Inhibition of xenoreactive natural antibody production by retroviral gene therapy

Science

Volumn 281, Issue 5384, 1998, Pages 1845-1847

  Bracy, Jennifer L ^a,b   Sachs, David H ^b   Iacomini, John ^b  

^a DREXEL UNIVERSITY COLLEGE OF MEDICINE   (United States)

^b MASSACHUSETTS GENERAL HOSPITAL   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ANTIBODY; CARBOHYDRATE; EPITOPE; GLUCOSYLTRANSFERASE; N ACETYLLACTOSAMINIDE ALPHA 1,3 GALACTOSYLTRANSFERASE; UNCLASSIFIED DRUG;

ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ANTIBODY PRODUCTION; ARTICLE; BONE MARROW CELL; CONTROLLED STUDY; GENE THERAPY; GENETIC ENGINEERING; GRAFT REJECTION; IMMUNOLOGICAL TOLERANCE; KIDNEY CELL; KNOCKOUT MOUSE; MOUSE; NONHUMAN; PRIORITY JOURNAL; RETROVIRUS; SWINE; XENOTRANSPLANTATION;

ANIMALS; ANTIBODY FORMATION; B-LYMPHOCYTES; BONE MARROW CELLS; BONE MARROW TRANSPLANTATION; CELL LINE; CLONING, MOLECULAR; ENZYME-LINKED IMMUNOSORBENT ASSAY; EPITOPES; GALACTOSYLTRANSFERASES; GENE TARGETING; GENE THERAPY; GENE TRANSFER TECHNIQUES; GENETIC VECTORS; GRAFT REJECTION; GRAFT VS HOST DISEASE; HUMANS; IMMUNE TOLERANCE; MICE; MICE, KNOCKOUT; RETROVIRIDAE; SWINE; TRANSPLANTATION, HETEROLOGOUS;

ANIMALIA; MURINAE; SUS SCROFA; UNIDENTIFIED RETROVIRUS;

EID: 0032544673     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.281.5384.1845     Document Type: Article

References (34)

1. M. E. Tumbleson, Swine in Biomedical Research (Plenum, New York, 1986).
2. D. H. Sachs, Pathol. Biol. 42, 217 (1994).
3. A. H. Good et al., Transplant. Proc. 24, 559 (1992).
4. R. Oriol, Y. Ye, E. Koren, D. K. Cooper, Transplantation 56, 1433 (1993).
5. J. L. Platt, Crit. Rev. Immunol. 16, 331 (1996).
6. _, Xenotransplantation, D. K. C. Cooper, E. Kemp, J. L. Platt, D. J. C. White, Eds. (Springer, Heidelberg, Germany, 1997), p. 8.
7. D. K. C. Cooper and A. D. Thall, World. J. Surg. 21, 901 (1997).
8. F. H. Bach et al., Nature Med. 1, 869 (1995).
9. Y.-G. Yang et al., J. Exp. Med. 187, 1335 (1998).
10. D. H. Sachs and T. Sablinski, Xenotransplantation 2, 234 (1995).
11. The pig αGT cDNA was isolated from pSAL3GT1 (a gift from K. Gustafsson, Institute of Child Health, London, UK) and subcloned into the Eco RI-Hinc II sites in pBluescript II KS minus (Stratagene, La Jolla, CA) to construct pKSαGT. A 550-bp Pst I fragment containing the murine phosphoglycerate kinase (PGK) promoter isolated from the plasmid SP95.3.5 (a gift from C. LeGuern, Massachusetts General Hospital, Boston) was then cloned into the Pst I site of pKSαGT to construct pPGKαGT. A Bam HI-Bss HII restriction fragment containing αGT under control of the PGK promoter isolated from pPGKαGT was cloned into the Bgl II-Mlu I sites in the 3′LTR of the N2A vector to construct LGTRV. The GN24 and NB5 retroviral producer cell lines were provided by C. LeGuern. The GN24 line is a GP+E86-derived (31) retroviral producer cell line carrying only the unmodified N2A retroviral vector, which contains the neomycin resistance gene. The NB5 tine is a PA317-derived (32) amphotropic retroviral producer line carrying the unmodified N2 retroviral vector (33). All virus supernatants were titered by infecting NIH 3T3 cells in the presence of 4 to 6 μg of polybrene per milliliter and determining the number of G418-resistant colonies obtained per milliliter of supernatant.
12. K. M. Strahan et al., Immunogenetics 41, 101 (1995).
13. P. A. Hantzopoulos, B. A. Sullenger, G. Ungers, E. Gilboa, Proc. Natl. Acad. Sci. U.S.A. 86, 3519 (1989).
14. D. Markovitz, S. Goff, A. Bank, Virology 167, 400 (1988).
15. C. Wood, E. A. Kabat, L. A. Murphy, I. J. Goldstein, Arch. Biochem. Biophys. 198, 1 (1979).
16. J. L. Bracy and J. Iacomini, unpublished data.
17. +/+ controls (Jackson Laboratory, Bar Harbor, ME). All mice were housed in viral antibody-free micro-isolator conditions in autoclaved cages and maintained on irradiated feed and autoclaved acidified drinking water.
18. A. D. Thall, P. Maly, J. B. Lowe, J. Biol. Chem. 270, 21437 (1995).
19. All mammals except humans, apes, and Old World monkeys express a functional αGT enzyme and αGal epitopes on all tissues. Therefore, most mammals, such as mice, are tolerant to αGal and do not produce antibodies that are reactive with this epitope.
20. A. Thall, H. Murphy, J. B. Lowe, Transplant. Proc. 28, 561 (1996).
21. J. Bagley, K. Aboody-Guterman, X. Breakefield, J. Iacomini, Transplantation 65, 1233 (1998).
22. DNA was analyzed using semi-nested PCR with primers specific for the PGK promoter (5′-CTTTCGACCTGCAGGAATTC forward primer) and pig αGT (5′-GGCGTTTCTCAGGATTAAACC reverse primer-1) for the first round of amplification. In the second round of amplification, the PGK-specific primer and a second αGT primer (5′-CCTTTTCGTTGCCTATAGCG reverse primer-2) were used. Primers specific for mouse β-actin (5′-AACCCCAAGGCCAACCGCGAGAAGATGACC forward primer; 5′-GGTGATGACCTGGCCGTCAGGCAGCTCGTA reverse primer) were used as controls.
23. r-transduced bone marrow was first observed at 5 weeks after transplantation. The level of αGal XNA reactivity in mice reconstituted with LGTA7-transduced bone marrow was indistinguishable from that observed by ELISA using lactosamine-conjugated BSA-coated plates. Lactosamine-BSA is identical to αGal-BSA except the terminal αGal carbohydrate residue is absent.
24. J. L. Bracy and J. Iacomini, unpublished data.
25. 4, 3% glycerol (pH 6.0)] containing 0.5 unit of α-galactosidase (Sigma) or buffer alone as a control for 1 hour at 40°C. The cells were washed in HBSS and analyzed by flow cytometry. Stained cells were analyzed on a Becton Dickinson FACscan. All staining data was analyzed using Win-List software (Verity Software House, Topsham, ME). All serum samples used in binding assays were obtained from sex-and age-matched mice.
26. r-transduced bone marrow together with rabbit complement in the presence of calcein-AM and ethidium homodimer. Live cells take up calcein-AM, but exclude ethidium homodimer. Upon entering live cells, calcein-AM is degraded and emits light at 535 nm. Dead cells do not degrade calcein-AM, but take up ethidium homodimer, which emits light at 650 nm. Thus, live cells appear green and nuclei of dead cells appear red when examined by fluorescent microscopy.
27. H. Auchincloss Jr. and D. H. Sachs, Annu. Rev. Immunol. 16, 433 (1998).
28. J. L. Platt, Nature 392, 11 (1998).
29. A group of mice (n = 4) reconstituted with LGTA7-transduced bone marrow was examined over a 51-week period after transplantation. No αGal XNA was detected by ELISA in the sera of these mice at 51 weeks after transplantation.
30. U. Galili, R. E. Mandrell, R. M. Hamadeh, S. B. Shohet, J. M. Griffiss, Infect. Immun. 56, 1730 (1988).
31. D. Markovitz, S. Goff, A. Bank, J. Virol. 62, 1120 (1988).
32. A. D. Miller and C. Buttimore, Mol. Cell. Biol. 6, 2895 (1986).
33. D. Armentano et al. J. Virol. 61, 1647 (1987).
34. We thank S. Pillai, G. Waneck, N. Cretin-Bühler, and J. Bagley for critical review of the manuscript, F. Neethling for help with the PK-15 assay, and D. K. C. Cooper and U. Galili for advice and encouragement. Supported in part by a sponsored research agreement between Massachusetts General Hospital and BioTransplant, Inc.