| RRC ID |
35166
|
| Author |
Park CG, Rodriguez A, Ueta H, Lee H, Pack M, Matsuno K, Steinman RM.
|
| Title |
Generation of anti-human DEC205/CD205 monoclonal antibodies that recognize epitopes conserved in different mammals.
|
| Journal |
J Immunol Methods
|
| Abstract |
DEC205/CD205 is a C-type multilectin receptor, expressed highly in dendritic cells (DCs). Previous efforts to generate anti-human DEC205 (anti-hDEC205) monoclonal antibodies (mAbs) from mice immunized with subdomain proteins of hDEC205 resulted in a few mAbs. Recently, we expressed and utilized a full-length extracellular domain protein of hDEC205 to successfully generate 5 strong anti-hDEC205 mAbs from mice. In this study, DEC205 knockout (KO) mice were immunized with this full-length extracellular domain protein of hDEC205. One of the 3 immunized DEC205 KO mice was chosen for the highest anti-hDEC205 titer by flow cytometric analysis of serum samples on CHO cells stably expressing hDEC205 (CHO/hDEC205 cells) and used for hybridoma fusion. From a single fusion, more than 400 anti-hDEC205 hybridomas were identified by flow cytometric screen with CHO/hDEC205 cells, and a total of 115 hybridomas secreting strong anti-hDEC205 mAb were saved and named HD1 through HD115. To characterize in detail, 10 HD mAbs were chosen for superior anti-hDEC205 reactivity and further subjected to cloning and purification. Interestingly, out of those 10 chosen anti-hDEC205 HD mAbs, 5 mAbs were also strongly reactive to mouse DEC205 while 8 mAbs were found to stain DEC205(+) DCs on monkey spleen sections. In addition, we also identified that HD83, one of the 10 chosen HD mAbs, stains DEC205(+) DCs in rat spleen and lymph node. Therefore, by immunizing DEC205 KO mice with a full-length extracellular domain protein of hDEC205, we generated a large number of strong anti-hDEC205 mAbs many of which are cross-species reactive and able to visualize DEC205(+) DCs in lymphoid tissues of other mammals.
|
| Volume |
377(1-2)
|
| Pages |
15-22
|
| Published |
2012-3-30
|
| DOI |
10.1016/j.jim.2011.12.009
|
| PII |
S0022-1759(12)00011-7
|
| PMID |
22273672
|
| PMC |
PMC3324822
|
| MeSH |
Animals
Antibodies, Monoclonal / biosynthesis*
Antibodies, Monoclonal / immunology
Antigens, CD / immunology*
CHO Cells
Cloning, Molecular
Cricetinae
Dendritic Cells / immunology*
Epitopes / immunology
Humans
Immunohistochemistry
Lectins, C-Type / immunology*
Mice
Mice, Inbred C57BL
Mice, Knockout
Minor Histocompatibility Antigens
Receptors, Cell Surface / immunology*
Recombinant Fusion Proteins / immunology
Specific Pathogen-Free Organisms
Transfection
|
| IF |
1.901
|
| Times Cited |
10
|
|
WOS Category
|
BIOCHEMICAL RESEARCH METHODS
IMMUNOLOGY
|
| Resource |
| Rats |
ACI/Nkyo |
