> In article <B19E4F226C2DD211BE8E0008C7FAB88F079FE5 at BHQROC1EX1>,
> Chen, Peter <ChenP at USA.REDCROSS.ORG> wrote:
> >
> >Hi, I was wondering if anyone knows a technique that eliminates
> >double-negative T cells while enriching for B cells from the lymph nodes of
> >gld Balb/b mice. I'm experimenting with magnetic sorting but there are few
> >ideal antibodies that would target DN T cells with high efficiency and some
> >antibodies (for instance anti-TCR) are not made in rat (the magnetic
> >particles are conjugated to anti-rat IgG). This technique will be an
> >enriching step before FACS cell sorting but I need to enrich a target
> >population that only constitutes about 2% of the total. Any advice would be
> >greatly appreciated!
>There is a rat mAb to mouse CD3:
@C Portoles, P.//Rojo, J.//Golby, A.//Bonneville, M.//Gromkowski,
S.//Greenbaum, L.//Janeway, C. A., Jr.//Murphy, D. B.//Bottomly, K.
Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one
of which may interact with CD4 during T cell activation J
Immunol 1989 142 12 4169-75 KT-3, KT3 ??? The TCR is
comprised of two variable chains that confer specificity, called
alpha:beta or gamma:delta, physically associated with five different
molecules that comprise the complex known as CD3. Antibodies to this
complex are very useful, as they react with all T lymphocytes. A rat mAb
to mouse CD3 has been prepared. It reacts with 100% of T cells in all
mouse strains tested but with no other cell type. It binds to the CD3
epsilon chain. This antibody activates cloned T cell lines and normal T
cells, provided suitable accessory cells and signals are present. This
antibody detects a determinant similar to but not identical with those
detected by two previously reported hamster anti-CD3 epsilon antibodies.
This antibody fixes C efficiently, and it is thus useful for depletion of
T cells from bulk populations. Activation of T cells by one of the three
different anti-CD3 epsilon antibodies was inhibited by the Fab fragment of
anti-CD4, similar to the effects of anti-CD4 Fab on two previously
reported anti-TCR V region antibodies that bind a CD3 epsilon-associated
epitope. This further defines a site involving TCR V regions and CD3
epsilon with which CD4 appears to associate during T cell activation.
Antibodies, Monoclonal biosynthesis; Antigen Presenting Cells immunology;
Antigenic Determinants immunology; Binding Sites, Antibody; Binding,
Competitive; Cell Line; Clone Cells immunology; Clone Cells metabolism;
Hamsters ; Immunoglobulins, Fab physiology; Mice ; Mice, Inbred AKR; Mice,
Inbred BALB C; Mice, Inbred CBA; Mice, Inbred C3H; Mice, Inbred C57BL;
Mice, Inbred DBA; Precipitin Tests; Rats ; Rats, Inbred Lew; Species
Specificity; T Lymphocytes metabolism
*Antibodies, Monoclonal immunology; *Antigenic Determinants analysis;
*Antigens, Differentiation, T Lymphocyte immunology; *Lymphocyte
Transformation; *Receptors, Antigen, T Cell immunology; *T Lymphocytes
immunology
Animal; Support, Non U.S. Gov't; Support, U.S. Gov't, Non P.H.S.; Support,
U.S. Gov't, P.H.S.
Antibodies, Monoclonal; Antigenic Determinants; Antigens, CD3; Antigens,
Differentiation, T Lymphocyte; Binding Sites, Antibody; Immunoglobulins,
Fab; Receptors, Antigen, T Cell
Good luck, jorg
