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D Gaucher

Publications and source records attributed to D Gaucher.

6 recordsLinked to original sources

Molecular cloning of gerbil interleukin 12 and its expression as a bioactive single-chain protein.

Complementary DNAs coding for gerbil interleukin 12 (IL-12) p40/p35 subunits were cloned by a combination of cross species reverse transcriptase-polymerase chain reaction (RT-PCR) and 3' rapid amplification of cDNA ends (RACE) techniques. IL-12 p40/p35 had 79% nucleotide identity and 81% amino acid homology to mouse IL-12 p40/p35. The p40/p35 subunits were expressed as a single polypeptide separated by a short hinge sequence that allowed for proper folding and assembly. COS-7 cells transfected with DNA encoding the single-chain gerbil IL-12 (pSCjIL12) secreted high levels of the protein which stimulated proliferation of ConA-activated gerbil spleen lymphoblasts in a dose-dependent manner.

Amino Acid Sequence↗

Hormonal induced lactation in transgenic goats.

The aim of this study was to hormonally induce lactation in prepubertal, nulliparous, and male goats both transgenic and non-transgenic. Analysis of milk quality, recombinant protein expression levels, total amount of recombinant protein produced, and the affect on long-term reproductive capability was assessed. Fifty-one goats (Saanen, Alpine, and Toggenburg), male and non-pregnant females, 2-31 months of age, either non-transgenic or transgenic were evaluated with a total of 10 transgenes (constructs) represented. Animals were given estradiol (0.25 mg/kg, i.m.) and progesterone (0.75 mg/kg, i.m.) on days 1, 3, 5, 7, 9, 11 and 13, while prednisilone (0.4 mg/kg, i.m.) was administered on days 14-16 with mammary massage occurring daily from day 5 onward. Forty of 51 animals, (36 of 38 females and 4 of 13 males) produced milk with total volumes in the 30-day experiment, ranging from 20 microl to 530 mls per day, or approximately 500 microl to 6.8 liters total. Milk composition was analyzed for various parameters (total protein, fat content, total solids and somatic cell count) with no significant differences found between induced and natural milk. Expression levels of recombinant proteins from transgenic animals that were analyzed during the induced lactation, and subsequently during normal lactations, were found to have no significant differences. Total amount of recombinant protein produced was evaluated at different expression levels with no statistical significance seen. While over 90% of the females placed in the regimen became pregnant, there was a correlation between increased age at time of induction and an increase in number of breedings, or reproductive cycles needed to establish a pregnancy after induction. For males, 100% placed in the regimen settled females after hormonal induction of lactation. Semen quality was evaluated prior to, during, and after hormonal treatments. Semen volume and sperm number did not differ; however, for a small percentage of males, there was a decrease in sperm and post thaw motility after hormonal treatments. These levels returned to normal within 4-5 weeks. Subsequent natural lactations showed total milk volumes within breed standards. These findings indicate that hormonal induction of lactation in the caprine species is a viable alternative to pregnancy for initiating lactation and milk production, does not adversely impact reproductive performance long-term, and can benefit the early assessment of recombinant proteins produced in a transgenic founder program.

Animals↗

Serum from Entamoeba histolytica-infected gerbils selectively suppresses T cell proliferation by inhibiting interleukin-2 production.

Suppression of T and B cell responses during invasive amebiasis may be serum mediated. The mechanism of serum-mediated suppression of spleen cell lymphoproliferation from gerbils with amebic liver abscess was examined. Compared with uninfected gerbil serum (10%), serum samples collected at days 10, 30, and 60, but not at day 20 after infection, augmented both concanavalin A (Con A; T cell mitogen)- and lipopolysaccharide (LPS; B cell mitogen)-induced proliferation of homologous spleen cells. Only day 20 serum (>5%) inhibited Con A- but not LPS-induced proliferation of spleen cells from uninfected gerbils. The suppressive mechanism was independent of nitric oxide and prostaglandin but involved reduced interleukin (IL)-2 production. Addition of exogenous IL-2 reversed the suppressive effect of day 20 serum on Con A-stimulated proliferation. These results identify a mechanism whereby serum may contribute to transient suppression of T cell responses during Entamoeba histolytica infections.

Animals↗

Synergism of IL-2-stimulated splenocytes and Pentostam enhances the killing of Leishmania donovani in vitro.

Control of infections by the obligate intramacrophage protozoan parasite Leishmania donovani is traditionally done with pentavalent antimonial drugs. It was of interest to determine if immunotherapy with IL-2-stimulated splenocytes would enhance drug action in an in vitro model system. It is confirmed that nontoxic doses of Pentostam decrease infection in a dose-dependent manner in vitro in terms of both the number of amastigotes/100 macrophages and the % of infected macrophages; the curative effect was most apparent when the drug was used on the 7th day of the infection, when the parasite was in its proliferative phase. It is also confirmed that recombinant IL-2-stimulated splenocytes induced infected macrophages to reduce significantly their parasite burden, especially when the infection was treated in the nonproliferative phase of the parasite, also in a dose-dependent manner. Leishmanicidal action in the infected macrophages was induced by cytokine(s) released from the lymphokine-stimulated cells. Immunotherapy in the nonproliferative phase, combined with drug treatment in the proliferative phase, reduced the infection to levels significantly below those produced by either treatment alone. Immunotherapy with IL-2-stimulated splenocytes in combination with Pentostam is, therefore, an excellent candidate treatment for the effective reduction of experimental infections by L. donovani.

Animals↗

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