Intralymphnode injection of human monocyte macrophage colony stimulating factor (CSF-1) as a method of obtaining high titer anti-CSF-1 antibodies.
We describe a rabbit intralymphnode immunization technique for obtaining a high titer antihuman CSF-1 antiserum with small amounts of antigen. This procedure provided a rapid (42 days after primo-injection), stable maximum immune response with a high titer antiserum precipitating 30% of 125I CSF-1 at a 1:25.000 dilution. The specificity of the immune serum was assessed by competitive binding experiments in RIA and neutralization of the CSF-1 biological activity in culture. The antiserum was also tested for its ability to detect CSF-1 in Western blotting, immunocytochemistry and immunohisto-chemistry. The results show that the immune serum specifically recognizes the biological active domain of human CSF-1 molecules from different origins and only detects dimeric forms. The potential uses of this anti CSF-1 antiserum are discussed.
Praloran
V
V
Laboratoire d'Hematologie, Centre Hospitalier Regional Universitaire, Nantes, France.
Martin-Thouvenin
V
V
Berrada
L
L
Moreau
A
A
Lanotte
M
M
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D001667:Binding, Competitive; D015153:Blotting, Western; D002460:Cell Line; D003114:Colony-Forming Units Assay; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007106:Immune Sera; D007114:Immunization; D007124:Immunoenzyme Techniques; D007272:Injections, Intralymphatic; D008198:Lymph Nodes / Q000737:chemistry / Q000276:immunology; D016173:Macrophage Colony-Stimulating Factor / Q000276:immunology / Q000494:pharmacology*; D008297:Male; D009500:Neutralization Tests; D011817:Rabbits; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000494:pharmacology
D000906:Antibodies; D000939:Epitopes; D007106:Immune Sera; D011994:Recombinant Proteins; D016173:Macrophage Colony-Stimulating Factor