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Biomedical subjects

A Bensaid

Publications and source records attributed to A Bensaid.

At least 55 records · Page 3Linked to original sources

Some properties of HU are modified after the infection of Escherichia coli by bacteriophage T4.

Escherichia coli HU, an abundant, nucleoid-associated, DNA-binding protein, plays a role in several biological processes including DNA replication. Many other bacteria have well-conserved HU homologs, and there are several more-distantly related members of the family, including TF1, encoded by Bacillus subtilis phage SPO1. We have asked whether coliphage T4, like SPO1, encodes an HU homolog or whether it alters the properties of host HU. We have been unable to detect a T4-specified HU homolog, but we have shown that E. coli HU extracted from phage-infected cells differs in some properties from that extracted from uninfected cells. First, HU from uninfected cells inhibits a reconstituted T4 DNA replication system, whereas HU from infected cells does not. Second, HU from infected cells appears to bind a T4-encoded polypeptide, as shown by coimmunoprecipitation. We propose that such binding alters HU function in T4-infected cells.

Bacterial Proteins↗

In vivo leukocyte tropism of bovine leukemia virus in sheep and cattle.

Bovine leukemia virus (BLV), an oncovirus related to human T-cell leukemia virus type I, causes a B-cell lymphoproliferative syndrome in cattle, leading to an inversion of the T-cell/B-cell ratio and, more rarely, to a B-cell lymphosarcoma. Sheep are highly sensitive to BLV experimental infection and develop B-cell pathologies similar to those in cattle in 90% of the cases. BLV tropism for B cells has been well documented, but the infection of other cell populations may also be involved in the BLV-induced lymphoproliferative syndrome. We thus looked for BLV provirus in other leukocyte populations in sheep and cattle by using PCR. We found that while B cells harbor the highest proviral load, CD8+ T cells, monocytes, and granulocytes, but not CD4+ T cells, also bear BLV provirus. As previously described, we found that persistent lymphocytosis in cows is characterized by an expansion of the CD5+ B-cell subpopulation but we did not confirm this observation in sheep in which the expanded B-cell population expressed the CD11b marker. Nevertheless, BLV could be detected both in bovine CD5+ and CD5- B cells and in sheep CD11b+ and CD11b- B cells, indicating that the restricted BLV tropism for a specific B-cell subpopulation cannot explain its expansion encountered in BLV infection. Altogether, this work shows that BLV tropism in leukocytes is wider than previously thought. These results lead the way to further studies of cellular interactions among B cells and other leukocytes that may intervene in the development of the lymphoproliferative syndrome induced by BLV infection.

Animals↗

In vitro infection of bovine brain endothelial cells by Cowdria ruminantium.

Endothelial cells from bovine brain capillaries (BBEC) were successfully infected with the Gardel stock of Cowdria ruminantium. Growth conditions of cowdria in BBEC and bovine umbilical endothelial cells (BUE) were identical with average intervals between passages of 8.6 and 8.9 days in BBEC (17 passages) and BUE (28 passages), respectively. The time required to complete the life cycle and the morphology of the parasite were identical in both types of cells. The demonstration that BBEC may be infected with C ruminantium offers the means to study the mechanism of infection of the blood brain barrier endothelium by this parasite.

Animals↗

An attempt to correlate cattle breed origins and diseases associated with or transmitted by the tick Amblyomma variegatum in the French West Indies.

By using biological data and historical research, we have tried to explain the difference between resistance and susceptibility to the diseases transmitted (cowdriosis) or associated (dermatophilosis) with the tick Amblyomma variegatum, in two cattle breeds of the French West Indies: the Creole crossbred cattle of Guadeloupe and the Brahman zebu cattle of Martinique. Have been studied the polymorphisms of 5 independent genetic systems (erythrocytic haemoglobin, serum albumin and transferrin, the class I region of the BoLA complex and the gamma S crystallin gene) in different breeds comprising Bos taurus cattle of Europe and Africa, Bos indicus of West and East Africa, as well as the Brahman of Martinique and the Creole crossbred of Guadeloupe. By comparing the different allele frequencies of these 5 non related polymorphic loci and by using the two different mathematical matrices of NEI and of CAVALLI-SFORZA, have been established the genetic distances between these breeds. It appears clearly that the Creole cattle of Guadeloupe are in an intermediate position between the Bos taurus N'Dama breed of West Africa and two Bos indicus zebu breeds, namely the West African Sudan zebu and the Brahman. Thanks to studies of different archieves in the Caribbean and in Europe, historical evidence have been accumulated on the geographical origins and on the chronology of the establishment of Creole and Brahman cattle in the French West Indies. The high resistance of the Creole cattle of Guadeloupe to diseases associated with or transmitted by the "Senegalese" tick Amblyomma variegatum seems to be due to the inheritance of a pool of genes from West African cattle and more particularly from the N'Dama breed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

An attempt to identify genetic markers of resistance or susceptibility to dermatophilosis in the zebu Brahman population of Martinique.

Dermatophilosis is a disease associated with the tick Amblyomma variegatum, and a genetic predisposition to the manifestation of symptoms has been demonstrated. Indeed, the Creole cattle of Guadeloupe constitute a population which is highly resistant to this disease, whereas the Brahman zebu cattle of Martinique seem very susceptible. However, in this Brahman population there is a gradient regarding the severity of symptoms depending on individuals. In several herds of these pure zebu Brahman, kept under the same farming conditions, we selected susceptible and resistant groups because of the simultaneous presence of animals affected or not by clinical dermatophilosis. In these animals we studied several highly polymorphic genetic systems such as haemoglobin, albumin, the BoLA Complex (class I and II) and the gamma S crystallin gene. Only exon 2 of the BoLA-DRB3 gene, investigated by PCR-RFLP technique, showed interesting polymorphisms. We have established a genotypic map showing at least 4 different alleles of which 1 seems particular to one animal susceptible to dermatophilosis. Before reaching any conclusion further investigations with more DNA samples of susceptible animals are needed.

Actinomycetales Infections↗

Somatic cell mapping of T-cell receptor CD3 complex and CD8 genes in cattle.

Bovine genes encoding T-cell receptor, CD3, and CD8 molecules have been mapped to syntenic groups using bovine x rodent hybrid somatic cells. T-cell receptor alpha and delta chains were assigned to bovine syntenic group U5, and the beta and gamma genes were syntenic with each other and with markers on U13. CD3E and CD3D genes were syntenic with each other and located to bovine syntenic group U19. CD8 was most concordant with markers of syntenic group U16, although the concordancy was only 85% and the assignment must be regarded as tentative. The comparative gene maps of human chromosome 7, bovine syntenic group U13, and mouse chromosomes 6 and 13 suggest extensive evolutionary conservation.

Animals↗

Identification of expressed bovine class I MHC genes at two loci and demonstration of physical linkage.

A cDNA library prepared from lymphocytes of a cow (E98), homozygous at major histocompatibility complex (MHC) loci (BoLA phenotype w10, KN104), was screened with a bovine MHC class I probe. Of the cDNA clones isolated, two, (2.1 and 5.1) were selected and showed divergence at both 5' and 3' termini. E98 DNA was digested with rare-cutter enzymes (Sfi I, Mlu I, Not I, and Cla I) and fragments were size-separated by field inversion gel electrophoresis (FIGE). Hybridization with an entire class I cDNA probe revealed multiple fragments generated by each enzyme. When the 3' untranslated regions (UT) of 2.1 and 5.1 were used as probes, only one fragment was revealed in each digested sample, showing locus specificity of these probes in cattle. Further, DNA of transfected mouse fibroblasts L4 (expressing KN104) and L10 (expressing w10) hybridized to the 3'UT regions of clones 2.1 and 5.1, respectively. Northern blot analysis of the mRNA of the L4 and L10 transfected cells provided further evidence that the cDNA clones 2.1 and 5.1 code for the BoLA-KN104 and BoLA-w10 class I molecules respectively, and thus these represent the products of two different genes. A long range physical mapping of the BoLA-w10 and KN104 genes was performed using FIGE analysis of DNA of an homozygous and an heterozygous animal. This analysis revealed that the BoLA-w10 and KN104 genes are separated by not more than 210 kilobases (kb) and that they are components of a multigene family spanning 1550 kb. As the w10 gene is at the BoLA-A locus we assign the KN104 gene to a B locus.

Animals↗

Analysis of the reactivity of anti-bovine CD8 monoclonal antibodies with cloned T cell lines and mouse L-cells transfected with bovine CD8.

Mouse L-cells transfected with bovine CD8 and two Theileria parva-infected cloned T cell lines expressing bovine CD8 were used to screen the panel of ten monoclonal antibodies (mAbs) submitted to the workshop. Eight of the ten mAbs reacted with the transfectant and both the cloned T cell lines. However, two mAbs CC58 and BAT82A did not recognise the transfectant and only reacted with one of the T cell lines. Further biochemical studies indicated that the eight mAbs react with both homo- and heterodimeric forms of bovine CD8 whilst the two mAbs CC58 and BAT82A react with only heterodimeric forms. These data suggest that bovine DC8 is encoded by two genes as is the case in mouse and man.

Animals↗

Cell surface phenotype of two cloned populations of bovine lymphocytes displaying non-specific cytotoxic activity.

Monoclonal antibodies specific for T cell differentiation antigens were tested on four cloned populations of lymphocytes derived from the peripheral blood mononuclear cells of an animal immunised with Theileria parva. The clones were defined functionally in terms of cytotoxic activity, MHC restriction and expression of messenger RNA for CD3 and T cell receptor (TCR). Two clones contained RNA transcripts for CD3, TCR-alpha and beta and were positive for CD2, CD5 and CD6; one of these was a typical CD4+ class II MHC-restricted non-cytotoxic clone while the other was a CD8+ class I MHC-restricted cytotoxic clone. By contrast, the remaining two clones had the characteristics of non-specific killer cells in that they exhibited moderate levels of non-MHC-restricted killing; they contained TCR-delta mRNA and a 1.2 kb truncated form of TCR-beta message, but they did not contain CD3 or TCR-alpha mRNA. One of these non-specific killer clones only expressed CD2 whereas the other clone only expressed CD8, but without the CD8 determinant recognised by monoclonal antibodies CC58 and BAT52. All four clones were negative for the WC1 antigen which is expressed on gamma/delta T lymphocytes.

Animals↗

Identification of a bovine surface antigen uniquely expressed on CD4-CD8- T cell receptor gamma/delta+ T lymphocytes.

In this study, two monoclonal antibodies, IL-A29 and CC15, are described that identify a novel bovine cell surface marker of 215/300 kDa. The antibodies reacted with a discrete population of resting lymphocytes in peripheral blood which, in young animals, constituted about 25% of the mononuclear cells. Thymus, lymph nodes and spleen contained less than 5% positive cells. These cells were negative for surface Ig, a monocyte/granulocyte marker, and the T lymphocyte antigens CD2, CD6, CD4 and CD8. Immunohistological analyses revealed the presence of IL-A29/CC15-positive lymphocytes in the thymic medulla, in the outer cortex of lymph nodes, in the marginal zones of the spleen, in the dermal and epidermal layers of the skin and in the lamina propria of the gut. The IL-A29/CC15+ cells in unfractionated blood mononuclear cells responded in autologous and allogeneic mixed lymphocyte cultures, and when purified they responded to concanavalin A in the presence of recombinant interleukin 2. These observations suggested this population of cells belonged to the T cell lineage. In order to unambiguously define their lineage, cDNA clones encoding bovine T cell receptor (TcR) and CD3 proteins were isolated. Northern blot analyses of IL-A29/CC15+ cell populations and of established cell lines of various lineages demonstrated that they expressed TcR delta and CD3 gamma, delta and epsilon mRNA: TcR alpha was not expressed, whereas only a truncated form of TcR beta mRNA was present. These results indicate that the IL-A29 and CC15 antibodies define a unique population of CD4-CD8-, gamma/delta T cells.

Animals↗

Biochemical characterization of activation-associated bovine class I major histocompatibility complex antigens.

Utilizing a 'sandwich' ELISA assay we have been able to demonstrate that mAb W6/32, B1G6 and IL-A19 are reactive with three different monomorphic determinants on bovine class I major histocompatibility complex (MHC) molecules. Sequential immunoprecipitations performed with the mAb revealed that class I molecules on PBM comprise a single population with respect to reactivity with the mAb in that the beta 2m-associated proteins bear all three epitopes. By contrast, TCGF-driven lymphoblasts and cells transformed by Theileria parva (Tp) additionally express molecules of Mr 45000 bound to beta 2m which are recognized by mAb B1G6 and IL-A19 but not by W6/32. These two subclasses of molecules were further distinguished on the basis that, when tunicamycin was added to cultures in the preparation of cells for analysis, mAb W6/32 precipitated class I heavy chains of Mr 39000 while the extra molecules detected only by mAb B1G6 and IL-A19 were of Mr 37000 and 39000. On thymocytes, the mAb W6/32-non-reactive class I molecules are present in low amounts and are expressed by cells in the medulla area, unlike BoT1 (analogous to human CD1) molecules which are expressed by the cortical cells. Our studies also revealed that the supposed beta 2m-specific mAb B1G6 does not recognize the beta 2m-associated molecules (BoT1) precipitated by mAb TH97A and thus the specificity of mAb B1G6 in cattle is for an epitope on bovine beta 2m which is strongly influenced by the nature of the heavy chain with which the beta 2m is associated.

Animals↗