TIP, a T-cell factor identified using high-throughput screening increases survival in a graft-versus-host disease model.
A coordinated effort combining bioinformatic tools with high-throughput cell-based screening assays was implemented to identify novel factors involved in T-cell biology. We generated a unique library of cDNAs encoding predicted secreted and transmembrane domain-containing proteins generated by analyzing the Human Genome Sciences cDNA database with a combination of two algorithms that predict signal peptides. Supernatants from mammalian cells transiently transfected with this library were incubated with primary T cells and T-cell lines in several high-throughput assays. Here we describe the discovery of a T cell factor, TIP (T cell immunomodulatory protein), which does not show any homology to proteins with known function. Treatment of primary human and murine T cells with TIP in vitro resulted in the secretion of IFN-gamma, TNF-alpha, and IL-10, whereas in vivo TIP had a protective effect in a mouse acute graft-versus-host disease (GVHD) model. Therefore, combining functional genomics with high-throughput cell-based screening is a valuable and efficient approach to identifying immunomodulatory activities for novel proteins.
Fiscella
Michele
M
Department of Preclinical Discovery, Human Genome Sciences, Inc., Rockville, MD 20850, USA.
Perry
James W
JW
Teng
Baiqin
B
Bloom
Michael
M
Zhang
Chen
C
Leung
Kam
K
Pukac
Laurie
L
Florence
Kimberly
K
Concepcion
Alice
A
Liu
Binjun
B
Meng
Ying
Y
Chen
Cecil
C
Elgin
Erika Cochrane
EC
Kanakaraj
Palanisamy
P
Kaufmann
Thomas E
TE
Porter
Joelle
J
Cibotti
Ricardo
R
Mei
Yun
Y
Zhou
Joe
J
Chen
Guoxian
G
Roschke
Viktor
V
Komatsoulis
George
G
Mansfield
Brian
B
Ruben
Steve
S
Sanyal
Indra
I
Migone
Thi-Sau
TS
D000276:Adjuvants, Immunologic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D020869:Gene Expression Profiling / Q000379:methods; D006086:Graft vs Host Disease / Q000188:drug therapy* / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000196:embryology / Q000276:immunology; D051379:Mice / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D020539:Sequence Analysis, Protein / Q000379:methods*; D013491:Suppressor Factors, Immunologic / Q000008:administration & dosage* / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D037182:T-Cell Antigen Receptor Specificity / Q000235:genetics; D013601:T-Lymphocytes / Q000737:chemistry / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D014162:Transfection / Q000379:methods
D000276:Adjuvants, Immunologic; D011993:Recombinant Fusion Proteins; D013491:Suppressor Factors, Immunologic