Search

Problem: Fas antigen (APO-1/CD95) can regulate the activity of various cells during adulthood. This study aimed at determining whether Fas may also be involved in the regulation of very early events such as the embryo preimplantation stage., Method of Study: We used mouse embryo stem (ES) cell line as a model for testing the effect of Fas crosslinking upon anti-Fas monoclonal antibody (MoAb) treatment. In addition, this treatment was also applied to in-vitro mouse-embryo culture., Results: Flow-cytometry analysis of cultured ES cells demonstrated an increase in Fas expression. unchanged in the presence of mouse interleukin-2, while greatly upregulated in the presence of lipopolysaccharide (LPS). As determined by various means, ES cells may undergo a Fas-mediated apoptosis, slightly but significantly intensified by the addition of LPS to cell cultures. We also report that anti-Fas MoAb directly inhibited two-cell stage mouse-embryo (preimplantation) development in in-vitro culture conditions., Conclusion: These data suggest a novel mechanism controlling the regulation of physiological cell turnover as well as blastocyst implantation in early embryo development.

The action of activated complement on the upper respiratory tract was studied using the registration of ciliary beating and the transmission and scanning electron microscopy of ciliated cell lesions. The alternative pathway of activation was done using (1) non-specific activators, i. e. zymosan, dextran sulfate and polymerised sIgA, and (2) specific activators, i. e. "Ag-sIgA" immune complexes on normal mucosa and contact between an antigen and the tracheal mucosa of an immunized animal. In all cases, a quick alteration of the ciliary beating rythm was registered as well as more or less extensive cytological destruction. Such results were obtained with or without adjunction of complement; this suggested the existence of complement locally. A new method to demonstrate the occurrence of complement was developed, and the results showed that complement was present in the trachea. The above results were corroborated (1) by EDTA inhibition of the two complement pathways which prevented stopping of the ciliary beating and eliminated most of the cytological lesions, whilst EGTA left the alternative pathway operative with consequent alterations, and (2) by using C6-deficient rabbits in which no alterations were found. This study demonstrated that under certain conditions, local sIgA may activate the complement, and this fact explains many aspects of these immunological reactions of the respiratory epithelium.

A peptide of simple chemical structure has demonstrated its efficiency in preventing the large cellular destruction that locally activated complement produced on the ciliary epithelium of the respiratory tract. Previously (1980), it was demonstrated by the authors that these cellular destructions after sensitization of the epithelium was due to the local activation of the complement (alternate pathway) by immune complexes with secretory IgA. The cellular protection afforded by Naaga was demonstrated by the persistance of a normal ciliary beating when the sensitized mucosa is in contact with the antigen; by electron microscopic studies both in transmission and scanning E.M. contrasting with the complete cellular destructions of the epithelium which appear obvious. The protection appear complete when Naaga (56 mM) is present in the testing solution (or instillated before the test). By in vitro human complement studies; study of the cytolytic sequence inhibition for the classical pathway 1,5.10(-3) M of Naaga produces a 50% inhibition of 1 H50 hemolytic unit. For the alternate pathway, the same inhibition is observed with 1,75.10(-3) M of Naaga; by two-dimensions immuno-electrophoresis: a dilution of 1/2 of C3 in Naaga reduced to 1/10 of its normal value the C3b profile; the "Rockets" technique demonstrated that the same 1/2 dilution of Naaga in complement prevents the clivage of factor B and that this peptide acts by inhibition of the alternate C3 convertase formation (see illustrations). If we consider the subject of this study i.e. the upper respiratory tract mucosa and knowing the physiopathological importance of the muco ciliary complex in preventing dust, microbs and other particulate foreign materiel to penetrate the epithelium, the therapeutic importance of such a simple non toxic and unharmful chemical compound must be stressed.