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M W Wang

Publications and source records attributed to M W Wang.

64 records · Page 4Linked to original sources

Immunofluorescent localization, by use of anti-idiotypic antibody, of monoclonal anti-progesterone antibody in the mouse uterus before implantation.

Mouse monoclonal anti-progesterone IgG1 antibody designated DB3 has an anti-fertility effect when injected into female mice shortly after mating. In BALB/c mice, pregnancy is blocked, probably as a result of progesterone withdrawal with inhibition of implantation. Rabbit polyclonal anti-idiotype raised against DB3 has been used in an indirect immunofluorescence method on frozen tissue sections to demonstrate the presence of DB3 on the surface of uterine luminal and glandular epithelia before implantation. DB3 was only detectable 30-60 h after a single parenteral injection (9 nmol antibody per mouse i.p. or i.v. at 32 h post coitum). Immunolocalization was both pregnancy-dependent and anti-progesterone antibody-specific, as it was not seen in pseudopregnant mice or mice treated with P3 (mouse myeloma IgG1 protein, using polyclonal anti-P3 anti-idiotype as a probe) or saline. The immunofluorescent reaction was completely blocked by addition of DB3 idiotype in vitro. The results indicate that anti-progesterone antibody binds to an antigen associated with luminal and glandular epithelia which may locally inhibit the uterine uptake of progesterone and disrupt the process of implantation.

Animals↗

Inhibition of pregnancy before and after implantation in rats with monoclonal antibody against progesterone.

A monoclonal antibody against progesterone completely blocked pregnancy when rats were injected intraperitoneally at 0.53 mumol/kg/day on days 1 and 2, or on day 11 of a first pregnancy. The antibody was equally effective when injected again during a second pregnancy. There was no evidence for anti-idiotypic antibody production even when the injection of a heterologous (mouse) antibody was repeated in a second pregnancy.

Animals↗

Anti-idiotypic antibody used for the localization of parenterally administered monoclonal anti-progesterone antibody in mice.

Affinity-purified rabbit and sheep anti-idiotypic antisera raised against mouse monoclonal anti-progesterone IgG1 antibody (DB3) or mouse myeloma IgG1 protein P3 (MOPC 21) showed high binding specificities to the respective idiotypes used for immunization as determined by RIA or ELISA. They have been used in an indirect immunofluorescent method to demonstrate the localization of parenterally administered idiotypes in pregnant or pseudopregnant BALB/c mouse frozen tissue preparations, at known stages post coitum after a single intraperitoneal or intravenous injection of DB3 or P3. DB3 was visualized on the surface of uterine luminal and glandular epithelia of pregnant mice 36 h after treatment; the localization was DB3-specific as it was not seen in mice treated with P3 (using sheep anti-P3 anti-idiotype as a probe) or saline. The fluorescent staining reaction in oviduct was weak and only appeared on the surface of the oviducal serosa (peritoneal side). Both DB3 and P3 were also localized in liver (granules of Kupffer cells), kidney (glomerular basement membrane), spleen (on the membrane surface of mononuclear cells in the white pulp), and peritoneal exudate cells (on the membrane surface). Staining could be completely blocked by the addition of the free idiotypes against which the anti-idiotypes were made but not by the unrelated idiotype. Anti-idiotypic labelling in vivo is more specific and selective than anti-whole immunoglobulin labelling.

Animals↗

Efficacy and specificity of monoclonal antibodies to progesterone in preventing the establishment of pregnancy in the mouse.

Anti-progesterone monoclonal antibody prevents the establishment of pregnancy in BALB/c mice by the prevention of implantation when injected i.p. 32 h after mating. To determine the specificity of this effect, mice were injected with immune and non-immune purified mouse immunoglobulins. The results show that anti-implantation efficacy was due to high-affinity antibody which bound progesterone since two further mouse immunoglobulin (Ig) G1 preparations, mouse IgA and mouse IgM which failed to bind the steroid, had no effect on pregnancy rates. From a panel of anti-progesterone monoclonal antibodies, six with a high affinity (affinity constant, 0.24-0.80 litres/nmol) and specificity for progesterone were selected for additional studies. Anti-implantation efficacy for five antibodies was similar, with a 50% effective dose within the range of 0.8-2.0 nmol. Antibody reached high concentrations in plasma within 12 h after i.p. injection, and declined with a half-life of about 80 h. Purified F(ab')2 fragments of antibody also bound progesterone, but were less effective than the native molecule in blocking pregnancy. The results show that implantation in the mouse can be blocked by a high-affinity antibody that binds progesterone and which is removed from the blood at a slow rate.

Animals↗

Idiotype-anti-idiotype interactions of VHIX-coded anti-progesterone and anti-arsonate antibodies. Comparison of passive haemagglutination and radioimmunoassays.

The reactivity and specificity of polyclonal and monoclonal anti-idiotypic antibodies raised against monoclonal anti-progesterone and anti-arsonate antibodies have been studied by solid phase radioimmunoassay (RIA) with immobilized idiotype and by passive haemagglutination with idiotype-coupled red cells. The sensitivity of the two methods was comparable, though some cross-reactions were only detected by RIA. Passive haemagglutination was found to be especially suitable in screening for monoclonal anti-idiotypes in hybridoma supernatants and ascites, and had advantages over RIA in detection of syngeneic anti-idiotypes. Demonstration of binding site-associated idiotopes was possible by haemagglutination inhibition. RIA and haemagglutination were used to investigate the idiotypic relationships between BALB/c antiprogesterone and anti-arsonate monoclonal antibodies which share heavy chains encoded by VHIX variable region genes.

Amino Acid Sequence↗

Pipecolic acid enhancement of GABA response in single neurons of rat brain.

Using unit recording and microelectrophoresis, influence of pipecolic acid (PA), a major metabolite of lysine in the brain, on GABA and glycine responses was studied in the cerebral cortical and hippocampal pyramidal neurons of rats. With small currents, PA had no effect on the single neuron activities but enhanced GABA response without affecting glycine response. The finding provides a new evidence that PA may have a connection with central GABA system.

Action Potentials↗