A sensitive immunoassay to detect the alpha-chemokine GRO in rabbit blood and lung fluids.
GRO-alpha, GRO-beta and GRO-gamma are closely related peptides that stimulate growth of tumor cells and activate leukocytes in acute inflammatory reactions. In order to study the biology of GRO peptides in the lungs of experimental animals, we have developed and characterized a sensitive and specific immunoassay for rabbit GRO, and used this assay to measure GRO in rabbit lung fluids and plasma. GRO was cloned from a rabbit cDNA library and expressed in Escherichia coli. Specific goat polyclonal antibodies were used to create an antigen-capture immunoassay. The assay is sensitive to approximately 30 pg/ml GRO and does not crossreact with rabbit IL-8 or MCP-1, or human GRO. The assay accurately measures GRO in rabbit bronchoalveolar lavage fluid, plasma and serum. Rabbit erythrocytes bind little GRO and do not interfere with the detection of GRO in lung fluids. Circulating GRO was detected in the plasma of 4 of 6 pathogen-free rabbits, but the function of circulating GRO in normal animals is uncertain. This immunoassay will facilitate the study of the biology of GRO in rabbits with acute and chronic inflammation in the lungs and other tissues.
Kajikawa
O
O
Medical Research Service, Seattle Veterans' Affairs Medical Center, WA 98108, USA.
Johnson
M C
MC
Goodman
R B
RB
Frevert
C W
CW
Martin
T R
TR
D000818:Animals; D001323:Autoantibodies / Q000097:blood; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry; D054360:Chemokine CXCL1; D018925:Chemokines / Q000032:analysis* / Q000097:blood* / Q000235:genetics; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000032:analysis* / Q000097:blood* / Q000235:genetics; D004912:Erythrocytes / Q000276:immunology; D005069:Evaluation Studies as Topic; D006133:Growth Substances / Q000032:analysis* / Q000097:blood* / Q000235:genetics; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000706:statistics & numerical data; D036341:Intercellular Signaling Peptides and Proteins; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D013045:Species Specificity
D001323:Autoantibodies; C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D018925:Chemokines; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D011994:Recombinant Proteins