Safety and efficacy of activated transfected killer cells for neutropenic fungal infections.
BACKGROUND
Invasive fungal infections cause considerable morbidity and mortality in neutropenic patients. White blood cell transfusions are a promising treatment for such infections, but technical barriers have prevented their widespread use.


METHODS
To recapitulate white blood cell transfusions, we are developing a cell-based immunotherapy using a phagocytic cell line, HL-60. We sought to stably transfect HL-60 cells with a suicide trap (herpes simplex virus thymidine kinase), to enable purging of the cells when desired, and a bioluminescence marker, to track the cells in vivo in mice.


RESULTS
Transfection was stable despite 20 months of continuous culture or storage in liquid nitrogen. Activation of these transfected cells with retinoic acid and dimethyl sulfamethoxazole enhanced their microbicidal effects. Activated transfected killer (ATAK) cells were completely eliminated after exposure to ganciclovir, confirming function of the suicide trap. ATAK cells improved the survival of neutropenic mice with lethal disseminated candidiasis and inhalational aspergillosis. Bioluminescence and histopathologic analysis confirmed that the cells were purged from surviving mice after ganciclovir treatment. Comprehensive necropsy, histopathology, and metabolomic analysis revealed no toxicity of the cells.


CONCLUSIONS
These results lay the groundwork for continued translational development of this promising, novel technology for the treatment of refractory infections in neutropenic hosts.
Lin
Lin
L
Division of Infectious Diseases, Los Angeles Biomedical Research Institute, Harbor-University of California at Los Angeles (UCLA) Medical Center, 1124 West Carson Street, Torrance, CA 90502, USA.
Ibrahim
Ashraf S
AS
Baquir
Beverlie
B
Fu
Yue
Y
Applebaum
David
D
Schwartz
Julie
J
Wang
Amy
A
Avanesian
Valentina
V
Spellberg
Brad
B
D019264:Adoptive Transfer; D000818:Animals; D001228:Aspergillosis / Q000150:complications / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D002177:Candidiasis / Q000150:complications / Q000401:mortality / Q000473:pathology / Q000628:therapy*; D005260:Female; D007694:Killer Cells, Natural / Q000276:immunology*; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D009503:Neutropenia / Q000628:therapy*; D016019:Survival Analysis; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection
D013937:Thymidine Kinase
J Leukoc Biol. 2005 Aug;78(2):338-44
15857941
N Engl J Med. 1977 Mar 31;296(13):701-5
320476
Curr Opin Infect Dis. 2008 Aug;21(4):409-14
18594294
Hybridoma. 1999 Feb;18(1):93-7
10211795
Am J Med. 1998 Dec;105(6):478-83
9870832
J Leukoc Biol. 2007 Mar;81(3):632-41
17158608
Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):7024-8
8346212
Clin Pharmacokinet. 2009;48(6):399-418
19650679
Biol Blood Marrow Transplant. 2006 Jun;12(6):635-40
16737936
Antimicrob Agents Chemother. 1989 Feb;33(2):147-51
2541654
Antimicrob Agents Chemother. 2006 Aug;50(8):2895-6
16870798
Hum Gene Ther. 1998 May 1;9(7):1083-92
9607419
Haematologica. 2006 Jul;91(7):986-9
16757415
Transfusion. 1994 Jul;34(7):630-46
8053044
Cancer. 2004 Dec 15;101(12):2859-65
15529309
J Infect Dis. 2001 Jan 15;183(2):321-328
11112098
Clin Infect Dis. 2002 Apr 1;34(7):909-17
11880955
Clin Infect Dis. 2009 Mar 1;48(5):503-35
19191635
J Clin Invest. 1976 Sep;58(3):705-15
956397
Rev Inst Med Trop Sao Paulo. 1995 Sep-Oct;37(5):397-406
8729749
J Clin Invest. 1965 May;44:778-88
14276135
N Engl J Med. 1977 Mar 31;296(13):706-11
320477
J Hematother Stem Cell Res. 2001 Aug;10(4):535-44
11522236
Blood. 1977 Dec;50(6):1145-50
922162
Infect Immun. 2003 Oct;71(10):5756-64
14500497
Anticancer Res. 1999 May-Jun;19(3B):2163-5
10472325
Science. 1992 Jun 12;256(5063):1550-2
1317968
Clin Infect Dis. 2007 Feb 1;44(3):402-9
17205448
Clin Infect Dis. 2008 Feb 1;46(3):327-60
18177225
Antimicrob Agents Chemother. 2004 May;48(5):1908-11
15105158
Curr Opin Pediatr. 1999 Feb;11(1):47-55
10084084
J Clin Invest. 1967 Dec;46(12):1943-53
6073999
Transplant Rev. 1972;9:3-53
4403119
Med Mycol. 2007 Jun;45(4):321-46
17510856