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CONCANAVALIN A AND INFLUENZA VIRUS INFECTED CELLS 227 Interactions of Concanavalin A with the Membrane of Influenza Virus Infected Cells and with Envelope Components of the Virus Particle R . R O T T , H . B E C H T , H . - D . KLENK, a n d C . SCHOLTISSEK Institut für Virologie, Justus Liebig-Universität, Gießen ( Z . Naturforsch. 27 b, 227—233 [1972] ; received December 12, 1971) Dedicated, to Prof. WERNER SCHÄFER on the occasion of his 60th birthday Treatment of host cells with Concanavalin A prevents the assembly or release of fowl plague virus from chick fibroblasts and inhibits fusion of BHK cells by the paramyxovirus SV5. Metabolic events of the host cells are not greatly impaired during the early phase of infection. There is evi dence that the carbohydrate moiety of the Con A receptor of fowl plague virus is associated with the viral neuraminidase. Agglutination by Concanavalin A (Con A ) is one Materials and Methods of the manifestations of the alterations which the membranes of the host cells undergo after they had Virus, tissue cultures and inhibition assay been infected with enveloped viruses. This pheno- Fowl plague virus as a representative of influenza viruses was grown in the allantoic cavity of 11-day chick embryos and used for the infection of chick embryo fibroblasts. After an adsorption period of 30 min. the cultures were washed and 2 ml of Eagle's Minimal Essential Medium containing 200 //g of Concanavalin A (Con A, purchased from Sigma) were added. At hourly intervals post infection two dishes were removed from the incubator, medium and cells were harvested and assayed for hemagglutinin, neuraminidase, RNP-antigen and plaque forming units according to standard procedures. Special treatments are described in results or in the legends of the respective tables. menon has been observed with a large number of RNA viruses and vaccinia virus in various cell systems, and it could never be observed with nonenveloped viruses 1-4 . Since parts of the cell mem- branes altered during the budding process f o r m the matrix of the viral envelopes, it was not surprising to find suspensions of completed virus particles to be agglutinable by Con A 2 ' 5 . This means that the specific receptors for the phytagglutinin are exposed at the surface of the infected cell and appear in the viral envelope. Agglutinability of fowl plague infected fibroblasts occurs very early during the re- plication cycle, even before hemagglutinin or infectious virus is released into the medium 1 . Exposure of Concanavalin A receptors seems to be among the earliest changes demonstrable in an infected cell. This observation prompted us to study the influence of Con A on viral replication, particularly on the synthesis of viral components and on viral release. Since it had been shown that removal of the outer layer of the viral envelope by bromelin left a spikeless particle which was not agglutinable by Con A any longer 5 , we tried in a further series of experiments to determine more precisely whether the specific receptor determinant is associated with hemagglutinin or neuraminidase, the components re- moved by the bromelin treatment. Requests for reprints should be sent to Dr. R. ROTT, Institut für Virologie d. Justus-Liebig-Univ., D-6300 Gießen, Frankfurter Str. 87. Agglutination of infected cells by Con A and inhibition by viral components Chide embryo fibroblasts infected with fowl plague virus were washed 6 hours post infection, and the cell layer was dispersed with EDTA as described before 2 . 0.1 ml of isolated hemagglutinin or neuraminidase in at twofold dilution series was mixed with 0.1 ml of 40 ^g/ml of Con A, corresponding to approximately 4 minimal cell agglutinating units. After 10 min at room temperature 0.2 ml of the cell suspension were added, the reaction was continuously observed and the final result scored after 10 minutes. Preparation of hemagglutinin and neuraminidase Hemagglutinin was obtained from virus concentrated from allantoic fluid. The preparation was split with Tween 80 (2.5mg/ml) and ether, and hemagglutinin was isolated by adsorption to erythrocytes at 4 °C and elution at 37 °C. The eluate was concentrated by pressure dialysis. Hemagglutinin and neuraminidase which were maximally segregated were prepared from the classical "viromicrosomes" 9 of infected chorioallantoic membranes. After the allantoic fluid had been harvested the membranes were removed from the eggs, washed in Unauthenticated Download Date | 8/11/17 10:15 PM R. ROTT, H. BECHT, H.-D. KLENK, AND C. SCHOLTISSEK 228 PBS, homogenized as a 20% suspension, centrifuged briefly to remove crude debris, and the "viromicrosomes" were finally spun down as described. The pellet was resuspended in 0.02 M Tris/HCl buffer, pH 7.8, containing 0.5 M KCl, homogenized with 0.5% of Triton X-100 for about 2 min and treated with V 2 Vol. of peroxide-free ethyl ether. Treatment of the suspension with the detergent and ether was done in an ice bath. The aqueous phase was removed after centrifugation at 4000 rpm for 20 min and residual ether was blown off with a stream of nitrogen. After a final centrifugation step at pi 25, the supernatant was used for the precipitation test with Con A. Neuraminidase was prepared by destroying the other virus components and most of the cellular proteins with pronase 10. Trials to precipitate hemagglutinin or neuraminidase with Con A 5 0 ( « g / m l of Con A had been added. This finding was not surprising, since it had been Experiments on cell fusion Inhibition of cell fusion was studied with the W3 strain of the parainfluenza virus SV5 grown in the MDBK line of bovine kidney cells as described by CHOPPIN 6 . Influence of Con A on RNA and protein synthesis ( 3 H) Incorporation of uridine (30 C/mM) or ( 3 H) leucine (1 C/mM) into chick fibroplasts in Petri dishes of 9 cm in diameter was measured according to SCHOLTISSEK u . Virus-specific RNA was distinguished from newly synthesized cellular RNA by specific hybridization with a surplus of non-labelled viral plus- and minus-strand R N A 1 2 . The activity of influenza RNAdependent RNA polymerase was measured in a cytoplasmic extract of infected cells 5 hours p.i. by incorporation of ( 3 H)GTP into virus minus-strand RNA in the presence of cofactors as described previously 1S. be amounts detected of in hemagglutinating the culture 128 128 64 2 <2 <2 Removal of Con A [hrs. p . i . ] H A units in m e d i u m Cell associated H A units PFU/ml 1 2 3 4 5 6 without Con A < 2 < 2 < 2 < 2 < 2 < 2 128 12 6 3 < 2 < 2 < 2 1000 5-105 2-105 1.5 • 10 5 1 • 10 5 1 • 105 7.5 • 10 5 1.2-107 1* 2* 3* 4* < < < < < < 4 8 4 2 2 2 1 • 105 2-105 2.7 • 10 5 3.5 • lO 5 7.5 • 10 4 5 - 10 4 5* 6* 2 2 2 2 2 2 < < < Table 2. Replication of FPV in the presence of Concanavalin A for different periods. Con A containing medium (50 jug/m\) was removed at the time after infection indicated and incubation was continued with normal medium until 8 hours p. i. * = Incubation with normal medium was continued for 6 hours. known f r o m previous experiments that virus particles are readily agglutinated by Con A . Absence of viral activities in the culture medium could have been due, therefore, to aggregate formation of released virus b y Con A . On the other hand, binding of Con A b y the altered surface of the infected cells, which becomes manifest by the agglutinability of such cells, could mediate a block in viral release. A series of experiments was set up to decide between these alternatives. Table 2 presents the results of exreplaced The dose response curve (Table 1) shows that no could 0 0.1 1 10 50 100 periments where the Con A-containing medium w as Results significant HA-units Table 1. Dose response of FPV release in the presence of Concanavalin A . The Tween/ether split-product from virions or the hemagglutinin/neuraminidase mixture prepared from "viromicrosomes" were mixed with equal amounts of Con A (100 jug/ml) at room temperature and centrifuged after 30 min at pi 5. The supernatant was assayed for residual hemagglutinating and neuraminidase activities. The pellet was assayed after ultrasonic dispersion. Monolayers of baby hamster kidney (BHK 21-F) cells 7 were propagated in reinforced Eagle's Medium 8 with 10% calf serum and 10% tryptose phosphate broth. Monolayers in plastic Petri dishes (diameter 6 cm) were inoculated with SV5 at a multiplicity of 10 PFU/ cell. After an adsorption period of 90 min 2 ml reinforced Eagle's Medium without calf serum and tryptose phosphate broth were added. A m o u n t o f Con A in culture m e d i u m Ug/ml] medium activity when by normal culture medium at different periods after infection. These data show that completion of the 8 hours replication cycle of FPV under the usual conditions or a continued incuba- Unauthenticated Download Date | 8/11/17 10:15 PM CONCANAVALIN A AND INFLUENZA VIRUS INFECTED CELLS 229 tion of another 6 hours after removal of Con A did not liberate any virus into the medium. than in controls, when the cell homogenates were Even treatment of the monolayer f o r 2 hours with 1 0 0 / / g / m l of Con A before the cultures were inoculated with infectious allantoic fluid prevented the appearance of any virus-specific material in the medium after 8 hours of incubation (Table 3 ) . and ether (equal volumes). Con A added Cell associated PFU/ml H A units + +* 512 <2 64 5-107 1.4 • 10 4 7 • 10 5 Neuraminidase units 5400 80 1360 HA-units in m e d i u m 256 <2 <2 Table 3. Binding of Concanavalin A to uninfected cells. Con A (50 jMg/ml) was added 2 hours prior to infection, the cells were washed twice, infected and incubated as usual. Cells were processed by freezing and thawing ( + , —) or by ultrasonication ( + * ) . Agglutinating titer o f Con A solution Con A nonabsorbed absorbed sonicated or treated with Tween 8 0 without 20 16 20 20 20 20 20 3-104 5-104 1.2 • 105 1.5 • 10 5 1 • 10 5 7.5 • 10 4 7.5 • 10 4 400 530 580 620 520 580 560 96 96 96 96 96 96 96 256 3.5 • 10 7 3600 96 Con A added 1/16 1/2 Firm binding of Con A by normal cells could also be demonstrated in absorption experiments. The reduced agglutination titers f o r FPV-infected cells in Table 4 show clearly that a considerable amount of Con A was absorbed by uninfected chick fibroblasts during the 2 hours incubation. + + + + + + + + The unexpected result of the experiments presented in Table 2 and 3 was the lack of any demonstrable hemagglutinin after the cells had been broken b y the commonly used three cycles of freezing and thawing. This finding, based solely on the conventional technique of processing infected cells, would have indicated that not only release of virus particles but also synthesis of viral components was RNPantigen units Table 5. Aggregation of cell associated activity by Concanavalin A . 50 jug/ml Con A was added at the time indicated. Cells were harvested 8 hours after infection and assayed after ultrasonication for 15". H o u r s after infection Table 4. Absorption of Concanavalin A by uninfected cells. Approximately 10 7 chick fibroblasts were suspended in 1 ml of PBS containing 100 /ig of Con A. After 2 hours at room temperature the cells were spun down and the agglutinating activity of the supernatant tested with FPV-infected cells. Neuraminidase units [fig N A N A / ml] Addition Cell associated o f Con A [hours p. i.] H A PFU/ml units 0 1 2 3 4 5 6 (12.5mg/ml) Cellassociated H A units Hemadsorption <2 < 2 < 2 < 2 48 < 2 420 < 2 512 < 2 750 < 2 512 < 2 750 4 Table 6. Hemadsorption by chick fibroblasts infected with fowl plague virus after treatment with Concanavalin A . 100 [Ag of Con A were added to the test plates immediately after the adsorption period. The monolayers were washed twice with PBS at hourly intervals and 2.5 ml of a 1% suspension of chick erythrocytes were added. After 15 min at room temperature the plates were washed again five times and the degree of hemadsorption (H + + + + ) was scored microscopically. blocked. However, presence of hemagglutinin and of could Aggregation of virus products and their firm at- fluorescence tachment to cell debris became obvious when Con A RNP-antigen in cells incubated with Con A be clearly demonstrated b y a brilliant in ultraviolet light after staining of these cells with was added at different periods post infection after fluorescein-conjugated virus synthesis had proceeded normally (Table 5 ) . anti-hemagglutinin and anti- RNP-serum. Hemagglutinin could be regained f r o m Hemadsorption Con A-treated cells, although in smaller quantities treated with Con A furnished further evidence that to infected cells which were pre- Unauthenticated Download Date | 8/11/17 10:15 PM R. ROTT, H. BECHT, H.-D. KLENK, AND C. SCHOLTISSEK 230 virus-specific products had been synthesized and introduced into the cell membrane (Table 6 ) . Microscopic examination of Con A-treated InCon fected A After 2 4 hours the Con A-treated cell layer was still adherent to the Petri dish. All cells had a granulated appearance and many cells had rounded and contracted. The monolayers of infected control culincubated without Con A were totally stroyed. However, the cells in the still Con A-treated monolayers were all de- adherent stained with trypan blue, indicating severe cell damage b y the infection. Uninfected cultures incubated in the presence of Con A had only few rounded and stained cells. Inhibition of virus-induced Concanavalin cell fusion Disintegrations per min • l O - 2 (3H) ( 3 H ) uridine TCAleucine RNA extract protein 0 0 6 6 6 6 15 15 15 15 23 23 23 23 780 860 270 570 555 1120 98 288 92 125 68 177 * 6150 6565 1720 4200 2230 3950 540 2360 668 1140 310 1150 83 533 mono- layers tures Start o f pulse hrs. p . i . by A Table 7 shows that the parainfluenza virus SV5 + — — — + — + + — + — + — + + — + + + — — — + + 1110 1150 282 1240 305 515 121 715 45 224 104 537 * * 212 Table 8. Effect of Con A on R N A - and protein metabolism of uninfected and infected tissue cultures. 100 y/ml Con A was added to 50% of the cultures immediately after infection. ( 3 H) uridine or ( 3 H) leucine (1.25/^C/culture) were added at different times after infection. The pulse length was 1 hr. Noninfected control cultures were treated and worked up in prallel. * = Monolayer completely destroyed. causes in BHK 21-F cells extensive cell fusion. Giant cell formation began about 10 hours after infection plication cycle. RNA-synthesis itself seems not to be and rapidly progressed to the formation by 2 4 hours greatly influenced since the radioactivity in the tri- of a single syncytium 7 . Con A had an inhibitory chloroacetic effect on cell fusion. Giant cell formation was com- derivatives) parallels the radioactivity incorporated acid-soluble pool (uridine phosphate into R N A . Concanavalin A added amount [hrs. p . i . ] [ / / g / d i s h ] 200 1.5 3 50 200 6 50 200 Control 0 8 10 11 14 Furthermore, the effect of Con A on the product- 24 [hrs. p . i . ] ion la 1 1 1 1 1 1 5 1 >100 -10 > 1 0 0 1 complete fusion pletely suppressed, if 2 0 0 jug Con A per dish were added 1.5, 3 or 6 hours after infection. At a concentration of 5 0 jug per dish there is still a significant delay in giant cell formation. of Con A on cell metabolism viral RNA influenza R N A polymerase was examined of influenza R N A polymerase in the presence of 10 Table 7. Inhibition of cell-fusion by Concanavalin A . a Nuclei per cell. The number of nuclei per cell is used as a parameter for cell fusion induced by SV5. Eßect of 5 hours after infection. As shown in Fig. 1 the yield Con A is still about 6 0 % of that without lectin. Since infected cells were still adherent to the Petri dish 15 hours after infection, the synthesis of viral R N A in these cells was examined. By specific hybridization with non-labelled viral plus- and minusstrand R N A it was found, that 15 hours after infection in Con A-treated cells less than \% of the newly synthesized R N A (pulse with ( 3 H ) uridine f r o m 15 to 16 hours p.i.) was virus-specific. At 4 hours p.i. virus-specific R N A is about 15% of the newly synthesized R N A Inhibition and on influenza synthesis viral 12. of cell agglutination by components of the envelope Hemagglutinin prepared by desintegrating FP\ As shown in Table 8 Con A has a stimulating ef- viria with Twreen 8 0 and ether inhibited agglutina- fect on the uptake of ( 3 H ) uridine and ( 3 H ) leucine tion of FPV-infected fibroblasts by Con A. Aggluti- into uninfected as well as into infected cells, even nation gradually reappeared upon serial dilution of 6 hours post infection, a very late stage of the re- the hemagglutinin (Table 9 ) . Unauthenticated Download Date | 8/11/17 10:15 PM CONCANAVALIN A AND INFLUENZA VIRUS INFECTED CELLS 74 231 Since it is known that this "hemagglutinin" contains at least two virus-specific components, the true hemagglutinating substance and viral neuraminidase, we tried to decide whether the Con A-specific carbohydrate is attached to one of these structures or to both. Since hemagglutinin is not available yet in pure f o r m and the purification procedure f o r isolated neuraminidase involves treatment with pronase, another approach was chosen f o r this question. ?o 5 Precipitation of neuraminidase by Con A Con A was added to a mixture of hemagglutinin/ neuraminidase solubilized f r o m viromicrosomes with 0.5 M KCl and 0.5% Triton X - 1 0 0 . The phytagglutinin precipitated almost exclusively neuraminidase. After centrifugation the supernatant was nearly de0 5 10 . min 15 prived of enzymatic activity, but it had retained vir- Fig. 1. Effect of Con A on the production of influenza R N A polymerase. 5 infected tissue cultures received 100 y/ml Con A immediately after infection, 5 infected cultures were incubated without Con A . 5 hours p. i. a cytoplasmic fraction (3.1 p g / m l protein) was prepared and tested for R N A polymerase activity by incubation at 32 ° C with ( 3 H) G T P plus cofactors 13 . o — o = plus Con A ; • — • = without Con A . Dilution of hemagglutinin ( - l o g 2) Cell agglutination 1 2 3 4 5 6 7 PBS tually all of its hemagglutinating capacity (Table 1 0 ) . The same experiment with a hemagglutinin prepared with Tween and ether f r o m viria had a somewhat different outcome; hemagglutinating a substantial amount activity was removed from supernatant in this case. When the complex dissociated by readdition of a detergent NP-40, of the was (Nonidet 1 % ) , hardly any hemagglutinin was pre- cipitated. A still different result was obtained when neuraminidase purified by treatment of viria or of 0 0 0 + ++ ++ ++ + + CAM-extracts with pronase was used in the precipitation test. A considerable amount of enzyme was not precipitated by Con A f r o m these preparations. Table 9. Inhibition of Concanavalin A mediated agglutina tion of FPV-infected cells by hemagglutinin. 0.1 ml of Con A (40 « g / m l ) was mixed with 0.1 ml of diluted hemagglutinin (30 000 H A units isolated from purified F P V by Tween/ether treatment). After 10 min at room temperature 0.2 ml of E D T A dispersed cells were added and the agglutination pattern read 15 min later. T y p e o f viral c o m p o n e n t s In each experiment total hemagglutinating and enzymatic activities could not be fully recovered f r o m the pellets, because binding of Con A entrapped these components so firmly within the precipitates that they could hardly be dispersed. Con A a d d e d Sediment 1800 512 64 1400 1600 108 < 24 8 760 48 16 52 200 13 • < 12 8 256 700 11 130 440 37 — — — — — 13 42 156 92 + Tween-ether treated F P V + Neuraminidase purified b y pronase t r e a t m e n t Neuraminidase [ u g N A N A / m l ] Sediment Supernatant 32 8 T r i t o n treated V i r o m i c r o s o m e s in K C l T w e e n - e t h e r treated F P V in the presence o f N o n i d e t N P - 4 0 H A units Supernatant — + Table 10. Precipitation of envelope components of F P V by Concanavalin A . 1 mg/ml of Con A were mixed with an equal volume of the virus preparations, left at room temperature for 30 min and centrifuged at a performance index of 7.5. Unauthenticated Download Date | 8/11/17 10:15 PM R. ROTT, H. BECHT, H.-D. KLENK, AND C. SCHOLTISSEK 232 after treating the cell homogenate with Tween and Discussion The absence of any virus-specific activities in the medium of FPV-infected cultures containing Con A was not primarily due to precipitation and subsequent removal of liberated particles. The media lacked virus because a block of viral release must have occurred by binding of Con A to the cell membrane. Since virus production was also prevented by treating the cultures prior to infection, a firm binding of Con A at the cell membrane must have taken place. Attachment of the lectin to the membrane of normal cells could also be shown b y the decrease in agglutination titers f o r infected after normal fibroblasts cells had been suspended in a Con A solution. These findings are consistent with reports f r o m other laboratories where it could be shown that Con A is bound by normal as well as b y transformed cells14'15. All these results indicate that Con A receptors are accessible f o r the phytagglutinin at the surface of normal cells. The appropriate arrangement and exposure of the receptors at the surface of an altered membrane seems to be the essential condition f o r the agglutinability of transformed cells or of cells infected with an enveloped virus. After reaction of the receptors with Con A formation of a lattice structure could cause a considerable loss of the membrane's flexibility and could impair its normal functions which are probably necessary f o r a normal synthesis of the components of the viral envelope and f o r an undisturbed budding process. Rigidity of the cell membrane by such a network would explain the prolonged adhesion of the monolayer to the Petri dish simulating the absence of a fully developed CPE. The increased uptake of ( 3 H ) uridine implicates that the membrane becomes more permeable, which is probably responsible f o r the stimulation of U T P and of R N A labelling. Such an alteration of the cell membrane seems also to be responsible f o r the inhibitory effect of Con A on cell fusion which is a c o m m o n feature of paramyxoviruses and requires an intact viral envelope. It is conceivable that Con A impairs the formation of the envelope at the cell membrane b y a mechanism outlined above. Alternatively, it could be possible that the cell-fusing activity of the viral envelope is inhibited by the mere adsorption ether is due to a depressed synthesis of these components under the influence of Con A or merely a result of the imperfectness of this method of solubilization. The relatively low degree of hemadsorption in Con A-treated cultures would favour the first possibility. The viral envelope components which had been synthesized were firmly tied to the cell debris by Con A after freezing and thawing of the cells. This is consistent with It cannot be fully assessed whether the relatively low recovery of hemagglutinin and neuraminidase observation that outer layer of the viral envelope. When hemagglutinin and neuraminidase were dissociated and were individually in solution with a minimal degree of complexing, Con A precipitated neuraminidase. This underlines the suggestions derived f r o m acrylamide gel analysis that neuraminidase is a glycoprotein 16 and it means that the Con A receptor substance is mainly associated with neuraminidase and not with hemagglutinin. Digestion with pronase had obviously removed part neuraminidase of the carbohydrate from the molecule which indicates that the cleavage site of the protease is closely adjacent to the area where the carbohydrate moiety is attached. Our results suggest that during the assembly of the components of the viral envelope carbohydrates are selectively incorporated into the viral protein structures to f o r m the two major glycoproteins hemagglutinin and neuraminidase, each carrying dif- ferent carbohydrate moieties. A final consideration is pertinent to the fact that Con A is a bifunctional reagent which binds firmly to neuraminidase via the carbohydrate moiety of this glycoprotein and is capable of blocking viral release. Comparable conditions are met in experiments with anti-neuraminidase antibodies which inhibit viral release 1 7 . Since Con A, a bifunctional reagent which binds to neuraminidase and to receptors of the cell membrane without influencing the enzyme's active site, is capable of eliciting the same effect, the conclusion reached with monovalent antibodies is stressed that inhibition of virus release by anti-neuraminidase antibodies is rather due to lattice formation than to a block of the active site of the neuraminidase molecule of Con A to the cell membrane. our previous Con A receptor substances are associated with the 18. We gratefully acknowledge the excellent assistance of MICHAELA ORLICH and BRIGITTE PIONTEK. The work was supported by the Sonderforschungsbereich 47 (Virologie). Unauthenticated Download Date | 8/11/17 10:15 PM SERUM HEPATITIS AND SEROPROPHYLAXIS 1 2 3 4 5 6 7 8 9 H. BECHT, R . ROTT, and H.-D. KLENK, Z . med. M i k r o b i o l . u. Immunol. 1 5 6 , 3 0 5 [ 1 9 7 1 ] . H. BECHT, R . ROTT, and H.-D. KLENK, J. gen. V i r o l o g y , 14, 1 [ 1 9 7 2 ] . J. M . ZARLING and S. S. TEVETHIA, V i r o l o g y 4 5 , 313 [1971]. J. D . ORAM, D . C. ELLWOOD, G. APPLEYARD, and J. L . STANLEY, Nature [ L o n d o n ] 2 3 3 , 5 0 [ 1 9 7 1 ] . H.-D. KLENK, R . ROTT, and H. BECHT, V i r o l o g y , in press. P . W . CHOPPIN, V i r o l o g y 3 9 , 1 3 0 [ 1 9 6 9 ] . K . V . HOLMES and P . W . CHOPPIN, J. e x p . M e d i c i n e 1 2 4 , 501 [ 1 9 6 6 ] . R . BABLANIAN, H . J. EGGERS, and I. TAMM, V i r o l o g y 2 6 , 100 [ 1 9 6 5 ] . R . ROTT and W . SCHÄFER, Z . Naturforsch. 16, 3 1 0 [ 1 9 6 1 ] . 10 11 12 13 14 15 16 17 18 OF MEASLES 233 J. T . SETO, R . DRZENIEK, and R . ROTT, Biochim. b i o p h y s i c a A c t a [ A m s t e r d a m ] 113, 4 0 2 [ 1 9 6 6 ] . C. SCHOLTISSEK, Biochim. b i o p h y s i c a A c t a [ A m s t e r d a m ] 145,228 [1971]. C. SCHOLTISSEK and R . ROTT, V i r o l o g y 4 0 , 9 8 9 [ 1 9 7 0 ] . C. SCHOLTISSEK and R . ROTT, J. gen. V i r o l o g y 4 , 125 [1969]. G . L . NICOLSON, Nature [ L o n d o n ] 2 3 3 , 2 4 4 [ 1 9 7 1 ] . L . MALLUCCI, Nature [ L o n d o n ] 2 3 3 , 2 4 1 [ 1 9 7 1 ] . F o r rev. see H.-D. KLENK ( 1 9 7 2 ) . M o s b a c h e r C o l l o q u i u m , Springer-Verlag, Berlin, H e i d e l b e r g , N e w Y o r k . J. T . SETO and R . ROTT, V i r o l o g y 3 0 , 731 [ 1 9 6 6 ] . H . BECHT, U. HÄMMERLING, and R . ROTT, V i r o l o g y 4 6 , 3 3 7 [1971]. Serum Hepatitis and the Seroprophylaxis of Measles A . P. WATERSON Department of V i r o l o g y , R o y a l Postgraduate M e d i c a l School, D u C a n e R o a d , L o n d o n , W . 12., E n g l a n d (Z. Naturforsch. 27 b, 233—241 [1972] ; received January 20, 1972) Dedicated, to Prof. WERNER SCHÄFER on the occasion of his 60th birthday T h e events leading to the recognition of serum hepatitis ( " h o m o l o g o u s serum j a u n d i c e " ) as a disease entity are d e s c r i b e d and reasons given w h y s e r o p r o p h y l a x i s against measles played an important role in this discovery. T h e i n j e c t i o n of human serum as a p r o p h y l a c t i c was practised on an increasing scale f r o m about 1920 to a b o u t 1950, by which time it had largely been superseded by i m m u n o g l o b u l i n (gamma g l o b u l i n ) . A p a r t f r o m the transfusion of b l o o d and b l o o d products the other m a j o r human to human transfer of material was vaccination against yellow fever in the years 1937 — 1 9 4 0 . T h e c o n s e q u e n c e s of this large scale interchange between human s u b j e c t s included the dissemination of serum hepatitis virus, and the possibility of transfer of other viruses is discussed. Serum hepatitis has recently become amenable to laboratory study, in spite of the fact that the virus which causes it has not yet been isolated 2 . Much of the interest in the disease has centred around the patients and staff of renal dialysis units 3 and those involved in renal transplantation, but the treatment of chronic renal failure is only one of a number of situations involving operative treatment of the patient and the transfusion of large volumes of blood, either as repeated small transfusions or as a large quantity given at once. Transplant surgery and major cardiac survery are, like renal dialysis, procedures which twenty years ago were performed on relatively few patients, but have now become routine measures practised widely on relatively large numbers of patients, and they too have contributed to the increase in the quantities of blood and plasma transfused. There has also been an increasing range of blood products, such as anti-haemophilic globu- Requests for reprints should b e sent to Dr. A . P . WATERSON, Department of V i r o l o g y , R o y a l Postgraduate M e d i c a l School, D u C a n e R o a d , London, W. 12 ( E n g l a n d ) . lin, cryoprecipitate, concentrated platelets and radioiodine-labelled fibrinogen, some of which are available as preparations pooled from several donors. In addition, some of these situations involve either artificial suppression of the immune response, as after transplants, or a natural diminution in this response, as in the uraemic patients on long-term renal dialysis. There has also been a steady increase in the number of surgical operations, and in elaborate diagnostic procedures such as cardiac catheterization and various radiological investigations, all entailing at least some spillage of the patient's blood and its contact with doctors and perhaps other patients. The application of these techniques has of course entailed an enormous increase in laboratory investigations involving the handling of samples of blood and serum. This increase in the scale of laboratory investigation extends also to non-surgical patients, for example to those undergoing intensive therapy for leukaemia or chorion-carcinoma, such therapy involving the combination of blood transfusion and immuno-suppression. Unauthenticated Download Date | 8/11/17 10:15 PM