Use of DNA immunization to produce polyclonal antibodies against the native human neurokinin-1 receptor for substance P.
Antibodies against the native form of the human NK1 receptor (hNK1R) for the neuropeptide substance P (SP), an important immunoregulator, are difficult to produce using classical immunization techniques. We show here that mice immunized with a plasmid harboring hNK1R cDNA developed antibodies recognizing extracellular epitopes of native hNK1R expressed on CHO cell membranes, as shown by FACS and immunofluorescence analysis, some antibodies being specifically directed against the second extracellular loop (E2) of the receptor. This original strategy, DNA immunization, thus efficiently generated new immunological tools to further analyse the role of SP in the regulation of immune cell functions.
Tymciu
Sylvie
S
CEA, Service de Pharmacologie et d'Immunologie, DSV/DRM, Bât 136, Centre d'Etudes de Saclay, 91191 Cedex, Gif-sur-Yvette, France.
Wijkhuisen
Anne
A
Sagot
Marie-Astrid
MA
Frobert
Yveline
Y
Grassi
Jacques
J
Créminon
Christophe
C
Couraud
Jean-Yves
JY
Boquet
Didier
D
D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000276:immunology*; D015153:Blotting, Western; D016466:CHO Cells; D019556:COS Cells; D002462:Cell Membrane / Q000235:genetics / Q000276:immunology; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics* / Q000276:immunology*; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D007114:Immunization / Q000379:methods*; D051379:Mice; D015213:Neuroimmunomodulation / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics / Q000276:immunology; D011489:Protein Denaturation / Q000276:immunology; D017434:Protein Structure, Tertiary / Q000235:genetics; D018040:Receptors, Neurokinin-1 / Q000235:genetics* / Q000276:immunology*; D013373:Substance P / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D018076:DNA, Complementary; D000939:Epitopes; D018040:Receptors, Neurokinin-1; D013373:Substance P