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Biomedical subjects

R Laub

Publications and source records attributed to R Laub.

At least 19 recordsLinked to original sources
PubMed2003

Prevalence of human erythrovirus B19 DNA in healthy Belgian blood donors and correlation with specific antibodies against structural and non-structural viral proteins.

BACKGROUND AND OBJECTIVES: Human parvovirus (erythrovirus) B19 is recognized as a major contaminant of blood and blood products. To reduce the risk of contamination, plasma-pool screening and exclusion of highly viraemic donations are recommended. The objectives of this study were to estimate the prevalence of B19 DNA in our blood-donor population, to determine the appropriate pool size to be tested (taking into account parameters such as prevalence, viral load, test sensitivity, and the efficacy of inactivation procedures), and to correlate viral loads with the serological status of donors as regards antibodies against different viral proteins. MATERIALS AND METHODS: Pools of different sizes were tested for B19, using a sensitive nested polymerase chain reaction (PCR) as well as an simple, un-nested, less sensitive PCR. Positive pools were resolved to the level of individual donations, and the viral load and serological markers were determined. RESULTS: Of 16,859 donations, 27 (one of 625) were found to be B19 DNA positive, with viral loads ranging from 10(2) to > 10(7) IU/ml. Twenty-five of the positive donations were tested for VP-specific anti-B19 antibodies, and eight (32%) were negative for both immunoglobulin (Ig)M and IgG. They were probably collected in the preseroconversion window period or from chronic carriers without detectable antibodies. We regarded the seven (28%) IgM-positive donors as being in the early phase of infection. The remaining 10 (40%) IgM-negative, IgG-positive donors were probably carriers of persistent infection (i.e. PCR positive despite the presence of IgG antibodies), as suggested by their low viral loads (< 10(4) IU/ml). Fifteen out of 36 major pools contained one or more contaminated donations. Among these, 12 tested positive by nested PCR and only three by un-nested PCR, this reflecting a viral load of > 10(4) IU/ml. CONCLUSIONS: By testing all donations as pools of 480 by un-nested PCR, and resolving positive pools to identify the responsible donations, it is possible to ensure that the viral load in fractionation pools (5000 donations) remains < 10(3) IU/ml, compatible with the efficacy of inactivation procedures and complying with Food and Drug Administration (FDA) recommendations.

Algorithms↗
PubMed2002

Polychlorinated biphenyls and organochlorine pesticides are eliminated from therapeutic Factor VIII and immunoglobulin concentrates and reduced in albumin by plasma fractionation.

BACKGROUND AND OBJECTIVES: Persistent organochlorine pollutants, such as polychlorinated biphenyls (PCBs) and organochlorine pesticides, are found in the general population and tend to accumulate in blood and tissues. Their distribution was examined in the starting plasma pools for fractionation, Cohn plasma fractions and therapeutic concentrates. MATERIALS AND METHODS: In each process fraction, total protein, cholesterol and triglycerides were measured as well as organochlorine pesticides and PCB congeners. RESULTS: Organochlorine compounds were undetectable in cryoprecipitate, Cohn fraction I, Factor VIII and immunoglobulin concentrates, and reduced in albumin preparations. CONCLUSION: Cohn plasma fractionation is very efficient for removing pollutants present in the starting material. Biological processing techniques should be analysed for their capacity to eliminate/reduce persistent organochlorine pollutants from the therapeutic proteins.

Adolescent↗
PubMed2000

A multiple transgenic mouse model with a partially humanized activation pathway for helper T cell responses.

Mice expressing human CD4 and human MHC II molecules provide a valuable model both for the investigation of the immunopathogenetic role of human autoantigens and for the development of therapeutic strategies based on modulating helper T cell activation in vivo. Here we present a novel mouse model expressing HLA-DR17 (a split antigen of HLA-DR3) together with human CD4 in the absence of murine cd4 (CD4/DR3 mice). Human CD4 accurately replaces murine cd4 within T cells. In particular, the preservation of cd8(+) and CD4(+) T cell subsets distinguishes CD4/DR3 mice from other multiple transgenic models in which the alternative T cell subsets are fundamentally disturbed. Moreover, human CD4 is also faithfully expressed on antigen presenting cells such as dendritic cells and monocyte/macrophages, so that the overall transgenic CD4 expression pattern resembles very closely that of humans. HLA-DR3 expression in the thymus correlates very closely to that of mouse MHC II. In contrast, only 70% of mouse MHC II positive cells in spleen, lymph node, and peripheral blood coexpress HLA-DR3. No significant bias was found with regard to particular leucocytes in this respect. The stimulation of helper T cells clearly depends on the interaction between the human transgene products, since mAbs to HLA-DR and/or CD4 completely blocked in vitro recall responses to tetanus toxoid. CD4/DR3 mice represent a partially humanized animal model which will facilitate studies of DR3-associated autoimmune responses and the in vivo determination of the therapeutic potential of mAbs to human CD4.

Animals↗
PubMed2000

PCR detects HCV RNA in a plasma pool contaminated by a single preseroconversion donation of genotype 5a.

BACKGROUND AND OBJECTIVES: To determine the prevalence of HCV-RNA-positive plasma pools in Belgium, to validate our PCR method and to increase the safety of the released blood products. MATERIALS AND METHODS: Plasma pools consisting each of about 5,000 donations from Belgian unpaid volunteer blood donors were analysed by PCR for the presence of HCV RNA. Two different extraction methods were compared and validated. RESULTS: Two out of 367 plasma pools were found to be HCV RNA positive and were discarded. For one of these two pools, the look-back procedure identified an anti-HCV-negative contaminated donation. The HCV genotype of both the contaminated pool and the donation was 5a, a genotype rare in Europe. The viral load of the preseroconverted donation was 2.9 x 10(7) gEq/ml according to the bDNA method. CONCLUSION: In the case of plasma derivatives, various important steps are already included to increase safety. Nucleic acid testing of manufacturing plasma pools ensures that viral load in the starting material is as low as possible.

Belgium↗
PubMed1999

Study of the protective effects of hyperimmune immunoglobulins G and M against endotoxin in mice and rats.

We prepared solutions of human IgM and IgG to various lipopolysaccharide (LPS) species. These were then tested, along with solutions of non-LPS specific human IgG or IgM, for their ability to confer passive immunity against experimental endotoxemia in two animal models. The immunoglobulins were first tested for an effect on the lethality induced by seven different LPSs in actinomycin-D sensitized mice, or by three different bacteria in normal mice. When the immunoglobulins were administered 1 h before challenge, a small protective effect was observed. This protection was dependent upon both the anti-LPS agent, the chemical composition of the LPS, or the strain of gram-negative bacteria used for injection. The anti-LPS IgM and IgG preparations reduced the mortality induced by Escherichia coli but not by Serratia marcescens or Klebsiella pneumoniae, indicating protection by strain-specific antibodies. When the antibodies were preincubated with LPS or bacteria for 30 min before administration, almost complete protection was seen. The influence of these immunoglobulin preparations or of human albumin (as a control) on the hypotensive and vascular-permeabilizing effects of LPS in rats was then studied. A dose-dependent inhibitory effect was observed with IgG preparations and albumin. At 200 mg/kg, anti-LPS IgG reduced the effects of LPS, while at 400 mg/kg, both anti-LPS and normal IgG preparations showed protection, as did human albumin used at the same dose. The IgM-enriched preparation worsened the initial hypotensive phase after LPS, whereas the anti-LPS IgM significantly reduced the second phase of the hypotension, but only at the largest dose of 400 mg/kg. In this second model using the rat, a clear difference between the activity of IgG and IgM was thus observed. We conclude that pretreatment with human immunoglobulins from large plasma pools modestly, but significantly, attenuated the effects of murine and rat Gram-negative sepsis, but that protection was incomplete. Our results suggest that single regimen intervention strategies may not be sufficient to influence the course of the disease.

Animals↗
PubMed1999

Influence of human anti-lipopolysaccharide immunoglobulins on tissue distribution and clearance of lipopolysaccharide in rats.

To examine the influence of passive immunization on the biological fate of injected lipopolysaccharide (LPS), we used a human IgG preparation (anti-LPS IgG) rich in antibodies to a large panel of smooth and rough purified LPS extracts as well as a normal IgG preparation (standard IgG). Our approach was to compare the uptake of 125I-labeled LPS by the tissues of saline or IgG-treated rats. After intravenous injection, one fraction of 125I-labeled Escherichia coli O55:B5 LPS is rapidly taken up by tissues, while another fraction remained in the blood. Uptake of 125I-labeled LPS was principally observed into the liver and spleen. In rats treated prophylactically with standard IgG, these tissues accumulated significantly larger amount of LPS than the tissues of rats treated with anti-LPS IgG. Nevertheless, both IgG preparations increased the specific binding of LPS by the liver and spleen. High levels of homologous unlabeled LPS decreased the uptake of LPS by the liver, presumably by occupying tissue receptors, whereas in the presence of E. coli O127:B8 LPS, an increase of the uptake of 125I-labeled LPS by the liver and lungs was observed. The pharmacokinetics and tissue distribution of LPS-IgG complexes pre-formed in vitro were compared. In the presence of standard IgG, a unexpected increase of the uptake of LPS by the tissues was recorded, whereas LPS-anti-LPS IgG complexes decreased the binding of 125I-labeled LPS to the tissues. On the other hand, the vascular effects induced by LPS did not appear to be modified in rats pretreated with either IgG preparation. In conclusion, although passive immunization against LPS slightly modified the uptake and clearance of LPS, neither in vitro nor in vivo formation of LPS-anti-LPS IgG complexes afforded a very significant protection against the toxic effects of LPS.

Animals↗
PubMed1999

Inhibitors in German hemophilia A patients treated with a double virus inactivated factor VIII concentrate bind to the C2 domain of FVIII light chain.

To reduce the risk of transmission of hepatitis A virus, an Octapharma produced factor VIII (fVIII) concentrate treated with solvent detergent (FVIII-SD) was further pasteurized after purification. This product, Octavi SDPlus (FVIII-SDP), was marketed in Europe in 1993 to 1995. Inhibitors appeared from September to October, 1995, in 12 of 109 previously treated German hemophilia A patients. A study of similarly treated Belgian patients, who also developed inhibitors, had shown antibodies to the fVIII light chain (domains A3-C1-C2) only. In the present study, the epitope specificity of 8 German inhibitor plasmas was also found to be restricted to the light chain. In radioimmunoprecipitation assays to localize the light chain epitope(s), antibody binding to heavy chain (domains A1-A2-B) was 11-148 fold lower than to the C2 domain, and binding to recombinant A3-C1 was barely detectable. These results were supported by >95% neutralization of a high responder inhibitor titer by the C2 domain.

Adolescent↗
PubMed1998

Modification of factor VIII in therapeutic concentrates after virus inactivation by solvent-detergent and pasteurisation.

The addition of a pasteurisation step to a solvent/detergent (SD) treated FVIII concentrate has recently resulted in enhanced inhibitor incidence in patients in Germany and Belgium. We have investigated the effect of virus inactivation procedures on FVIII function by preparing experimental concentrates from the same starting cryoprecipitate with the following procedures: none (N); dry heat (DH); pasteurisation (P); solvent/detergent (SD); solvent detergent + dry heat (SDDH); solvent detergent + pasteurisation (SDP). In addition, several clinical SD concentrates with and without pasteurisation were studied. There were no significant differences in fibrinogen and vWF content and in the ratio of one-stage/chromogenic FVIII activity among any of the samples studied. In thrombin proteolysis and FXa generation experiments, there were no differences in results on samples N, DH, P, and SDDH from those on sample SD. However sample SDP gave markedly different results from sample SD in the following respects: slower thrombin proteolysis (t(1/2) = 12.0 min vs 1.9 min); more rapid FXa generation (rate 2.5 times that of SD); enhanced phospholipid binding (K(D) = 3.89 x 10(-11) M vs 5.53 x 10(-10) M). Similar differences between SDP and SD were seen in the clinical samples. The observed changes in the FVIII activity occurred in combination with SD and pasteurisation, but not with either treatment alone. These results suggest that SDP treatment may enhance exposure of the phospholipid binding site in the C2 domain of FVIII, and since inhibitors to the SDP product are predominantly against C2, these findings could be relevant to the enhanced immunogenicity of the SDP product.

Detergents↗
PubMed1997

Heating lyophilised factor VIII does not alter its recognition by specific antibodies.

BACKGROUND AND OBJECTIVES: Alterations of factor VIII (FVIII) during preparation procedures can potentially affect its immunogenicity. One method evaluating such alterations could be by determining the reactivity of FVIII with specific antibodies. MATERIALS AND METHODS: Since heat treatment is currently used to reduce the risk of viral transmission, we evaluated the immunoreactivity of plasma-derived FVIII before and after heating at different temperatures and for different periods. Freeze-dried FVIII was used for these experiments as part of the validation procedure of a novel FVIII preparation. RESULTS: Heating FVIII for up to 72 h at 80 degrees C does not alter its reactivity with specific rabbit antibodies or mouse monoclonal antibodies, although some loss of FVIII activity occurred after 72 h. After heating for 2 h at 100 degrees C, a procedure that reduced FVIII activity by about 50%, there were still no significant effects on FVIII reactivity with monoclonal antibodies. CONCLUSIONS: Freeze-dried preparations of plasma-derived FVIII seem to be resistant to heat-induced structural denaturation.

Animals↗
PubMed1997

Factor VIII inhibitors in previously treated haemophilia A patients with a double virus-inactivated plasma derived factor VIII concentrate.

Antibodies to factor VIII (inhibitors) are usually produced at the beginning of treatment with factor VIII and are rare in multitransfused patients. Such antibodies are deemed to be patient-related, as supported by the description of a number of associated risk factors. However, a second category of inhibitors has recently been identified, namely antibodies occurring in multitransfused patients as a result of exposure to a particular factor VIII concentrate. A first outbreak of product-related inhibitors was recently described. The present paper describes the second well-documented occurrence of such inhibitors. Eight out of 140 multitransfused patients with severe haemophilia A developed an inhibitor to factor VIII shortly after changing treatment to a double-virus inactivated plasma-derived factor VIII concentrate. In addition to solvent-detergent treatment, this concentrate was pasteurised at 63 degrees C for 10 hours. Exposure to the pasteurised product before inhibitor detection ranged from 9 to 45 days. Inhibitor titers varied between 2.2 and 60 Bethesda Units and recovery of transfused factor VIII ranged from 0.21 to 0.68 (expressed as i.u./dl factor VIII rise per i.u./kg administered). In contrast to usual inhibitors in haemophilia A patients, these product-related inhibitors showed complex inhibition kinetics. They were found specific for the factor VIII light chain. The inhibitors gradually declined when exposure to the pasteurised product was stopped, despite further treatment with other factor VIII concentrates. The present data stress the importance of carefully monitored clinical studies, both in previously treated and previously untreated patients, before introduction of a new or modified clotting factor concentrate.

Adolescent↗
PubMed1996

Screening and characterization of specific anti-lipopolysaccharide antibodies in Belgian blood donors by enzyme-linked immunosorbent assays.

The goal of this project was to find and collect high concentrations of endotoxin-specific antibodies for therapeutic IgG- or IgM-enriched preparations. Various enzyme-linked immunosorbent assays (ELISAs) were developed to perform longitudinal studies of the serological response to a large panel of smooth and rough purified lipopolysaccharide (LPS) extracts in a population of healthy blood donors. To accomplish this, 1612 human serum samples from volunteer blood donors collected by seven different blood banks in Belgium were screened and specific IgM and IgG activities were measured. Approximately 17% of the donors had anti-LPS concentrations higher than 40 mg L-1. Of these, 10.9% had anti-smooth LPS antibodies, 3.7% had anti-rough LPS antibodies and 2.8% were found to be positive towards both types of LPS. The mean anti-LPS antibody concentration was 8 mg L-1 for rough LPS and 14 mg L-1 for smooth LPS. Age- and sex-related distributions of the activities indicated that the greatest prevalence of high anti-LPS concentration was in women aged 40-49 years and in men older than 60 years. Differential absorption experiments showed that the pooled serum of selected blood donors contained a mixture of specific and cross-reacting antibodies. We detected predominantly anti-LPS activities due to the IgG1 and IgG2 subclasses. The range of specificities to different LPS was increased by the pooling of selected sera. It was concluded that pools of naturally occurring specific anti-LPS immunoglobulin antibodies may be obtained in Belgium by screening blood donors using ELISAs that we have developed.

Adolescent↗
PubMed1991

[Practical experiences in the application of the BL-series of monoclonal antibodies for phenotyping of lymphocytes].

Determination of peripheral lymphocytes has increasing importance in laboratory examinations. However, evidence concerning differential diagnosis is low. For experimental questions and therapeutical monitoring application of this method is more distributed. This paper transfers practical experiences concerning the detection of cellular surface antigens. For the investigations, monoclonal antibodies from the Karl-Marx-University Leipzig were used in immunofluorescence microscopy. Limits of the test are demonstrated (in skin diseases and HIV-infection f.e.).

Antibodies, Monoclonal↗
PubMed1990

[Anti-epidermal cytotoxic reactions in the effector stage of lichen ruber planus].

Immunohistochemical investigations on skin lesions of lichen ruber planus indicate HLA-DR-expression by keratinocytes, Langerhans cells (LC), and infiltrated mononuclear cells. In the early stage of this condition, the lymphocytic infiltrate consists of a considerable part of (CD8+) cytotoxic/suppressor- and (LGL+) natural killer cells. Evidently, the cytotoxic potential of these lymphocyte subpopulations is directed against basal keratinocytes and epidermal LC. As a result of cytotoxic/cytolytic activities, non-cell associated CD 1-reactivity accumulates in the dermoepidermal junction zone, thus indicating damaged LC.

Antibodies, Monoclonal↗
PubMed1990

[Experiments for physiologic improvement of disinfectants].

The use of customary trade disinfectants lead to occupational disease no. 80 in many employers of public health. For that reason our aim is to improve the skin compatibility of this disinfectants. Its possible to achieve a pH-shift to a more physiological level eighter by buffer salt addition or by neutralization. Following this, disinfectants were investigated in a microbiological test.

Dermatitis, Contact↗
PubMed1990

[Mechanism of action and therapeutic use of cyclosporin A. Experimental and clinical experiences in dermatology].

The clinical application of cyclosporine A caused many success in transplantation surgery as well as in the treatment of (auto-) immune disorders. The biological highly active substance differs from conventional immunosuppressive drugs (i.e. steroids, azathioprine) by it's selective action on T-lymphocytes. However, current experimental findings indicate a direct influence on epidermal keratinocytes, too. First therapeutical experiences, concerning systemic and topical application in different skin diseases are summarized.

Animals↗
PubMed1990

[The effect of disinfectants on epidermal Langerhans cells--possibilities for the improvement of skin tolerance].

The aim of this paper is the characterization of the skin tolerance to several desinfectants. For that reason, guinea pig epidermis was treated for 1, 7 and 14 days with conventional working dilution of peracetic acid-containing and phosphoric acid-containing desinfectants. In addition, buffered desinfectants were applied. In all cases, the exposure to desinfectants causes a disappearance of histochemically detectable Langerhans cells in the treated epidermis. The lowest number of Langerhans cells was encountered after the application of Wofasteril. The buffering of the working dilution causes significantly lower (p = 0.01) damages in the Langerhans cell population. Possibilities for improving the skin tolerance of this desinfectant in normal use could result.

Animals↗
PubMed1990

[Changes in the functional surface markers of lymphocytes in short-term culture].

Cultured human lymphocytes are a suitable system for testing pharmacological, allergological, and toxicological effects of substances in vitro. For such investigations, the knowledge about the behaviour of the test system under standardized conditions is an important prerequisite. In the 72 h short-time-culture, no changes in T-lymphocytes were found. Small decreases occurred in the subpopulations of cytotoxical/suppressor cells, and inducer/helper cells, respectively. Number of cells expressing class II antigens of the main histocompatibility complex decreases, too (to 62%). Tritium thymidine incorporation rates inform about DNA-synthesis.

Antibodies, Monoclonal↗