Identification of unique truncated KC/GRO beta chemokines with potent hematopoietic and anti-infective activities.
SK&F 107647, a previously described synthetic immunomodulatory peptide, indirectly stimulates bone marrow progenitor cells and phagocytic cells, and enhances host defense effector mechanisms in bacterial and fungal infection models in vivo. In vitro, SK&F 107647 induces the production of a soluble mediator that augments colony forming cell (CFU-GM) formation in the presence of CSFs. In this paper we purified and sequenced the stromal cell-derived hematopoietic synergistic factors (HSF) secreted from both murine and human cell lines stimulated with SK&F 107647. Murine HSF is an N-terminal 4-aa truncated form of the CXC chemokine, KC, while human HSF was identified as an N-terminal 4-aa truncated form of the CXC chemokine, GRO beta. In comparison to their full-length forms, truncated KC and truncated GRO beta were 10 million times more potent as synergistic growth stimulants for CFU-GM. Enhanced potency of these novel truncated chemokines relative to their full-length forms was also demonstrated in respiratory burst assays, CD11b Ag expression, and intracellular killing of the opportunistic pathogen, Candida albicans. Administration of truncated KC significantly enhanced survival of mice lethally infected with C. albicans. The results reported herein delineate the biological mechanism of action of SK&F 107647, which functions via the induction of unique specific truncated forms of the chemokines KC and GRO beta. To our knowledge, this represents the first example where any form of KC or GRO beta were purified from marrow stromal cells. Additionally, this is the first demonstration of in vivo efficacy of a CXC chemokine in an animal infectious fungal disease model.
King
A G
AG
Department of Molecular Virology, Project Management, Microbial Infectivity, Molecular Genetics, SmithKline Beecham Pharmaceuticals, Collegeville, PA 19426, USA. Andrew_G_King@sbphrd.com
Johanson
K
K
Frey
C L
CL
DeMarsh
P L
PL
White
J R
JR
McDevitt
P
P
McNulty
D
D
Balcarek
J
J
Jonak
Z L
ZL
Bhatnagar
P K
PK
Pelus
L M
LM
D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D000935:Antifungal Agents / Q000097:blood / Q000276:immunology / Q000302:isolation & purification*; D001854:Bone Marrow Cells / Q000737:chemistry / Q000276:immunology; D002177:Candidiasis / Q000276:immunology / Q000401:mortality / Q000517:prevention & control; D002460:Cell Line; D054360:Chemokine CXCL1; D019743:Chemokines, CXC / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D002630:Chemotactic Factors / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004357:Drug Synergism; D005260:Female; D006133:Growth Substances / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D007274:Injections, Intraperitoneal; D036341:Intercellular Signaling Peptides and Proteins; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D018375:Neutrophil Activation / Q000276:immunology; D009842:Oligopeptides / Q000008:administration & dosage / Q000494:pharmacology; D010446:Peptide Fragments / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry; D017154:Stromal Cells / Q000737:chemistry / Q000276:immunology
D000276:Adjuvants, Immunologic; D000935:Antifungal Agents; C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; C475895:Cxcl1 protein, mouse; D006133:Growth Substances; D007106:Immune Sera; D036341:Intercellular Signaling Peptides and Proteins; D009842:Oligopeptides; D010446:Peptide Fragments; D011994:Recombinant Proteins; C086049:SK&F 107647