bluuebunny/crossref_metadata_2025 huggingface dataset:
Transfer of Immunity Against
Listeria monocytogenes
by T Cells Purified by a Positive Selection Technique
Authors: McGregor D. D. , Crum E. D. , Jungi T. W. , Bell R. G.
Published: January 2020

Affinity columns prepared with rabbit antibody to the F(ab′)
2
fragment of rat immunoglobulin were used to separate rat thoracic duct lymphocytes into sub-populations that differ with respect to the density of their surface membrane immunoglobulin. Using this technique, it was shown that lymphocytes in the DNA synthetic (S) phase of the mitotic cycle are added in increased number to the lymph of rats infected with
Listeria monocytogenes.
The great majority of these S-phase cells lacked a high density of surface immunoglobulin as indicated by their failure to bind to the immunoabsorbent. Cells which can protect recipient rats against a challenge infection with
L. monocytogenes
also segregated with nonadherent thoracic duct lymphocytes obtained from
Listeria
-immune donors. These protective cells realized their full immunological potential only in recipients that shared histocompatibility-gene-coded structures with the immune lymphocyte donors. The above findings accord with the view that immunity to
L. monocytogenes
is mediated in rats by activated T cells which are formed as part of the animal's cell-mediated response to infection. Although
Listeria
-protective lymphocytes concentrate in the nonadherent, T-cell-enriched fraction, it was consistently observed that the adherent, B-cell-enriched fractions of immune donor thoracic duct lymphocytes also could transfer a low level of antimicrobial resistance. This immunity was restricted in allogeneic recipients, a finding which implies that the protection afforded by the adherent population is related to its content of T cells. Nonadherent S-phase lymphoblasts moved in substantial numbers from the blood into peritoneal inflammatory exudates induced by
L. monocytogenes.
The above finding encourages the belief that recently activated T cells realize their protective function locally in centers of infection where they have secondary effects on macrophages.
Degradation of Levan by
Actinomyces viscosus
Authors: Miller Chris H. , Somers Penelope J. B.
Published: January 2020

Actinomyces viscosus
ATCC 15987 was examined for its ability to hydrolyze its own levan. Washed whole cells and an ammonium sulfate fraction from cell-free culture fluids were shown to possess levan hydrolase activity. Analyses of reaction mixtures by gel filtration and thin-layer chromatography demonstrated that the product of levan hydrolysis was free fructose. The cell-associated and extracellular enzyme preparations also hydrolyzed inulin and the levans synthesized by
Aerobacter levanicum
and
Bacillus subtilis.
Growth of
A. viscosus
in media supplemented with 0.1%
A. viscosus
levan resulted in a 33-fold increase and a 7-fold increase in the specific activities of the respective extracellular and cell-associated enzymes when compared with those from 55 mM glucose cultures. Growth in the presence of 29.2 mM sucrose resulted in a 28-fold increase and a 5-fold increase in the specific activities of the respective enzymes when compared with those from the glucose cultures. The extracellular enzyme exhibited high activity over a wide pH range, with 87 and 89% of its pH 6.0 optimum activity at pH 5.0 and 7.0, respectively. The cell-associated enzyme also exhibited optimum activity at pH 6.0, but this was decreased to 10 and 20% at pH 5.0 and 7.0, respectively. Analysis for the presence of extracellular levan during growth of
A. viscosus
in sucrose broths demonstrated that peak levan concentrations occurred during the mid-exponential to late-exponential phase of growth followed by a rapid decline in extracellular levan as a result of levan hydrolase activity.
Demonstration of eosinophil degranulation on the surface of opsonized schistosomules by phase-contrast cinemicrography
Authors: Densen P , Mahmoud A A , Sullivan J , Warren K S , Mandell G L
Published: January 2020

Phase-contrast cinephotomicrography was used to examine eosinophil-schistosomule interaction. Eosinophil degranulation against the surface of schistosomules was observed in the presence of immune serum.
Activation of the alternative complement pathway by a streptococcal lipoteichoic acid
Authors: Fiedel B A , Jackson R W
Published: January 2020

A glycerol lipoteichoic acid antigen from Streptococcus pyogenes 1-RP41 was found by rabbit erythrocyte hemolytic assay to activate the alternative complement pathway in human sera. Over a narrow concentration range of the teichoic acid, complement consumption was dose dependent, whereas at higher concentrations of the acid complement consumption could not be detected.
Biological properties of equine herpesvirus type 1 DNA: transfectivity and transforming capacity
Authors: Allen G P , Randall C C
Published: January 2020

DNA extracted from purified virions of equine herpesvirus type 1 (EHV-1) was examined for its transfectivity and transforming ability. The infectivity of the herpesvirus DNA was demonstrated by addition of calcium phosphate-DNA coprecipitates to monolayers of permissive horse cells, with resultant plaque formation. The efficiency of transfection (50 to 100 plaque-forming units/microgram of DNA) was reduced by treatment of the viral DNA with deoxyribonuclease or sonication but not with Pronase or antivirus neutralizing serum. When nonpermissive mouse 3T3 Cells lacking the enzyme thymidine kinase (TK-) were transfected with intact EHV-1 DNA, clones of cells transformed to the TK+ phenotype were isolated in selective HAT medium (hypoxanthine, aminopterin, thymidine), which prevents growth of the TK- parental phenotype. The efficiency of transformation ranged from one to five transformants per microgram of EHV-1 DNA. The TK activity of the biochemically transformed cells was characterized by biochemical, electrophoretic, and immunological techniques. By these criteria, the TK activity was identical to the EHV-1 TK and different from the host wild-type enzyme. In contrast to the parental TK+ 3T3 cells, the EHV-1-transformed TK+ cells were unable to grow in the presence of arabinosylthymine, a drug selectively phosphorylated by herpesvirus TKs. These results indicate that stable transfer of EHV-1 genes into nonpermissive cells can be achieved with purified viral DNA.
Gonococcal color and opacity variants: virulence for chicken embryos
Authors: Salit I E , Gotschlich E C
Published: January 2020

Previous studies have noted that the prevalence of certain gonococcal colony types is influenced by the physiological state of the human host. Consequently eight different gonococcal strains were passed on clear typing medium, and opaque (Op) and transparent (Tr) variants of heavily pilated (P++) bacteria were selected. P++Op and P++Tr were injected into 11-day-old chicken embryos, and the resulting mortality was determined. In every case the Tr variants were more lethal to the embryos than were the Op variants (P = 0.01). P++Tr were stable in vivo, but chicken embryos injected with P++Op strains developed bacteremia with P++Tr organisms. However, chicken blood was not bactericidal for either colonial variant. These results indicate that transparent pilated bacteria may be more virulent than opaque pilated bacteria and that in vivo selection of transparent forms occurs.
Role of bacterial products in periodontitis: humoral immune response to Eikenella corrodens
Authors: Johnson D A , Behling U H , Listgarten M , Nowotny A
Published: January 2020

Eikenella corrodens can induce periodontitis-like disease in gnotobiotic rats. Some components of this bacterial cell elicit measurable humoral immune response during the development of the disease, but in this system endotoxin is not among the efficient immunogens. Because no humoral immune response could be seen to the endotoxin of Eikenella corrodens it is assumed that this endotoxin can act uncontrolled in monoinfected rats. Accordingly, the lack of protective humoral immune response to pathogenic components of Eikenella corrodens may be the major factor permitting the development of the disease described here. The possibility that both cell-mediated immunity and uncontrolled endotoxic action are parts of the pathomechanism of the disease is supported by our observations.
Role of motility in experimental cholera in adult rabbits
Authors: Yancey R J , Willis D L , Berry L J
Published: January 2020

The role of motility in the pathogenesis of cholera was evaluated in ligated ileal loops of adult rabbits. Four strains of Vibrio cholerae (including both Inaba and Ogawa serotypes of both classical and El Tor biotypes) were compared with their aflagellated, but fully toxigenic and prototrophic, isogenic derivatives as to their ability to produce fluid accumulation in the rabbit gut. The nonmotile mutants required an at least 100-fold-higher dose than their respective wild-type strains to produce comparable fluid accumulation responses. The decreased ability of nonmotile strains to produce a fluid response was not due to their failure to multiply in vivo, since they increased in numbers in the rabbit ileum at the same rate as the wild-type strains, but probably was related to their inability to associate with the intestinal mucosa. After 3 h of incubation, 45 to 53% of motile, [35S]-labeled cells adsorbed to the intestinal wall, whereas only 3 to 15% (depending upon the strain) of the nonmotile bacteria were associated.
Virulence and immunity of Staphylococcus aureus BB and certain deficient mutants
Authors: Hasegawa N , San Clemente C L
Published: January 2020

Coagulase-negative and deoxyribonuclease-negative mutants were isolated from Staphylococcus aureus BB by treatment with N-methyl-N'-nitro-N-nitrosoguanidine. Comparison of virulence (50% lethal dose) to mice of these six mutant strains and S. aureus BB was determined by both intravenous and intraperitoneal routes. The ratios of the 50% lethal dose of coagulase-negative mutants to that of the parental strain S. aureus BB ranged from 201 to 403 for intravenous infection and 30.7 to 52.7 for intraperitoneal infection. The virulence of deoxyribonuclease-negative mutants was essentially the same as that of S. aureus BB. When mice were immunized subcutaneously with live S. aureus BB or its deoxyribonuclease-negative mutants, the resulting protection against the intravenous challenge of S. aureus BB was remarkable. The ratios of the 50% lethal dose for the mice that were immunized by these strains to that for the untreated mice extended from 40.1 to 60.6 for intravenous infection and 6.61 to 11.5 for the intraperitoneal route. However, no effect against S. aureus BB challenge was shown in the mice that were immunized with coagulase-negative mutants.
Dynamics of Escherichia coli infection and meningitis in infant rats
Authors: Bortolussi R , Ferrieri P , Wannamaker L W
Published: January 2020

Escherichia coli strains isolated from newborn infants were injected intraperitoneally into infant rats. Strains possessing the K1 capsular polysaccharide antigen were significantly more virulent than strains lacking this antigen. When 5-day-old animals were injected with 1.2 X 10(1) colony forming units of a K1 E. coli strain (serotype O18ac:K1:H7), about 80% had bacteria isolated from their blood. Forty-eight percent of bacteremic animals had positive cerebrospinal fluid cultures. The development of bacteremia with greater than 10(4) colony-forming units per ml of blood correlated with positive cultures of cerebrospinal fluid. Some animals, studied with serial blood cultures, were able to clear bacteria spontaneously from their blood, whereas others succumbed to infection within 48 h of challenge. The susceptibility of infant rats to E. coli infection was age dependent and appeared related to the K1 antigen.
Complement activation by cell wall fractions of Micropolyspora faeni
Authors: Smith S M , Burrell R , Snyder I S
Published: January 2020

The ability of several cell wall fractions of Micropolyspora faeni, a thermophilic actinomycete associated with farmer's lung disease, to activate complement is reported. Cell walls, obtained by mechanical disruption, were purified by enzyme treatment and chemical extractions. Fractions containing the most purified cell walls were most active in consuming complement, as measured by reduction of hemolytic complement levels of normal human serum. Cell wall fractions activated the alternative complement pathway, as shown by monitoring the conversion of C3 proactivator (factor B) to C3 activator (activated factor B) in the presence of specific cation chelators. Selective degradation of cell walls by lysozyme resulted in a decreased ability to consume complement and implicated peptidoglycan as the major complement-reactive component. The role of this nonspecific complement activation in relation to farmer's lung disease is discussed.
Mucopolysaccharide Material Resulting from the Interaction of
Treponema pallidum
(Nichols Strain) with Cultured Mammalian Cells
Authors: Fitzgerald T. J. , Johnson R. C. , Wolff E. T.
Published: January 2020

During incubation of
Treponema pallidum
(Nichols strain) with cultured mammlian cells derived from normal rabbit testes (NRT), an amorphous material accumulated at the surface of the cultured cells. This material was randomly distributed on all tissue cells within the culture chambers. The amount of amorphous material was dependent on the treponemal inocula. With 3 × 10
8
organisms per ml, this material was readily apparent within 2 days; with 4 × 10
7
organisms per ml, this material was detectable within 4 to 5 days; with lower inocula, the accumulation of amorphous material was far less apparent. Deposition of this surface-associated material required attachment of treponemes to the cultured cells, and the amount deposited was related to the number of treponemes attached per cell. This amorphous material was not detected when NRT cells were incubated with preparations of
T. pallidum
that were heat or air inactivated. In addition, the accumulaton of amorphous material was not due to a soluble component from host testicular tissue or to a soluble component developing during treponemal infection. This was demonstrated by the inability of membrane filtered preparations of
T. pallidum
to induce the deposition of amorphous material at the surface of the cultured cells. The nature of this material appeared to be acidic mucopolysaccharide as indicated by its metachromatic staining properties, its stainability with ruthenium red, and its partial degradation by bovine and streptomyces hyaluronidase. This amorphous material that accumulated in vitro at the surface of cultured cells may be similar to the mucoid material that accumulates in vivo during syphilitic infection.
Response of C3H/HeJ and C3H/HeN mice and their peritoneal macrophages to the toxicity of Chlamydia psittaci elementary bodies
Authors: Ivins B E , Wyrick P B
Published: January 2020

Intravenous injection of toxic doses of Chlamydia psittaci elementary bodies into endotoxin-responsive C3H/HeN mice or endotoxin-nonresponsive C3H/HeJ mice resulted in essentially identical time intervals to death. Inoculation of monolayer cultures of thioglycolate-stimulated peritoneal macrophages from the two strains of mice with 250 elementary bodies per macrophage resulted in immediate host cell toxicity, although the C3H/HeJ macrophages were somewhat less sensitive to elementary body toxicity than were the C3H/HeN macrophages.
Enhancement of the immunogenicity of phase I antigen of Coxiella burnetii
Authors: Wachter R F , Briggs G P , Pedersen C E
Published: January 2020

The immunogenicity of the soluble phase I antigen of Coxiella burnetii for guinea pigs was enhanced by a nuclease-resistant complex of polyriboinosinic-polyribocytidylic acid, poly-L-lysine, and carboxymethyl cellulose.
Immunity to antigenically related salmonellae: effects of humoral factors on the bactericidal activity of normal and immune peritoneal exudate cells
Authors: Meléndez M , González M C , Reid M , Fuentes C , Castillo D
Published: January 2020

Immunity against Salmonella enteritidis and Listeria monocytogenes was studied by measuring in vitro the bactericidal activity of peritoneal exudate cells (PEC) of control (normal PEC) and S. typhi Ty2-immune (immune PEC) mice. Specific immune serum, anti-S. tyhphi Ty2, heat inactivated at 56 degrees C for 30 min, significantly inhibited the growth of S. enteritidis only with immune PEC. These opsonic factors had no effect upon the activity of normal PEC. That such inhibition could not be demonstrated in Listeria experiments, either with immune or normal PEC, suggests that S. enteritidis was specifically recognized, in vitro, by the thermostable opsonin anti-S. typhi Ty2 and that macrophages from immune PEC were more efficient in inhibiting bacterial growth than those from normal PEC. Thus, the interaction between macrophages and the microorganism seems to play an essential role in cell-mediated as well as humoral immunity.
Selective Binding of Blood Group-Reactive Salivary Mucins by
Streptococcus mutans
and Other Oral Organisms
Authors: Gibbons R. J. , Qureshi J. V.
Published: January 2020

Strains of
Streptococcus mutans
of four genetic groups and five serotypes, and strains of
S. sanguis, S. mitis, S. salivarius, Actinomyces naeslundii
, and
A. viscosus
, were found to bind blood group-reactive (BGR) mucin isolated from whole human saliva. The bacteria studied bound mucins with blood type A or B reactivity to a similar extent, suggesting that the carbohydrate moieties responsible for the A and B antigenic determinants were not involved. The organisms studied appeared to bind different fractions of BGR mucin molecules because preparations absorbed with cells of a given oral species no longer contained BGR molecules which bound to homologous organisms but still possessed BGR components which bound to varying degrees to other bacteria. Differences were even noted among
S. mutans
strains belonging to different genetic groups and serological types. Immunoglobulins could not be detected in the mucin preparations, and addition of anti-human immunoglobulin A (IgA), IgG, or IgM serum to reaction mixtures did not affect binding. Mucin pretreated with periodate or iodoacetate no longer bound to
S. mutans
H12 cells, suggesting that carbohydrate moieties and sulfhydryl groups played an essential role. Active cell metabolism was not required for BGR mucin binding; however, pretreatment of H12 cells with periodate or heat (100°C for 15 min) reduced binding. Mucin labeled with [
14
C]phenyl isothiocyanate appeared to bind to
S. mutans
H12 cells comparably to untreated mucin; the binding also appeared to be specific because less than 15% of the labeled material became bound when incubated with an excess of streptococci. Binding of [
14
C]phenyl isothiocyanate-labeled mucin was not affected by neutral sugars tested or by preparations of c antigen, glycerol teichoic acid, dextran, or crude glucosyltransferase. However, binding was inhibited by several amines. BGR salivary mucins are present in the acquired pellicle covering teeth; the ability of bacteria to selectively bind such components suggest that they may serve as receptor molecules involved in the attachment of bacteria to teeth.
Influence of oxygen tension, sulfhydryl compounds, and serum on the motility and virulence of Treponema pallidum (Nichols strain) in a cell-free system
Authors: Norris S J , Miller J N , Sykes J A , Fitzgerald T J
Published: January 2020

The motility and virulence of Treponema pallidum (Nichols strain) were monitored during incubation in a modified tissue culture medium to study the effects of oxygen tension and medium composition on survival of the organism. A basal medium of Eagle minimal essential medium with 50% fresh, heat-inactivated normal rabbit serum was used inasmuch as better survival occurred with 50% normal rabbit serum than with lower concentrations. Addition of 0.5 to 2.0 mM dithiothreitol or 2.0 mM dithioerythritol to the basal medium led to significantly longer retention of T. pallidum viability in the presence of 3% oxygen than under aerobic or anaerobic conditions. The results of this investigation lend support to the classification of T. pallidum as a microaerophilic organism and provide direction for the design of potentially successful culture systems, with or without tissue culture cells.
Suppression of in vitro growth of virulent and avirulent herpes simplex viruses by cell-mediated immune mechanisms, antibody, and interferon
Authors: Shimizu F , Satoh J , Tada M , Kumagai K
Published: January 2020

A rounding cell-forming--GC strain, which is a variant of a syncytial giant cell-forming herpes simplex virus (+GC Miyama strain), was highly attenuated for Swiss, BALB/c nu/nu, and nu/+ mice, whereas +GC was highly virulent to all the mice tested. +GC and -GC were antigenically indistinguishable from each other by cross-neutralization and cross-immunization. Immunosuppression induced by cyclophosphamide converted the nonlethal -GC infection of mice into a fatal infection. -GC replication in tissue culture was more effectively suppressed by spleen cells immunized with either +GC or -GC than was the +GC replication. -GC replication was also inhibited more effectively by antibody or the antibody-dependent cell-mediated system than was the +GC replication. -GC is highly sensitive to mouse interferon, but +GC was relatively resistant. These findings indicate that attenuation of this avirulent -GC strain may be due to a high susceptibility of its replication to humoral and cell-mediated defense factors. The probable roles of each defense factor in recovery from the infection with virulent and attenuated herpes simplex virus are also discussed.
Intestinal Colonization and Virulence of
Salmonella
in Mice
Authors: Hohmann Arthur W. , Schmidt Günter , Rowley Derrick
Published: January 2020

Within 3 h after oral challenge of mice with
Salmonella typhimurium
, foci of infection developed in the Peyer's patches of the small intestine. The numbers of organisms in the cecum, although in excess of those found in the small intestine, were not firmly associated with the cecal wall but were present largely in the cecum's contents. The Peyer's patches at first were remarkably incapable of eliminating even small numbers of
Salmonella
, but at about 7 days after infection developed the ability to eliminate a less virulent strain of
S. typhimurium.
Selected strains of
Salmonella
of varied virulence, and hybrid
Escherichia coli/Salmonella typhimurium
with varied O-antigens, revealed that those of low virulence could multiply within the intestinal Peyer's patches at nearly the same rate as a virulent strain, and the ability to multiply within the Peyer's patches was not dependent upon O-antigen type or smooth lipopolysaccharide. The ability of these strains to adhere to intestinal mucosa in vitro did not reflect on their ability to colonize the Peyer's patches, although strains of high in vitro adhesive ability appeared in greater numbers initially after oral challenge. Anti-O serum, ineffective in reducing the in vitro adhesive ability of virulent
S. typhimurium
, when given with the oral challenge prevented Peyer's patch colonization but was unable to prevent the appearance of a systemic infection. Anti-H serum, although effective in vitro in preventing adherence, had no effect in vivo. These experiments suggest that adhesiveness is neither essential nor sufficient for the virulence of
Salmonella
and that the usual development of a systemic infection after colonization of the small intestinal Peyer's patches may be subverted by the presence of O-antibody.
Interferon production in glia and glioma cell lines
Authors: Larsson I , Landström L E , Larner E , Lundgren E , Miörner H , Strannegård L
Published: January 2020

Interferon (IF) was produced in glia and glioma cell lines in titers comparable to those produced by human fibroblasts. It was inducible by both Sendai virus and polyriboinosinic:polyribocytidylic acid. "Superinduction" resulted in up to 500-fold-higher titers of IF. The IF appeared to be of the fibroblast type, as revealed by experiments using heat treatment, assay of antiviral activity in heterologous cell lines, and neutralization with specific antisera. Since large amounts of IF may easily be produced with glioma cell lines, such cells may be suitable for mass production of IF.
Heightened Resistance of Athymic, Nude (nu/nu) Mice to Experimental
Pseudomonas aeruginosa
Ocular Infection
Authors: Hazlett L. D. , Berk R. S.
Published: January 2020

BALB/c-derived athymic, nude (nu/nu) mice exhibited a heightened natural resistance to experimental corneal infection with
Pseudomonas aeruginosa
when compared with their heterozygote (nu/+) littermates. Stereomicroscopic examination of the eyes of nu/nu mice 24 h after corneal trauma and topical bacterial application revealed slightly cloudy corneas (iris visible), whereas nu/+ littermate corneas were opaque (iris not visible). Nu/+ mice failed to resolve the infection, and endophthalmitis and shrinkage of the infected eye occurred in these mice within 2 weeks after experimental
Pseudomonas
infection, as in the parent BALB/c strain. However, nu/nu mice, similarly infected, resolved the infection within 24 h and never exhibited full corneal opacity or eye shrinkage. Histological examination of the corneas of nu/nu mice 24 h after experimental wounding and bacterial application demonstrated subepithelial capillaries and a few polymorphonuclear neutrophils (with numerous intracellular bacteria) in the central cornea. In contrast, the equivalent corneal areas of infected nu/+ littermates, examined similarly, showed a more striking neutrophilic response (but with few intracellular bacteria) to similar bacterial infection, as well as a lack of blood vessels within the central cornea. The central corneas of uninfected and saline control nu/nu mice also were observed. This area in nu/nu mice exhibited an infrequent polymorphonuclear neutrophil (with no intracellular bacteria) and capillaries similar in size and location to those described for experimentally infected nu/nu mouse corneas. Untreated and saline control nu/+ mice, on the other hand, lacked both vessels and polymorphonuclear neutrophils in the central cornea.
Microscopic characterization of rabbit lung damage produced by Pseudomonas aeruginosa proteases
Authors: Gray L , Kreger A
Published: January 2020

The intratracheal administration of highly purified Pseudomonas aeruginosa proteases (ca. 10 to 100 microgram) elicited extensive, grossly observable rabbit lung damage by 3 h postinjection. Light and electron microscopic characterization of the lesions revealed: (i) progressive injury and necrosis of type I epithelial cells and capillary endothelial cells from 3 h to 1 day postinjection, and progressively increasing accumulations of erythrocytes, plasma proteins, fibrin, and released type II epithelial cell lamellar bodies in alveolar lumina during that time period; (ii) progressively increasing accumulations of macrophages, but not of polymorphonuclear leukocytes, in alveolar lumina from 3 h to 6 days postinjection; (iii) progressive hyperplasia of type II epithelial cells from 12 h to 4 days postinjection; (iv) progressive infiltration of alveolar septa by mononuclear inflammatory cells (interstitial pneumonitis) from 2 to 6 days postinjection; (v) no loss of alveolar septal connective tissue and no damage to pulmonary arterioles and venules; and (vi) almost normal alveolar structure by ca. 8 days postinjection. The study revealed that the intra-alveolar hemorrhage, the injury and necrosis of alveolar septal cells, and the infiltration by mononuclear cells that have been reported to occur during human pseudomonas pneumonia can also be elicited by the experimental administration of pseudomonas proteases. Thus, the results support the idea that in vivo production and activity of P. aeruginosa proteases is important, at least in part, in eliciting the lung damage characteristic of pseudomonas pneumonia.
Mitomycin C stimulates production of a toxin in Shigella species that causes morphological changes in Chinese hamster ovary cells
Authors: Takeda Y , Okamoto K , Miwatani T
Published: January 2020

Shigella dysenteriae produced a toxin that caused morphological changes in Chinese hamster ovary cells which was different from the neurotoxin (Y. Takeda, K. Okamoto, and T. Miwatani, Infect. Immun. 18:546--548, 1977). It was found that the toxin was produced not only by S. dysenteriae but also by S. flexneri, S. boydii, and S. sonnei. The production of the toxin was stimulated by the addition of 1 microgram of mitomycin C per ml to the culture in the early logarithmic growth phase.
Binding of antigen-enzyme complex by antigen-binding cells as an approach for immunogdiagnosis
Authors: Modabber F Z , Bahr G M
Published: January 2020

An immunoenzyme method capable of detecting specific antigen-binding cells is developed. The method is based on the chemical coupling of the antigen to beta-galactosidase and binding of the conjugate to the corresponding antigen-binding cells. Human hydatid fluid was used as an antigen model, and cells derived from rabbits immunized with human hydatid fluid showed specific binding to human hydatid fluid-beta-galactosidase conjugate. The method is specific and reproducible and could be used as an immunodiagnostic test for various diseases in which little or no antibody is produced.
Large-Scale Production and Partial Purification of Mouse Immune Interferon
Authors: Osborne L. C. , Georgiades J. A. , Johnson H. M.
Published: January 2020

Large-scale production of high-titered (10
2.2
to 10
4
U/ml) immune interferon (type II) was carried out in roller cultures of mouse spleen cells by using the T-cell mitogen staphylococcal enterotoxin A. Precipitation of 90% of this interferon by 55 to 80% saturated ammonium sulfate resulted in a 20-fold concentration and a two- to sixfold purification. After application of this interferon to either bovine serum albumin (BSA)-Affi-Gel 10 or hydroxylapatite columns, 100% of the interferon activity was recovered. By BSA-Affi-Gel 10 chromatography, 7% of the recovered activity was not bound, 45% was eluted with pH gradient 5 to 7, and 48% was eluted with 1 M NaCl. The pH- and salt-eluted interferons from the BSA-Affi-Gel 10 column were purified 62- and 390-fold, respectively, when compared with the starting materials. Rechromatography of the pH- and salt-eluted interferon peaks from the BSA-Affi-Gel 10 column did not alter their elution patterns. Stepwise elution of interferon from the BSA-Affi-Gel 10 columns with buffers of various pH and salt contents also resulted in greater than 300-fold purification. Specific activities of up to 2 × 10
5
U of interferon per mg of protein were attained with either elution procedure from BSA-Affi-Gel 10 columns. By hydroxylapatite chromatography, 5% of the recovered activity was not bound, 20% was eluted with a salt gradient, and 75% was eluted with 30% glycerin. Purification was 107- and 16-fold, respectively, for the two fractions. Ultrogel AcA 34 chromatography of the interferon resulted in two peaks of activity, a major one with a molecular weight of approximately 40,000 and a minor peak of molecular weight 70,000 to 90,000. Thus, by different types of chromatography, immune interferon was found to be heterogeneous.
Antibody responses in the cerebrospinal fulid of cynomolgus monkeys after intracerebral inoculation with paramyxoviruses
Authors: Yamanouchi K , Sato T A , Kobune F , Shishido A
Published: January 2020

Cynomolgus monkeys with or without measles antibody were intracerebrally inoculated with measles or canine distemper viruses, and antibody responses in the cerebrospinal fluid (CSF) were investigated. In measles antibody-free monkeys, natural infection with wild measles virus or intracerebral inoculations with two attenuated measles vaccines evoked primary antibody responses to measles virus in the sera but not in the CSF. In measles-immune monkeys, intracerebral inoculation with the TYCSA strain of measles virus produced a significantly high titer of measles antibody in the CSF with a minimal rise in the serum antibody and resulted in a significant decrease in serum/CSF antibody ratios. Intracerebral inoculation of a neurotropic canine distemper virus, the Onderstepoort strain, into measles-immune monkeys caused production of both measles and distemper antibodies in the CSF. Inoculation of measles-immune monkeys intravenously with measles virus or intracerebrally with rubella virus, which has no antigenic relation to measles virus, failed to evoke a measles antibody response in the CSF. These results indicated that local production of measles antibody in the CSF was caused by a stimulus within the central nervous system of measles virus antigen or canine distemper virus antigen that partially cross-reacted with measles virus antigen as a secondary antibody response.
Effect of Growth Rate and Glucose Concentration on the Activity of the Phosphoenolpyruvate Phosphotransferase System in
Streptococcus mutans
Ingbritt Grown in Continuous Culture
Authors: Ellwood D. C. , Phipps P. J. , Hamilton I. R.
Published: January 2020

Streptococcus mutans
Ingbritt was grown anaerobically in a chemostat with a glucose limitation, as well as with an excess of glucose (amino acid limitation) at dilution rates (
D
) between 0.05 and 0.4 h
−1
(mean generation time = 12 to 1.5 h). The glucose-limited culture produced cells having 1.5- to 6.0-fold greater glycolytic activity than the cells from the glucose-excess culture. The preferred substrate for these cells was glucose, with the glycolytic rate for sucrose being only slightly lower; the rate for fructose was half that of glucose. The glycolytic rate of the glucose-limited cells was maximum at
D
= 0.1 h
−1
, with a decline in rate as the growth rate approached
D
= 0.4 h
−1
. A comparison of the activity of phosphoenolpyruvate phosphotransferase system (PTS) in the two types of cells showed that the glucose-limited cells had 1.7- to 5.6-fold greater PTS activity for the three sugars than the glucose-excess-grown cells. Whereas little difference was seen between the three sugars with the latter cells, the glucose-PTS had the greatest activity with glucose-limited cells, with the maximum in cells grown at
D
= 0.1 h
−1
. Comparison of the rate of sugar uptake in the chemostat with the rate of PTS transport activity in the cells at each growth rate demonstrated that only under conditions of slow growth with a glucose limitation was the PTS system capable of supporting growth on glucose. Furthermore, PTS activity in cells grown with an excess of glucose was insignificant when compared with glucose uptake during growth in the chemostat. This evidence supports the observation that
S. mutans
possesses at least one other system, in addition to the PTS, for the transport of glucose into the cell. The organism was, however, devoid of glucose-proton symport transport activity.
Replication of herpes simplex virus type 1 in macrophages from resistant and susceptible mice
Authors: Lopez C , Dudas G
Published: January 2020

Studies were carried out to determine whether the in vitro capacity of adherent peritoneal cells to replicate herpes simplex virus type 1 (HSV-1) might correlate with the in vivo susceptibility of mice genetically resistant, moderately susceptible, or very susceptible to HSV-1 infection. Unstimulated and proteose peptone-stimulated monolayers restricted viral replication when infected immediately, but replicated HSV-1 when infected after 3 to 7 days of culture. Macrophages from resistant C57Bl/6 mice restricted HSV-1 replication significantly better than cells from susceptible mice. This function did not segregate with resistance, since macrophages from resistant F1 mice failed to restrict HSV-1 replication. Induction of peritoneal exudate cells with thioglycolate yielded cells capable of replicating HSV-1 when infected immediately after plating and after 4 days of culture.
Strain specificity of opsonins for group B streptococci types II and III
Authors: Shigeoka A O , Hall R T , Hill H R
Published: January 2020

Strains of types II and III group B streptococci do not appear to be uniformly susceptible to opsonization by antibody-containing human sera, as studied using both a chemiluminescence and a radiolabeled bacterial uptake technique. We could not demonstrate a correlation of serum-sensitive or resistant strains with capsular antigen quantities, although serum absorption studies with whole organisms and HCl, trichloroacetic acid, and saline extracts indicated that the antibody to type-specific capsular polysaccharide is important in opsonizing both serum-resistant and serum-sensitive strains. Since trypsin treatment produced significantly enhanced opsonization of serum-resistant and serum-sensitive strains, proteins present on some group B streptococci may be important antiphagocytic factors.
Polymyxin B sulfate modification of bacterial endotoxin: effects on the development of endotoxin shock in dogs
Authors: From A H , Fong J S , Good R A
Published: January 2020

The effects of endotoxin (lipopolysaccharide [LPS]) on the pathogenesis of canine endotoxin shock were compared with those of LPS which had interacted with polymyxin B sulfate prior to administration. Both LPS and polymyxin B-modified LPS caused comparable early decreases in aortic blood pressure, leukocyte and platelet numbers, and serum complement levels. However, in dogs receiving polymyxin B-modified LPS the late hypotensive phase was significantly ameliorated and lethality was significantly decreased. These data indicate that polymyxin B-modified LPS, though significantly less lethal than unmodified LPS, was capable of major interactions with several components of the humoral defense system, and support the concept that such interactions are not determinative in the pathogenesis of canine endotoxin shock.
Serological studies on chemostat-grown cultures of Lactobacillus fermentum and Lactobacillus plantarum
Authors: Knox K W , Campbell L K , Broady K W , Wicken A J
Published: January 2020

Lactobacillus fermentum NCTC 6991 and Lactobacillus plantarum NCIB 7220 were grown in a chemostat in the diffusible fraction of complex medium at pH 6.0 with glucose limitation. Organisms grown at different dilution rates (D) were injected into rabbits, and the resultant antisera were examined for reactivity with antigens previously isolated from batch-grown organisms. For L. fermentum, antisera obtained on injecting cells grown at D = 0.5 h-1 contained a significantly higher level of antibody reacting with lipoteichoic acid and a lower level of antibody reacting with wall polysaccharide than did antisera obtained with slower-growing cells (D = 0.05 and 0.033). Antibodies to the cell wall polysaccharide were alpha-D-glucosyl specific and cross-reacted with dextran and alpha-D-glucosyl ribitol teichoic acid from L. plantarum. The immunogenicity of the ribitol teichoic acid and lipoteichoic acid components of L. plantarum was not influenced by injecting organisms grown at different rates. However, chemical and serological studies indicate that growth of L. plantarum in the diffusible fraction of complex medium results in a wall teichoic acid of lower glucose substitution. This apparently influences the specificity of the resultant antibodies so that some sera react much less with glucosyl-substituted lipoteichoic acid and dextran.
Depression of the antibody response in Pseudomonas aeruginosa-injected mice
Authors: Garzelli C , Colizzi V , Campa M , Bozzi L , Falcone G
Published: January 2020

Heat-killed Pseudomonas aeruginosa inhibit antibody response in C57BL/6 mice. The depression of this response is dependent on the dose of bacteria injected, on the time interval between microorganism injection and antigen administration, and on the nature of the antigen used. Cell transfer experiments provide evidence that suppressor cells are not operative in this model. Furthermore, the results show that P. aeruginosa induces a marked dose-dependent proliferation of spleen cells in vivo, and the in vitro targets of this proliferative effect are B lymphocytes. It is suggested that whole, heat-killed P. aeruginosa in vivo also behave as cell mitogens on B lymphocytes which, when strongly stimulated to proliferate, temporarily lose their capacity to mount a normal antibody response.
Effects of BCG infection on the susceptibility of mouse macrophages to endotoxin
Authors: Peavy D L , Baughn R E , Musher D M , Musher D M
Published: January 2020

Mice infected intravenously with Mycobacterium bovis (BCG) are 100 to 1,000 times more sensitive to the lethal effects of bacterial lipopolysaccharides (LPS). Since BCG infection results in macrophage activation and LPS may cause pathophysiological effects through interaction with this cell type, it was of interest to determine whether macrophages from BCG-infected animals were more susceptible to the toxic effects of LPS in vitro. When LPS-susceptible, C57BL/6 mice were infected with BCG, a significant reduction in the 50% lethal dose of LPS was first observed after 7 days and persisted for several weeks. Macrophages from these animals had greatly increased susceptibility to LPS in vitro, which correlated with the development of acquired cellular resistance as determined by their ability to inhibit the growth of Listeria monocytogenes. In contrast, BCG infection of C3H/HeJ mice, a strain resistant to LPS, did not alter the 50% lethal dose of LPS for these animals or increase the sensitivity of their peritoneal macrophages to LPS in vitro. These results indicate that susceptibility of BCG-infected mice to the lethal effects of LPS parallels the susceptibility of their macrophages in vitro; release of vasoactive substances from LPS-susceptible activated macrophages in vivo may be, in part, responsible for lethality.
Lecithin Changes in Murine
Mycoplasma pulmonis
Respiratory Infection
Authors: Pollack J. D. , Weiss H. S. , Somerson N. L.
Published: January 2020

We examined the lipid content of bronchoalveolar (BA) washes from both mice and rats infected with
Mycoplasma pulmonis,
an etiological agent of murine pneumonia. During a 30-day period after intranasal inoculation, the total lipid content from infected and control rats (in milligrams per animal) remained relatively equal and unchanged. The saturated, unsaturated, and total lecithin contents in infected rats (in milligrams per animal) all increased. The maximum lecithin values were detected at 7 to 10 days after infection; later, the levels fell to control values. There was essentially no change in any lecithin value from uninfected animals. Although in BA washes from infected animals the mass of disaturated lecithins increased, the percentage of this fraction in the total lecithin pool decreased. The fatty acids of the lecithins from BA washes of infected mice had significantly less palmitic and significantly more oleic and linoleic acids than the lecithins isolated from the BA washes of control animals. Both the relative decrease in the mass of disaturated lecithins in the BA washes and the increase in the percentage of esterified unsaturated fatty acids in the lecithins may be directly related to the reduced lung function reported to occur during the course of murine
M. pulmonis
pneumonia.
Inhibition of macrophage migration by muramyl peptides
Authors: Nagao S , Tanaka A , Yamamoto Y , Koga T , Onoue K , Shiba T , Kusumoto K , Kotani S
Published: January 2020

In the capillary tube migration system a synthetic muramyl dipeptide (MDP; N-acetylmuramyl-L-alanyl-D-isoglutamine), a part of bacterial cell wall peptidoglycans, inhibited the migration of peritoneal exudate macrophages from normal guinea pigs or rats. The migration inhibition was also caused by some MDP-containing peptidoglycan fragments from cell walls of Lactobacillus plantarum and Staphylococcus epidermidis. The migration inhibition could not be explained on the basis of macrophage migration inhibitory factor. A stereochemically highly specific structure of MDP required for its adjuvant activity was also required for the macrophage migration inhibition. These findings suggest that MDP and MDP-containing cell wall fragments may activate macrophages and that this activation may be important in the exertion of their adjuvant activity.
Suppression of local intestinal immunoglobulin A immune response to cholera toxin by subcutaneous administration of cholera toxoids
Authors: Hamilton S R , Yardley J H , Brown G D
Published: January 2020

Cholera toxin has been shown to modulate immune responses, generally producing enhancement when administered simultaneously with antigen and suppression when administered a day or more earlier. In a previous study using chronically isolated ileal loops in rabbits, we found that two subcutaneous (s.c.) "priming" and "boosting" doses of biologically active cholera toxin suppressed the local intestinal immunoglobulin A response to intraloop doses of cholera toxin. In the study reported here, two different biologically inactive but antigenically intact cholera toxoids, glutaraldehyde toxoid and choleragenoid, where administered s.c. by the same immunization schedule as for toxin in the earlier experiment. Suppression of local immune response to intraloop cholera toxin as compared with animals receiving no s.c. inoculations was again found. The results suggest that in this model suppression was immunological (mediated by an immunological mechanism) rather than toxigenic (mediated by biological activity of cholera toxin). In addition, the occurrence of suppression of local intestinal immune response after systemic immunization suggests that suboptimal protection against enteric infections could occur after s.c. vaccination.
Purification and properties of rabbit alveolar macrophage lysozyme
Authors: Carroll S F , Martinez R J
Published: January 2020

Lysozyme was isolated from Bacillus Calmette-Guerin-elicited rabbit alveolar macrophages by acid extraction and purified to homogeneity by a single-column procedure. Yields of the purified enzyme averaged between 20 and 30 mg per rabbit, values far in excess of those obtained with previously published methods. Rabbit lysozyme has a molecular weight of 14,300 and exhibits optimal lytic activity against Micrococcus lysodeikticus at an ionic strength of 0.04, pH 6.5. Our results indicate that lysozyme and other granule components can be fractionated from elicited alveolar macrophages by using simple techniques, suggesting methods for the bulk purification of lysosomal constituents.
Kinetics of different specific immunological parameters after rabies vaccination in mice
Authors: Blancou J , Andral L , Lagrange P H , Tsiang H
Published: January 2020

A significant protection to an intracerebral challenge of 70 mean lethal doses of a standard live rabies virus strain was obtained in BCG-pretreated mice or in normal mice which had been immunized with a single subcutaneous injection of a beta-propiolactone-inactivated rabies vaccine. Concomitantly, levels of delayed-type hypersensitivity (measured in vivo by the footpad test) and serum-neutralizing activity were evaluated at various times after immunization. All immune criteria were significantly augmented in the BCG-pretreated, rabies-immune mice as compared to normal, rabies-immune mice. However, peak levels of protection, delayed-type hypersensitivity, and serum-neutralizing activity did not occur at the same times. For instance, in the BCG-pretreated, rabies-immune mice, delayed-type hypersensitivity peaked on day 7, protection peaked on day 21, and serum-neutralizing activity peaked on day 60. In BCG-pretreated mice, which did not receive the rabies vaccine, positive delayed-type hypersensitivity, some protection, and serum neutralizing activity were observed 4 to 5 weeks after BCG pretreatment. The possible relationships between specific and nonspecific immunity provoked by rabies virus antigens, tissue culture cell-associated antigens (derived from the bovine fetal kidney cells in which the rabies virus was grown, and BCG are discussed.