Introduction: Human-cl rhFVIII (Nuwiq®), a new generation recombinant factor VIII (rFVIII), is the first rFVIII produced in a human cell-line approved by the European Medicines Agency.
Aims: To describe the development, upscaling and process validation for industrial-scale human-cl rhFVIII purification.
Methods And Results: The purification process involves one centrifugation, two filtration, five chromatography columns and two dedicated pathogen clearance steps (solvent/detergent treatment and 20 nm nanofiltration).
Immunization with a recombinant glycoprotein 160 envelope immunogen derived from a virus of genetic subtype B induced strong specific T-helper cell responses in asymptomatic human immunodeficiency virus (HIV) carriers infected with subtypes B to G. This indicates that the HIV-specific T-helper immunity, which is the basis for development of antibodies and cytotoxic T lymphocytes, can be improved by both homologous and heterologous antigens. It also suggests that a particular immunogen can be effective against many different HIV strains.
View Article and Find Full Text PDFForty asymptomatic HIV-infected individuals with CD4+ lymphocyte levels above 400x10(6)/l were immunized over 5 years with recombinant envelope glycoprotein gp160 (rgp160). After 5 years there was a trend towards more non-progressors in the immunized group as compared to the matched controls. Since immunizations could activate HIV, the first 6 immunizations were followed by 2 weeks of zidovudine or placebo, double-blind.
View Article and Find Full Text PDFAIDS Res Hum Retroviruses
July 1999
The outer envelope glycoprotein (gp120) from subtypes A-E of HIV-1 was purified using a specific high mannose-binding lectin, Galanthus nivalis agglutinin. All isolates were grown in peripheral blood lymphocyte cells in order to avoid selection in cell lines. A comparison of the reactivities of the envelope proteins was made using sera from patients infected with the different subtypes.
View Article and Find Full Text PDFObjectives: To induce recovery of HIV-1-specific immune responses by combining immunization with antiviral chemotherapy.
Design: Forty HIV-infected patients entered a double-blind study with recombinant gp160 in combination with zidovudine or placebo. The pretreatment observation period was around 2 years and the treatment period 5 years.
Methods Mol Med
October 2012
The human immunodeficiency viruses (HIV-1 and HIV-2) are the etiologic agents of the acquired immunodeficiency syndrome (AIDS) and related disorders (1-3) Simian immunodeficiency virus(SIV) is the corresponding virus for nonhuman primates. SIVmac has been isolated from rhesus monkeys (Macaca mulatta) with immunosuppression and malignant lymphomas (4).
View Article and Find Full Text PDFPeptides were synthesized which combined HIV-1 B-epitopes from gp41, p34pol and heterologous T-cell epitopes from hepatitis B virus (HBV) core or tetanus toxoid. Mixtures of these composite peptides and peptides representing single HIV-1 B-epitopes were used to immunize rabbits in an adjuvant-free immunization regimen. Fusion to T-cell epitopes made the HIV-1 B-epitopes immunogenic and high titers of anti-HIV-1 antibodies were reached.
View Article and Find Full Text PDFJ Acquir Immune Defic Syndr (1988)
September 1994
Mouse monoclonal antibodies with high human immunodeficiency virus type 1 (HIV-1) neutralizing titers were used for passive immunotherapy of eleven late-state HIV-infected patients. In five patients the serum level of the core protein p24 decreased, while in five cases it remained unchanged. The level of viral RNA in plasma as measured by quantitative polymerase chain reaction (PCR) decreased in four cases, was stable in another four, and increased in three cases.
View Article and Find Full Text PDFTwo-phase extraction in a system composed of dextran and polyethylene glycol was used to purify simian immunodeficiency virus, SIVMAC251 (32H isolate) from 25 l of culture supernatant. The virus partitioned to the interphase with 80% recovery of gag peptide p27 and reverse transcriptase and an about 25% recovery of the external env glycoprotein, gp148. The virus was treated with octylglycoside and its subcomponents separated.
View Article and Find Full Text PDFStrong specific T-cell responses to human immunodeficiency virus type 1 (HIV-1) gp160 were induced by immunization with recombinant gp160 (rgp160). It was given as postinfection vaccination to 40 asymptomatic HIV-1 seropositive patients. The participants received 6 doses of 160 micrograms rgp160 administered intramuscularly at 0, 1, 4, 8, 17, and 26 weeks and were monitored for 1 year.
View Article and Find Full Text PDFAIDS Res Hum Retroviruses
May 1993
Lectin affinity chromatography was used to purify in a single step the envelope glycoproteins of HIV-1, HIV-2, and SIV. Envelope glycoproteins carry the major determinants essential for protection by the humoral immune response. The purification of these proteins has previously been a laborious procedure.
View Article and Find Full Text PDFHuman monoclonal antibodies specific for human cytomegalovirus (CMV) antigens have been established using peripheral blood lymphocytes from a seropositive donor. Immortalization of antigen-specific B cells was achieved by Epstein-Barr virus transformation followed by somatic cell fusion of antigen-specific lymphoblastoid cells. Four clones producing high-affinity antibodies (0.
View Article and Find Full Text PDFJ Clin Lab Anal
March 1992
A mouse monoclonal hybridoma cell line producing IgG 1k to human immunodeficiency virus (HIV-1) gp120 envelope protein was cultured in several systems. A small-scale flask culture was essential for characterizing the culture variables of the hybridoma. A dialysis tubing culture appeared to be an excellent alternative to in vivo cultures of ascitic fluid, and gave high mouse monoclonal antibody (Mab) concentrations.
View Article and Find Full Text PDFAIDS Res Hum Retroviruses
December 1990
The conventionally applied centrifugation protocols for the concentration and purification of human immunodeficiency virus type 1 (HIV-1) result in a low recovery of the external glycoprotein, gp120. This is consistent with what has been found with other retroviruses. In the search for a method allowing the copurification of the gp120 with the virion we have applied two-phase extraction based on water-soluble polymers.
View Article and Find Full Text PDFA cloned Bam H1 fragment of genomic Bordetella pertussis DNA which recognized a frequently repeated sequence in the genome of B. pertussis was used as a probe in a DNA hybridization assay for the detection of Bordetella. Extensive studies on cross-reactivity were carried out in standardized strains and in cultured swab specimens from patients without suspected pertussis.
View Article and Find Full Text PDFOne hundred nucleoside analogs with fluorine substitutions at various positions on the pentose ring were evaluated for inhibitory activity against human immunodeficiency virus type 1 (HIV-1). Nine compounds emerged as inhibitors of HIV-1 replication, with various degrees of selectivity; the most active of these was 3'-fluoro-3'-deoxythymidine, followed by 5'-amino-3'-fluoro-3'-deoxyadenosine. Substitution of fluorine at the 2'-deoxy or 3'-deoxy position resulted in increased antiviral activity of the thymidine analogs, whereas the activity of adenosine or cytidine analogs was not increased by fluorination at either position.
View Article and Find Full Text PDFThe IgG subclass of antiviral antibodies to human cytomegalovirus (CMV) is mainly of IgG1 type. Most CMV seropositive sera also have virus-specific IgG3, but of a lower titre as measured by enzyme-linked immunosorbent assay (ELISA). We studied the reactivity pattern of these two IgG subclasses to CMV structural polypeptides in order to define how virus-specific IgG1 and IgG3 contribute to the neutralization of CMV.
View Article and Find Full Text PDFAntimicrob Agents Chemother
May 1989
We describe a synergistic effect of combinations of foscarnet and 3'-azido-3'-deoxythymidine against human immunodeficiency virus type 1 multiplication in cell culture, an additive effect of foscarnet and 3'-azido-3'-deoxythymidine triphosphate against human immunodeficiency virus type 1 reverse transcriptase, and a low toxicity in cell culture of combinations of the two drugs.
View Article and Find Full Text PDFSome 3'-blocked pyrimidine analogs were synthesized and tested as inhibitors of replication of human immunodeficiency virus (HIV) and Moloney-murine leukemia virus (MuLV). The analogs were of 3 kinds: (1) analogs of 3'-azido-3'-deoxythymidine (AZT) in which the C-5 CH3 of the base was exchanged for H (AZU) or C2H5 (AZEU); (2) 3'-fluoro-3'-deoxythymidine (FLT) and analogs thereof, in which the C-5 CH3 of the base was exchanged for H (FLU), C2H5 (FLEU) or nC3H7 (FLPU); (3) the threo analogs of AZT (AZT increases) and AZU (AZU increases). All analogs were less active inhibitors of HIV replication than AZT, except FLT, which was as active as AZT.
View Article and Find Full Text PDFWe compared four different procedures for the purification and concentration of nucleoside triphosphates in cell extracts prior to HPLC analysis. Two methods involved precipitation, with either acetonitrile or calcium fluoride. The acetonitrile procedure yielded reasonable recovery and sufficient purity for the subsequent HPLC analysis.
View Article and Find Full Text PDFThe subclass distribution of antiviral antibodies to three herpes viruses was studied in a population of healthy blood donors. Subclassification by monoclonal antibodies led to the identification of certain viral IgG patterns. IgG1 appeared to be formed in response to almost all CMV, HSV-1 and VZV infections.
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