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M Letellier

Publications and source records attributed to M Letellier.

32 records · Page 2Linked to original sources
PubMed1988

A molecular-genetic analysis of cytotoxic T lymphocyte function.

Two genes that are specifically expressed in T cells with cytolytic activity were isolated from a CTL cDNA library by differential screening. Both appear to encode serine proteases, thus suggesting a cascade mechanism, similar to complement, in activated CTL. Both CTL-specific proteases have a number of unusual structural features that suggest that they will have novel substrate specificities. One of the proteins (CCPI) has been oriented to the granules found in the cytoplasm of CTL. Taken together, these data strongly suggest that these molecules play an important role in target-cell lysis by CTL. Furthermore, we believe that the detailed molecular knowledge being accumulated through these studies may lead to the development of innovative forms of immunotherapy.

Amino Acid Sequence↗
PubMed1987

A serine protease (CCP1) is sequestered in the cytoplasmic granules of cytotoxic T lymphocytes.

Based upon the use of a new predictive algorithm, three peptides were synthesized that correspond to likely antigenic sites of the cytotoxic T cell-specific protease cytotoxic cell protein 1 (CCP1). Antibodies raised against these peptides, under reducing conditions, bound to a single protein of m.w. 29,000 found in two actively cytotoxic T cells. This protein was absent from a "cytotoxic" T cell line that had lost its cytolytic properties. By using immunocytologic techniques, the protein was localized within cytoplasmic granules. In addition, granules purified from the cytoplasm of cytotoxic T cells were shown, by Western blot analysis, to contain the protein. The molecule detected by the antibodies behaves, upon reduction, similarly to a diisopropylfluorophosphate-binding protein. Thus, here we provide evidence that CCP1, a protein whose expression correlates with cytotoxicity, is contained within organelles which have been implicated in killing. These findings strongly suggest that CCP1 plays a key role in T cell-mediated target cell lysis.

Cell Line↗
PubMed1987

Pinealectomy or superior cervical ganglionectomy do not alter reproduction in the wolf (Canis lupus).

Twelve wolves (6 male and 6 female) were used to study the role of the pineal in photoperiodic mediation of seasonal reproduction. Eight wolves were pinealectomized (PNX) or sham-pinealectomized (S-PNX) at 5 mo of age, and 4 were superior cervical ganglionectomized (SCGX) at 16 mo of age (2 males and 2 females per treatment). All attained puberty at the species-typical time, during their second breeding season, except 2 SCGX males that did not survive. Reproductive cycles of an additional male that was SCGX as an adult and the PNX and S-PNX wolves, followed for a minimum of 3 yr, did not differ from each other or from those of unoperated colony wolves on measures of serum testosterone and luteinizing hormone for males, or of serum estradiol and progesterone for females. Nor was the range of dates for ovulation different for treated vs. untreated females. Surgical transection of the olfactory tracts of 1 male and 1 female PNX wolf, inducing anosmia to control for the possibility of pheromonally synchronized cycles, also failed to alter the seasonality of these reproductive parameters. These results do not conform to the model of pineal mediation of sexual cycles for photoperiod-sensitive species. In spite of evidence for photoperiod influence, the wolf apparently relies on a system other than the pineal for seasonal control of reproduction.

Animals↗
PubMed1987

B lymphocytes respond specifically to phytohaemagglutinin after liposome-dependent transfer of purified phytohaemagglutinin receptors.

Phytohaemagglutinin (PHA) receptor glycoproteins purified by affinity chromatography from porcine splenic lymphocytes, were reconstituted into vesicles made of phosphatidylcholine and phosphatidylserine, concomitantly with Sendai virus fusogenic proteins HN and F. The vesicles were used as a vehicle to insert the PHA receptor glycoproteins into a highly enriched population of porcine B lymphocytes. Fluorescence analyses showed that 52 +/- 2% of the reconstituted B cells had incorporated the lectin receptors. The modified B lymphocytes were assayed for their response (tritiated thymidine incorporation into nucleic acids) to PHA, concanavalin A (Con A), or to lipopolysaccharide (LPS). The results showed that porcine B cells fused with vesicles containing only viral fusogenic proteins failed to respond to either PHA or Con A. Tritiated thymidine incorporation was similar to background values. The cells did, however, respond to LPS with values of label incorporation similar to those observed in the case of pre-fused B lymphocytes. When purified B lymphocytes were fused with vesicles containing PHA receptors and viral fusogenic proteins, assays of thymidine incorporation showed a statistically significant (P less than 0.001) and specific response of the modified cells to PHA stimulation. Reconstituted cells cultured in the presence of PHA incorporated approximately nine times more radioactive label than pre-fused cells or cells fused with vesicles containing only fusogenic viral proteins. In marked contrast, reconstituted B lymphocytes did not show any significant label incorporation above background level in response to Con A, but they retained their ability to respond to LPS. Our findings suggest that B lymphocytes can be made to respond specifically to PHA by insertion of appropriate lymphocyte-derived receptors.

Animals↗
PubMed1987

Functional incorporation and preferred orientation of phytohemagglutinin receptor glycoproteins in phospholipid vesicles.

Porcine lymphocyte Phaseolus vulgaris phytohemagglutinin (PHA) receptor glycoproteins purified by affinity chromatography have been reassembled into vesicles made of phosphatidylcholine and phosphatidylserine by detergent (dodecyltrimethylammonium bromide) dialysis. The receptor glycoproteins were incorporated into the lipid vesicles in a nonselective manner with a yield of 65-70%. Vesicles containing the glycoproteins were sealed as evidenced by their impermeability to calcium ions, using quin 2 trapped inside the vesicles. The vesicles were agglutinated by PHA, suggesting that the saccharidic moiety of the reconstituted glycoproteins was, at least in part, oriented towards the extravesicular medium. This observation was further supported by the fact that the vesicles bound 125I-labeled PHA in a specific and saturable manner. At maximum amount of lectin bound, a ratio of 1.01 +/- 0.05 microgram of PHA per microgram glycoprotein incorporated was measured. When the binding data were analyzed by Scatchard plot, a downward concave profile was observed, suggestive of a positive cooperativity at low concentrations of lectin. The orientation of the reconstituted lectin receptor glycoproteins was determined by proteolytic treatments of labeled glycoproteins. The combined action of trypsin and chymotrypsin released, in the 120,000 X g supernatant, approximately 80% of label when 125I-tagged PHA receptor glycoproteins were incorporated into the vesicles. When the oligosaccharidic moieties of the receptor glycoproteins were specifically labeled, the simultaneous action of the two enzymes released approximately 70% of tritium labeling present in the reconstituted system. Taken together, these results suggest that the reconstituted PHA receptors are preferentially oriented into the phospholipid vesicles. The reconstituted PHA receptor glycoproteins competed effectively with cellular receptors in the assay of PHA-induced porcine lymphocyte activation. A 50% inhibition of [3H]thymidine incorporation was observed when 1 microgram of glycoproteins in vesicles was added to the cultured cells, whereas vesicles alone had no effect at this (equivalent) concentration.

Animals↗
PubMed1987

Phosphate transport and alkaline phosphatase in confluent MDCK cell monolayers.

Several studies have suggested that the brush border membrane alkaline phosphatase is involved in the renal reabsorption of inorganic phosphate along the proximal convoluted tubule. However, other studies on the influence of l(-)bromotetramisole, an inhibitor of alkaline phosphatase, upon phosphate transport into brush border membrane vesicles have failed to show an involvement of alkaline phosphatase. The present experiments were designed to demonstrate that the MDCK (Madin, Darby, canine, kidney) cell line can be used as an alternative model to study phosphate transport, to examine the effect of l(-)bromotetramisole and the role of alkaline phosphatase. MDCK cell monolayers were concomitantly used for alkaline phosphatase activity measurement and phosphate transport analysis. While alkaline phosphatase activity increases by 37-fold from day 2 to day 8 of culture, reaching a plateau at day 10, the sodium-dependent phosphate transport into the cell monolayers decreases by 2-fold during that same period. The phosphate transport was also studied in the presence of l(-)bromotetramisole at pH 8.5. The sodium-dependent phosphate uptake of inorganic phosphate is reduced by 43% in the presence of l(-)bromotetramisole at day 3 of culture but is not reduced in the 7-day-old culture. Our results suggest a participation of alkaline phosphatase upon phosphate transport in MDCK cell monolayers and indicate that this cell line constitutes a good model to study the relationship between alkaline phosphatase and phosphate transport.

Alkaline Phosphatase↗
PubMed1985

Phytohemagglutinin (PHA)-resistant Chinese hamster ovary cell mutants acquire sensitivity to the lectin after fusion with liposomes containing PHA receptor glycoproteins.

Phytohemagglutinin receptor glycoproteins have been inserted into phospholipid vesicles and these have been fused with phytohemagglutinin-resistant chinese hamster ovary cells. Our results show that the fused cells acquire "neoreceptors" for the lectin phytohemagglutinin. Fluorescence activated cell sorting analyses show that approximately 40% of the cells fused with the receptor-containing vesicles. Studies with 125I-labelled lectin showed that fused cells bound three times more ligand than untreated mutant cells. Furthermore, lectin receptors were functionally inserted in the mutant cell plasma membrane. Fused cells cultured in the presence of lectin (200 micrograms ml-1) lost rapidly (8 hours or less) their ability to incorporate [3H] thymidine. Whereas mutant cells cultured for 16 hours in the presence of 50-400 micrograms ml-1 of lectin remained viable, fused cells showed a 45% decrease in 3H-labelled nucleotide incorporation. The method described here should be of general applicability for the study of lectin-dependent cytotoxicity in chinese hamster ovary cell lines.

Animals↗
PubMedDate not supplied

Freezing D2O clay gels.

To obtain the T1 surface value in smectites/D2O diluted suspensions or gels, as was obtained on a monolayer deuterated clay, we freeze them. The broad Pake's doublets similar to ice doublets and with the same T1 show that we can separate frozen from unfrozen D2O. The latter exhibits a narrower line and a single T1 and is attributed to the liquid surface water layer in rapid exchange with the nearby supercooled water, the quantity of which diminishes with the lowering of the temperature depending on the gel porosity. It is possible to measure the supercooled water quantity and to correct the T1 measured values to extract the T1 surface. The value extrapolated at room temperature allows the complete clay surface area measurement. The example of a montmorillonite is given and a comparison with laponite is made.

Aluminum Silicates↗