Differential gene expression profile of first-generation and second-generation rapamycin-resistant allogeneic T cells.
OBJECTIVE
We completed a phase II clinical trial evaluating rapamycin-resistant allogeneic T cells (T-rapa) and now have evaluated a T-rapa product manufactured in 6 days (T-rapa(6)) rather than 12 days (T-Rapa(12)).


METHODS
Using gene expression microarrays, we addressed our hypothesis that the two products would express a similar phenotype. The products had similar phenotypes using conventional comparison methods of cytokine secretion and surface markers.


RESULTS
Unsupervised analysis of 34,340 genes revealed that T-rapa(6) and T-rapa(12) products clustered together, distinct from culture input CD4(+) T cells. Statistical analysis of T-rapa(6) products revealed differential expression of 19.3% of genes (n = 6641) compared with input CD4(+) cells; similarly, 17.8% of genes (n = 6147) were differentially expressed between T-rapa(12) products and input CD4(+) cells. Compared with input CD4(+) cells, T-rapa(6) and T-rapa(12) products were similar in terms of up-regulation of major gene families (cell cycle, stress response, glucose catabolism, DNA metabolism) and down-regulation (inflammatory response, immune response, apoptosis, transcriptional regulation). However, when directly compared, T-rapa(6) and T-rapa(12) products showed differential expression of 5.8% of genes (n = 1994; T-rapa(6) vs. T-rapa(12)).


CONCLUSIONS
Second-generation T-rapa(6) cells possess a similar yet distinct gene expression profile relative to first-generation T-rapa(12) cells and may mediate differential effects after adoptive transfer.
Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, MD 20892-1288, USA. castiellol@cc.nih.gov
Luciano
L
Castiello
Miriam
M
Mossoba
Antonella
A
Viterbo
Marianna
M
Sabatino
Vicki
V
Fellowes
Jason E
JE
Foley
Matthew
M
Winterton
David C
DC
Halverson
Sara
S
Civini
Ping
P
Jin
Daniel H
DH
Fowler
David F
DF
Stroncek
D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D064987:Cell- and Tissue-Based Therapy; D005786:Gene Expression Regulation / Q000187:drug effects; D006086:Graft vs Host Disease / Q000188:drug therapy* / Q000276:immunology* / Q000473:pathology; D006801:Humans; D007166:Immunosuppressive Agents / Q000008:administration & dosage; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000302:isolation & purification*; D020123:Sirolimus / Q000008:administration & dosage; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000187:drug effects; D050378:T-Lymphocytes, Regulatory / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D059467:Transcriptome
D007166:Immunosuppressive Agents; D012313:RNA; D020123:Sirolimus