J. Hasbold et al., PROPERTIES OF MOUSE CD40 - CELLULAR-DISTRIBUTION OF CD40 AND B-CELL ACTIVATION BY MONOCLONAL ANTI-MOUSE CD40 ANTIBODIES, European Journal of Immunology, 24(8), 1994, pp. 1835-1842
We describe here the derivation of a rat monoclonal antibody (mAb) aga
inst mouse CD40 (designated 3/23), which stains 45-50 % of spleen cell
s of adult mice, approximately 90 % of which are B cells. Interestingl
y, some 5-10 % of both CD4(+) and CD8(+) T cells in the spleens of (so
me, but not all) adult, unimmunized mice are also CD40(+), whereas CD4
0(+) cells were not detectable in the thymus, even following collagena
se digestion. Some 35-40 % of lymphoid cells in the bone marrow of adu
lt mice are CD40(+) and virtually all of these are B220(+), and hence
of the B cell lineage: triple-color flow cytometry showed that CD40 is
expressed at low levels on some 30 % of pre-B cells, at intermediate
levels on 80 % of immature B cells and on essentially all mature B cel
ls in the bone marrow. These results, therefore, suggest that in the m
ouse CD40 is expressed relatively late during the process of B cell di
fferentiation. The mAb induced marked up-regulation of major histocomp
atibility complex class II molecules, CD23 and B7.2 antigens on mature
B cells. It also stimulated modest levels of DNA synthesis in mature
B cells by itself: this was markedly enhanced by suboptimal concentrat
ions of mitogenic (but not non-mitogenic) anti-mu and anti-delta mAb,
and moderately enhanced by co-stimulation with interleukin-4. Hypercro
ss-linking of CD40 (using biotinylated mAb and avidin) also enhanced t
he proliferative response to anti-CD40.