Inactivation of an X-linked transgene in murine extraembryonic and adult tissues.

Transgenes located on the X chromosome have been used to study the mechanisms involved in X-chromosome inactivation. Analysis of the transgenic mouse strain M-TKneo1 carrying a neomycin resistance gene inserted in the X chromosome showed that, in adult somatic tissues, this transgene is subject to X-inactivation and to de novo methylation as other endogenous X-linked genes. During mouse embryogenesis, X-linked genes show a preferential paternal inactivation in extraembryonic tissues, whereas these genes are subject to random inactivation in embryonic tissues. It has been suggested that, in the mouse, the extraembryonic tissues carry a parental imprint at the time of inactivation. The study of the neo transgene expression in extraembryonic endoderm has shown not only that neo is inactivated but also that, at the RNA level, paternal inactivation of the transgene seems essentially complete. The differences between our results and previously obtained results with a mouse alpha-fetoprotein transgene, which was only inactivated in neonatal tissues but not in extraembryonic tissues, are discussed.

Mapping of the mouse X chromosome using random genomic probes and an interspecific mouse cross.

Two libraries enriched in murine X chromosome material have been constructed in the lambda vector NM 1149 from flow-sorted chromosomes. Inserts of unique genomic sequence DNA were purified and their X chromosome specificity characterised by hybridisation to a panel of somatic cell hybrid lines. Of the first five such X chromosome-specific probes characterised, all detect restriction fragment length polymorphisms (RFLPs) between inbred mouse laboratory strains such as C57BL/6 and BALB/c and the SPE/Pas mouse strain established from a wild Mus spretus mouse, when their DNAs are digested with the restriction enzyme TaqI. Taking advantage of these RFLPs, all five probes have been localised on the X chromosome using an interspecific backcross between the B6CBARI and SPE/Pas mouse strains segregating the X chromosome markers hypoxanthine phosphoribosyl transferase (Hprt) and Tabby (Ta). Three of the probes map to the region between the centromere and Hprt, and two distal to Ta. Since such X-specific sequence probes detect RFLPs between M. spretus and M. musculus domesticus DNAs with high frequency, a large panel of well localised probes should soon be available for studies of biological problems associated with the X chromosome which can best be approached using the murine species.

Differential Hm antigen expression on EC cells and early differentiated derivatives.

Differences in the expression of minor histocompatibility (Hm) alloantigens on two mouse embryonal carcinoma (EC) cell lines and the PYS-2 and T.D.M.-1 differentiated derivatives have been demonstrated by their ability to elicit a cytolytic T-lymphocyte (CTL) response. Experiments involving the use of various responder-target strain combinations on the one hand and Recombinant Inbred (RI) mice strains on the other have shown that: (i) there are major differences in Hm expression on the EC cells compared with the differentiated derivatives whose Hm expression appears more akin to that of adult splenocytes; (ii) although both EC cell lines show reduced Hm immunogenicity compared with adult splenocytes, major differences in the expression and possibly presentation between the F9 and PCC3 EC cell lines can be detected both by in vivo priming and by in vitro cold competition target experiments. These results are discussed in connection with the unexpected finding that some EC cell lines are capable of specific competition effects for appropriate CTL effectors despite their inability to stimulate such effectors in vitro and the absence of major histocompatibility complex (MHC) products.

Isolation and characterization of a monoclonal antibody specific for fibrinogen and fibrin of human origin.

A hybridoma cell line secreting a monoclonal antibody directed against human fibrinogen has been isolated. The antigenic determinant recognized is present on both the fibrinogen and fibrin molecules but is apparently absent from the D and E fragments and from fibrinopeptides A and B. The antibody seems to recognize a conformational structure present in native fibrinogen and in which several or possibly all the fibrinogen polypeptide chains may participate.

The significance of natural killer activity in the resistance of allogeneic hosts to the embryonal carcinoma cell line F9.

A number of mouse strains of known F9 resistance phenotype have been tested for their in vitro natural killer activity against both F9 and YAC target cells. The pattern of in vitro natural killer activity observed does not correlate with the pattern of in vivo F9 resistance. The relative efficiencies of in vitro lysis of the F9 and YAC targets by endogenous natural killer effectors from the 129/Sv, SJL/J, C57Bl/6J, and BALB/cJ strains do, however, parallel each other, even though overall levels of F9 target lysis are very low. Since cold competition experiments indicate that the embryonal carcinoma cell lines tested, F9, PCC4/Aza, and PCC3/A/1, can compete efficiently with YAC as target, this low lysis of embryonal carcinoma cells may be due to an intrinsic lysis resistance. The finding that the parietal endoderm cell line PYS-2 and the trophoectodermal cell line TDM1 also compete with YAC targets in cold competition experiments is discussed in relation to previous reports suggesting that such differentiated derivatives lack natural killer target structures.

The genetics of teratocarcinoma transplantation: tumor formation in allogeneic hosts by the embryonal carcinoma cell lines F9 and PCC3.

Two cultured lines of murine embryonal carcinoma, F9 and PCC3, have been grafted to a variety of allogeneic hosts. The host strains have been classified by their resistance or sensitivity to these carcinomas. Resistance seems to be immunological in nature.Allograft rejection does not correlate withH-2 haplotype, and seems to be controlled by a limited number of recessive factors, presumably histocompatibility genes. We infer that these factors have limited polymorphism in the mouse species. Recombinational analysis of strain A/He has revealed the presence of a recessive factor linked to theH-2 locus. Tumor resistance of strains C57BL/6 and AKR appears to result from the interaction of dominant or semi-dominant factors in theH-2 region with other recessive elements in the genetic background.Though F(1) hybrids between resistant mouse strains and the syngeneic strain 129 are largely tumor-sensitive, a low level of hybrid resistance to F9 has been observed and shown to be eliminated by X-irradiation.