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Biomedical subjects

P Perrin

Publications and source records attributed to P Perrin.

At least 217 records · Page 12Linked to original sources
PubMed1979

[Glycosylation and isoelectric properties of complete and defective rabies viruses (author's transl)].

From a structural point of view an essential distinction between complete and defective rabies viruses is difference in size. In addition, isoelectric properties differ. The complete virus has an isoelectric point approaching neutrality, whereas the defective virus focuses between pH 3-4.7. The isoelectric points of the glycoprotein from complete and defective viruses differ in a corresponding fashion. The Pasteur virus cultivated on BHK21C13 cells, contains glycoproteins, the glycopeptides of which have a structure containing the following five monosaccharides: sialic acid, D-galactose, (N-acetyl)D-glucosamine, D-mannose and L-fucose. The glycosylation of the glycoproteins is different, at least in so far as the relative sialic acid/glucosamine ratio is concerned.

Acetylglucosamine↗
PubMed1976

Forty years of transurethral prostatic resections.

The 4 consecutive series of transurethral prostatic resections compared were done by the senior authors during a period of nearly 40 years: 420 cases in 1937 to 1938, 520 cases in 1940 to 1945, 480 cases in 1950 to 1955 and 270 cases in 1972 to 1973. The most notable improvements in the recent series are the fewer patients with temperatures more than 100 degrees (69 virsus 29 per cent) and the improved mortality rate (5.5 verus 0.7 per cent). In our opinion these improvements are owing to antibacterial medication, improved instruments, more skillful performance of the operation, the use of isotonic irrigating fluid and improved postoperative care.

Fever↗
PubMed1976

[Rabies glycoprotein purification by isoelectric focusing (author's transl)].

A glycoprotein was extracted with Triton X100 from rabies virus grown in primary foetal bovine kidney cells. This glycoprotein was further purified by iso electro focusing and showed a major peak at pH 7,0 and a smaller peak at pH 4,6. Purified fractions were migrated on polyacrylamide gels and assayed for immunogenicity.

Glycoproteins↗
PubMedDate not supplied

Production, purification and analysis of an experimental DNA vaccine against rabies.

BACKGROUND: The basic and applied research efforts devoted to the development of DNA vaccines must be accompanied by manufacturing processes capable of being scaled up and delivering a clinical-grade product. This work describes a rapid process of this kind, based on hydrophobic interaction chromatography (HIC) for the production of milligram quantities of an experimental DNA rabies vaccine. Its properties and protective activity are tested in comparison with the same plasmid DNA purified with a commercial kit. METHODS: The experimental DNA vaccine encoding the rabies virus glycoprotein was amplified in vivo in Escherichia coli. The plasmid was isolated by alkaline lysis, pre-purified and concentrated by isopropanol and (NH4)2SO4 precipitation, and purified by HIC and dialysis. Product quality was controlled by using high-performance liquid chromatography (HPLC), Southern slot blotting, agarose gel electrophoresis, the kinetic-QCL Limulus amoebocyte lysate assay, and protein assays. The expression of the rabies virus glycoprotein was tested in vitro in neuroblastoma cells. The production of rabies-virus-neutralising antibodies and the protection against an intracerebral virus challenge were tested in mice. RESULTS: One hundred and forty-two milligrams of the plasmid, with an HPLC purity greater than 99% were obtained from 4.5 l medium. Control analysis showed that the vaccine conforms to specifications in terms of impurities (endotoxins, genomic DNA, RNA, proteins). Furthermore, the final experimental vaccine induces rabies-virus-neutralising antibodies and protects mice against a rabies virus challenge. CONCLUSIONS: This study demonstrates that the method developed for the purification of milligram amounts of plasmid delivers an endotoxin-free, experimental rabies DNA vaccine, with protective activity similar to that obtained with the vaccine purified using a commercial kit.

Animals↗
PubMedDate not supplied

The antigen-specific cell-mediated immune response in mice is suppressed by infection with pathogenic lyssaviruses.

Responsiveness of T cells (RTC) was studied in BALB/c mice intramuscularly infected with various lyssaviruses. After infection by this peripheral route, two types of viruses could be classified according to their effects: 1) pathogenic viruses, including fixed rabies Pasteur virus (serogenotype 1) and wild viruses belonging to serogenotype 1 (from a rabid fox in France and from a cow infected by a vampire bat in Brazil) or to serogenotype 5 (European bat lyssavirus 1); and 2) non-pathogenic viruses, including Mokola virus (serogenotype 3). RTC was tested by analysing in vitro the capacity of splenic T cells from infected BALB/c mice to produce cytokines after antigenic (purified lyssavirus antigens) or polyclonal stimulation (concanavalin A). Cytokine production was followed by assaying the biological activity of interleukin-2 and by testing for interleukin-2, interleukin-4 and interferon-gamma (IL2, IL4 and IFN gamma ) messenger RNAs (mRNA) by transcription into complementary DNA and amplification by the polymerase chain reaction. The initial biologically active IL2 and cytokine mRNA production was observed in mice infected with pathogenic or non-pathogenic lyssaviruses. Only mice with symptoms (infected with pathogenic viruses) lost the capacity to produce cytokines in vitro after antigen-specific stimulation. No such loss was observed after polyclonal stimulation. In mice peripherally infected with non-pathogenic viruses, no loss was observed after stimulation with lyssavirus antigens. Thus, infection with pathogenic lyssaviruses by the peripheral route induces in BALB/c mice a loss of T-cell responsiveness after antigen activation, but not after polyclonal activation.

Animals↗
PubMedDate not supplied

Evaluation of the induction of specific cytotoxic T lymphocytes following immunization of F1 hybrid mice with rabies antigens.

The measurement of the production of cytotoxic T lymphocytes (CTL) induced in mice by rabies antigens currently uses spleen cells from immunized A/J mice as effector cells and infected neuroblastoma syngeneic cells as target cells. For several reasons, including difficulties in obtaining A/J mice, as well as genetic analysis of immune responses, it would be advantageous to use strains of mice other than the A/J mice. However, cell lines other than the neuroblastoma Neuro-2a line are difficult to infect by the rabies virus. Therefore, using the same target cells expressing the major histocompatibility complex H-2kd, we have developed an experimental system based on the induction of CTL in F1 BALB/c X C3H/HeJ hybrid H-2kd mice. Splenocytes from F1 hybrid mice primed with inactivated purified rabies virus (IPRV) exhibited cytotoxic activity specific for syngeneic infected target cells (Neuro-2a). High amounts of IPRV were required for the induction of CTL following in vivo priming. The antigen dose required for CTL induction was reduced by in vitro restimulation. In addition to specific CTL, a high level of natural killer cell activity was induced in F1 hybrid mice by priming with IPRV. Among rabies antigen preparations tested (IPRV, purified glycoprotein and ribonucleoprotein), only IPRV induced strong CTL stimulation.

Animals↗