bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhanced superoxide anion release from phagocytes by muramyl dipeptide or lipopolysaccharide
Authors: Kaku M , Yagawa K , Nagao S , Tanaka A
Published: January 2020

Muramyl dipeptide (MDP) and lipopolysaccharide (LPS) from Escherichia coli were tested for the ability to influence superoxide anion (O2-) release from guinea pig phagocytes. Both MDP and LPS alone did not, by themselves, stimulate O2- release by macrophages and polymorphonuclear leukocytes. However, the preincubation of macrophages with MDP or LPS primed the macrophages to release an enhanced amount of O2- when stimulated by cytochalasin E and wheat germ agglutinin. When polymorphonuclear leukocytes were treated in the same way, only LPS showed an enhancing effect. MDP enhanced NADPH oxidase activity of macrophages, which is probably the reason for enhanced O2- release by MDP.
Tumor-dependent resistance of rat peritoneal macrophages to herpes simplex virus
Authors: Bonina L , Iannello D , Merendino R , Arena A , Mastroeni P
Published: January 2020

By their position at sites of initial infection and their wide distribution in major organs of the body, macrophages may be decisive in determining the susceptibility or resistance of the host to virus infection. Macrophage restriction of virus replication has been shown to be closely related to virus strains or virus types and to the age of the infected host. We report the effects of the development of a solid tumor in rats on intrinsic in vitro macrophage activity against herpes simplex virus type 1. The results obtained with the infectious center assays and the analysis of single-cycle growth curves of herpes simplex virus type 1 in macrophages obtained from normal and tumor-bearing rats showed a depression of antiviral activity of macrophages from tumor-bearing rats. The possibility of immunomodulation by bacterial adjuvants on tumor-bearing rats and the effects on the antiviral activity of peritoneal macrophages were furthermore demonstrated.
Structure and biological properties of solubilized envelope proteins of Bordetella pertussis
Authors: Robinson A , Hawkins D C
Published: January 2020

The structure and biological properties of solubilized envelope proteins of Bordetella pertussis have been examined. Several envelope proteins were found to be specific for phase I strains of B. pertussis and could be isolated by selective detergent extraction. These proteins had molecular weights of 90,000, 86,000, 81,000, 33,000, 31,000, and 30,000 and were reduced or absent in envelope preparations from Bordetella bronchiseptica, Bordetella parapertussis, or phase IV strains of B. pertussis. When the envelope preparations from phase I B. pertussis were assayed in the mouse intracerebral protection test they were found to be highly protective, and there was a strong correlation between the protective potency and the lymphocytosis-promoting factor (LPF) content of different preparations. Treatment with glutaraldehyde reduced the LPF activity, toxicity, and protective potency of the envelope extracts. Similarly affinity chromatography of envelope proteins on columns of haptoglobin coupled to Sepharose 4B reduced both the LPF content and the protective potency. The addition of a small amount of purified LPF to the haptoglobin-treated proteins restored the protective potency. The LPF by itself was nonprotective, indicating a potentiating role of LPF in the mouse intracerebral challenge test.
Induction of bacteremia in newborn rats by Escherichia coli K1 is correlated with only certain O (lipopolysaccharide) antigen types
Authors: Pluschke G , Mercer A , Kusećek B , Pohl A , Achtman M
Published: January 2020

A total of 95 Escherichia coli strains (O1:K1, O7:K1, or O18:K1), obtained from different sources of human infections and from healthy individuals, were analyzed for the ability to cause bacteremia after colonizing the gut of newborn rats. Strains of all three serotypes were able to multiply extensively in the gut after oral inoculation and to translocate (in small numbers) to the mesenteric lymph nodes. With only few exceptions, O7:K1 and O18:K1 strains were able to cause bacteremia, while O1:K1 strains could not. Mixed-infection experiments revealed that the bacteria present in the blood during a case of bacteremia are in most cases the descendants of one cell that has multiplied extraintestinally after translocation to the mesenteric lymph nodes. It appears that virulent O7:K1 and O18:K1, but not avirulent O1:K1, bacteria are able to multiply directly in the bloodstream of the newborn rats. No correlation between virulence and the source of isolation of the different strains was observed. Disease isolates thus do not seem to differ from fecal isolates of the same serotype in special virulence properties. The differences in virulence among different O serotypes of K1 E. coli observed in the rat model were comparable to their relative frequency of isolation from meningitis in newborn children.
Humoral immune response in chinchillas to the capsular polysaccharides of Streptococcus pneumoniae
Authors: Giebink G S , Schiffman G
Published: January 2020

Vaccines made from the capsular polysaccharides of Streptococcus pneumoniae have been shown to reduce the incidence of pneumococcal disease in certain populations and have recently been evaluated for their ability to elicit protection against experimental pneumococcal otitis media in a chinchilla model. In this study, chinchillas were vaccinated with a dodecavalent preparation of pneumococcal capsular polysaccharides (PCP) to obtain more information on the immunogenicity of these polysaccharide antigens. All 12 PCP types elicited an antibody response, but the optimum PCP dose and the kinetics of the antibody response varied among types. Immunological paralysis was demonstrated with an immunogenic dose of PCP after primary immunization with a large PCP dose (25 micrograms or more). Pertussis vaccine acted as neither an immunoadjuvant nor an immunosuppressant in the serum antibody response to type 7F PCP in chinchillas.
Platelet aggregation by Streptococcus pyogenes
Authors: Kurpiewski G E , Forrester L J , Campbell B J , Barrett J T
Published: January 2020

Heat-killed group A Streptococcus pyogenes induced platelet aggregation in platelet-rich plasma. Aggregation was dependent upon the ratio of platelets to bacteria, with maximal aggregation occurring at 0.8 platelets per bacterium (final concentration, 300,000 per microliter). Inhibition of the reaction by 3 mM EDTA indicated it was a true aggregation and not merely adhesion and agglutination. Lactic acid dehydrogenase assays indicated lysis of platelets did not occur during a 6-min incubation period. Aggregation was inhibited in a dose-dependent manner by acetylsalicylic acid (100 microM to 10 mM) and quinacrine (15.6 to 250 microM), with no decrease in aggregation at the lowest concentration of inhibitor tested. S. pyogenes induced the release of [14C]serotonin, which was maximal (50%) at 2.4 min, when aggregation was nearly complete. Gel-filtered platelets were not aggregated unless fibrinogen (final concentration, 1.8 mg/ml) was included in the reaction mixture. Staphylococcus aureus, a group B streptococcus, and Escherichia coli were unable to induce aggregation in platelet-rich plasma under the conditions used for S. pyogenes.
Inhibition of Streptococcus mutans by the lactoperoxidase antimicrobial system
Authors: Thomas E L , Pera K A , Smith K W , Chwang A K
Published: January 2020

Inhibition of bacterial metabolism by the lactoperoxidase (LP)-hydrogen peroxide (H2O2)-thiocyanate system was studied with representatives of serotypes a through g of Streptococcus mutans. The aims were to determine whether the amount of H2O2 released from these catalase-negative bacteria is sufficient to activate the LP system and whether these oral bacteria are resistant to inhibition by the LP system, which is active in human saliva. When the washed, stationary-phase cells were incubated aerobically with LP, thiocyanate, and glucose (Glc), greater than 90% inhibition of Glc utilization and lactate production was obtained with strains that released large amounts of H2O2 (BHT, FA-1, OMZ-176); 20 to 50% inhibition was obtained with strains that released about half as much H2O2 (B-13, Ingbritt); and no inhibition was obtained with strains that released only small amounts of H2O2 (AHT, HS-6, GS-5, LM-7, OMZ-175, 6715-15). Inhibition was most effective at pH 5, whereas release of H2O2 and accumulation of the inhibitor (hypothiocyanite ion) were highest at pH 8. With H2O2-releasing cells from early stationary phase, preincubation with Glc abolished inhibition, though it did not influence H2O2 release. Cells harvested 24 h later were depleted of sulfhydryl compounds. Inhibition of these cells was abolished by preincubation with Glc and certain sulfhydryl or disulfide compounds (reduced or oxidized glutathione, cysteine or cystine). This preincubation increased cell sulfhydryl content but had no effect on H2O2 release. All strains were inhibited when incubated with LP, thiocyanate, and added (exogenous) H2O2. Smaller amounts of H2O2 were required to inhibit at pH 5, and larger amounts were required to inhibit cells preincubated with Glc or with Glc and the sulfhydryl or disulfide compounds. The results indicate that pH, amount of H2O2, cell sulfhydryl content, and stored-carbohydrate content determine susceptibility to inhibition.
Role of Adherence in the Pathogenesis of Neonatal Group B Streptococcal Infection
Authors: Broughton Robert A. , Baker Carol J.
Published: January 2020

The ability of group B streptococci to attach to buccal epithelial cells from adult volunteers, healthy neonates, and infants with invasive group B streptococcal infection was assessed by using
3
H-labeled bacteria incubated at a bacteria-to-cells ratio of 1,000:1. Type III group B streptococcal clinical isolates adhered significantly better to the epithelial cells of healthy neonates than to those of adults (mean bacteria per cell of 31 versus 7, respectively;
P
< 0.005). In contrast, no statistically significant differences in adherence of type Ia or type II strains to cells of neonates and adults were noted. The adherence of strains isolated from 15 infants with invasive group B streptococcal infection was significantly greater to the cells of infected infants than to those of age-matched controls (mean bacteria per cell of 39 versus 18, respectively;
P
< 0.005). In contrast, no significant difference was noted in the adherence of a usually adherent type Ia strain and a nonadherent type III strain to the cells of infected infants compared with control infants. These results indicate that the serotype of group B streptococci with the greatest virulence for neonates (type III) adheres better to neonatal than to adult epithelial cells. Infants who develop invasive infection may have an increased number of epithelial cell surface receptor sites for attachment of group B streptococci, the bacteria may elaborate products which unmask receptor sites, or both.
Repression of the Virulence of
Yersinia pestis
by an F' Plasmid
Authors: Zsigray Robert M. , Lawton William D. , Surgalla Michael J.
Published: January 2020

An F-
lac
plasmid from
Escherichia coli
was transferred to virulent
Yersinia pestis
, resulting in the repression of virulence. The
Y. pestis
F-
lac
clones retained all of the known virulence traits but were avirulent and calcium independent. Every
lac
segregant derived from the F-
lac
clones was fully virulent and calcium dependent.
Chemical characterization of extracellular polysaccharides produced by Actinomyces viscosus T14V and T14Av
Authors: Imai S , Kuramitsu H
Published: January 2020

The human isolates Actinomyces viscosus T14V and T14Av produced extracellular polysaccharides in the absence of sucrose. In contrast to strain T14V, strain T14Av produced abundant viscous slime polysaccharide in the culture supernatant fluids when grown in a chemically defined medium containing glucose. After resolution of the T14Av polysaccharides into seven fractions, it was demonstrated that two of these exhibited viscous properties and lacked methylpentose and muramic acid. The major slime polysaccharide purified by gel filtration and ion-exchange column chromatography contained 39% (moles percent carbohydrate) galactose, 37% N-acetylglucosamine, 19% glucose, and 5% mannose. Only trace amounts of protein and phosphorus were detected in this preparation. On the other hand, strain T14V produced negligible slime polysaccharide under the same culture conditions. The major extracellular polysaccharide fraction from this strain contained methylpentoses, hexoses, hexosamines, muramic acid, protein, and phosphorus, suggesting that this fraction might be derived from the cell wall.
Attachment of enterotoxigenic Escherichia coli to human intestinal cells
Authors: Deneke C F , McGowan K , Thorne G M , Gorbach S L
Published: January 2020

An adherence test was developed with human ileal cells isolated from patients with a long-standing ileostomy by saline lavage through the stoma. Enterotoxigenic Escherichia coli isolated from humans bound to the human ileal cells to a greater extent (1.2 x 10(6) bacteria bound) than did control E. coli strains, including K99 pili antigen-producing strains (whether originally isolated from pigs or calves), the rabbit pathogen RDEC-1, the laboratory-derived nontoxigenic strain 334LL, or human normal fecal strains. However, K88 strains, either K88ab or K88ac, bound to the human ileal cells as well as did enterotoxigenic E. coli. Ileal cells isolated from two donors with different blood types behaved similarly. These cells remained viable and retained their binding ability for at least 3 days when stored in tissue culture medium at 4 degrees C.
Chemical composition and biological functions of Listeria monocytogenes cell wall preparations
Authors: Hether N W , Campbell P A , Baker L A , Jackson L L
Published: January 2020

A crude Listeria cell wall fraction, a purified fraction (PF) with demonstrated biological activity, as well as a third fraction of base-hydrolyzed PF (BHPF) were analyzed for chemical composition and activities not previously described. Listeria cell wall fraction and PF contained significant quantities of lipid, whereas BHPF was lipid depleted. Fatty acid compositions were typical of gram-positive bacteria. PF and BHPF were depleted in protein. Alanine, glutamic acid, diaminopimelic acid, glucosamine, and muramic acid were found in all fractions, in enhanced concentration in PF and BHPF, and with molar ratios typical of bacterial peptidoglycans. Major neutral sugars were rhamnose, ribose, ribitol, and glucose. The concentrations of rhamnose, ribose, and glucose were increased in BHPF. Differences in chemical composition of the fractions reflected differences in their biological activities: Listeria cell wall fraction induced resistance to Listeria infection, whereas PF did not. Mitogenic and adjuvant activities were demonstrated for Listeria cell wall fraction and PF but were lost in BHPF. BHPF retained the ability to induce macrophage-mediated tumoricidal activity and decrease resistance to Listeria infection.
Purification and some properties of a non-o1 Vibrio cholerae enterotoxin that is identical to cholera enterotoxin
Authors: Yamamoto K , Takeda Y , Miwatani T , Craig J P
Published: January 2020

Cholera-like enterotoxin was isolated and purified from the culture supernatant of a non-O1 strain of Vibrio cholerae, E8498, isolated from the environment. Enterotoxin was purified by aluminum hydroxide absorption and elution and successive gel filtrations on Sephadex G-100, Bio-Gel A-5m, and Sephadex G-75. Purified enterotoxin gave a single stained band on polyacrylamide gel disc electrophoresis, and the mobility was the same as that of cholera enterotoxin. The specific biological activity of the purified enterotoxin was almost the same as that of cholera enterotoxin in the Chinese hamster ovary cell assay, fluid accumulation in mouse ligated intestine, increase in vascular permeability in rabbit skin, and passive immune hemolysis. Sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis showed that the purified enterotoxin consisted of subunits A and B, identical to those of cholera enterotoxin, and Ouchterlony double gel diffusion tests indicated that the two toxins were immunologically identical. Enterotoxins prepared from several non-O1 strains isolated from human patients were also immunologically identical to cholera enterotoxin.
Effect of highly purified coagulase and culture filtrate on virulence and immunity of a coagulase-negative mutant of staphylococcus aureus BB
Authors: Hasegawa N , Kondo I , Hoshina S , Kurosaka K , Igarashi H
Published: January 2020

The virulence of the coagulase-deficient mutant BB-Cgl- 1301 (50% lethal dose [LD50] for mice by the intravenous route) was compared with that of its parental strain, Staphylococcus aureus BB. The BB strain produced free coagulase of serotype I, whereas the mutant 1301 did not. Mice were infected with strain 1301, alone or in combination with a highly purified coagulase type I or type II solution, or with concentrated culture filtrates of parent strain BB or mutant strain 1301. The ratios of the LD50S of 1301 and its combinations to that of BB ranged from 34.9 to 461. Combining strain 1301 with a concentrated culture filtrate of BB (BB-CF2.5) was the most effective for enhancement of its virulence. When mice were infected with a combination of strain 1301 and BB-CF2.5, the LD50 of strain 1301 (1.72 mg of cells [wet weight]) was decreased to 0.13 mg (1.3 x 10(8) CFU). This LD50 yielded the smallest ratio, 34.9, as compared with the LD50 of BB (0.00373 mg). In contrast, when the mice subcutaneously immunized with strain 1301 and BB-CF50 were intravenously challenged by strain BB, the LD50 for the immunized mice was 17.4 times the LD50 for the unimmunized control mice (0.0429 mg as compared with 0.00246 mg), indicating that combination was the most effective for enhancement of mouse immunization with strain 1301. However, combining strain 1301 with the highly purified sample of coagulase increased neither the virulence nor the immunizing power of mutant strain 1301.
Purification and characterization of a wall protein antigen from Clostridium botulinum type A
Authors: Takumi K , Takeoka A , Kawata T
Published: January 2020

A wall surface protein, designated antigen S, was extracted from Clostridium botulinum type A strain 190L with 0.1% Brij 58-2 M LiCl and purified sequentially by acetone pecipitation, ion-exchange chromatography, hydroxyapatite chromatography, chromatofocusing, and gel filtration. Crossed immunoelectrophoresis of the purified antigen S preparation against homologous multispecific antiserum to whole cells revealed only a single precipitin line. Antigen S had an apparent molecular weight of about 195,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigen was composed predominantly of acidic amino acid residues, and the isoelectric point was estimated to be at pH 4.75. Tryptic digestion of antigen S destroyed antigenic activity and produced one major polypeptide fragment with a molecular weight of about 44,000. Indirect immunoferritin labeling showed that antigen S was located on the outer layer of the cell wall.
Characteristics of Complement-Dependent Release of Phospholipid from
Escherichia coli
Authors: Wilson Lawrence A. , Spitznagel John K.
Published: January 2020

The release of
32
P-labeled bacterial phospholipid from a smooth
Escherichia coli
by serum components depends on complement activated by antibody. Phospholipid release in excess antibody tends to be proportional to the concentration of complement as does the release of other cellular constituents. Phospholipids are not simply stripped off during cell lysis. Whereas 94% of the total phospholipid freed from
E. coli
by mechanical lysis sediments at centrifugal forces sufficient to sediment molecules of 10
6
molecular weight, similar centrifugation sediments only 50% of the phospholipid released by antibody-complement. In fact, after mechanical lysis more than 50% of the phospholipid sediments at velocities sufficient to bring down cell envelopes. Although the bulk of the bacterial phospholipid is located in the cell envelopes, isolated
32
P-labeled cell envelopes and phenol-extracted lipopolysaccharide fails to release phospholipids in the presence of antibody-complement. Moreover, ethylenediaminetetraacetic acid, which like antibody-complement causes loss of cellular selective permeability and prepares
E. coli
cell walls for the action of lysozyme, releases only small amounts of phospholipid from
E. coli
and these are sedimentable. The most likely mechanism of phospholipid release caused by antibody-complement appears to be the activation directly or indirectly of an enzyme which is present only in the intact cells.
Epicillin: In Vitro Laboratory Studies
Authors: Basch H. , Erickson R. , Gadebusch H.
Published: January 2020

A new semisynthetic penicillin, structurally related to ampicillin, has been assigned the generic name epicillin, 6-[
d
-2-amino-2-(1, 4-cyclohexadienyl) acetamido]-penicillanic acid. The antimicrobial spectrum and level of activity of epicillin in vitro are similar to those of ampicillin. In studies with recent clinical isolates, these two antibiotics, when compared with carbenicillin, showed consistently higher antimicrobial activity against
Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli
, and
Proteus
species. When tested against
Pseudomonas aeruginosa
isolates, on the other hand, epicillin exhibited a level of intrinsic activity superior to that of ampicillin but less than that of carbenicillin. Epicillin is an amphoteric substance that is sensitive to penicillinase, is acid-stable, and is minimally, but reversibly, bound to human serum protein.
Effects of Intrarenal Inoculation of
Staphylococcus aureus
on Mice
Authors: Kondo Isamu , Masuda Shōgo , Kimura Kōzō , Kurosaka Kōsei , Hasegawa Noriko
Published: January 2020

The extreme susceptibility of the mouse kidney to infection with
Staphylococcus aureus
was confirmed by direct intrarenal (i.r.) inoculation with this organism. By applying Poisson's distribution formula to the results from infection of mice with small inocula (as few as one coccus or less per kidney), it was estimated that even three organisms could multiply in situ to produce abscess lesions, if the organisms were inoculated directly into the kidneys. This susceptibility of the mouse kidney for staphylococcal infection was not uniformly manifested with every strain, but correlated well with the virulence found through intravenous infection of the strains tested. The i.r. inoculation method by which infection of mice was easily established, using a small inoculum of
S. aureus
, similar to the amount suspected to occur in natural infections of man, is applicable for the analysis of the mechanisms of staphylococcal infections and any resulting immunity in man. The present paper describes the details and some experimental results obtained by our method.
Effect of Endotoxin on Pyruvate Kinase Activity in Mouse Liver
Authors: Snyder Irvin S. , Deters Michael , Ingle James
Published: January 2020

The pyruvate kinase (PK) activity of mouse liver increases after injection of endotoxin. It decreases in animals given cortisone alone and remains essentially unchanged in those given cortisone and endotoxin at the same time. CCl
4
causes an increase in liver PK activity, but neither it nor endotoxin changes the activity of muscle PK. Addition of octonoate to liver homogenates inhibits the activity of liver PK. These results suggest that the rapid depletion in liver glycogen after administration of endotoxin or CCl
4
may be related to increased PK activity. Induction of tolerance does not prevent the increase in liver PK activity in challenged animals.
Utilization of Palmitic Acid by
Mycobacterium avium
Authors: McCarthy Charlotte
Published: January 2020

Mycobacterium avium
accumulates
14
C-palmitic acid with saturation kinetics; the process is both temperature dependent and
p
H sensitive. The fatty acid is incorporated into triglyceride in vivo and the conversion is detectable within 5 min after exposure of the cells to
14
C-palmitic acid. The triglyceride is rapidly utilized because
14
CO
2
evolution from it begins within 30 min after
14
C-palmitic acid accumulation. Data from silicic acid column chromatography of extracts of cultures that have divided many times in medium containing
14
C-palmitic acid indicate that a large proportion of the cell lipid is triglyceride, but the radioactivity is widely dispersed among the other lipids. It is estimated that about 5% of the cell dry weight is triglyceride in a postexponential culture.
Effect of Adrenocorticosteroid on Glucose Metabolism in BCG-Sensitized Alveolar Macrophages
Authors: McCall Charles E. , Volk James V. , Cooper M. Robert , Dechatelet Lawrence R.
Published: January 2020

The increases in oxidation of glucose-
1
-
14
C
and glucose-
6
-
14
C
in BCG-sensitized alveolar macrophages are inhibited by administration of methylprednisolone sodium succinate. Both resting and phagocytizing cells are affected.
Hemopoietic Colony-Forming Cell Responses in Mice Infected with Ectromelia Virus
Authors: McNeill T. A. , Killen M.
Published: January 2020

The responses of in vitro colony-forming cells (granulocyte-macrophage progenitor cells) were studied in the bone marrow, spleen, and blood of mice after intraperitoneal injection of 10
3
plaque-forming units of ectromelia virus. This study showed that a colony-forming cell response occurred during infection and was accompanied by an increase in the serum level of colony-stimulating factor and, at a later stage, colony-inhibiting factor. Changes in the proportions of colony-forming cells were not due to relative changes in other cell populations. The relationships between colony-forming cell responses and levels of infection were complex, higher levels of infection being associated with good colony-forming cell responses in bone marrow and apparently poor colony-forming cell responses in blood and spleen.
Adjuvant Action of Bacterial Endotoxin and Colchicine on Antibody Formation in the Hamster
Authors: Merritt Katharine
Published: January 2020

Endotoxin was demonstrated not to act as an adjuvant in the primary antibody response in the Syrian hamster. Minimal adjuvant action was demonstrable in antibody formation to a second injection of bovine gamma globulin. The immunological status of the hamster towards the endotoxin did not alter this weak adjuvant action. It was confirmed that the antimitotic drug colchicine will act as an adjuvant in the primary antibody response to sheep red blood cells but not to bovine gamma globulin or endotoxin in the Syrian hamster.
Influence of
Escherichia coli
Lipopolysaccharide on Histamine-Forming Capacity and Antibody Formation in Lymphoid Tissues
Authors: Covert J. B. , Zarkower A.
Published: January 2020

Mice were injected intraperitoneally with 50 μg of
Escherichia coli
lipopolysaccharide (LPS). The spleen tissue exhibited increased histamine-forming capacity (HFC) 4 hr later. When this increased HFC was produced within 12 hr after intraperitoneal injection of sheep red blood cells (SRBC), there was an enhanced anti-SRBC antibody response in the spleen. In mice experiencing an increased spleen HFC at 36 hr after injection of SRBC, there was a significant decrease in the number of antibody-producing cells. The injection of histidine decarboxylase inhibitors simultaneously with the LPS greatly reduced the stimulation of HFC and also the adjuvant activity of the LPS.
Passive Protection of Mice Against Intraperitoneally Injected
Vibrio cholerae
by γG and γM Antibody
Authors: Pike Robert M. , Chandler Cleo H.
Published: January 2020

Fractions of rabbit anti-
Vibrio cholerae
serum containing γG antibodies were compared with fractions containing γM antibodies for their ability to protect mice against lethal infection resulting from the intraperitoneal injection of organisms suspended in mucin. About twice as much γG as γM (estimated by quantitative precipitation) was required to protect against approximately 1,000 50% lethal doses when the antibody was given intraperitoneally 4 hr before challenge. When protective serum fractions were given subcutaneously, however, the amount of γM required to protect was increased about 40-fold, whereas γG was about equally effective subcutaneously and intraperitoneally.
Biochemistry of
Vibrio cholerae
Virulence II. Skin Permeability Factor/Cholera Enterotoxin Production in a Chemically Defined Medium
Authors: Callahan Lynn T. , Ryder Richard C. , Richardson Stephen H.
Published: January 2020

A chemically defined medium capable of eliciting high titers of skin permeability factor (PF)/enterotoxin from three different strains of
Vibrio cholerae
has been developed. Toxin/antigen elaboration in synthetic and in complex media was monitored by a specific passive hemagglutination-inhibition test. A distinct temporal difference in the pattern of toxin/antigen elaboration was noted when the two types of media were compared. In complex media, PF activity and corresponding antigen release were coincident, whereas in the defined medium a biphasic pattern resulting in elaboration of nontoxic antigen during the second phase was seen. Possible reasons for the latter observation are discussed, and several experiments illustrating the unique utility of the defined medium are presented.
Immunity in Experimental Salmonellosis II. Basis for the Avirulence and Protective Capacity of
gal
E Mutants of
Salmonella typhimurium
Authors: Germanier R. , Fürer E.
Published: January 2020

Salmonella typhimurium
strains which are deficient in uridine diphosphate (UDP)-galactose-4-epimerase (
gal
E mutants) owe their outstanding protective capacity when used as live vaccine to the fact that when galactose is supplied exogenously, such as occurs in vivo, smooth cell wall lipopolysaccharides are synthesized. The mutants lose most of their protective capacity when this phenotypic curing is prevented by a second mutation of the kind found in strains LT
2
M
1
A (deficient in galactokinase) or E
32
(deficient in UDP-galactose-lipopolysaccharide transferase). Despite such phenotypic reversion, the
gal
E mutants are rendered avirulent as a result of galactose-induced bacteriolysis. Secondary mutants have been isolated which differ from each other with respect to the extent of galactose-induced lysis. The differences in galactose sensitivity are attributable to different activities of the other Leoloir pathway enzymes, namely, galactokinase and galactose-1-phosphate-uridyl transferase. The influence of these enzymes on lipopolysaccharide composition and galactose sensitivity and thus on virulence and immunogenicity of
gal
E mutants has been studied.
Characteristics of Streptolysin O Action
Authors: Oberley Terry D. , Duncan James L.
Published: January 2020

A study of the lysis of rabbit erythrocytes by streptolysin O (SO) revealed at least two steps in the hemolytic process. The initial interaction between SO and erythrocytes is the adsorption of the toxin molecule to the cell surface. Adsorption occurred at 4 C and was independent of ionic strength and
p
H; these results suggest that hydrophobic interactions between SO and the cell may be important in this process. Cholesterol was shown to prevent the adsorption of toxin to the cell, and it is proposed that cholesterol in the red cell membrane may be the site of toxin adsorption. The concept of a lipid attachment site is supported by the findings that proteolytic enzymes and sulfhydryl inhibitors known to affect external erythrocyte proteins did not affect SO hemolysis. Although the number of toxin molecules that will adsorb to a cell is limited, more than one toxin molecule was required for hemolysis. The step(s) following adsorption was dependent on temperature, ionic strength, and
p
H. Thus, it is evident that this step(s) is readily separable from adsorption, suggesting that an ionic interaction occurs between toxin and an erythrocyte membrane molecule. The step(s) following adsorption was also inhibited by divalent cations. Since
N
-ethyl maleimide will also inhibit lysis after toxin adsorption, it is possible that divalent cations may prevent SO hemolysis by reacting with free sulfhydryl groups on the toxin molecule.
Immunogenicity of Various Mycobacteria and the Corresponding Levels of Cross-Protection Developed Between Species
Authors: Collins F. M.
Published: January 2020

Seven mycobacterial species were tested for their ability to immunize mice against subsequent tuberculous challenge. The phylogenetic relationships between the vaccine and challenge strains had little direct influence on the level of resistance; a better correlation was found between the ability of the vaccine to survive or multiply in vivo and the immunogenicity of the organism.
Mycobacterium tuberculosis, M. bovis
(except for an SM
R
variant),
M. avium
, two of three
M. kansasii
strains, and one strain of
M. intracellulare
all survived or multiplied in vivo. They induced an effective resistance against a subsequent
M. tuberculosis
(Erdman) or BCG challenge. The SM
R
BCG, the Bostrom strain of
M. kansasii, M. scrofulaceum
, three strains of
M. intracellulare
, and one of
M. terrae
and
M. fortuitum
were unable to survive in vivo and were nonallergenic and nonimmunogenic. No evidence was found for an interference phenomenon between the atypical mycobacteria and the BCG vaccine sufficient to explain the inadequate levels of immunization reported in some field trials.
Antibodies Produced by Rabbits Immunized with Visna Virus
Authors: Karl Seung C. , Thormar Halldor
Published: January 2020

Immunization of New Zealand White rabbits with purified visna virus elicited antibody activity demonstrated by passive hemagglutination (PHA), complement fixation (CF), and indirect immunofluorescent tests. The antibody activities of hyperimmune sera and Sephadex G-200 fractions of the sera were studied. It was found that the PHA test was 10 to 100 times more sensitive than the CF test in detecting visna antibodies in rabbits. It was also found that the immunoglobulin M fractions from Sephadex G-200 filtration displayed greater PHA activity than did the immunoglobulin G fractions. Although neutralizing antibody was demonstrated in the serum of the natural host (sheep), our attempts to demonstrate neutralizing antibody in the sera from hyperimmunized rabbits (non-natural host) so far have failed.
Immunopathology of Experimental
Mycoplasma pneumoniae
Disease
Authors: Clyde Wallace A.
Published: January 2020

Experimental
Mycoplasma pneumoniae
pneumonia in the Syrian hamster resembles natural disease of man and is characterized by peribronchial round cells as well as exudates in the air passages. The nature of these intralumenal exudates was analyzed to assess host-parasite interaction at the respiratory epithelial surface, where infection with the mycoplasma is concentrated. Cells recovered by tracheobronchial lavage revealed changing patterns during the course of experimental disease; free macrophages decreased, polymorphonuclear leukocytes increased, and rosettes of epithelial cells surrounded by leukocytes appeared. Studies on formation of the rosettes in vitro with
M. pneumoniae
, sera, and cells suggested that complement-dependent immune phenomena were involved. The cellular events were maximal by 2 weeks after inoculation of normal hamsters. When previously infected animals were challenged, the course of infection was shortened, peribronchial infiltrates appeared rapidly, and development of exudates was both exaggerated and accelerated. These findings suggest that leukocytes entering the pulmonary parenchyma and lumen participate in the immune response to
M. pneumoniae
and contribute to host protection.
Mediation of Staphylococcal Adherence to Mucosal Cells by Lipoteichoic Acid
Authors: Carruthers Mary M. , Kabat William J.
Published: January 2020

Staphylococcal lipoteichoic acid markedly reduced adherence by
Staphylococcus aureus
to buccal cells in vitro, suggesting that lipoteichoic acid mediates adherence by that bacterium. Adherence inhibition by lipoteichoic acid was lost after deacylation of the preparation, suggesting that fatty acids on the molecule are essential to binding.
Immune-specific production of gamma interferon in human lymphocyte cultures in response to mumps virus
Authors: Nakayama T
Published: January 2020

The production of interferon (IFN) was investigated in human peripheral lymphocyte cultures stimulated by mumps virus. IFN-alpha and IFN-gamma were produced in lymphocyte cultures from immune donors. However, no IFN-gamma was produced in lymphocyte cultures from nonimmune donors. IFN-alpha was produced by B lymphocytes, and IFN-gamma was produced by T lymphocytes. An increase in the production of IFN-gamma resulted from the mixture of autologous macrophages in T-lymphocyte cultures. The production of IFN-gamma by T lymphocytes was found to depend on prior, possibly cellular, immunity.
Effect of methylamine and monodansylcadaverine on the susceptibility of McCoy cells to Chlamydia trachomatis infection
Authors: Söderlund G , Kihlström E
Published: January 2020

We used inhibitors of receptor-mediated endocytosis to study the mechanisms of infectivity, especially the uptake mechanism, of Chlamydia trachomatis for cultured cells. The effect of methylamine and monodansylcadaverine on the different stages of the chlamydial growth cycle in McCoy cells was examined. There was a dose-related decrease in the number of chlamydial inclusions in the presence of these agents. Monodansylcadaverine also decreased the chlamydia-dependent uptake of radioactive amino acids. The agents did not affect the attachment of chlamydiae to the cells, but they increased the protease-removable fraction of cell-bound chlamydiae. The amines reduced the number of inclusions when added at different times during the first 24 h of infection. However, this effect was influenced by host cell density, so that the effect of the amines at the early infectious phase was nullified in confluent monolayers, whereas, during later phases, the effect was comparatively independent of host cell density. This indicates that the amines have different modes of action at different infectious stages. The effect of the amines was reversible, and they had no effect on the infectivity of pretreated chlamydial elementary bodies. These experiments suggest that methylamine and monodansylcadaverine inhibit both the internalization of chlamydiae into McCoy cells and their intracellular development. These results are consistent with the hypothesis that chlamydiae utilize a constitutive cellular process, such as receptor-mediated endocytosis, to enter cells.
Sendai virus glycoproteins are T cell-dependent B cell mitogens
Authors: Kizaka S , Goodman-Snitkoff G , McSharry J J
Published: January 2020

UV-inactivated Sendai virus is mitogenic for murine splenocytes, whereas infectious Sendai virus kills spleen cells in vitro. The isolated hemagglutinin-neuraminidase (HN) and fusion (F) glycoproteins of Sendai virus are also mitogenic for cultured mouse spleen cells. A mixture of these glycoproteins (1 microgram/well) gives maximum stimulation 96 h after culture initiation. Viral proteins remaining insoluble after Triton X-100 extraction are also mitogenic for mouse spleen cells, with maximum stimulation occurring at 72 h after culture initiation with 1 to 5 microgram/well. On the basis of protein concentration, the HN and F glycoproteins are approximately three times more mitogenic than the Triton X-100-insoluble material. The mitogenic response of the HN and F glycoproteins has two components, a T cell-independent B cell proliferation, which is less than one-half of the total stimulation observed, and a T cell-dependent B cell proliferation. In contrast, the Triton X-100-insoluble material is a T cell-dependent B cell mitogen. Purified T lymphocytes do not respond to the mitogenic signal of either HN-F or Triton X-100-insoluble material.
Down regulation of hepatic glucocorticoid receptors after endotoxin treatment
Authors: Stith R D , McCallum R E
Published: January 2020

The mechanism by which endotoxin administration results in hypoglycemia was evaluated by characterizing [3H]dexamethasone binding and phosphoenolpyruvate carboxykinase activity in hepatic cytosol preparations from treated and control mice. Starved mice were given Escherichia coli O111:B4 endotoxin or saline intraperitoneally on day 3 after bilateral adrenalectomy. [3H]dexamethasone binding was measured by the charcoal method after the incubation of cytosol preparations with [3H]dexamethasone in the presence or absence of unlabeled dexamethasone. Changes in [3H]dexamethasone binding were found to be time and dose dependent in treated mice. When mice given different doses of endotoxin reached the same stage of morbidity, as indicated by the average time of death, significantly lower glucocorticoid binding was measured. Scatchard analysis of binding isotherms defined a single class of binding sites. Association and dissociation rate constants and the equilibrium dissociation constant (Kd) were not altered, but the maximum number of binding sites was depressed by endotoxin. The rank order of potency of competitors for [3H]dexamethasone binding, dexamethasone greater than hydrocortisone = corticosterone greater than deoxycorticosterone greater than progesterone greater than testosterone = estradiol, was consistent with a glucocorticoid receptor, although the competition was not altered by endotoxin. Endotoxin treatment prevented the glucocorticoid-induced increase in hepatic phosphoenolpyruvate carboxykinase activity. We conclude that the hypoglycemia of endotoxin poisoning is effected, in part, by the inhibition of the glucocorticoid-mediated induction of phosphoenolpyruvate carboxykinase via the down regulation of hepatic glucocorticoid receptors.
Activation of macrophages by quinonyl-N-acetylmuramyl dipeptide
Authors: Saiki I , Tokushima Y , Nishimura K , Yamamura Y , Azuma I
Published: January 2020

The effect of 6-O-QS-10-N-acetylmuramyl-L-valyl-D-isoglutamine methyl ester (quinonyl-MDP-66) on various functions of macrophages was examined. Mouse peritoneal macrophages, when treated either in vitro or in vivo with quinonyl-MDP-66 suspended in phosphate-buffered saline, showed a capacity for cytolysis and cytostasis against tumor targets and released H2O2 in the presence of phorbol myristate acetate. The macrophages induced by quinonyl-MDP-66 also had both antibody-dependent cell-mediated cytotoxicity and phagocytic activity against erythroid targets. The fact that synthetic quinonyl-MDP-66 stimulates the macrophages to become more cytotoxic than do other MDP analogs suggests that the lipophilic residue (QS-10) in quinonyl-MDP-66 may be important for the development of this activity.
Modification of the lipid moiety of the enterobacterial common antigen by the "Pseudomonas factor"
Authors: Kuhn H M , Neter E , Mayer H
Published: January 2020

Pseudomonas aeruginosa produces a factor (PF) which affects the enterobacterial common antigen (ECA); resulting in failure of the antigen to modify erythrocytes for hemagglutination by ECA antibodies. In the present study the nature of PF was determined. Pronase treatment abolished its activity, indicating the protein nature of PF. PF-treated ECA no longer coated erythrocytes but still reacted with ECA antibodies in immunoelectrophoresis tests with monospecific antiserum to ECA, although differences were noted between the precipitation patterns of PF-treated and untreated ECA. Therefore, PF does not significantly affect the antigenic determinant of ECA but rather affects its lipid carrier, an L-glycerophosphatide. Accordingly, differences in the sugar chain could not be detected by high-voltage paper electrophoretic examinations of partial hydrolysates of PF-treated and untreated ECA. PF liberates all fatty acids from ECA, similarly to commercial lipases, as evidenced by the liberation of unsubstituted glycerol upon HF degradation at 0 degrees C of PF-treated ECA. The lipase activity of PF is indicated also by the observation that a strain of P. aeruginosa with reduced lipase production and an exolipase-negative strain affect ECA either less or not at all. We conclude that PF is a lipase acting on the lipid moiety of ECA, which is responsible for the coating of erythrocytes, but not significantly on the serological determinant, the amino sugar chain.
In vitro propagation of antigen-specific T lymphocytes that adoptively transfer resistance to Listeria monocytogenes
Authors: Kearns R J , DeFreitas E C
Published: January 2020

Murine T cells generated against heat-killed Listeria monocytogenes or Listeria intracellular product (LIP) were propagated in a source of Interleukin 2. Both T-cell cultures were greater than 98% Lyt 1+, 2/3- and proliferated specifically against LIP and L. monocytogenes crude whole-cell antigen in vitro. Proliferation of both T-cell cultures required the presence of antigen and accessory cells syngeneic to the T cells at the left end of the major histocompatibility complex. The ability of these cultures to adoptively transfer protection against challenge with viable Listeria cells was dramatically different. As few as 10(6) LIP-specific T cells conferred significant protection against a lethal challenge of Listeria cells, whereas cultures induced against crude whole-cell antigen showed little or no protective function. The resistance conferred by LIP-specific T cells was specific in that the cells did not reduce the mortality seen after challenge with Salmonella typhimurium.
Cytomegalovirus infection of murine testicular interstitial Leydig cells
Authors: Baskar J F , Stanat S C , Huang E S
Published: January 2020

We studied the susceptibility of mouse testicular interstitial Leydig cells to cytomegalovirus both in vivo and in vitro. The in vivo studies included intratesticular and intraperitoneal infection of 6-week-old mice with murine cytomegalovirus (MCMV); the in vitro studies involved an MCMV-Leydig cell interaction using a Leydig tumor cell line (I-10). MCMV-specific antigens were detected in interstitial Leydig cells in sections of MCMV-inoculated testes by an indirect immunofluorescence test. MCMV DNA was also localized in the same testes cells derived from mice, which received intratesticular and intraperitoneal MCMV inoculations, respectively, by in situ DNA-RNA hybridization. Cytopathic effects were seen in MCMV-infected I-10 cell cultures 2 or more days after exposure to MCMV. The infected cells showed intranuclear inclusions characteristic of cytomegalovirus when stained with May-Grunwald-Giemsa stain. The indirect immunofluorescence test was also positive with MCMV-infected I-10 cells. MCMV DNA was detected in these cells by in situ DNA-RNA cytohybridization, and the presence of viral particles in MCMV-infected I-10 cells was confirmed by electron microscopy. Thus, we conclude that the interstitial Leydig cell is susceptible to MCMV infection both in vivo and in vitro.
Confirmation of association of protein I serotype of Neisseria gonorrhoeae with ability to cause disseminated infection
Authors: Cannon J G , Buchanan T M , Sparling P F
Published: January 2020

Previous work indicates that strains of Neisseria gonorrhoeae isolated in Seattle, Wash., and Atlanta, Ga., show an association between serotypes 1 and 2 of protein I of the outer membrane and the ability to cause disseminated infection (T.M. Buchanan and J.F. Hildebrandt, Infect. Immun. 32:985-994, 1981). By using the same serotyping system, we confirmed the association between those serotypes and both disseminated infection and serum resistance in strains from North Carolina. Some strains of the same serotype had protein I species with different apparent molecular weights.