: 349–350, 1996

    Google Scholar 

  21. Hoerr I, Obst R, Rammensee HG, Jung G: In vivo application of RNA leads to induction of specific cytotoxic T lymphocytes and antibodies. Eur J Immunol 30: 1–7, 2000

    Google Scholar 

  22. Martinon F, Krishnan S, Lenzen G, Magné R, Gomard E, Guillet JG, Lévy JP, Meulien: Induction of virus-specific cytotoxic T lymphocytes in vivo by liposome-entrapped mRNA. Eur J Immunol 23: 1719–1722, 1993

    Google Scholar 

  23. Souza LR, De Lucca FL: Transfer of cellular immunity to a synthetic peptide of HIV-1 to human lymphocytes with exogenous RNA: Is the transcription factor NF-κB involved? In: Proceedings of Fourth European Conference on Experimental AIDS Research, Tampere, Finland, June 18-21, 1999. Monduzzi Editore, Bologna, 1999, pp 173–176

    Google Scholar 

  24. Souza LR, Watanabe MAE, De Lucca FL: RNA-mediated activation of the transcription factor NF-êB in human lymphocytes: Implications for HIV immunopathogenesis? In: Proceedings of Fifth European Conference on Experimental AIDS Research, Madrid, Spain, June 16-19, 2000. Monduzzi Editore, Bologna, 2000, pp 39–42

    Google Scholar 

  25. Eddy SR: Noncoding RNA genes. Curr Opin Genet Develop 9: 695–699, 1999

    Google Scholar 

  26. Steele G Jr, Wang BS, Richie JP, Ervin T, Yankee R, Mannick JA: Results of xenogeneic I-RNA therapy in patients with metastatic renal cell carcinoma. Cancer 47: 1286–1288, 1982

    Google Scholar 

  27. Richie JP, Wang BS, Steele GD, Wilson RE, Mannick JA: In vivo and in vitro effects of xenogeneic immune ribonucleic acid in patients with advanced renal cell carcinoma: A phase I study. J Urol 126: 24–28, 1981

    Google Scholar 

  28. Powderly WG, Landay A, Lederman MM: Recovery of immune system with antiretroviral therapy. J Am Med Assoc 280: 72–77, 1998

    Google Scholar 

  29. Siess DC, Vedder CT, Merkens LS, Tanaka T, Freed AC, McCoy SL, Heinrich MC, Deffebach ME, Bennett RM, Hefeneider SH: A human gene coding for a membrane-associated nucleic acid-binding protein. J Biol Chem 275: 33655–33662, 2000

    Google Scholar 

  30. Watanabe MAE, Sawan FM, De Lucca FL: The immunological activity of RNA isolated from the spleens of mice immunized with B16 melanoma cells is enhanced by the calcium phosphate-mediated uptake method. Proceedings of XVI International Cancer Congress 269–273, 1994

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Fernando L. De Lucca.

Rights and permissions

Reprints and permissions

About this article

Cite this article

De Lucca, F.L., Sawan, F.M., Watanabe, M.A.E. et al. Effect of the calcium phosphate-mediated RNA uptake on the transfer of cellular immunity of a synthetic peptide of HIV-1 to human lymphocytes by exogenous RNA. Mol Cell Biochem 228, 9–14 (2001). https://doi.org/10.1023/A:1013305708539

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1013305708539

Navigation