bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of a streptococcal lipoteichoic acid on host responses in mice
Authors: Miller G A , Urban J , Jackson R W
Published: January 2020

A teichoic acid (TA) extracted from Streptococcus pyogenes 1-RP41 was previously shown to be an immunosuppressant under certain conditions (Miller and Jackson, 1973). The TA has now been shown to be a lipoteichoic acid composed of 40% glycerol, 20% alanine, 13% phosphorus, and 8% glucose, with a variable content of fatty acids. Teh presence of the polyglycerol phosphate backbone and fatty acid was required for maximum immunosuppression of the primary immunoglobulin M response to sheep cells. A complex, nonlinear, time-dependent dosage relationship in suppression of the anti-sheep erythrocyte response in mice was observed. TA depressed the anamnestic response to sheep cells in the mouse and could affect this response whether administered before the primary antigen challenge or immediately before the secondary challenge. In distinct contrast, TA enhanced antibody production to Escherichia coli O55:B5 lipopolysaccharide when assessed by counting plaque-forming cells or measuring antilipopolysaccharide serum titers. The TA failed to stimulate a large uptake of [3H]TdR by murine spleen cells; however, it significantly enhanced the clearance of carbon by the reticuloendothelial system.
Antibody-dependent cell-mediated cytotoxicity in cows: comparison of effector cell activity against heterologous erthrocyte and herpesvirus-infected bovine target cells
Authors: Rouse B T , Wardley R C , Babiuk L A
Published: January 2020

Bovine peripheral blood leukocytes (PBL) and cells collected from the bovine mammary gland were assayed for antibody-dependent cell-mediated cytotoxicity (ADCC) against chicken erythrocyte (CRBC) and bovine herpesvirus-infected bovine kidney cell targets. Bovine antisera were used to sensitize target cells. Both PBL and mammary leukocytes expressed ADCC, with the latter cell population having greater activity against both target cells. Only the CRBC target cells were killed by nonadherent PBL and phagocyte-depleted PBL. Nonadherent mammary leukocytes, rich in monocytes and macrophages, did kill virus-infected target cells. Carbonyl iron-treated mammary leukocytes failed to kill virus-infected targets but could destroy CRBC targets. Antimacrophage serum inhibited lysis of both CRBC and virus-infected targets, but antilymphocyte serum only inhibited CRBC killing. These observations indicated that at least two kinds of cells could mediate ADCC against CRBC but only cells of the mononuclear phagocytic series could kill virus-infected target cells. The herpesvirus-infected target cells became susceptible to ADCC 9 h after virus infection. A case is made for investigating the phenomenon of ADCC using in vitro systems that closely mimic the in vivo situation. The possible role of the ADCC mechanism as instrumental in causing recovery from herpesvirus infections is discussed.
Cytomegalovirus infection and immunity in renal allograft recipients: assessment of the competence of humoral immunity
Authors: Rytel M W , Balay J
Published: January 2020

Cytomegalovirus (CMV) infections are prevalent in renal allograft recipients. The purpose of this ongoing study is to attempt to elucidate the mechanism(s) responsible for the enhanced susceptibility to CMV infections on the part of transplant patients and for their apparent inability to eradicate the infection once it starts. The present report assesses the competence of humoral immunity to CMV in renal allograft recipients. The total study population was comprised of 41 renal allograft recipients (10 followed prospectively) and 38 age-matched control subjects. The overall CMV infection rate in renal allograft recipients was 90.2%, and in 11 cohort control subjects it was 45.5%. Active infection was present in 61.0% of transplant patients (24.0% of these had CMV disease) and in 18.2% of the cohorts. THESE DIFFERENCES ARE SIGNIFICANT. CMV complement fixation and neutralization antibody prevalence was similar in 10 patients with renal failure undergoing hemodialysis before transplantation and in 23 control subjects. There was similarly no difference in antibody response between allograft recipients in whom the infection was primary (eight subjects) or secondary (17 subjects). We conclude that despite immunosuppressant therapy (with azathioprine and corticosteroids), humoral immunity to CMV is not impaired in transplant patients with either a primary or secondary infection.
Rosette-forming cells during immune response to Toxoplasma gondii in mice
Authors: Masihi K N , Werner H
Published: January 2020

The rosette-forming cell (RFC) response of mice immunized with varying doses of Toxoplasma gondii was studied by immunocytoadherence (ICA). The specificity of ICA in the present system was tested by passive sensitization with hyperimmune serum in vivo and in vitro. A slight increase in RFC was observed with the latter. Prior treatment of spleen cells from immunized animals with rabbit anti-mouse immunoglobulin resulted in total inhibition of ICA. During the primary and the secondary response after 10 days, the number of RFC rose rapidly to reach the peak on the 3rd day. With secondary immunization 30 days later, the peak shifted to the 2nd day. Mice infected with a lower dose of Toxoplasma had a greater number of RFC during the secondary response after 10 days than with a larger dose.
Partial purification and characterization of a heat-labile enterotoxin of Escherichia coli
Authors: Schenkein I , Green R F , Santos D S , Maas W K
Published: January 2020

A partially purified enterotoxin was obtained from the growth medium of Escherichia coli strain 711 (P307), a derivative of E. coli K-12, by ultrafiltration, precipitation with ammonium sulfate, molecular sieving, and anion exchange column chromatography. The active moiety, which is heat-labile, behaved like a protein particle of 180,000 to 200,000 daltons during molecular sieving and ultracentrifugation. During polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE), it dissociated into two subunits with apparent molecular weights of 68,000 to 70,000 and 14,000 to 15,000. SDS-PAGE after heating in SDS changed the larger subunit to an apparent molecular weight of about 40,000; the smaller subunit did not change. The intact particle induced rounding of the cells in Y-1 mouse adrenal tumor cells used for assay. The detergent-dissociated molecules were not active. Proteolysis of the purified toxin by tolylsulfonyl phenylalanyl chloromethyl ketone-trypsin appeared to enhance its activity. The addition of serum to the assay medium resulted in partial depression of the activity. Activity was also abolished by preincubation of the toxin with either a rabbit antiserum to it or solutions containing GM1 ganglioside. The length of time needed to evoke a response in the assay system by fractions from different stages in the purification of the enterotoxin was a useful parameter in the evaluation of specific activity.
Regulation and extracellular glucosyltransferase production and the relationship between extracellular and cell-associated activities in Streptococcus mutans
Authors: Janda W M , Kuramitsu H K
Published: January 2020

The regulation of extracellular glucosyltransferase production in Streptococcus mutans GS-5 has been studied using a chemically defined medium. Most of the glucosyltransferase activity produced by cells grown in the chemically defined medium was extracellular, in contrast with the distribution between cell-associated and extracellular glucosyltransferase activity when cells were grown in complex medium. The production of extracellular glucosyltransferase activity coincided with the logarithmic growth phase, and further accumulation ceased when glucose was exhausted from the medium. Accumulation of extracellular glucosyltransferase activity was inhibited immediately by chloramphenicol and rifamycin, added either at the beginning of growth or during mid-logarithmic growth. Low concentrations of chloramphenicol inhibited both cellular protein synthesis and the accumulation of extracellular glucosyltransferase activity to the same extent, indicating a close coupling between glucosyltransferase synthesis and secretion. Experiments using cell lysates showed that no intracellular accumulation of glucosyltransferase activity occurred in the presence of the inhibitors and that the intracellular activity is very low relative to the cell-surface activity. The utilization of cells depleted of cell-associated glucosyltransferase activity indicated that most of the cell-associated glucosyltransferase activity does not act as a precursor for the extracellular enzyme. Sugar analogue inhibitors of glycoprotein synthesis did not have any specific effects on the synthesis or secretion of extracellular glucosyltransferase activity.
In vitro activity of guinea pig transfer factor released into plasma
Authors: Paquet A , Olson G B , Jeter W S
Published: January 2020

Plasma fractions and plasma dialysate from 2,4-dinitrochlorobenzene- and tuberculin-sensitive guinea pigs that had been treated with either antilymphocytic serum or normal control serum were analyzed for their ability to transfer lymphocyte transformation, passive cutaneous anaphylaxis, and macrophage migration inhibition, as well as delayed hypersensitivity in vivo. Antilymphocytic serum caused rapid release of material, which has characteristics of transfer factor, into the plasma. It was dialyzable, migrated electrophoretically with the alpha globulins and albumin, possessed a 280/260 (nm) optical density ratio of 0.7, and caused in vitro lymphocyte transformation in the presence of the specific antigen. Passive cutaneous anaphylaxis antibodies were also present in the plasma of sensitive animals, but they were isolated in electrophoretic or dialysis fractions separate from those containing transfer activity.
Inhibition of human neutrophil chemotaxis and chemiluminescence by amphotericin B
Authors: Björkstén B , Ray C , Quie P G
Published: January 2020

The effect of the antifungal drugs amphotericin B, fluocytosine, miconazole, griseofulvin, and nystatin on the chemotactic responsiveness of human neutrophils was studied. Amphotericin B in a concentration of 2 mug/ml inhibited chemotatic responsiveness, and in a concentration of 5 mug/ml it also inhibited chemiluminescence. The inhibition of chemotaxis could be reversed by washing the cells. The other antifungal drugs did not inhibit chemotaxis even in concentrations much higher than those obtained in human serum during treatment.
Interaction between Coxiella burnetii and guinea pig peritoneal macrophages
Authors: Kishimoto R A , Walker J S
Published: January 2020

The phagocytosis and subsequent degradation of phase I and II Coxiella burnetii by macrophages obtained from immune and nonimmune guinea pigs were compared. Phase I rickettsiae were more resistant to phagocytosis than were phase II organisms. There was no significant difference in the percentage of phagocytosis of either phase of rickettsiae by macrophages from immune or nonimmune animals. After ingestion, phase I and II organisms pretreated with normal serum multiplied and destroyed normal macrophages as well as macrophages obtained from guinea pigs immunized with phase II rickettsiae. In contrast, only phase I organisms were degraded by macrophages from phase I-immunized animals in the presence of normal serum. Immune serum rendered rickettsiae more susceptible to phagocytosis and also potentiated the destruction of organisms by all types of macrophages. The specificity of macrophages from phase I animals to degrade only phase I rickettsiae was demonstrated by the ability of Rickettsia rickettsii to replicate in these macrophages.
Serum-free culture of hamster lymphoid cells and differential inhibition of lipopolysaccharide stimulation by isologous serum
Authors: Streilein J S , Hart D A
Published: January 2020

The optimal conditions for serum-free cultures of hamster lymphoid cells were determined. The cells responded to both the nonspecific thymus-derived lymphocyte mitogen concanavalin A (Con A) and the nonspecific bone marrow-derived lymphocyte mitogens lipopolysaccharide (LPS) and dextran sulfate (DxS). Normal MHA hamster serum was shown to specifically inhibit the response to LPS and DxS by 95% without inhibiting the response to Con A. The serum also did not inhibit one-way mixed lymphocyte reaction. Incubation of cells in 5% serum for short periods of time, followed by serum-free culture, did not lead to inhibition. The serum inhibited the LPS response by 65% even when added 24 h after initiation of the culture. The activity was associated with the high-molecular-weight material on Sephadex G-200 fractionation of serum. The inhibitory fraction did not possess antibody activity to LPS. The possibility that the material is an alpha2-macroglobulin is discussed.
Interferon production in mice by components of Salmonella minnesota R595 lipid A
Authors: Schiller J G , Ribovich R , Feingold D S , Youngner J S
Published: January 2020

The lipid A precipitate obtained by partial hydrolysis in 1% acetic acid of the glycolipid of a mutant strain of Salmonella minnesota (rough strain R595) was separated into a number of components by thin-layer chromatography. When tested as bovine serum albumin complexes, the minimal interferon-inducing dose in mice of the parent glycolipid was 0.1 mug, whereas that of lipid A was 1.0 mug. The minimal dose of the separated components ranged from 0.1 to 20 mug. The components did not differ significantly in regard to glucosamine-phosphate-fatty acid ratios. The results further emphasize that lipid A is a heterogeneous substance that contains a variety of components of differing biological activity.
Requirements for ingestion of Chlamydia psittaci by mouse fibroblasts (L cells)
Authors: Byrne G I
Published: January 2020

Ingestion of 14C-amino acid-labeled Chlamydia psittaci (6BC) by mouse fibroblasts (L cells) was inhibited when the host cells were incubated for 30 min at 37 degrees C in Earle salts containing 10 mug of crystalline trypsin per ml. Tryptic digestion also inhibited the ingestion of 1-mum polystrene latex beads. Trypsin-treated L cells almost completely recovered their ability to ingest chlamydiae after 4 h at 37 degrees C in medium 199 with 5% fetal calf serum. Cycloheximide (10 mug/ml) blocked this recovery. Heating 14C-amino acid-labeled C. psittaci for 3 min at 60 degrees C inhibited its ingestion by L cells, whereas inactivating it with ultraviolet light was without effect on the ingestion rate. These results show that efficient ingestion of C. psittaci by L cells involves trypsin-labile sites on the host and heat-sensitive sites on the parasite. The failure of excess unlabeled infectious C. psittaci to promote the ingestion of 14C-labeled heat-inactivated chlamydiae suggests that direct interaction between these two sites must occur for uptake to proceed normally.
Susceptibility of mice treated with cyclophosphamide to lethal infection with Leptospira interrogans Serovar pomona
Authors: Adler B , Faine S
Published: January 2020

Mice not normally susceptible to infection with Leptospira interrogans serovar pomona were rendered susceptible to lethal infections by treatment with a single dose of 300 mg of cyclophosphamide (Cy) per kg administered optimally from 4 days before to 1 day after infection. Cy-treated mice with either passively or actively acquired antibody were protected from death. Blood levels of leptospires in infected untreated and in Cy-treated mice remained similar until 2 days after infection, when untreated mice cleared the leptospires. Soon afterwards, opsonizing and agglutinating antibody appeared. Cy-treated mice given spleen cells from other normal or specifically immune mice were protected from infection. An important factor in the natural resistance of mice to leptospiral infection appears to be their capacity to produce circulating antibody within 48 to 72 h. Applications are suggested for this animal model in vaccination and protection studies.
Purification and properties of streptococcal hyaluronate lyase
Authors: Hill J
Published: January 2020

Hyaluronate lyase (hyaluronidase) has been purified and characterized from a group A type 4 Streptococcus. Production of the enzyme was favored by growth in trypsinized veal infusion in the presence of hyaluronate oligosaccharide and tetrasaccharide. Detectable enzymatic activity was diminished in the presence of N-acetylglucosamine and glucuronic acid. Purification of hyaluronate lyase consisted of 40 to 60% ammonium sulfate precipitation, diethylaminoethyl A-50 Sephadex ion-exchange chromatography, gel filtration with G-200 Sephadex, and adsorption to Sepharose 6B. Purified enzyme was antigenically homogeneous and free of proteinase, deoxyribonuclease, streptolysin 0, and streptokinase. Active hyaluronate lyase was recovered from neutral polyacrylamide gels, and it appeared to be a glycoprotein. A single band was detected by sodium dodecyl sulfate-acrylamide electrophoresis, which had a molecular weight of approximately 50,000. A molecular weight of 70,000 was observed by gel filtration. The purified enzyme had a Km of 3.8 x 10(-4) and a pH optimum of 6.0. Reducing agents increased the activity of crude enzyme at least threefold and were necessary to prevent inactivation of the purified enzyme.
Model delineating the effects of a salivary pellicle on the adsorption of Streptococcus miteor onto hydroxyapatite
Authors: Gibbons R J , Moreno E C , Spinell D M
Published: January 2020

A model describing the adsorption isotherms for Streptococcus miteor strain 26 to untreated and saliva-treated hydroxyapatite was developed. The strengths of the adsorption bonds in the two systems were similar, however, the presence of selectively adsorbed salivary components increased the number of available binding sites fourfold.
Immunity to Mycobacterium leprae infections in mice stimulated by M. leprae, BCG, and graft-versus-host reactions
Authors: Shepard C C , Van Landingham R , Walker L L
Published: January 2020

Infections of mice with Mycobacterium leprae in one rear foot pad immunized them against a second infection in the other rear foot pad. Purified bacilli harvested from the first infection also produced immuniy when injection into the foot pads of previously uninfected mice. Injections of BCG afforded similar protection, but had no adjuvant effect on M. leprae. M. duvali, a cultivable mycobacterium that is reported to be more closely related antigenically to M. leprae than BCG is, provided much less protection against M. leprae challenge than BCG did. Moreover, when M. duvali was mixed with BCG, it was not any more effective than BCG alone. Graft-versus-host reactions, induced by injections of parental spleen cells into F1 hybrids, provided no protection against M. tuberculosis and M. marinum challenge. They gave moderate protection against M. leprae in one experiment but not in another with a different schedule. Allogenic spleen cells had a protective effect when injected locally into the infected foot pad. The effect produced by these injections of spleen cells was a delay in the appearance of bacterial growth; however, there was no decrease in the rate of logarithmic growth when it did appear and no reduction in the eventual plateau level.
In vivo distribution of Pseudomonas aeruginosa slime glycolipoprotein: association with leukocytes
Authors: Lynn M , Sensakovic J W , Bartell P F
Published: January 2020

Intraperitoneal injection of slime glycolipoprotein (GLP) from Pseudomonas aeruginosa induced leukopenia and death of mice, similar to the effect of infection with viable organisms. Differential counts established that the leukopenia was characterized by a decrease in the number of polymorphonuclear leukocytes, followed by death of mice. Mice immunized with GLP survived challenge and responded with a leukocytosis that had a substantial increase in circulating polymorphonuclear leukocytes. Leukocytes from GLP-injected mice were agglutinated by anti-GLP serum, indicating an association between GLP and leukocytes. Other results indicated that 14C-labeled GLP is deposited mainly in the liver. Normal leukocytes labeled with 51Cr were injected intravenously into mice receiving an intraperitoneal injection of GLP. As with GLP, the 51Gr-labeled leukocytes were sequestered in the liver. These results indicate that GLP enters the blood stream and becomes associated mainly with neutrophils, and that the neutrophil-GLP complex is deposited in the liver, possibly accounting for the leukopenia in mice.
Adjuvant activity of synthetic N-acetylmuramyl peptides in rats
Authors: Tanaka A , Saito R , Sugiyama K , Morisaki I , Kotani S
Published: January 2020

The minimal adjuvant-active structure in bacterial cell wall peptidoglycan for the induction of delayed hypersensitivity in rats (WKA) was the N-acetylmuramyl dipeptides (N-acetylmuramyl L-alanyl-D-isoglutamine and N-acetylmuramyl L-alanyl-D-glutamic acid).
Magnesium-dependent adenosine triphosphatase as a marker enzyme for the plasma membrane of human polymorphonuclear leukocytes
Authors: Harlan J , DeChatelet L R , Iverson D B , McCall C E
Published: January 2020

The adenosine triphosphatase (ATPase) activities of human polymorphonuclear leukocytes (PMNL) were studied with an assay that monitored the release of 32P-labeled inorganic pyrophosphate (32P1) from gamma-[32P]adenosine 5'-triphosphate (ATP). In cell homogenates, (Na+ + K+)-sensitive, ouabain-inhibitable ATPase comprised an insignificant fraction of the total ATPase activity. Additions of p-nitrophenyl phosphate and beta-glycerophosphate (substrates for nonspecific acid and alkaline phosphatases) and of tartrate (inhibitor of acid phosphatase) gave no indication of inhibition. This suggested that the assay was relatively specific for ATP hydrolysis. The activity was found to have a pH optimum of 8.7 and a Km for ATP of 0.6 mM. There was an absolute requirement for Mg2+, with other divalent cations substituting less efficiently. When the Mg2+-dependent ATPase activity of intact cells was compared with that in homogenized cells, no significant difference was observed. The activity in intact cells was linear with respect to incubation time up to at least l0 min. Trypan blue staining and lactate dehydrogenase assays revealed that greater than 92% of the PMNL remained intact and viable during the assay. No soluble ATPase was released from the cells under assay conditions. In following the distribution of gamma[32P]ATP and 32P2 counts became cell associated. Since the experimental evidence supports the observation that PMNL remain intact and viable and that ATP does not penetrate the cell under assay conditions, it is proposed that greater than 90% of the Mg2+-dependent ATPase of the human PMNL is associated with a plasma membrnae enzyme. This would qualify the enzyme for the role of a plasma membrane marker for future fractionation and isolation attempts.
Relationship of pili to colonial morphology among pathogenic and nonpathogenic species of Neisseria
Authors: McGee Z A , Dourmashkin R R , Gross J G , Clark J B , Taylor-Robinson D
Published: January 2020

Growth in colonies with type 1 morphology and the presence of pili are characteristics that have been associated with virulence of gonococci for humans. To determine whether the presence of pili per se might be responsible for colony type 1 morphology, the relationship of pili to colony type was examined in various species of Neisseria. Short pili (175 to 210 nm in length) were seen only on nonpathogenic neisseria, whereas long pili (up to 4,300 nm) were seen on organisms of both nonpathogenic and pathogenic species. Although long pili, similar to those found on organisms from high-domed, type 1 colonies of gonococci, were observed on organisms from high-domed, type 1 colonies of nonpathogenic Neisseria species, they were also observed on low-convex, type 4 colonies of meningococci and nonpathogenic neisseria. Among meningococci there was no difference in the morphology of colonies consisting of organisms with many long pili and colonies consisting of organisms that completely lacked pili. Thus, there was no consistent relationship of pili to colonial morphology. Unless the pili of N. gonorrhoeae are unique among Neisseria species in their influence on colonial morphology, it is likely that factors other than pili determine colony type 1 morphology of gonococci. Whether these same factors, either alone or in conjunction with pili, are also responsible for gonococcal virulence warrants further investigation.
JC virus, a human polyomavirus associated with progressive multifocal leukoencephalopathy: additional biological characteristics and antigenic relationships
Authors: Padgett B L , Rogers C M , Walker D L
Published: January 2020

JC virus, a human polyomavirus, failed to grow or produce cytopathic effects in any of a variety of cells tested other than primary human fetal glial (PHFG) cells. Cells tested included other primary human cells and glial cells from other animals. Only a rare cell in inoculated insusceptible human cell cultures produced T or virion antigen. In PHFG cell cultures JC virus produced subtle cytopathic effects, and the majority of progeny remained cell associated. Only a few cells in the heterogenous PHFG cell cultures contained T antigen at 24 h postinoculation, and virion antigen was not detected until 48 h postinoculation. The infectivity of JC virus was resistant to inactivation by ether and by heating at 50 degrees C for 1 h. A three-way minor antigenic relationship was demonstrated among the virion antigens of JC virus, BK virus, and simian virus 40 by neurtralization and/or hemagglutination inhibition tests. Serological evidence is presented for the existence of JC virus as a distinct entity before the use of simian virus 40-contaminated poliovirus vaccines and for the nonexistence of an animal reservoir for JC virus infection.
Antibacterial product of peritoneal exudate cell cultures from guinea pigs infected with mycobacteria, listeriae, and rickettsiae
Authors: Sharma S D , Middlebrook G
Published: January 2020

In an in vitro model of cellular immunity, the antibacterial product of immunologically mediated mononuclear cell activation was studied from guinea pigs infected with listeriae and rickettsiae and compared with the product previously described from animals infected with mycobacteria. We found that this product, active against gram-positive bacilli, appeared to be identical in the three different infections with regard to its heat stability, its chromatographic adsorption and elution pattern, its susceptibility to inactivation by proteolytic enzymes, and its antibacterial spectrum
Nutrition and enterotoxin synthesis by enterotoxigenic strains of Escherichia coli: defined medium for production of heat-stable enterotoxin
Authors: Alderete J F , Robertson D C
Published: January 2020

A defined medium has been developed which supports synthesis of heat-stable enterotoxin (ST) by porcine and bovine strains of enterotoxigenic (ENT+) Escherichia coli in levels equivalent or better than a complex Casamino Acids-salts medium. The medium components did not support production of heat-labile enterotoxin (LT) but were similar for ST synthesis by ENT+ strains producting only ST and those which produced ST in addition to LT. The amino acids in Casamino Acids found to be necessary for growth and enterotoxin synthesis were proline, serine, aspartic acid, and alanine. Maximal growth and toxin levels were obtained after 8 h of incubation. Improved growth, but not an increase in synthesis of ST, was observed in the presence of Mg2+, Mn2+ and Fe3+ compared with Mg2+ alone. A chelator, tricine, was necessary for maximal cell densities,, probably to solubilize trace ions and make them more available to the bacteria. Increased growth was observed upon addition of glucose to both complex and defined media; however, glucose as well as gluconate and pyruvate appeared to cause repression of toxin synthesis. Addition of vitamins, oleic acid, or DL-lactic acid to the defined medium slightly increased levels of ST.
Mitogenic activity of cell wall components from smooth and rough strains of Brucella abortus
Authors: Kreutzer D L , Scheffel J W , Draper L R , Robertson D C
Published: January 2020

On the basis of [3H]thymidine incorporation by normal mouse spleen cell cultures, cell wall preparations from a smooth (45/0) strain and a rough (45/20) strain of Brucella abortus were strongly mitogenic. On the other hand, none of several subcomponents extracted from the cell wall preparations, including aqueous and phenolic lipopolysaccharides, was active. These results contrast with the marked mitogenic activity of lipopolysaccharides isolated from other gram-negative bacteria such as Salmonella typhimurium.
Cellular immunity to toxoplasma and besnoitia in hamsters: specificity and the effects of cortisol
Authors: Lindberg R E , Frenkel J K
Published: January 2020

The inhibitory effects of cortisol on cellular immunity were studied in vitro by using hamster peritoneal exudate cells. Two obligate, intracellular protozoa--Toxoplasma gondii and Besnoitia jellisoni-- were used to control for specificity of effects. Results indicate that immune lymphocytes specifically confer immunity to (or "arm") macrophages that specifically express immunity. This arming can be inhibited by 5 microng of cortisol per ml. Macrophages that have been armed already will continue to express immunity (by limiting parasite growth specifically) in the presence of 5 microng of cortisol per ml. Cortisol levels of 20 microng/ml are required to inhibit the expression of immunity by armed macrophages. It was also found that lymphocytes, from hamsters given 20 mg of cortisol subcutaneously 2 days before the harvest of cells, did not arm macrophages, whereas macrophages from these same animals could be armed by immune lymphocytes from untreated hamsters. Therefore, it was concluded that in relation to cellular immunity, lymphocytes are more sensitive to cortisol than are macrophages. Since antibody to these parasites is almost always present in vivo, we also tested the effects of cortisol on the disposition of antibody-modified organisms by activated (not armed) macrophages, and found that 50 microng of cortisol per ml was needed to inhibit macrophage effects on antibody-treated organisms.
Susceptibility of Lipopolysaccharide Mutants to the Bactericidal Action of Human Neutrophil Lysosomal Fractions
Authors: Rest Richard F. , Cooney Marjorie H. , Spitznagel John K.
Published: January 2020

Acetate extracts of purified human neutrophil granules (a mixed population containing specific and azurophil granules) were dialyzed against phosphate-buffered saline (pH 7.0) and tested for bactericidal activity against smooth parent and rough mutant, gram-negative bacteria. Rough (Re) mutants of
Escherichia coli, Salmonella typhimurium,
and
Salmonella minnesota
were exquisitely more sensitive to extracts of human polymorphonuclear leukocyte granules than were their smooth (S) parents. The mean lethal dose (LD
50
) for the parent strains was 25 to 50 μg of granule protein per ml. As much as 500 μg of extract per ml failed to kill 100% of the S parents. The LD
50
for the rough mutants was 1.5 to 2.0 μg of the same granule extract per ml; 100% killing occurred with 5 to 10 μg of lysosomal protein per ml. Conditions affecting the growth of the bacteria greatly affected their sensitivity to the granule extracts. Granule extract killed bacteria grown with aeration to log phase 10 to 15 times more efficiently than the same bacteria grown to stationary phase under static conditions. The bactericidal incubation mixture also influenced results, in that greater killing occurred with tryptone than with phosphate or
N
-2-hydroxyethyl piperazine-
N
′-2-ethanesulfonic acid-buffered saline. Bactericidal activity depended on lysosomal protein concentration, time, and temperature. Boiled lysosomal fractions failed to kill the S parents but retained 20 to 50% of their ability to kill the Re mutants. Parents (smooth) were killed more efficiently at pH 5 to 6, whereas their Re mutants were killed more efficiently at pH 7 to 8.
Effect of treatment with BCG on the course of visceral leishmaniasis in BALB/c mice
Authors: Smrkovski L L , Larson C L
Published: January 2020

Intravenous inoculation of BCG was found to be both prophylactic and therapeutic in BALB/c mice against challenge with amastigotes of Leishmania donovani. Spleens and livers of mice inoculated with BCG maintained total parasite burdens at significantly lower levels when compared to controls. BCG administered intravenously 14 days prior to and on the same day of protozoan challenge was more protective than vaccine given 30 and 14 days prior to challenge. A level of 10(7) viable units of BCG provided more protection against challenge with parasites than did 10(6) viable units. BCG given the same route as the challenge dose of amastigotes provided more protection than if administered via some other route. BCG given to mice with an already established infection was shown to significantly reduce their parasite burdens.
Bacteriolytic activity in staphylococci
Authors: Satta G , Varaldo P E , Grazi G , Fontana R
Published: January 2020

The bacteriolytic activity of Staphylococcus aureus and Staphylococcus albus strains was tested with various media. Whereas S. aureus strains were found to be active under all conditions, the percentage of active S. albus strains was significantly influenced by the composition of the medium. Ionic strength and concentration of the organic nitrogen source were found to be the main factors affecting the expression of bacteriolytic activity of straphylococci. Virtually all of 318 S. aureus and 603 S. albus strains were active on a medium containing 3% peptone, 0.3% glucose, 0.2% yeast extract, 0.1% disodium phosphate, 2.2% sodium chloride and 0.9% agar. The optimal conditions for the bacteriolytic activity of S. aureus strains were different from those of most S. albus strains. Within S. albus, optimal conditions differed also from strain to strain. It is suggested that further studies on this subject may prove useful for the identification and taxonomy of staphylococci. A possible relationship between the production of extracellular bacteriolytic enzymes and pathogenic properties of staphylococci is also considered.
New models of chronic synovitis in rabbits induced by mycoplasmas: microbiological, histopathological, and immunological observations on rabbits injected with Mycoplasma arthritidis and Mycoplasma pulmonis
Authors: Cole B C , Griffiths M M , Eichwald E J , Ward J R
Published: January 2020

A dose-dependent chronic synovitis was induced in rabbit knees after the intra-articular injection of both Mycoplasma arthritidis and Mycoplasma pulmonis. The inflammation progressed from an initial acute phase at 1 week characterized by edema, infiltration of the synovium with monocytes and heterophils, and desquamation of lining cells, to a more chronic phase at 1 and 3 months, in which villus hyperplasia, lymph "nodules," mononuclear cell infiltration, fibroplasia, and collagen deposition were prominent. With one exception, mycoplasmas could no longer be cultivated from the joints 1 month postinoculation. Both mycoplasma species evoked a humoral antibody response that was more marked in synovial fluids than in peripheral blood. A cell-mediated immune reaction, as evidence by enhanced uptake by [3H]thymidine by sensitized blood, spleen, or node lymphocytes in the presence of homologous antigen, was detected only in rabbits injected with M. pulmonis. Lymphocytes taken from arthritic rabbits were no more cytotoxic toward synovial cells derived from normal or arthritic rabbits than were normal lymphocytes. The models of synovitis described in this study offer a convenient probe for determining the mechanisms of mycoplasma-induced inflammation, since they require only a single injection of the initiating agent and, in addition, utilize an animal host large enough for detailed investigation into the nature of mycoplasma/synovium interactions.
Biological Activities of Lipopolysaccharides and Lipid A from
Rhodospirillaceae
Authors: Galanos C. , Roppel J. , Weckesser J. , Rietschel E. T. , Mayer H.
Published: January 2020

The lipopolysaccharides and free lipid A from several strains of
Rhodospirillaceae
were assayed comparatively with those of
Enterobacteriaceae
in a number of biological tests. Free lipid A's from
Rhodopseudomonas gelatinosa
and
Rhodospirillum tenue
exhibited strong serological cross-reactions with each other and with free lipid A from
Salmonella.
Lipid A's from
Rhodopseudomonas viridis
and
Rhodopseudomonas palustris
, although cross-reacting with each other, did not do so with either the lipid A of
R. gelatinosa
or
R. tenue
or with that of
Salmonella.
The presence or absence of the above cross-reactions agreed with corresponding similarities or differences in the chemical structure of the lipid A preparations. The lipopolysaccharide of
R. gelatinosa
was highly toxic for adrenalectomized mice and pyrogenic for rabbits; however, it exhibited no anti-complementary activity. The activity of the
R. tenue
lipopolysaccharide was very low in both the lethality and pyrogenicity tests. Its corresponding free lipid A also exhibited low pyrogenic activity; however, its lethal toxicity for adrenalectomized mice was considerably higher than that of the intact parent lipopolysaccharide. Both intact lipopolysaccharide and, unexpectedly, the free lipid A exhibited no anti-complementary activity. The lipopolysaccharides of
R. viridis
and
R. palustris
were virtually nontoxic for mice and nonpyrogenic for rabbits. Both lipopolysaccharides were highly potent in their interaction with complement. They therefore represent the first example of nontoxic lipopolysaccharides exhibiting high anti-complementary activity.
Solid-phase radioimmunoassay for the detection of rotavirus
Authors: Middleton P J , Holdaway M D , Petric M , Szymanski M T , Tam J S
Published: January 2020

A solid-phase radioimmunoassay method has been developed for the detection of rotavirus in the form of a purified antigen and in stool. The parameters of the radioimmunoassay were examined and optimized to give high sensitivity and same-day results. Compared with electron microscopy, the assay is up to 10 times as sensitive for detection of the virus in stool and up to 128 times as sensitive for detection of a purified virus antigen. In a field study on stool specimens it was at least as efficient as electron microscopy.
Common antigen of Mycobacterium leprae, M. lepraemurium, M. avium, and M. fortuitum in comparative studies using two different types of antisera
Authors: Kronvall G , Closs O , Bjune G
Published: January 2020

No. 21 mycobacterial antigens of Mycobacterium lepraemurium, M. avium, M. fortuitum, and M. leprae were compared in crossed immunoelectrophoresis using two different antibody sources, a serum pool from lepromatous leprosy patients (LSII) and a rabbit anti-M. smegmatis antiserum. M. lepraemurium, like M. avium, was found to contain the 21 A and 21 C determinants. M. fortuitum contained in addition a new type of determinant, 21 D.M. leprae antigen no. 21 carried the A as well as the B dertminants, the latter found so far only in the leprosy bacillus. The separate taxonomic position of M. leprae, suggested by earlier studies of the no. 21 antigen, is further supported by the present results, which also demonstrate the potential use of submolecular heterogeneity for such investigations.
Humoral immune response to the heat-labile enterotoxin of Escherichia coli in naturally acquired diarrhea and antitoxin determination by passive immune hemolysis
Authors: Evans D J , Ruiz-Palacios G , Evans D E , DuPont H L , Pickering L K , Olarte J
Published: January 2020

Acute- and convalescent-phase sera from 132 students attending a university in rural Mexico were assayed for antibody against the heat-labile enterotoxin (LT) of Escherichia coli by neutralization of LT activity in the Y-1 adrenal cell assay and by passive immune hemolysis of LT-sensitized sheep erythrocytes. The two titration methods produced comparable results with respect to antitoxin responses detected. An inverse relationship was found between acute geometric mean antitoxin titer and the occurrence of diarrhea associated with LT-producing E. coli, especially in newly arrived students from the U.S.A. A significant correlation (P less than 0.00 5) was found between a rise in antitoxin titer detectable by the passive immune hemolysis technique and diarrhea with LT-producing E. coli isolated. Thus, humoral antitoxin titers appear to be a useful indicator of immune status with respect to enterotoxigenic (LT) E. coli diarrhea.
Preparation of arthritogenic hydrosoluble peptidoglycans from both arthritogenic and non-arthritogenic bacterial cell walls
Authors: Kohashi O , Pearson C M , Watanabe Y , Kotani S
Published: January 2020

Cell wall lytic enzyme (Kyowa lytic no. 2 enzyme) liberated arthritogenic hydrosoluble peptidoglycans from both arthritogenic and non-arthritogenic bacterial cell walls. From these cell walls, mutanolysin (peptidoglycan-degrading enzyme) also liberated hydrosoluble peptidoglycans which, however, lacked arthritogenicity. Based on the chemical composition of these peptidoglycans, it was suggested that their arthritis-inducing ability depends on a relatively long chain of glycan units that consists of repeated units of N-acetylglucosaminyl-N-acetylmuramic acid. However, the glycan chain lengths on these peptidoglycans appeared to be related to their adjuvancy rather than to an antigen(s) responsible for development of arthritis in rats.
Gastrointestinal antibody responses in axenic mice to topically administered Escherichia coli
Authors: Ebersole J L , Molinari J A
Published: January 2020

Immunoglobulin levels were determined in gastrointestinal secretions of both conventional and axenic mice, as was the stability of the immunoglobulin classes in these secretions. Axenic animals were then administered nonviable Escherichia coli O111:B4 by topical application into the oral cavity. Immunoglobulin (Ig) A, IgG1, AND IgG2 were detected in the gastrointestinal secretions, with an apparent gradation in stability (IgA greater than IgG1 greater than IgG2) under the conditions investigated. Specific antibodies (IgA and IgG) to both the somatic and the flagellar antigens of E. coli were demonstrated. The time course of the responses to the polysaccharide and protein antigens suggested that a secondary response to lipolysaccharide was being observed in the secretions. Examination of the autoclaved diet fed to the germfree animals indicated the presence of antigenically intact polysaccharide materials (i.e., lipopolysaccharides), whereas antigenic bacterial proteins were uniformly undetectable. Therefore, endotoxin in the diet was apparently causing a primary type of sensitization in the mice, whereas the active oral immunization regimen induced a secondary antibacterial response in the secretions.
Biological and physicochemical properties of the lipopolysaccharide of Chromatium vinosum
Authors: Hurlbert R E , Hurlbert I M
Published: January 2020

The lipopolysaccharide (LPS) of Chromatium vinosum has anticomplementary activity. This anticomplementary activity is destroyed by alkaline digestion of the LPS and is suppressed by both Mg2+ and Ca2+ ions. Treatment of the LPS with ethylenediaminetetraacetic acid, sodium deoxycholate, or dimethyl sulfoxide did not affect its toxicity toward mice; however, alkaline-treated LPS was not toxic. Treatment of the LPS with sodium deoxycholate, dimethyl sulfoxide, or sodium dodecyl sulfate resulted in reversible dissociation into subunits. Aggregation of the subunits into the original form was achieved by removing the dispersing agent by dialysis against distilled water followed by freezing and thawing. Electron micrographs of phenol-extracted LPS showed long filaments. Electron micrographs of sodium deoxycholate- and sodium dodecyl sulfate-treated and dialyzed LPS showed a mixture of small subunits and short filaments, whereas dimethyl sulfoxide-treated and dialyzed LPS contained only small ovoid spheres. The LPS produced an ordered series of multiple bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A similar banding pattern was observed for Salmonella abortus-equi and Proteus mirabilis LPS. The C. vinosum LPS appears to be mitogenic for mouse spleen cells.
Tubular structures associated with acute nonbacterial gastroenteritis in young children
Authors: Kimura T , Murakami T
Published: January 2020

We clearly demonstrated a fine morphology of the tumbular structures antigenically related to the reovirus-like particles associated with acute nonbacterial gastroenteritis in young children. The tubular structures with a caplike structure on each end seemed to be complete forms, 75 to 80 nm in width and approximately 1,000 nm in length.
Lung Bacterial Clearance in Murine Pneumococcal Pneumonia
Authors: Ansfield Michael J. , Woods Donald E. , Johanson W. G.
Published: January 2020

We studied the bactericidal capacity of the rat lung during the development of pneumococcal pneumonia. Pneumonia was produced in a lower lobe by the intrabronchial instillation of 10
4
Streptococcus pneumoniae
cells in buffer. Lung bacterial counts progressively increased, reaching 10
7
per lung within 48 h, and the increase was associated with localized atelectasis and consolidation. Bacterial multiplication was inhibited with tetracycline at various intervals after infection, and the subsequent clearance of pneumococci was determined. Viable pneumococci were rapidly killed by lung defenses if bacterial multiplication was inhibited within 12 h of the onset of infection. No change occurred in the bacterial populationif tetracycline was delayed until 24 h after infection, indicating that pneumococcal killing by lung defenses had ceased. This effect could be reproduced with the addition of pneumococcal capsular polysaccharide to the inoculum, which produced a dose-related inhibition of pneumococcal clearance. The clearance of
S. epidermidis
was not impaired in the presence of pneumococcal pneumonia or by administration of exogenous capsular polysaccharide. These data indicate that pneumococcal pneumonia causes a marked impairment in lung antipneumococcal defenses within 24 h of the onset of infection. This acquired defect in antibacterial defenses may be due to the accumulation of pneumococcal capsular material in the lungs of infected animals.
Clostridium botulinum type D toxin: purification, molecular structure, and some immunological properties
Authors: Miyazaki S , Iwasaki M , Sakaguchi G
Published: January 2020

Clostridium botulinum type D progenitor toxin was purified. The addition of ribonucleic acid to the whole culture helped initial acid precipitation of the toxin. As with type B, both L (16S) and M toxins (12S) obtained from a hemagglutinin-positive strain, whereas M toxin only was produced by a hemagglutinin-negative strain. M toxin (molecular weight, 300,000) consisted of one molecule each of a toxic (molecular weight, 170,000) and a nontoxic component (molecular weight, 130,000); L toxin consisted of both components plus hemagglutinin. The specific toxicity of M toxin was 5 X 10(8) mean lethal doses per mg of N; that of L toxin was 2.4 X 10(8) mean lethal doses per mg of N. These toxins were fully or nearly fully active, but in un-nicked form. Trypsinization caused nicking in the toxic component, forming a molecule made up of two peptide chains with molecular weights of 110,000 and 60,000; there was little or no increase in toxicity. The toxic component of type D was not antigenically related to that of type C, whereas the nontoxic component was antigenically indistinguishable from that of type C. The toxicities of both L nad M toxins of the hemagglutinin-positive strain were increased twofold by trypsinization. Neither toxin contained the C2 toxic factor elaborated by C and D strain.
Direct evidence for the presence of lipopolysaccharide components in Pseudomonas ribosomal vaccine
Authors: Lieberman M M
Published: January 2020

The presence of sugars specific to lipopolysaccharide, glucose, and rhamnose was demonstrated in a Pseudomonas ribosomal vaccine. The detection of these sugars was accomplished by radiological means after paper chromatography of the neutral fraction of acid-hydrolyzed vaccine.