A soluble form of the human Fc receptor Fc gamma RIIA: cloning, transcript analysis and detection.
The Fc gamma receptors (Fc gamma R) are glycoproteins that bind the Fc region of immunoglobulin G. Human hematopoietic cells express three biochemically distinct classes of Fc gamma receptors: Fc gamma RI (CD64), Fc gamma RII (CD32) and Fc gamma RIII (CD16). Complementary DNA (cDNA) clones for each of the human Fc gamma receptors have been isolated from myeloid and lymphoid cells. We describe the isolation and characterization of four Fc gamma RII clones from a cDNA library obtained from a megakaryocyte-like cell line, human erythroleukemia (HEL). Three clones encode the Fc gamma RIIA transmembrane (TM) form, while one novel clone lacks the TM region but retains the cytoplasmic domain. By conducting reverse transcription coupled to polymerase chain reaction (PCR), we found transcripts coding for this unique form of receptor in RNA from platelets, HEL cells and a second megakaryocyte-like cell line, CHRF-288-11. These results were confirmed by RNase protection analysis of RNA from HEL cells. The structure of the novel cDNA suggested that it codes for a soluble form of Fc gamma RIIA. A soluble Fc gamma RII protein was detected in the conditioned medium from HEL cells but not from the Fc gamma RII-negative T cell line, Jurkat, by immunoprecipitation with the anti-Fc gamma RII monoclonal antibody (mAb), IV.3. The immunoprecipitated protein was of the expected size for a soluble Fc gamma RII lacking the TM region but retaining the cytoplasmic domain. Soluble Fc gamma RIIA may be important in modulating the interaction between immune complexes and membrane-associated Fc gamma RII.
Division of Hematology, Children's Hospital of Philadelphia, PA 19104.
E F
EF
Rappaport
D L
DL
Cassel
D O
DO
Walterhouse
S E
SE
McKenzie
S
S
Surrey
M A
MA
Keller
A D
AD
Schreiber
E
E
Schwartz
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned; D006801:Humans; D007163:Immunosorbent Techniques; D004915:Leukemia, Erythroblastic, Acute; D008533:Megakaryocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012326:RNA Splicing; D011961:Receptors, Fc / Q000032:analysis / Q000235:genetics*; D012995:Solubility; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D017077:Culture Media, Conditioned; D011961:Receptors, Fc