Approaches to enhancing the retroviral transduction of human synoviocytes

Maria A. Del Vecchio, Helga I. Georgescu, James E. McCormack, Paul D. Robbins, Christopher H Evans

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

This report concerns a clinical trial for rheumatoid arthritis (RA), approved by the US National Institutes of Health and the Food and Drug Administration. An amphotropic retrovirus (MFG-IRAP) was used ex vivo to transfer a cDNA encoding human interleukin-1 receptor antagonist (IL-1Ra) to synovium. The protocol required the transduced cells to secrete at least 30 ng IL-1 Ra/106 cells per 48 h before reimplantation. Here we have evaluated various protocols for their efficiency in transducing cultures of human rheumatoid synoviocytes. The most reliably efficient methods used high titer retrovirus (approximately 108 infectious particles/ml). Transduction efficiency was increased further by exposing the cells to virus under flow-through conditions. The use of dioctadecylamidoglycylspermine (DOGS) as a polycation instead of Polybrene (hexadimethrine bromide) provided an additional small increment in efficiency. Under normal conditions of static transduction, standard titer, clinical grade retrovirus (approximately 5 × 105 infectious particles/ml) failed to achieve the expression levels required by the clinical trial. However, the shortfall could be remedied by increasing the time of transduction under static conditions, transducing under flow-through conditions, or transducing during centrifugation.

Original languageEnglish (US)
Pages (from-to)259-263
Number of pages5
JournalArthritis Research
Volume3
Issue number4
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Retroviridae
Hexadimethrine Bromide
Clinical Trials
Interleukin-1 Receptors
Synovial Membrane
Replantation
National Institutes of Health (U.S.)
United States Food and Drug Administration
Centrifugation
Interleukin-1
Rheumatoid Arthritis
Complementary DNA
Viruses
Synoviocytes

Keywords

  • Arthritis
  • Flow-through
  • High-titer retrovirus
  • Interleukin-1 receptor antagonist

ASJC Scopus subject areas

  • Rheumatology

Cite this

Del Vecchio, M. A., Georgescu, H. I., McCormack, J. E., Robbins, P. D., & Evans, C. H. (2001). Approaches to enhancing the retroviral transduction of human synoviocytes. Arthritis Research, 3(4), 259-263. https://doi.org/10.1186/ar311

Approaches to enhancing the retroviral transduction of human synoviocytes. / Del Vecchio, Maria A.; Georgescu, Helga I.; McCormack, James E.; Robbins, Paul D.; Evans, Christopher H.

In: Arthritis Research, Vol. 3, No. 4, 2001, p. 259-263.

Research output: Contribution to journalArticle

Del Vecchio, MA, Georgescu, HI, McCormack, JE, Robbins, PD & Evans, CH 2001, 'Approaches to enhancing the retroviral transduction of human synoviocytes', Arthritis Research, vol. 3, no. 4, pp. 259-263. https://doi.org/10.1186/ar311
Del Vecchio, Maria A. ; Georgescu, Helga I. ; McCormack, James E. ; Robbins, Paul D. ; Evans, Christopher H. / Approaches to enhancing the retroviral transduction of human synoviocytes. In: Arthritis Research. 2001 ; Vol. 3, No. 4. pp. 259-263.
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