R-phycoerythrin-cyanine 5 tandem discerns CD72 polymorphism.
By fluorescence analysis, we demonstrated that a fluorochrome tandem composed of R-phycoerythrin and cyanine 5 specifically recognized B cells from SJL, AKR, MRL/Mp, and NOD mouse strains, whereas B cells from C57BL/6, DBA/2, SWR, 129/Sv, and BALB/c were not stained. A strict correlation was observed between the fixation of the fluorochrome and the pattern of expression of the pan-B cell marker CD72, i.e., early expression in B-cell lineage development and downregulation on the terminally differentiated activated B cells. Three allelic forms, CD72a, CD72b and CD72c have been well characterized, and show a high number of amino acid substitutions concentrated in the membrane-distal extracellular domain. Using a PCR approach, we determined that all mouse strains positive for fluorochrome staining display the CD72c allelic form. Moreover, a genetic analysis showed that the fixation of the fluorochrome on the B cell is exclusively dependent on the presence of the CD72c allele. Together, these results strongly suggest that the tandem binds a molecular complex comprising the CD72c molecule or recognizes directly the CD72c molecule itself.
Doucet
M
M
Laboratoire d'Immunobiologie, Université Denis Diderot, Paris, France.
Soussi
N
N
Crain-Denoyelle
A M
AM
Gendron
M C
MC
Sanchez
P
P
D000483:Alleles; D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics*; D001402:B-Lymphocytes; D002232:Carbocyanines; D005456:Fluorescent Dyes; D008040:Genetic Linkage; D051379:Mice; D008815:Mice, Inbred Strains / Q000235:genetics*; D010799:Phycoerythrin; D011110:Polymorphism, Genetic; D013154:Spleen / Q000166:cytology; D013194:Staining and Labeling
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C064228:CD72 protein, human; D002232:Carbocyanines; D005456:Fluorescent Dyes; C085321:cyanine dye 5; D010799:Phycoerythrin