bluuebunny/crossref_metadata_2025 huggingface dataset:
Localization of the third component of complement on the cell wall of encapsulated Staphylococcus aureus M: implications for the mechanism of resistance to phagocytosis
Authors: Wilkinson B J , Sisson S P , Kim Y , Peterson P K
Published: January 2020

Encapsulated Staphylococcus aureus strains are more virulent than unencapsulated staphylococci, and this phenomenon has been associated with decreased opsonization of encapsulated bacteria by normal human serum. Peptidoglycan, a major cell wall component of S. aureus, has been shown to promote opsonization of this bacterial species by certain components of the serum complement system. However, when the processes of complement activation and opsonization of encapsulated staphylococci have been studied, it has been found that encapsulated bacteria activate complement efficiently and C3 is bacteria associated, yet these organisms are not efficiently phagocytized by human polymorphonuclear leukocytes. In this study, the hypothesis was tested that opsonically active molecules are not on the true external surface of encapsulated organisms but rather are cell wall associated and thus "hidden" from human polymorphonuclear leukocytes. By using immunoelectronmicroscopy, C3 was found to be localized on the cell wall of an encapsulated S. aureus strain after incubation of the organism in normal human serum. When shrinkage of the capsule was prevented by treatment with anticapsular antibody, the C3 was again shown to be cell wall associated and located beneath the bacterial capsule. These results suggest that encapsulated S. aureus may resist phagocytosis because opsonically active C3 molecules are not exposed at the true external surface of the bacterium.
Antigens of Streptococcus mutans: cellular localization of the serotype-specific polysaccharide of strain AHT and release during exponential growth
Authors: Craig R A , Riege D H , Bleiweis A S
Published: January 2020

The serotype-specific polysaccharide of Streptococcus mutans AHT (serotype a) was shown to be loosely associated with the cell surface of this organism. The antigen was extracted from whole cells by boiling in sodium acetate buffer, pH 4.0, for 10 min. The purified product was found to be a diheteroglycan of galactose and glucose (3.6:1, molar ratio). The antigen possessed serological characteristics similar to the a antigen previously extracted from purified cell walls with hot formamide. Its physicochemical structure was identical to the previously studied wall antigen. Electron micrographs, developed after immunocytological labeling of this antigen on whole cells, revealed it to compose a dense microcapsule surrounding the microbe. Analyses of spent culture fluids indicated that the antigen was released during exponential growth at a rate directly proportional to the increase in culture biomass. It is concluded that the serotype-specific antigen may be a prime immunogen due to its surface localization at both capsule and wall sites.
Bactericidal Effect of Oleic Acid on Group A Streptococci: Mechanism of Action
Authors: Speert David P. , Wannamaker Lewis W. , Gray Ernest D. , Clawson C. Carlyle
Published: January 2020

In contrast to
Staphylococcus aureus
and coagulase-negative staphylococci, group A streptococci are infrequently present on normal human skin, except in certain populations with endemic impetigo. This has been attributed to differences in susceptibility to the bactericidal effect of skin surface lipids, particularly unsaturated fatty acids. When an M type 6 strain group A streptococcus was exposed to 500 μg of oleic acid per ml, viable counts decreased by 4 logs in 5 min. The rank order of killing was 35 > 20 > 4°C. Oleic acid did not kill a strain of
S. aureus
, a strain of coagulase-negative staphylococcus, or a strain of
Escherichia coli
, but bound rapidly to these bacteria as well as to the group A streptococcus. The loss of [
3
H]uridine from labeled oleic acid-treated group A streptococcal cells was greater than 100 times that of controls. There was no loss of [
3
H]-thymidine from group A streptococci or of [
3
H]uridine or [
3
H]thymidine from identically exposed coagulase-negative staphylococci. When [
3
H]uridine was added to group A streptococci during mid-log-phase growth, cessation of uptake occurred within 5 min of addition of 50 μg of oleic acid per ml. Electron microscopic changes seen within 5 min included condensation of the nucleoid and distortion of the streptococcal surface by numerous clumps and blebs. Coagulase-negative staphylococci,
S. aureus
, and
E. coli
similarly exposed showed no comparable electron microscopic changes. We propose that oleic acid kills group A streptococci by altering the integrity of the cell membrane with resulting loss of ribonucleic acid but not deoxyribonucleic acid.
Use of antibodies to purified Newcastle disease virus glycoproteins for strain comparisons and characterizations
Authors: Avery R J , Niven J
Published: January 2020

The H/N and F1 glycoproteins responsible for hemagglutinin/neuraminidase (H/N) activity and fusion (F1) activity were purified from several Newcastle disease virus strains and used to produce antisera. These antisera can be used to clearly distinguish the virus strains by a number of techniques including radioimmunoassay, and so provide a means of classifying new virus isolates. The variations detected between strains are not related to field virulence.
Comparison of two antigen detection techniques in a primate model of Haemophilus influenzae type b infection
Authors: Scheifele D W , Daum R S , Syriopoulou V P , Siber G R , Smith A L
Published: January 2020

Rapid diagnosis of Haemophilus influenzae type b meningitis is possible using immunological tests for capsular antigen (polyribophosphate, PRP), such as countercurrent immunoelectrophoresis (CIE) and latex particle agglutination (LPA). We compared two tests in monkeys with evolving, serially quantitated H. influenzae type b bacteremia (n = 23) and meningitis (n = 21). In vitro, the LPA test was sensitive to 0.5 ng of PRP/ml of saline, and the CIE test was sensitive to 1.0 ng/ml; in serum, however, CIE detected 5.0 ng of PRP/ml, whereas the sensitivity of LPA was unchanged. LPA detected PRP earlier in the course of bacteremia (mean, 12 h after onset; range, 4 to 36 h) than did CIE (mean, 45 h; range, 4 to 168 h) (P less than 0.01). A positive LPA test required greater than or equal to 100 bacteria per ml of blood, whereas CIE required greater than or equal to 1,000/ml. PRP accumulated with continuing blood stream infection, aiding detection of low-grade bacteremia. LPA detected antigen in cerebrospinal fluid (CSF) earlier in the course of meningitis and at a lower bacteria density than did CIE. Both methods detected antigen reliably with greater than or equal to 1,000 bacteria per ml of CSF. A close correlation existed between CSF concentrations of capsular antigen and bacteria (r = 0.90, P less than 0.001). We conclude that the LPA method permits earlier diagnosis of H. influenzae type b infection in part because of its greater sensitivity.
Effect of cell-mediated immune factors on the replication of an attenuated temperature-sensitive mutant of vaccinia virus
Authors: Keller F , Drillien R , Kirn A
Published: January 2020

Studies of the in vivo multiplication of an attenuated temperature-sensitive strain of vaccinia virus (ts2) indicated that temperature sensitivity alone could not account for the attenuation. Immunodepressive treatment of intracerebrally inoculated mice had a dramatic stimulatory effect on the multiplication of the attenuated strain and suggested that establishment of ts2 infection in the mice was hindered by host defense mechanisms mediated by cellular elements. Experiments carried out in vitro with the 51Cr release assay showed that cells infected with the ts2 mutant represented more susceptible targets to the cytotoxic action of immune spleen cells than cells infected with the parent vaccinia strain. Moreover, it appeared that ts2 replication, compared with wild-type replication in vitro, was more inhibited by the immune spleen cells. Although this work did not evaluate the role of specifically sensitized lymphocytes within the lesions, indirect evidence suggests that lymphocytes could cause a more effective halt in ts2 virus multiplication than in wild-type multiplication.
Effect of Growth Rate and Glucose Concentration on the Biochemical Properties of
Streptococcus mutans
Ingbritt in Continuous Culture
Authors: Hamilton I. R. , Phipps P. J. , Ellwood D. C.
Published: January 2020

A comparison was made of the properties of
Streptococcus mutans
Ingbritt grown in continuous culture under conditions of excess glucose (nitrogen limitation) and limiting glucose at mean generation times of 1.7 to 14 h. Only low levels of glucoamylase-specific glycogen were formed in cells from either culture, and the total carbohydrate content of the cells under excess glucose was only at most 1.6-fold higher than in the glucose-limited culture. A negligible amount of cell-free polysaccharide was formed in either culture, although a significant level of glucosyltransferase activity was observed in both, with the highest activity at
D
= 0.2 and 0.4 h
-1
with a glucose limitation. Other differences were observed. (i) Lactate was the main end product of the glucose-excess culture, whereas acetate, formate, and ethanol were the main products of the glucose-limited culture except at a mean generation time of 1.5, when lactate represented 30% of the products. (ii) The yield (in grams per mole of glucose) of the latter culture was 2.6- to 4.0- fold higher than the yield of the glucose-excess culture. (iii) Washed cells from the glucose-limited culture were much more acidogenic (1.7- to 6.2-fold) than the glucose-excess cells when incubated with glucose, sucrose, and fructose. Endogenous glycolytic activity by the latter cells was significant, being 31 to 92% of the exogenous glucose rate at the four dilution rates. (iv) Cells from the glucose-excess culture were more insensitive to fluoride than cells from the glucose-limited culture. The NaF 50% inhibition dose values for the effect of fluoride on the metabolism of glucose, sucrose, and fructose were calculated for the four dilution rates at four pH values. This analysis indicated that rapidly metabolizing cells were more sensitive to fluoride than cells that metabolized the sugars more slowly.
Antigenic heterogeneity of the non-serogroup antigen structure of Neisseria gonorrhoeae lipopolysaccharides
Authors: Apicella M A , Gagliardi N C
Published: January 2020

Studies of the antigenic structure of the polysaccharide component of gonococcal lipopolysaccaride (LPS) indicated that the non-serogroup antigen structure is antigenically heterogeneous. Immunodiffusion studies of Gc4 strain 8551 indicated that in addition to the Gc serogroup determinent, this polysaccharide contains two other sets of determinants, one which is shared with the other five Gc serogroups and a second which is shared by the Gc1 and Gc3 serogroups. Enzyme-linked immunosorbent assay (ELISA) studies confirmed the immunodiffusion, documented qualitative differences, and suggested that quantitative differences may exist in the common determinant from serogroup to serogroup. As measured by ELISA inhibition, native LPS showed the same antigenic arrangement as the purified LPS polysaccharides. Studies with LPS-derived polysaccharides from nonprototype strains in immunodiffusion gave results identical to the prototype strains. This antigen variation, although distinct from serogroup specificity, was related to the serogroup determinant.
Enzymatic induction of interferon production by galactose oxidase treatment of human lymphoid cells
Authors: Dianzani F , Monahan T M , Scupham A , Zucca M
Published: January 2020

Human lymphocyte cultures produced large amounts of interferon after treatment with the enzyme galactose oxidase. Interferon production was detectable as early as 3 h after enzymatic treatment and reached a level of about 10(4) reference units 20 to 24 h later. Galactose oxidase-induced interferon appeared to be immune interferon on the basis of acid lability, lack of neutralization by antibody to leukocyte interferon, and slow kinetics of activation of the cellular antiviral state. Interferon production was inhibited to the same extent (99%) by pretreatment of the cells with beta-galactosidase or with neuraminidase followed by beta-galactosidase, suggesting that the critical event for activation of interferon production is the oxidation of exposed galactose residues on lymphocyte membrane.
Generation of specfic T-cell suppressor function induced by Streptococcus mutans in monkeys and mice
Authors: Lamb J R , Kontiainen S , Lehner T
Published: January 2020

Specific suppressor cells in mice and monkeys can be induced with high doses of Streptococcus mutans antigen. When these cells are further cultured with a low dose of S. mutans antigen in vitro, they secrete specific suppressor factors which decrease the cooperative responses to this antigen. In view of the sensitivity of these cells to anti-Thy 1 and complement, and as they are not retained on nylon-wool, these suppressor cells are T cells. The suppressor factor exerts its effect on T helper cells and not on B cells. Suppressor cells and factors may regulate antibody responses to S. mutans and might be of significance in determining the dose and frequency of immunization and the type of adjuvant to be used.
Leishmanial Excreted Factor: Protein-Bound and Free Forms from Promastigote Cultures of
Leishmania tropica
and
Leishmania donovani
Authors: Slutzky Gerald M. , El-On Joseph , Greenblatt Charles L.
Published: January 2020

Leishmania
spp. growing in culture produce an immunologically active substance called excreted factor (EF), which precipitates antibodies raised against intact cells and has been implicated as the conditioning agent for parasite infection of host macrophages. An improved method for isolation of the material is described, based on Sephadex column chromatography of growth medium which had been boiled at pH 5.0. This procedure allows the detection of differences among the EF molecules of different species, and it overcomes previous shortcomings through the monitoring of immunological activity throughout. Analysis of the products of this procedure revealed that EFs from
Leishmania tropica
and
Leishmania donovani
share a common carrier protein, identified as rabbit serum albumin, and are chemically quite similar. Growth medium from
L. tropica
boiled at acidic pH contains primarily an EF-albumin complex of 75,000 molecular weight. Treated growth medium from
L. donovani
, on the other hand, contains both the albumin complex and a smaller molecule (less than 27,000 molecular weight) that is not associated with rabbit protein. This material accounts for nearly 20% of the EF of one
L. donovani
strain, but constitutes only a minute fraction of
L. tropica
EF. Treatment of the EF-albumin complex with trichloroacetic acid separates the molecule into two major subunits, one having a molecular weight of about 61,000 (without anti-
Leishmania
activity) and the other having a molecular weight of about 18,000 (with no anti-rabbit activity). The protein-free EF of
L. tropica
differs from that released by trichloroacetic acid extraction in that it is capable of precipitating antisera of nonhomologous serotypes, whereas the albumin complex and the trichloroacetic acid-treated EF fragment are not. EFs from both species display pH-dependent affinity for certain lectins.
Interaction of diphtheria toxin with phosphorylated molecules
Authors: Proia R L , Hart D A , Eidels L
Published: January 2020

The binding of diphtheria toxin to 125I-labeled cell surface glycoproteins from hamster thymocytes was shown to be inhibited by nucleotides. The relative effectiveness of the nucleotides (at 5 mM) was found to be thymidine triphosphate greater than adenosine triphosphate greater than guanosine triphosphate greater than uridine triphosphate greater than cytidine triphosphate. When adenine-containing compounds were used, the relative effectiveness was determined to be adenosine tetraphosphate greater than adenosine triphosphate greater than adenosine diphosphate greater than adenosine monophosphate. In addition, tetrapolyphosphate, tripolyphosphate, inositol hexaphosphate (phytic acid), and the highly phosphorylated proteins casein and phosvitin were also shown to be potent inhibitors of the binding of diphtheria toxin to 125I-labeled cell surface glycoproteins. Diphtheria toxin was shown to bind directly to 125I-casein; this binding was also inhibited by the highly phosphorylated compounds and was decreased by pretreatment of the 125I-casein with alkaline phosphatase. These results suggest that diphtheria toxin binds to regions of high phosphate density and raise the possibility that the site on the cell surface glycoproteins to which diphtheria toxin binds might be polyanionic in nature.
Induction of an inhibitor of interferon action in a mouse lymphokine preparation
Authors: Fleischmann W R , Georgiades J A , Osborne L C , Dianzani F , Johnson H M
Published: January 2020

An inhibitor of interferon action was identified in mouse lymphokine preparations. The inhibito was first detected in the supernatant fluid of mouse spleen cells at 72 h after stimulation by staphylococcal enterotoxin A. Inhibitor was not detected in supernatant fluids from unstimulated cultures. This inhibitor blocks the antiviral activity of both immune and fibroblast interferons. The inhibitor was purified 1,000-fold by two-step column chromatography. The partially purified inhibitor blocked the antiviral activity of up to 400 U of interferon. The immunosuppressive effect of interferon was also blocked by the inhibitor, suggesting that the inhibitor may modulate the immunoregulatory function of interferon.
Action of bacterial lipopolysaccharide on the respiration of mouse liver mitochondria
Authors: McGivney A , Bradley S G
Published: January 2020

Escherichia coli O127:B8 lipopolysaccharide (LPS) inhibited oxygen consumption by isolated mouse liver mitochondria at 10 micrograms of LPS per mg of protein when glutamate + malate was the substrate and adenosine 5'-diphosphate had been added (state 3 respiration), but had little effect when adenosine 5'-diphosphate was not added (state 4 respiration). LPS stimulated state 4 respiration at 10 micrograms/mg of mitochondrial protein when succinate was the substrate but had little effect on state 3 respiration. Lipid A from Shigella sonnei at 2 micrograms/mg of mitochondrial protein also stimulated state 4 respiration but did not affect state 3 respiration with succinate as the substrate. Lipid A, unlike LPS, caused a decrease in the adenosine 5'-diphosphate/O ratio. LPS at 100 micrograms/mg of mitochondrial protein impaired the reduction of cytochromes aa3, c, and b when succinate was the substrate but not when reduced nicotinamide dinucleotide, dithionite, or glutamate was the substrate.
Purification and characterization of a rhamnose-containing cell wall antigen of Streptococcus mutans B13 (serotype d)
Authors: Prakobphol A , Linzer R , Genco R J
Published: January 2020

A rhamnose-containing polysaccharide (RCP) was extracted and purified from cell walls of Streptococcus mutans B13 (serotype d) and was chemically and immunologically characterized. Walls were initially extracted with 5% trichloroacetic acid at 4 degrees C to remove the serotype antigen and were then sequentially extracted with increasing concentrations of hot acid. Extracts lacking galactose were combined and chromatographed on a column of diethylaminoethyl--Sephadex A25. The purified RCP contained 90% carbohydrate, 1.4% protein, and 0.16% phosphorus. Analysis by gas chromatography indicated that the RCP was composed of rhamnose and glucose in a 1.6:1 ratio. RCP was immunogenic in rabbits when animals were immunized with whole cells or cell walls. Antisera prepared against partially extracted cell walls of B13 appeared specific for RCP. These sera were not reactive with purified serotype d antigen or lipoteichoic acid in passive hemagglutination assays or by agar gel diffusion. The RCP appeared to be a cell wall polysaccharide that was both chemically and immunologically distinct from the serotype d antigen.
Different susceptibilities of skin to type 1 and type 2 herpes simplex viruses in newborn rabbits
Authors: Oh J O , Minasi P
Published: January 2020

Skin infections with type 1 herpes simplex virus (HSV-1) were compared with skin infections with type 2 virus (HSV-2). Five strains each of HSV-1 and HSV-2 were tested by injecting 10(3) 50% tissue culture infective doses of each strain subcutaneously into 1-day-old New Zealand white rabbits. All five strains of HSV- 2 produced severe skin lesions that resulted in wide dissemination of the infection to many organs, paralysis of the hind legs, and finally death. The virus could be isolated frequently from skin lesions, from various organs (liver, lungs, adrenal glands, brain, and eyes), and from circulating leukocytes and plasma. In contrast, all five strains of HSV-1 failed to produce significant skin lesions or dissemination of virus, only half of the skin lesions yielded HSV, and no virus could be isolated from the blood. These results indicate that HSV-1 dose not grow well in the skin of newborn rabbits and fails to disseminate, whereas HSV-2 is dermatotropic and disseminates readily to many organs by hematogenous routes.
Dengue-2 vaccine: preparation from a small-plaque virus clone
Authors: Eckels K H , Harrison V R , Summers P L , Russell P K
Published: January 2020

The S-1 clone of dengue type 2 virus was used for the preparation of a live-attenuated vaccine after passage in DBS-FRhL-2 cell culture. The vaccine virus had a relatively higher replicative capacity at superoptimal temperatures than its precursor virus, S-1, passaged in primary green monkey kidney cells (S-1 PGMK). There was also a tendency for the S-1 vaccine virus to exhibit leakiness at increased temperatures. Another in vitro marker, replication in monkey peripheral blood leukocytes, indicated less host restriction for the S-1 vaccine in comparative assays with S-1 PGMK virus. Mouse virulence appeared to remain stable on passage in DBS-FRhL-2 cells, whereas monkey immunogenicity decreased. Cautious trials of the dengue type 2 S-1 vaccine in humans are indicated.
Immunogenicity of the ribosomal fraction of Salmonella typhimurium: analysis of humoral immunity
Authors: Kita E , Kashiba S
Published: January 2020

The ribosomal fraction prepared from Salmonella typhimurium LT2 was further purified by gel filtration of Sepharose 4B and afforded excellent protection against homologous challenge. The highly effective immunogens were composed of several fractions which could give different types of protection to mice. The first type of protection was heat-labile antigens which could induce humoral immunity, and the second type of protection was heat-stable antigens capable of evoking cellular resistance in mice. The former were different from O-antigens and the latter were free of endotoxin and rich in ribonucleic acid. The third type of protection was heat-resistant substances of cell wall components, which were mainly composed of O-antigens. The high immunogenicity observed in this study could be obtained only by the heat-stable antigens rich in ribonucleic acid, and the immunity conferred by this kind of antigen was due to the cellular type of protection.
Vaccination of cows with a K99 extract to protect newborn calves against experimental enterotoxic colibacillosis
Authors: Nagy B
Published: January 2020

Pregnant cows and heifers were vaccinated with a crude K99 extract prepared from an Escherichia coli K-12, K99 strain. A similar group, injected with an extract prepared from the K-12 parent strain, served as a control. Eleven calves were born to both groups and challenged orally with a mean of 1.6 x 10(11) bacteria of enterotoxic E. coli B41 (O101:K99+, ST+) at the time of first colostrum uptake (1 to 7 h after birth). As a result of challenge, no death occurred in the vaccine group, but four calves died in the control group. Six calves in the vaccine group and all calves in the control group developed diarrhea. Colostral anti-O101 titers were very similar in both groups. Anti-K99 titers of colostral samples from the vaccinated dams were, however, significantly higher as compared to those of the controls. It is suggested that colostral antibodies, raised against the crude K99 extract vaccine, exerted a protective effect on newborn calves against the challenge enterotoxigenic E. coli O101:K99+, Ent+ strain.
Immunochemical studies of the lipopolysaccharides of Vibrio cholerae: constitution of O specific side chain and core polysaccharide
Authors: Raziuddin S
Published: January 2020

Lipopolysaccharides from various strains of Vibrio cholerae have been found to consist of distinct O-specific side chain and core polysaccharide regions in their degraded polysaccharides. The major identifiable components in the core polysaccharide were phosphorus, glucose, heptose, fructose, and ethanolamine phosphate, with small amounts or traces of mannose, rhamnose, and D-perosamine. On the other hand, glucose, fructose, mannose, rhamnose, glucosamine, D-quinovosamine, and D-perosamine were identified from O-specific side chain polysaccharide. The amounts of each component have been determined from O-specific side chain and core polysaccharides of the lipopolysaccharides from these bacterial strains.
Immunofluorescent characterization of lymphocytes in lungs of rats infected with Mycoplasma pulmonis
Authors: Davis J K , Maddox P A , Thorp R B , Cassell G H
Published: January 2020

Immunofluorescence was used to determine the relative percentages of T and B lymphocytes found in the lungs of normal and Mycoplasma pulmonis-infected F344 rats. Lymphocytes recovered from controls were approximately 25% T, 25% B, and 50% unclassified mononuclear cells. Infected animals had a 2.6-fold greater number of T cells and IgA-bearing cells, and a 1.6-fold greater number of unclassified mononuclear cells. These studies show that M. pulmonis infection significantly alters lung lymphocyte populations both quantitatively and in subpopulation distribution. Therefore, future studies of rat lung lymphocytes should utilize animals known to be free of this ubiquitous respiratory pathogen.
Concanavalin A-mediated affinity film for Treponema pallidum
Authors: Baseman J B , Zachar Z , Hayes N S
Published: January 2020

Freshly extracted Treponema pallidum bound to glass cover slips preexposed to specific lectins, permitting biological testing in the absence of complex tissue fluids.
Difference in antigen-presenting ability of macrophages between high- and low-responder mice in delayed-type hypersensitivity to Mycobacterium bovis BCG
Authors: Nakamura R M , Tokunaga T , Yamamoto S
Published: January 2020

Purified protein derivative-pulsed spleen macrophages of Mycobacterium bovis BCG high-responder mice stimulated BCG-primed lymphocytes of F1 (low x high) mice well in vitro, but those of BCG low-responder mice did not.
Ganglioside and monosaccharide inhibition of nonspecific lymphocyte mitogenicity by group A streptococcal pyrogenic exotoxins
Authors: Schlievert P M , Schoettle D J , Watson D W
Published: January 2020

Group A streptococcal pyrogenic exotoxins (SPE) types A, B, and C are potent nonspecific lymphocyte mitogens. The mitogenicity of these exotoxins was inhibited by gangliosides and sialic acid, whereas concanavalin A was unaffected. The capacity of both concanavalin A and SPE-A to stimulate lymphocytes was suppressed by alpha-methyl-D-mannopyranoside. Galactose reduced the activity of SPE-C. The sugars, glucose, N-acetylglucosamine, alpha-methyl-D-glucopyranoside, and fucose, did not affect SPE mitogenicity.
Demonstration of specific binding sites for human serum albumin in group C and G streptococci
Authors: Myhre E B , Kronvall G
Published: January 2020

A total of 297 bacterial strains belonging to 27 species was tested for quantitative uptake of radiolabeled human serum albumin. Specific binding sites with high affinity for human serum albumin were found exclusively in group C and G streptococci. The albumin binding was found to be a time-dependent, saturable, and displaceable process which obeyed simple kinetic equations. Scatchard analysis revealed that human serum albumin bound to a homogeneous population of receptors with an affinity in the order ot 10(7) liters/mol and that the average bacterial cell carried more than 80,000 binding sites. The albumin receptor is a heat-stable component susceptible to proteolytic digestion. It has a surface localization separate from the receptors for immunolgobulin G, fibrinogen, aggregated beta 2-microglobulin, and haptoglobin. In individual strains, albumin reactivity was also detected independently of these other types of interactions with human proteins.
Resistance to listeriosis in mice that are deficient in the fifth component of complement
Authors: Petit J C
Published: January 2020

Infection with Listeria monocytogenes was studied in strains of mice with genetic absence of the fifth component of complement (C5). Mice deficient in C5 consistently showed an increased growth of Listeria in their spleens as compared to normal mice. This increased growth was not corrected by administration of plasma containing C5. Furthermore, depletion of C5 and terminal complement components by administration of cobra venom factor did not impair the resistance to Listeria infection of normal mice. No phagocytic defect could be detected in macrophages from strains lacking C5. Transfer of bone marrow cells from C5+ but not from C5- mice corrected the marked increase of Listeria growth in mice having blockade of the reticuloendothelial system. We hypothesize that the defect of mice lacking C5 lies not in the absence of serum C5 but somewhere at the level of the macrophage.
Failure of synthetic muramyl dipeptide to increase antibacterial resistance
Authors: Finger H , Wirsing von König C H
Published: January 2020

Synthetic muranyl dipeptide, which potentiates antibody production and cellular immune responses at a dosage of 100 to 500 micrograms, did not enhance resistance to intravenous infection with a sublethal dose of 2 X 10(3) to 4 X 10(3) viable Listeria monocytogenes cells in mice when intraperitoneally injected either 20 min or 5 days before infection. Similarly, blockade of the mononuclear phagocyte system by dextran sulfate 500 could not be overcome by pretreatment with muramyl dipeptide. In contrast, dextran sulfate 500-induced loss of antibacterial resistance was found to be completely abolished by intraperitoneal injection of 3 X 10(9) killed Bordetella pertussis organisms when given 4 days before injection of dextran sulfate 500, i.e., 5 days before infection. B. pertussis were also effective in enhancing antibacterial resistance when administered 5 days before infection. The different behavior of the two adjuvants tested is assumed to be due to their different nonspecific proliferative capacities. Thus, B. pertussis are assumed to act by direct stimulation of the mononuclear phagocyte system whereas muramyl dipeptide does not.
In vitro synthesis of anti-mycobacterial antibodies in biopsies from skin lesions of leprosy patients
Authors: Lai A Fat R F , Chan Pin Jin J , van Furth R , Harboe M
Published: January 2020

To demonstrate local synthesis of anti-Mycobacterium leprae antibodies, biopsies from skin lesions of leprosy patients were cultured in vitro in a medium containing 14C-labeled lysine and isoleucine, and the culture fluids were analyzed by crossed immunoelectrophoresis with intermediate gel and autoradiography. The results show that anti-M. leprae antibodies were synthesized in vitro in the biopsies from the skin lesions of leprosy patients and that the specificity of the locally produced antibodies varied from patient to patient.
Replication of rubella virus in human mononuclear blood cells
Authors: van der Logt J T , van Loon A M , van der Veen J
Published: January 2020

Rubella virus was capable of replicating in both unstimulated and phytohemagglutinin-stimulated cultures of human mononuclear blood cells. Monocyte-derived macrophages were the main cell type responsible for viral replication. The susceptibility of macrophages increased during cultivation. Phytohemagglutinin-stimulated lymphocytes were able to support replication to a limited degree. No viral replication was detected in unstimulated lymphocytes. Both stimulation and viral replication in phytohemagglutinin-treated lymphocyte cultures were enhanced by the addition of murine macrophages. Human leukocyte interferon depressed the production of virus in these combined cultures. The finding that rubella virus is able to replicate in human lymphocytes as well as in macrophages may contribute to understanding the mechanisms of the suppressive effect of the virus on in vitro lymphocyte phytohemagglutinin responsiveness and in vivo immune functions.
Significance of bursa of Fabricius as target organ in infectious bursal disease of chickens
Authors: Käufer I , Weiss E
Published: January 2020

The highly virulent strain Cu-1 of infectious bursal disease virus caused 100% mortality in 4-week-old specific pathogen-free chickens. In contrast, chickens infected after bursectomy did not become sick and only showed some discrete and transient necrosis in lymphatic tissues. However, these chickens contained infectious virus and, subsequently, produced specific antibodies. The virus concentrations in the organs studied reached their maximum 2 days postinfection, but were about 1,000 times lower in non-bursectomized animals. It may be assumed that in bursectomized chickens the early events of infection are the same as in non-bursectomized ones. Virus is spread in varius organs, but due to the absence of a sufficient number of susceptible cells, virus multiplication is moderate and can be kept in check by the host defense mechanism. With the occurrence of circulating specific antibodies the virus can be rapidly eliminated. The studies particularly stress that the availability of a large number of highly susceptible cells is a crucial point in acute viral infections.
Bacteriostatic enterochelin-specific immunoglobulin from normal human serum
Authors: Moore D G , Yancey R J , Lankford C E , Earhart C F
Published: January 2020

Heat-inactivated normal human serum produces iron-reversible bacteriostasis of a number of microorganisms. This inhibitory effect was abolished by adsorption of serum with ultraviolet-killed cells of species that produce the siderophore enterochelin. Bacteriostasis also was alleviated by adsorption of serum with 2,3-dihydroxy-N-benzoyl-L-serine, a degradation product of enterochelin, bound to the insoluble matrix AH-Sepharose 4B. The adsorption process did not add iron or enterochelin to serum, nor did it remove transferrin. The immunoglobulin fraction from normal human serum was isolated; when added to a defined medium (M199) prepared so as to mimic normal human serum, the immunoglobulin rendered the medium inhibitory to an enterochelin-defective strain of Salmonella typhimurium. Adsorption of this medium with AH-Sepharose 4B-2,3-dihydroxy-N-benzoyl-L-serine removed the inhibition. Our results indicate that enterochelin-specific immunoglobulins exist in normal human serum. These immunoglobulins may act synergistically with transferrin to effect bacteriostasis of enterochelin-producing pathogens.
Correlation of the duration and magnitude of protection against Salmonella infection afforded by various vaccines with antibody titers
Authors: Angerman C R , Eisenstein T K
Published: January 2020

Groups of mice were immunized with optimal doses of the following vaccines of Salmonella typhimurium W118-2: acetone-killed cells, lipopolysaccharide, ribosomes, and live cells. At 3 weeks, 1 month, 2 months, 4 months, or 6 months postimmunization, sera were collected from control and vaccinated animals, and the anti-lipopolysaccharide and whole-cell agglutination titers of the sera were determined. Other groups of similarly vaccinated mice were tested for resistance to infection by challenging with live W118-2 and scoring the number of survivors 30 days postinfection. It was found that only ribosomes and live cells afforded significant protection 6 months after immunization. Thus, in duration of protection ribosomes were superior to the other nonviable vaccines tested. At all time intervals tested, purified lipopolysaccharide was the least effective vaccine. Protection afforded by the acetone-killed cell and ribosomal vaccines correlated better with the whole-cell agglutination titers than with the anti-lipopolysaccharide titers. However, the longer duration of protection afforded by the ribosomal vaccine, as compared with the acetone-killed vaccine, could not be accounted for by differences in whole-cell agglutination titers. These studies show that ribosomal vaccines are equal in all parameters to acetone-killed cells and have the advantage of providing longer-lasting immunity.
Comparative stimulation of granulocyte adherence and chemotaxis by bacterial products
Authors: Spagnuolo P J , Ellner J J
Published: January 2020

Many inflammatory stimuli increase both in vitro granulocyte adhesiveness and motility. The present investigation compared the effects of bacterial products on in vitro granulocyte adherence to nylon fiber and granulocyte motility. Escherichia coli endotoxin and Staphylococcus aureus and E. coli culture filtrates enhanced both motility and granulocyte adherence to nylon. Column chromatography of S. aureus culture filtrate demonstrated that similar molecular weight components stimulated both motility and granulocyte adherence. In contrast, endotoxin stimulated granulocyte adherence but not chemotaxis in plasma-free medium. In addition, a group D streptococcal culture filtrate enhanced motility but not adhesiveness of plasma-free granulocytes. These studies indicate that inflammatory stimuli may interact with granulocytes in concert to enhance both adhesiveness and motility, or they may interact independently to stimulate one in vitro function preferentially.
Evidence for an immunological relationship between Streptococcus mutans and human cardiac tissue
Authors: Hughes M , Machardy S M , Sheppard A J , Woods N C
Published: January 2020

Two-dimensional immunoelectrophoresis, indirect immunofluorescence, and radioimmunoassay were used to demonstrate that antisera from rabbits immunized with some strains of Streptococcus mutans contain antibodies that cross-react with human cardiac tissue. These rabbits were sensitized to a shocking dose of human heart antigen, and anaphylactic deaths were sometimes produced. Myocarditis was also a result of the immunization procedure. Data obtained with all five techniques were comparable. Cross-reactivity could be associated with three antigens designated ID, IF, and HL. Antigens ID and IF were major immunogens of S. mutans Ingbritt, but HL antibodies were produced only after hyperimmunization. Corss-reactivity was of an immunological nature and not the result of nonspecific factors such as bacterial Fc reactive components or antibody elicited to growth medium constituents. These findings support the hypothesis that immunization with S. mutans can induce autoimmune reactions and indicate that antigens must be selected with caution before formulating any dental caries vaccine.
Antitoxic immunity to cholera in dogs immunized orally with cholera toxin
Authors: Pierce N F , Cray W C , Engel P F
Published: January 2020

Colera toxin was evaluated as an oral immunogen against experimental canine cholera. Dogs were immunized orally with 100-microgram doses of purified cholera toxin or comparable doses of crude toxin. Both doses caused moderate diarrhea in most nonimmune dogs. Repeated oral doses (12 doses in 54 days) gave marked protection against the diarrheal effect of oral toxin, provoked a vigorous antitoxic response in jejunal mucosa, and gave nearly complete protection against subsequent oral challenge with living virulent Vibrio cholerae. Protection appeared to be due largely to the antitoxic response in intestinal mucosa. The effectiveness of cholera toxin as an oral vaccine contrasts with the previously described ineffectiveness of toxoid given orally. This study provides an example of mucosal immunity due to a nonreplicating vaccine given orally and suggests that cholera toxin may be useful as a component of an oral vaccine for cholera.
New type of antibody-enzyme conjugate which specifically kills Candida albicans
Authors: Okuda K , Ishiwara K , Noguchi Y , Takahashi T , Tadokoro I
Published: January 2020

A new type of antibody-enzyme conjugate was made, and its possible application to Candida infection was studied. Both lactoperoxidase and xanthine oxidase were conjugated to specific antibody against Candida albicans. In vitro microbiocidal activity of the new antibody-enzyme conjugate, when incubated together with xanthine and minute amount of halides, showed a remarkable level of candidacidal ability. When the new antibody-enzyme conjugate was given to Candida-infected mice, followed by injecting xanthine and a minute amount of halides, about 50% of these heavily infected mice survived, whereas all control nontreated mice died. These data suggest that the further eleboration of this new antibody-enzyme conjugate might lead us to improve our therapeutic methods of clinical medicine.
Alteration of polymorphonuclear leukocyte activity by viable Candida albicans
Authors: Hilger A E , Danley D L
Published: January 2020

The response of human polymorphonuclear leukocytes (PMN) to blastospores and pseudo-hyphae of the opportunistic fungus Candida albicans has been studied in vitro and in vivo. Of the fungicidal mechanisms elucidated thus far, the myeloperoxidase-hydrogen peroxide-halide system appears to be most effective against cells of this fungus. In our studies on the interaction between murine PMN and blastospores, we assayed the release of H2O2 by PMN incubated with viable or killed, unopsonized or opsonized blastospores by using two assay systems, lysis of murine erythrocytes and oxidation of scopoletin. Our results showed that PMN released increasing amounts of H2O2 when incubated with increasing numbers of opsonized or unopsonized killed blastospores, but released decreasing amounts of H2O2 when incubated with increasing numbers of opsonized or unopsonized viable blastospores. The oxidative metabolic burst by PMN in the presence of viable or killed blastospores was also measured by using reduction of nitroblue tetrazolium and chemiluminescence. Viable blastospores stimulated a stronger metabolic burst than killed blastospores, suggesting that PMN respond to live blastospores more vigorously than killed blastospores; however, live blastospores appear to alter or inhibit the release of H2O2 by PMN.
Isolation and Characterization of Melanin-Producing (
mel
) Mutants of
Vibrio cholerae
Authors: Ivins Bruce E. , Holmes Randall K.
Published: January 2020

Vibrio cholerae
strain Htx-3, a hypertoxinogenic mutant of
V. cholerae
569B Inaba, produces a dark brown pigment under certain growth conditions, whereas strain 569B does not. We investigated the biochemical basis for this pigment production and the possible relationships between pigmentation and other phenotypic properties in
V. cholerae.
After mutagenesis of
V. cholerae
569B with
N
-methyl-
N
′-nitro-
N
-nitrosoguanidine, 28 independently derived pigment-forming (
mel
) mutants were isolated and characterized. The
mel
mutants frequently differed from wild type in toxinogenicity or motility and occasionally differed in other phenotypic traits. Individual
mel
mutants differed from wild type both in the amount of toxin produced and in the growth conditions optimal for toxin production. It has not yet been established whether multiple phenotypic changes in individual
mel
mutants represent pleiotropic effects of single mutations or induction of multiple mutations by
N
-methyl-
N
′-nitro-
N
-nitrosoguanidine or both. Production of pigment by
mel
mutants occurred at temperatures from 22 to 40°C, was inhibited by anaerobiosis, and was stimulated by supplementation of growth media with the amino acid precursors of melanin (
l
-phenylalanine,
l
-tyrosine, or
l
-tyrosine plus
l
-cysteine). The pigment possessed several other properties reported for microbial melanins. We conclude that a biochemical pathway for melanin production is present in
V. cholerae
, that this pathway cannot be fully expressed in wild-type strain 569B, and that mutations in the gene(s) which we have designated
mel
can permit hyperproduction of melanin under appropriate conditions.
Growth of
Mycobacterium lepraemurium
in Cell-Impermeable Diffusion Chambers
Authors: Rightsel Wilton A. , Wiygul William C.
Published: January 2020

Successful growth of
Mycobacterium lepraemurium
has been achieved by use of a specialized diffusion chamber technique. The cell-impermeable porous chambers were maintained in animals for periods up to 50 days with and without macrophages and LM cells. A generation time of 6 to 8 days was found for the acid-fast bacilli in chambers containing macrophages when maintained in the mouse. Also, cell-free chambers maintained in the mouse gave a generation time of 11 days for
M. lepraemurium.
There was no doubt that chambers maintained in a susceptible host provided greater yields of bacilli than chambers maintained in a nonsusceptible host such as the guinea pig. In fact, better yields were obtained when the chambers were maintained in monolayer petri plate cultures of mouse peritoneal macrophages than when held in the guinea pig. The most pertinent observation was that living cells are not essential for growth of
M. lepraemurium
, and the results suggest that multiplication can occur in a cell-free environment within a susceptible host. These studies give evidence that the use of porous chambers has promising possibilities for further investigations on the cultivation of other fastidious mycobacteria.
Amino Acid Analysis and Molecular Weight Determination of Tetanus Toxin
Authors: Holmes Marlee J. , Ryan Wayne L.
Published: January 2020

A procedure for the isolation of tetanus toxin was developed. The toxin was precipitated with ammonium sulfate and further purified by use of molecular sieve chromatography and preparative disc gel electrophoresis. The degree of purification resulting from these fractionation procedures was monitored by analytical poly-acrylamide gel electrophoresis, immunodiffusion, and immunoelectrophoresis, all three of which demonstrated a single component. There was a 50-fold increase in specific activity between the crude culture filtrate and the final purified product, which had an average specific activity of 300 × 10
6
mouse minimal lethal doses per mg of N. A molecular weight of 140,000 was estimated by gel filtration on Bio-Gel P-200. A complete amino acid analysis of tetanus toxin is presented.
Post-Transfusion Cytomegaloviremia and Persistence of Cytomegalovirus in Blood
Authors: Armstrong Donald , Ely Mathew , Steger Lynn
Published: January 2020

Cytomegaloviremia was documented in seven patients. Three patients were renal transplant recipients and two were liver transplant recipients. One was a postcardiac surgery patient, and one had acute myelogenous leukemia. The transplant patients had received only banked blood or fresh frozen plasma and developed evidence of infection about 1 month after surgery. Illness varied from no apparent signs to fever alone, fever and a morbilliform rash, hepatitis and an infectious mononucleosis syndrome. The virus was isolated from the blood erythrocyte layer, the leukocyte layer, and the plasma and serum, in that order of frequency. In vitro studies demonstrated persistence of inoculated cytomegalovirus in the presence of erythrocytes (and tissue culture media) for up to 21 days. In whole blood under banking conditions, inoculated virus was recovered after 28 days and, in fresh frozen plasma, after 97 days.