Since their creation, leukocyte depletion filters, used to prepare leukocyte-poor red cell or platelet concentrates, have been continuously evoluting: their volume has been reduced and their composition has been modified, polyester fibers have replaced cotton wool or cellulose acetate. In the same way, their capacity to remove leukocytes has increased. The mechanisms of cell separation are still unclear. Through published studies, different factors seem to be implicated, such as wetable surface of the filter, surface tension or charge density of fibers. It was also observed that removal of lymphocytes from red cell concentrates is rather based on trapping in the fiber network, and that removal of granulocytes, monocytes and platelets is partly due to activation and subsequent adhesion to the fibers. There have been few studies of the mechanism of leukocytes depletion from platelets concentrates.
A l'Hôpital Régional de Saint-Louis, le risque transfusionnel et les fréquentes pénuries de sang nous ont conduits à mettre en pratique la TAD malgré les diverses difficultés rencontrées. Notre étude porte sur la recherche d'une technique la plus adaptée au contexte d'un hôpital périphérique en Afrique et sur son utilisation pendant quatre mois (d'avril à juillet 92). Nous exposons nos premiers résultats qui nous incitent à persévérer dans cette voie.
In France, HIV seroprevalence in blood donations still decreased from 1990 to 1992 with an important diminution in 1992, in first time donors as well as in regular donors. During the last 1992 semester, this prevalence reached 0.051 parts per thousand : 0.19 parts per thousand and 0.026 parts per thousand in donations from first time and regular donors respectively. The residual risk was estimated to be 1 for 580 000 donations in 1992 (CI 95 % = 1/2 000 000 to 1/330 000).Sex ratio was 3.7 and 3.4 in 1990 and 1991 and increased to 5.5 in 1992 due to a very small number of seropositive women, which seems to be an important epidemiological indicator. The major at-risk group is still represented by homo-bisexual men.
Les résultats de 35 882 recherches d'anticorps irréguliers érythrocytaires réalisées par la méthode du gel-test (gels et hématies DiaMed) ont été analysés. Les échantillons provenaient de différents services hospitaliers et cliniques.
The bulky production of recombinant proteins can be achieved by procaryotes or eucaryotes cells. Cells from higher eucaryotes may be required when proteins have to be modified post-transcriptionally (glycosylation phosphorylation, cleavage, folding...). Cells from higher vertebrates in culture are used to prepare proteins like human factor VIII and erythropoietin. The use of transgenic organism has been suggested to reach the same goal. Indeed a whole living organism allows a very potent amplification, the number of cells involved in the biosynthesis of the recombinant proteins being very numerous and in the best metabolic conditions. Biological fluids (blood, milk, insect hemolymph, egg white...) and possibly organs from transgenic animals are a priori the best sources of recombinant proteins. Blood is abundant and it is a by-product of slaughter house. Its composition is relatively complex and the circulating recombinant proteins may heavily alter health of animals. Milk is very abundant, its composition is relatively simple, it is poor in proteolytic enzymes and it can be collected easily. Hemolymph from insects is relatively scarce. Egg white will be a possible source of recombinant proteins, when transgenesis has become more accessible in birds. Organs from transgenic animals should be solicited only when a particular cell type is required for the biosynthesis of the recombinant proteins. Milk appears therefore, presently, as the best source of recombinant proteins from transgenic animals. About 15 public and private laboratories try to use these techniques. They consist in preparing vectors containing regulatory regions of one of the milk proteins genes and the coding part (cDNA or gene) of the corresponding proteins to be produced. The transfer of these gene constructs to mouse, rabbit, sheep, goat, pig, shows that these techniques are indeed very promising. A single protein, human alpha 1-antitrypsin produced in milk of transgenic sheep, has presently reached the preparation at an industrial scale. This method has two theoretical limitations: 1) some of the proteins secreted in milk may be not matured as their native counterparts. Experiments carried out so far (about 20 proteins has been produced at an experimental scale) indicate that the mammary cell is able to achieve glycosylation in a correct way; 2) a significant proportion of the recombinant proteins migrate from the alveolar compartment of the mammary gland to blood circulation and they can alter health of lactating animals.(ABSTRACT TRUNCATED AT 400 WORDS)
Murine monoclonal antibodies (MoAbs) directed against DAF (Decay Accelerating Factor, CD55 antigen) and MIRL (Membrane Inhibitor of Reactive Lysis, CD59 antigen) were used to identify the affected red cells (CD55-ICD59-) of PNH patients. MoAbs NaM16-4D3 (CD55, IgG2a) and NaM77-1E5 (CD59, IgG3) weakly agglutinate red cells and represent powerful tools to quantitate normal (PNHI) and abnormal (PNHII and PNHIII) cells from PNH patients by indirect flow cytometry. MoAbs NaM125-7H10 (CD55) and NaM123-6Gl2 (CD59), both IgM, were selected for their agglutinating properties and used for the separation of PNHI from PNHII and PNHIII red cells by the gel test technology. From analysis of artificial mixtures of DAF+ and DAF- cells, a direct relationship was established between fluorescent cells detected by flow cytometry, and erythrocytes agglutinated in microtyping cards. The method was further confirmed by analysis of ten blood samples from PHN patients and represent an alternative to classical hemolysis tests. On the basis of our experience we propose the following for the diagnosis of PNH: 1) agglutination test with NaCl microtyping cards using IgM CD55 and CD59; 2) flow cytometry analysis for accurate quantitation of CD55-/CD59- red cells.
Leukodepletion of red blood cell or platelet concentrates decreases significantly the number of febrile nonhemolytic transfusion reactions. Filtration appears to be very efficient on red blood cell transfusion reactions while its effect with platelet concentrates remains variable.
Evolution of HI v infection was studied in 480 hemophiliacs A and 78 hemophiliacs B treated in the << Centre-West >> a Region. 23,3 % hemophiliacs A and 46, 1 % hemophiliacs B were contaminated by HIV In this region, HIV seropvevalence in hemophiliacs A was lower than the prevalence noted at the national level (51,2 %); this is certainly due to the use of frozen cryoprecipitates in the treatment of a high number of hemophiliacs A.A higher number of hemophiliacs B developed the disease : 12,5 % hemophiliacs A versus 22 % hemophiliacs B.Moreover hemophilic B patients had a more vapid evolution towards the disease since 6 out of 14 hemophiliacs and 7 out of 8 hemophiliacs B with AIDS died.The fact that hemophiliacs B were significantly older than hemophiliacs A might be one of the reasons, but it must be noted that the contamination often occurred earlier in hemophiliacs B and was perhaps move important.The more severe evolution in the hemophiliac B group noted in our region is not found in American studies, which may be due to the different ways of preparing Factor IX concentrates in France and the United States.
A survey carried out on 4129 Tunisian blood donors permitted to evaluate the distribution of Rhesus blood group in Tunisia.The haplotypes frequency were as follow, R1: 0,367 r: 0,284 R0: 0,2082 R2: 0,1218 R': 0,00188 r'': 0,0001
The efficiency of strored platelet transfusion was evaluated in terms of clinical status in 136 thrombocytopenic patients. ln a paired prospective study in which fresh platelets were used as controls, clinical efficiency was assessed on the basis of the ability to increase platelet count (recovery) and the interval to the next transfusion (D). In 48 clinically stable patients, recovery of fresh and stored platelets was similar (47 % and 41 % respectively) and the interval to the next transfusion was D4 and D3. In contrast, 27 patients who had bacterial infections showed significantly different recoveries (24 %/ 5 %) and the interval to the next transfusion was D3/D1 for fresh and stored platelets respectively. Similarly, in 16 patients who were treated concurrently with Amphotericin B, 18 other patients with graft-versus-host disease, 5 with splenomegaly and 3 with veno-occlusive disease (VOD), fresh platelets performed better than stored platelets, showing recoveries of 27 %/ 18 %, 29 %/ 15 %, 15 %/ 1 %, 22 %/ 3 %. Furthermore, the need for retransfusion within 24 hours was significantly increased with stored platelets. In 19 patients with anti-HLA allo-immunization who were transfused with HLA-matched fresh and stored APC, efficiency was similar (38 %/ 36 % and D4/D3). This study indicates that the storage has a major detrimental effect on platelet recovery and survival in patients with certain clinical conditions.
L'allo-immunisation, chez les drépanocytaires homozygotes peut conduire à des situations d'impasse transfusionnelle, posant des difficultés thérapeutiques majeures. Nous présentons le cas d'une jeune femme drépanocytaire homozygote, porteuse d'ulcères de jambe invalidants, chez laquelle un programme transfusionnel régulier a été débuté, afin d'envisager des greffes cutanées. L'apparition d'une immunisation multiple, posant le problème de la sélection d'unités compatibles, a conduit à l'arrêt des transfusions.
Post-transfusion hepatitis C incidence was studied in a series of patients with bone marrow allograft. The risk of HCV seroconversion was evaluated according to the date of grafting and the screening tests carried out in blood donors at this time. Anti-HCV antibodies weve screened using Elisa tests of 2d generation and confirmed by Riba tests of 2d generation. Results were analysed. Out of 181 allografted patients from January 1987 to December 1991, 120 patients found anti-HCV negative prior to grafting, with at least six month post-transfusion follow-up were considered as evaluable in terms of HCV seroconversion.All these patients had received leucodepleted blood products and the most of them platelet unit concentrates.Prior to implementation of screening tests for non-A, non-B hepatitis, 14 % of patients had seroconverted (0,44 % per transfused product); after introduction of the screening for indirect markers (ALAT) and for antibodies directed against the antigen of hepatitis B virus core (anti-HBc), the seroconversion incidence was 4 % (0,26 % per product). At the present time, since the implementation of anti-HCV screening tests, the risk has reached 1,6 % (0,03 % per transfused product). 6 patients out of 7 having seroconverted have been developing chronic hepatitis.
The technique of filtration which has constantly been improved over past years makes it possible to obtain highly pure blood products (the rate of leukocyte depletion can reach 3 to 5 log). Several study groups (BEST, PSL) and international commitees of experts have defined a number of standards with which leukocyte depleted blood products must comply. Assessment of filration procedures is thus made necessary, it applies to the different steps of the filtration procedure: preparation techniques of red cell or platelet suspension, priming and rinsing of the filter. Likewise a number of parameters have to be checked: filtration time, temperature, age of suspensions. Moreover quality control procedures must be implemented, using adapted and validated measuring methods (Nageotte hemacytometer, for instance). The parameters to be monitored include the number of residual leukocytes, (mean value : 1 x 10(6) i.e. about 4 WBCs/mul), and the rate of hemoglobin or platelet recovery. Any new filtering equipment or material must satisfy strict requirements and standards including clinically acceptable limits as part of the same quality approach. Validation, in this case, consists in determining the maximal leukocyte content which the filter can absorb (capacity) and the average rate of leukocyte removal (efficacy).The constant monitoring of filtration performance contributes to improving the quality of red blood cell or platelet suspensions a nd thus meet clinicians' requirements for their patients.
Polymerase chain reaction (PCR) was applied to detect HCV-RNA in 75 hemodialyzed patients. Anti-HCV status was determined by ELISA-2 and by RIBA-2 for reactive samples by ELISA. ALT levels were monthly determined during the year preceding the end of the study. For 60 patients, anti-HCV serology was known since 1989 and 39 of them were tested for the presence of HCV-RNA at least four times during the 2 preceding years. The 9 patients who were negative for anti-HCV antibodies were negative by PCR. Of the 7 patients with an indeterminate profile by RIBA-2, 3 were positive by PCR : 1/1 with C-33c band only and 2/6 with C22-3 band only. Of the 59 patients reactive by RIBA-2, 57 were HCVRNA positive. Of the 2 HCV-RNA negative patients, one had been PCR positive before interferon therapy. Of the 38 patients without acute hepatitis tested by PCR on 5 successive samples, all the specimens of 11 and 23 patients were HCV-RNA negative and HCV-RNA positive respectively. In 4 patients, a transient viremia was observed. The group of HCV-RNA positive patients had mean ALT levels greater than those who were negative. A correlation was established between HCV infection and both the time on dialysis and the number of blood transfusions. A high concordance (97 %) was observed between antibodies to HCV and HCV-RNA.
Immunological ReviewsVolume 130, Issue 1 p. 41-68 Building Antibodies from their Genes Hennie R. Hoogenboom, Hennie R. Hoogenboom MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this authorJames D. Marks, James D. Marks MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this authorAndrew D. G Riffiths, Andrew D. G Riffiths MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this authorGreg Winter, Greg Winter MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK. MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this author Hennie R. Hoogenboom, Hennie R. Hoogenboom MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this authorJames D. Marks, James D. Marks MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this authorAndrew D. G Riffiths, Andrew D. G Riffiths MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this authorGreg Winter, Greg Winter MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK. MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.Search for more papers by this author First published: December 1992 https://doi.org/10.1111/j.1600-065X.1992.tb01520.xCitations: 104AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. 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Forty-nine human anti-D (Rho) monoclonal antibodies of the IgG and IgM classes were tested with red blood cells of various nonhuman primates, from anthropoid apes to Prosimians, and significant differences in reactivity were observed among antibodies of two classes depending on taxonomic position of primate animals. By and large, higher percentage of IgM mAbs gave positive reactions with nonhuman primate red cells and, particularly, with blood of lower monkeys : Old and New Worlds monkeys and Prosimians, than did those of IgG class. Allotypic reactions with red cells of African apes were produced by majority of IgG mAbs but by very few IgM reagents. Some of those reactions defined epitopes related to human D, such as chimpanzee Rc and gorilla Dgor. By contrast, individual differences among Old World monkey species were revealed almost exclusively in tests using anti-D mAbs of IgM class. Some of the epitopes detected by these antibodies on the red cells of macaques are related to human D alloantigen, as confirmed by absorption experiments. Differences among mAbs evidenced in tests with nonhuman primate red cells reflect the complexity of the immune reactions to the human D antigen.
The bulky production of recombinant proteins can be achieved by procaryotes or eucaryotes cells. Cells from higher eucaryotes may be required when proteins have to be modified post-transcriptionnally (glycosylation phosphorylation, cleavage, folding...). Cells from higher vertebrates in culture are used to prepare proteins like human factor VIII and erythropoietin. The use of transgenic organism has been suggested to reach the same goal. Indeed a whole living organism allows a very potent amplification, the number of cells involved in the biosynthesis of the recombinant proteins being very numerous and in the best metabolic conditions.Biological fluids (blood, milk, insect hemolymph, egg white...) and possibly organs from transgenic animals are a priori the best sources of recombinant proteins. Blood is abundant and it is a by-product of slaughter house. Its composition is relatively complex and the circulating recombinant proteins may heavily alter health of animals. Milk is very abundant, its composition is relatively simple, it is poor in proteolytic enzymes and it can be collected easily. Hemolymph from insects is relatively scarce. Egg white will be a possible source of recombinant proteins, when transgenesis has become more accessible in birds. Organs from transgenic animals should be solicited only when a particular cell type is required for the biosynthesis of the recombinant proteins. Milk appears therefore, presently, as the best source of recombinant proteins from transgenic animals.About 15 public and private laboratories try to use these techniques. They consist in preparing vectors containing regulatory regions of one of the milk proteins genes and the coding part (cDNA or gene) of the corresponding proteins to be produced. The transfer of these gene constructs to mouse, rabbit, sheep, goat, pig, shows that these techniques are indeed very promising. A single protein, human alpha1-antitrypsin produced in milk of transgenic sheeps, has presently reached the preparation at an industrial scale. This method has two theoretical limitations: 1) some of the proteins secreted in milk may be not matured as their native counterparts. Experiments carried out so far (about 20 proteins has been produced at an experimental scale) indicate that the mammary cell is able to achieve glycosylation in a correct way; 2) a significant proportion of the recombinant proteins migrate from the alveolar compartment of the mammary gland to blood circulation and they can alter health of lactating animals. Moreover, this method has still technical limitations which will be likely alleviated in a near future : ill-controlled expression of the transgene which is often not restricted to the mammary gland, low expression with vectors containing cDNA rather than gene, relative difficulty to obtain large transgenic animals.According to the amount of recombinant proteins to be prepared, several animal species can be used. Transgenic mouse can provide 100 mg-1g of recombinant proteins (for structural and pharmaceutical studies) by collecting the milk which is spontaneously secreted from isolated mammary gland of lactating animals, incubated in cold. Transgenic rabbits must allow the preparation of 1 kg of recombinant proteins per year from their milk collected with a milking machine. For higher quantities of proteins, goat, sheep, pig and cow are of course the best candidates.If the feasibility of the method has no longer to be demonstrated, it remains uneasy to evaluate its real cost at an industrial scale. This is of course due to the fact that only a small number of projects have been led to term and that the technical details of the method is still permanently being improved. An analysis done by an american company predicts that, by the end of this century, 10% of the recombinant proteins for pharmaceutical use will come from milk of transgenic animals. This corresponds to an annual market of 25 millions dollars.
