bluuebunny/crossref_metadata_2025 huggingface dataset:
Surface proteins of typhus and spotted fever group rickettsiae
Authors: Osterman J V , Eisemann C S
Published: January 2020

Six proteins, previously established as major constituents of intact organisms, were identified in cell envelopes obtained from intrinsically radiolabeled Rickettsia prowazekii. Extrinsic radioiodination of intact organisms conducted at 0.5 micronM iodide indicated that protein 4 was the most peripheral, although protein 1 also had reactive groups exposed on the surface of the organisms. A 10-fold increase in iodide concentration resulted in labeling of protein 2, and at 50 micronM iodide, all six major proteins were radiolabeled. Similar selective labeling was not achieved with R. conorii. Analysis of both typhus and spotted fever group organisms radiolabeled with galactose suggested that carbohydrate was associated with proteins 1, 3, and 4. Typhus soluble antigen included all major proteins except protein 2, which remained attached to particulate rickettsiae after ether extraction. Protein 4 appeared to be prominent in the surface topography of R. prowazekii, was a component of soluble antigen and may have an important role in rickettsiae-host interactions.
Antigens of Streptococcus sanguis: purification and characterization of the b antigen
Authors: Appelbaum B , Rosan B
Published: January 2020

The antigen defining Streptococcus sanguis serotype 2 has been designated the b antigen. This antigen can be detected in extracts, obtained from whole cells by autoclaving (Rantz and Randall extraction), as a single precipitin band using a reference antiserum (M-5). However, the extract can also be shown to contain a teichoic acid using anti-polyglycerol phosphate serum. This teichoic acid does not contain the antigenic determinant for group H specificity. Studies of the b antigen have been hampered because of the difficulty in separating the b antigen from the teichoic acid using ion-exchange and molecular sieve chromatography. However, a relatively pure preparation has been obtained by affinity chromatography using anti-polyglycerol phosphate serum coupled to Sepharose. The isolated b antigen is a typical streptococcal cell wall polysaccharide composed of glucose, rhamnose, and N-acetylglucosamine in a molar ratio of 2.5:1.0:0.1. The antigen appears to have a single antigenic determinant closely related to isomaltose (glucose alpha-1,6-glucoside) based upon hapten inhibition studies.
Purification of lipoteichoic acids by using phosphatidyl choline vesicles
Authors: Silvestri L J , Craig R A , Ingram L O , Hoffmann E M , Bleiweis A S
Published: January 2020

Lipoteichoic acid (LTA) is a component of nearly all gram-positive membranes and recently has been found to be excreted into growth media by certain lactic acid bacteria. Cell-free extracts of LTA are usually contaminated with proteins, polysaccharides, and nucleic acids, thus causing problems to investigators studying the true biological function(s) of LTA. This report describes the preparation of purified extracellular LTA of Streptococcus mutans BHT and intracellular LTA of S. mutans AHT by three techniques: gel filtration, hydrophobic interaction chromatography, and adsorption to phospholipid vesicles. Gel filtration, the most commonly employed method for LTA purification, was found to remove nucleic acids, teichoic acids, and much polysaccharide while greatly concentrating LTA. But gross amounts of antigenic carbohydrate and protein remained associated with the LTA preparation. Hydrophobic interaction chromatography employing octyl Sepharose-4B allowed the separation of protein but not polysaccharide from partially purified BHT LTA preparations. By means of a new technique described in this paper, synthetic membranes (vesicles) were found to effectively separate all contaminants from the intracellular (AHT) and extracellular (BHT) LTA of S. mutans. This rapid method, on a comparative basis, proved to be the most effective approach for the purification of LTA from two widely differing sources.
Binding of aggregated human beta2-microglobulin to surface protein structure in group A, C, and G streptococci
Authors: Kronvall G , Myhre E B , Björck L , Berggård I
Published: January 2020

A novel mammalian-microbial "short circuit" has been demonstrated between aggregated human beta2-microglobulin and group A, C, and G streptococci. Bacteria belonging to nine gram-positive and three gram-negative species were tested for binding of radiolabeled beta2-microglobulin. All 10 individual strains of group A streptococci showed a high degree of reactivity with aggregated human beta2-microglobulin. Among 27 group C and 28 group G streptococci, 9 and 6 strains, respectively, were highly reactive, whereas the remaining strains showed a lower, but definite level of beta2-microglobulin binding. Of 11 group B streptococci, 4 were slightly positive. All strains among the other eight species were completely negative. Simultaneous testing of A, C, and G streptococci for immunoglobulin binding showed a lack of correlation between type II and III Fc reactivity and beta2-microglobulin binding. There was no inhibition of uptake of aggregated beta2-microglobulin to reactive strains when excess amounts of human immunoglobulin were added. The beta2-microglobulin-binding surface structure was found to be markedly sensitive to trypsin digestion. The relative trypsin resistance of the immunoglobulin-binding protein in the digestion experiments further demonstrated the dissociation between these two reactivities.
External layers of Rickettsia prowazekii and Rickettsia rickettsii: occurrence of a slime layer
Authors: Silverman D J , Wisseman C L , Waddell A D , Jones M
Published: January 2020

Using a simple specific-antibody stabilization procedure on organisms gently liberated from their host cells, we have demonstrated by electron microscopy that Rickettsia prowazekii and Rickettsia rickettsii possess a coat of variable thickness, external to the outer leaflet of the cell wall and the structure designated by others as a "microcapsule," which corresponds most closely to the slime layer of certain other bacteria. Reactions in the methenamine silver and ruthenium red staining procedures and the failure to be visualized by standard procedures suggest that the slime layer is largely polysaccharide in nature. It is postulated that this slime layer accounts in large part for the large, electron-lucent, halo-like zone which is found by electron microscopy to surround organisms of the typhus and spotted fever groups in the cytoplasm of their host cells, that it may be the locus of some major group-specific antigens, and that it may function as an antiphagocytic mechanism, as an aid for attachment of rickettsiae to potential host cells, or both. Moreover, because the attenuated E strain of R. prowazekii has been shown to possess a substantial slime layer, the basis for attenuation is not likely to be a simple smooth-to-rough variation.
Interaction of human serum and neutrophils with Escherichia coli strains: differences between strains isolated from urine of patients with pyelonephritis or asymptomatic bacteriuria
Authors: Björkstén B , Kaijser B
Published: January 2020

Strains of Escherichia coli isolated from the urine of children with pyelonephritis (n = 18) or asymptomatic bacteriuria (ABU) (n = 20) were serotyped and compared with respect to opsonic requirements, sensitivity to serum, amount of chemiluminescence produced by normal neutrophils during phagocytosis of the various strains, and amount of capsular antigen. Strains isolated from cases of ABU more often had the capacity to activate the alternative complement pathway than had strains causing pyelonephritis. The ABU strains were also better opsonized in fresh human serum and were more often serum sensitive than the pyelonephritic strains. There were individual variations in neutrophil chemiluminescence during phagocytosis of the various strains, but the two groups of strains did not differ significantly. The strains from pyelonephritis cases produced a greater amount of capsular antigens than the ABU strains. There was no correlation between presence of a particular O or K antigen and capacity to activate the alternative complement pathway. The observed differences between strains isolated from patients with ABU and those isolated from patients with pyelonephritis may be relevant for further studies on differences in bacterial virulence or host defense factors.
Characterization of the immunosuppression accompanying virus-induced avian osteopetrosis
Authors: Smith R E , Van Eldik L J
Published: January 2020

Infection of chickens with a myeloblastosis-associated virus which induced a high incidence of osteopetrosis was accompanied by immunosuppression. The immunosuppression was manifested in the following ways. The weight of the bursa, spleen, and thymus was depressed in infected chickens. Infected animals had a diminished capacity to form hemolytic plaques in a direct assay. Spleen cells from osteopetrotic animals did not respond to phytohemagglutinin, and the spleen and bursa had a decreased proportion of cells possessing surface immunoglobulin. Osteopetrotic animals failed to show an age-dependent increase in the proportion of cells demonstrating surface immunoglobulin that was observed in normal animals. However, several individual chickens with heavy osteopetrosis responded to antigenic stimulation in a normal fashion, indicating that although immunosuppression usually accompanies avian osteopetrosis, it may not contribute directly to abnormal bone proliferation.
Plasmid-dependent attachment of Agrobacterium tumefaciens to plant tissue culture cells
Authors: Matthysse A G , Wyman P M , Holmes K V
Published: January 2020

Kinetic, microscopic, and biochemical studies show that virulent Ti (tumor inducing)-plasmid-containing strains of Agrobacterium attach to normal tobacco and carrot tissue culture cells. Kinetic studies showed that virulent strains of A. tumefaciens attach to the plant tissue culture cells in increasing numbers during the first 1 to 2 h of incubation of the bacteria with the plant cells. Five Ti-plasmid-containing virulent Agrobacterium strains showed greater attachment to tobacco cells than did five avirulent strains. Light and scanning electron microscopic observations confirmed that virulent strains showed little attachment. Bacterial attachment was blocked by prior incubation of the plant cells with lipopolysaccharide extracted from A. tumefaciens, but not from A. radiobacter, suggesting that bacterial lipopolysaccharide is one of the components involved in the attachment process. At least one other bacterial product may be required for attachment in tissue culture because the virulent A. tumefaciens NT1, which lacks the Ti plasmid, does not itself attach to tobacco cells, but its lipopolysaccharide does inhibit the attachment of virulent strains.
Immunological relatedness of ribosomes from mycobacteria, nocardiae and corynebacteria, and microorganisms in leprosy lesions
Authors: Laub R , Delville J , Cocito C
Published: January 2020

Serological relatedness of ribosomes from microorganisms of the Mycobacterium, Nocardia, and Corynebacterium genera has been analyzed by the microplate immunodiffusion technique. Mycobacterium and Nocardia proved homogeneous and closely related taxa, whereas Corynebacterium was found to be a heterogeneous phylum connected by remote links to the others. The taxonomic position of "diphtheroid microorganisms" (non-acid-fast, gram-positive bacteria morphologically similar to corynebactria), which were found together with Mycobacterium leprae in human leprosy lesions, was also investigated. Ribosomes of diphtheroid bacteria strongly cross-reacted with antisera against several mycobacteria and nocardiae but not against corynebacteria. Moreover, ribosomes from independently isolated diphtheroid strains proved serologically related and yielded strong cross-reactions with antisera against M. leprae as well as with sera from leprosy patients. Hence, diphtheroid microorganisms represent a homogeneous group immunologically related to mycobacteria in general and more specifically to M. leprae.
Demonstration of shared and unique immunological determinants in enterotoxins from Vibrio cholerae and Escherichia coli
Authors: Clements J D , Finkelstein R A
Published: January 2020

Immunodiffusion and biological neutralization studies demonstrated that the heat-labile enterotoxin (LT) from Escherichia coli has antigenic determinants in common with each of the isolated subunits (A and B) of the enterotoxin (choleragen) from Vibrio cholerae. Each of the enterotoxins also possesses unique antigenic specificities. Monospecific antiserum to LT was prepared by immunization with antigens derived by immune precipitation of E. coli cell-free supernatant with isolated specific anticholeragenoid antibodies. This antiserum neutralized the biological acitivity of both LT and cholera enterotoxin and recognized antigens of both in immunodiffusion. This antiserum was adsorbed with choleragenoid to remove antibodies directed against the shared "B" immunological determinants. The neutralizing effect of the antiserum on cholera toxin was completely removed, but the neutralizing activity against the E. coli preparations was retained, although somewhat reduced. Antisera to the isolated subunits (A and B) of cholera enterotoxin neutralized the biological activity of cholera enterotoxin and LT. These antisera also recognized the homologous and heterologous antigens in immunodiffusion. Multiple forms or conformations of LT and its components may explain the diversity of the properties which have been reported for it.
Host defense mechanisms against influenza virus: interaction of influenza virus with murine macrophages in vitro
Authors: Wells M A , Albrecht P , Daniel S , Ennis F A
Published: January 2020

The interaction of mouse macrophages with influenza virus was examined as part of a study into the defense mechanisms against influenza infection. Macrophages exposed to A/Port Chalmers/1/73 virus produced infectious foci on susceptible indicator cell monolayers. Sampling of supernatant fluids and cells from infected macrophage cultures showed release of virus adsorbed to the cell surface. Active virus replication in macrophages could not be demonstrated. Exposing macrophages to specific antibody before or after virus infection resulted in a significant decrease in the number of infectious macrophages. The results suggest that although macrophages are not the source of replicating influenza virus, they are able to spread the infection by having virus attaching to their surface. This activity is interfered with by the presence of specific antibody.
Depression of cell-mediated immunity in cholera
Authors: Palmer D L , Zaman S N
Published: January 2020

Cholera toxin may depress cell-mediated immunity by stimulation of adenyl cyclase and production of cyclic AMP in cellular systems or when given parenterally to experimental animals. Whether or not similar effects might be found during clinical infection with Vibrio cholerae was the subject of this study. Delayed hypersensitivity reactions to skin test antigens were found to be markedly depressed in Bengali patients with cholera 24 h after fluid repletion. Skin test response rates were lower in children and in adults with the disease than in both normal adults and children or in adults with an equivalent degree of malnutrition. Patients with equal degrees of dehydration due to noncholera diarrhea were significantly less immunosuppressed. Concurrent depression of other manifestations of cell-mediated immunity was not found.
Antitumor activity of Listeria monocytogenes on a guinea pig fibrosarcoma
Authors: Dustoor M M , Fulton A , Croft W , Blazkovec A A
Published: January 2020

Listeria monocytogenes-mediated tumor inhibition was studied in strain 13 guinea pigs by using a methylcholanthrene-induced fibrosarcoma (MCA-1). Mixtures of Listeria and tumor cells in ratios of 1:100, 1:200, or 1:400 (Listeria:MCA-1 cells) led to significant suppression of tumor growth. Intralesional injection of tumors on day 6 posttransplantation led to the regression of a highly significant number of tumors. Animals receiving injections of Listeria, either in a mixture with tumor cells or intralesionally, displayed enhanced skin test reactivity to a tumor extract. Tumor regressors were resistant for at least 2 to 3 months after the initial transplant to rechallenge with MCA-1 cells. Thus, with this particular tumor-host system, Listeria was successfully employed as an antitumor agent with no visibly detrimental side effects to the host.
Dissociation of leukocyte alkaline phosphatase from the bactericidal activity of neutrophils
Authors: DeChatelet L R , Shirley P S , McCall C E , Bass D A
Published: January 2020

We have identified an apparently healthy individual with no history of repeated infections whose neutrophils were virtually devoid of alkaline phosphatase activity by either spectrophotometric or histochemical assay. His cells showed normal bactericidal activity toward both Staphylococcus aureus and Escherichia coli when tested in vitro. We have also demonstrated that L-p-bromolevamisole, a potent inhibitor of alkaline phosphatase in many tissues, is likewise an effective inhibitor of the neutrophilic enzyme. Concentrations of this compound which caused nearly complete inhibition of the neutrophilic enzyme did not impair the ability of intact cells to kill bacteria. These results suggest that leukocyte alkaline phosphatase is not required for the normal bactericidal activity of neutrophils.
Host defenses in herpes simplex infections of the nervous system: effect of antibody on disease and viral spread
Authors: McKendall R R , Klassen T , Baringer J R
Published: January 2020

BALB/c mice passively immunized with antibody to herpes simplex virus type 1 and challenged in the footpad with 10(5.7) plaque-forming units of herpes simplex virus type 1 were shown to be protected from neurological disease and death compared with control mice treated with normal serum or antibody to Sindbis virus. One hundred percent of untreated mice had virus recoverable from dorsal root ganglia by 48 h after infection. Whereas amputation of the infected limb at 48 h had no effect, antibody administration (resulting in titers of 1:8 and 1:16) was found to prevent acute neurological disease if administered no later than 48 h after infection. Antibody also restricted the extent of latent infection in the lumbosacral ganglia. The data provide strong evidence that antibody is effective in preventing spread of virus both in the peripheral nervous system and in central nervous system (spinal cord) tissue.
Cell envelope of Neisseria gonorrhoeae CS7: peptidoglycan protein complex
Authors: Hebeler B H , Wong W , Morse S A , Young F E
Published: January 2020

Treatment of cells grown to exponential phase with 4% sodium dodecyl sulfate for 3 h at 100 degrees C resulted in solubilization of all cellular components except for peptidoglycan. In most strains, cells cultured in liquid gonococcal broth at pH 7.2 yielded a peptidoglycan composed primarily of N-acetylmuramic acid N-acetylglucosamine, alanine, glutamic acid, and diaminopimelic acid in a molar ratio of 1:1:2:1:1. The peptidoglycan in these cells accounted for 1 to 2% (dry weight) of the cells. However, in cells cultured at pH 6.0, the dry weight of peptidoglycan increased to 4 to 13%. Preliminary investigations indicated that the apparent increase in weight is strain dependent and is due in part to associated protein(s). Neisseria gonorrhoeae strain CS7 had elevated amounts of protein associated with the peptidoglycan regardless of growth pH. The peptidoglycan-protein complex could not be dissociated by additional extraction with sodium dodecyl sulfate, 10 M LiCl2, or ethylenediaminetetraacetate or by 7.5% polyacrylamide gel electrophoresis. The complex could be degraded by lysozyme, trypsin, chymotrypsin, Pronase B, and Chalaropsis sp. muramidase.
Preparation of glucosyltransferase from Streptococcus mutans by elution from water-insoluble polysaccharide with a dissociating solvent
Authors: Smith D J , Taubman M A , Ebersole J L
Published: January 2020

Glucosyltransferase (EC 2.4.1.5) was obtained by dissociation from water-insoluble polysaccharide in the presence of 6 M guanidine-hydrochloride. Water-insoluble polysaccharide was synthesized by cell-free culture supernatants from Streptococcus mutans strain 6715. Gel filtration of the glucosyltransferase on a column of 8% agarose in phosphate buffer, followed by filtration on a column of 4% cross-linked agarose in 6 M guanidine-hydrochloride, gave a 23-fold enrichment of the enzyme. The enriched glucosyltransferase preparation contained 22% carbohydrate and eluted at a position corresponding to a molecular weight of 422,000. Polyacrylamide gel (5%) electrophoresis of this preparation revealed two regions which stained for protein, formed water-insoluble polysaccharide in the presence of sucrose, and precipitated with antisera directed to crude glucosyltransferase preparations. The guanidine-eluted enzyme could be primed by 5 X 10(-5) M dextran T10 (molecular weight, 10,000). High-molecular-weight glucan and a possible glucan-binding protein were also obtained after the final gel filtration step (4% cross-linked agarose) in addition to glucosyltransferase.
Immunofluorescent studies of the local immune response in the mammary glands of rats
Authors: Lee C G , Ladds P W , Watson D L , Goddard M E
Published: January 2020

Two irritants, phosphate-buffered saline and alcohol, and antigens including killed Brucella abortus, live B. abortus, Staphylococcus aureus, and rat parvovirus were separately infused into rat mammary glands during pregnancy, and by using immunofluorescent techniques, the numbers of immunoglobulin-containing cells in glands during lactation and involution were determined. The study provided basic information on the local immune response of the mammary gland to antigens of various types. In all experiments, the number of immunoglobulin M (IgM) cells present was small and no trends were apparent. IgA cells were always more prevalent than IgG cells. Fewer IgA cells were in the glands of rats infused with phosphate-buffered saline and alcohol than in normal rats. IgA cell prevalence was greatest in response to infusion of live B. abortus. Responses to live S. aureus and parvovirus were less pronounced, and infusion of killed B. abortus did not induce an elevation in IgA cell prevalence. IgG cell prevalence was greatest in response to infusion of live B. abortus or S. aureus and was decreasingly less pronounced in response to killed B. abortus and rat parvovirus. With the exception of parvovirus infusion, in regard to IgA cells, all glands locally infused with antigen had elevated IgA and IgG cell numbers when compared with noninfused glands in the same animal.
Staphylococcus aureus cell surface: capsule as a barrier to bacteriophage adsorption
Authors: Wilkinson B J , Holmes K M
Published: January 2020

Encapsulated Staphylococcus aureus strains M and Smith diffuse bound phage 84 and 52A much less efficiently than their unencapsulated counterparts, M variant and Smith compact. It is proposed that the capsule acts as a barrier excluding phage from interaction with their receptor (cell wall peptidoglycan and teichoic acid). Inefficient phage adsorption by encapsulated staphylococci may explain, in part, the poor phage typing of such strains.
Protection of cotton rats against experimental Echinococcus multilocularis infections with BCG cell walls
Authors: Reuben J M , Tanner C E , Portelance V
Published: January 2020

Previous works has indicated that cotton rats treated with Mycobacterium bovis strain BCG are effectively protected against an infection with the metastatically proliferating metacestodes of Echinococcus multilocularis. In an attempt to induce a similar protection in the absence of tubercular granulomatous lesions, cotton rats were treated with BCG cell walls. A single injection of 150 micrograms of cell walls, emulsified in mineral oil-Tween-saline, 2 weeks before the inoculation of the parasite completely protected the animals against infection with E. multilocularis. This protection was correlated with an increase in the numbers of monocytes and, as judged by acid phosphatase activity, an activation of these cells. This study shows that BCG cell walls are as effective in protecting animals against E. multilocularis as the viable organism.
Erythrocyte Binding Properties of Streptococcal Lipoteichoic Acids
Authors: Beachey Edwin H. , Dale James B. , Simpson W. Andrew , Evans Judith D. , Knox Kenneth W. , Ofek Itzhak , Wicken Anthony J.
Published: January 2020

The lipoteichoic acids (LTA) of gram-positive bacteria are known to bind spontaneously to a variety of animal cell membranes. We investigated the biological and biochemical characteristics of the binding of LTA of
Streptococcus pyogenes
and
S. faecalis
to human and sheep erythrocytes. The kinetics of the binding of the radiolabeled LTA ([
3
H]LTA) from each of these organisms to erythrocytes was similar. The dissociation constants for sheep and adult human erythrocytes were 1.6 μM and 4.5 μM, respectively, whereas that of human cord blood erythrocytes was approximately 10-fold higher, 31 μM. The number of binding sites for sheep erythrocytes was calculated to be 7.2 × × 10
6
per cell, and that of human erythrocytes, 29 × 10
6
per cell. Binding was reversible. More than 50% of bound [
3
H]LTA was displaced from erythrocytes by a 50-fold excess of unlabeled LTA. LTA prepared from heterologous species of gram-positive bacteria were all inhibitory to the binding of [
3
H]LTA whether derived from
S. pyogenes
or from
S. faecalis
. Among a number of potential receptor analogues and other inhibitors tested, including serum albumin, gangliosides Gm
2
and Gm
3
, lipopolysaccharide of gram-negative bacteria, and various sugars, only albumin and the gangliosides significantly inhibited LTA binding. Trypsin or neuraminidase treatment of erythrocytes had no effect on LTA binding. Deacylation of [
3
H]LTA abolished binding ability and binding was restored by esterification of the deacylated material with stearoyl chloride, indicating that ester-linked lipids are necessary for membrane binding.
Modification of antibody response to type III pneumopolysaccharide by route of injection of pertussis vaccine
Authors: Speirs R S , Benson R W , Roberts D W
Published: January 2020

Pertussis vaccine (PV) or diphtheria toxoid-PV-tetanus toxoid (DPT) altered the antibody response of BALB/c female mice to type III pneumococcal polysaccharide antigen (S3). The key factor affecting the magnitude of the response to S3 was the route of injection of PV or DPT, whereas the route of injection of S3 was not crucial. Subcutaneous injections of DPT augmented the antibody response to low, optimal, and tolerogenic (high) doses of S3 injected either subcutaneously or intraperitoneally. This enhancement was persistent and was observed both when S3 and DPT were mixed and injected subcutaneously and when S3 and DPT were injected concurrently at separate subcutaneous sites. When either PV or DPT was injected intraperitoneally, the antibody response to subcutaneously or intraperitoneally injected S3 was significantly decreased. These experiments demonstrate a dichotomy of effect dependent on the route of administration of PV. Most studies in mice utilizing PV employ the intraperitoneal injection route, and it is important to consider whether PV treatment by this route may have unique effects. Our data suggest that intraperitoneal injection of PV suppresses B cells, possibly by influencing regulatory cell function.
Behavior of Escherichia coli K antigens K88ab, K88ac, and K88ad in immunoelectrophoresis, double diffusion, and hemagglutination
Authors: Guinée P A , Jansen W H
Published: January 2020

Porcine enteropathogenic Escherichia coli strains were found to possess a variant of the K88 antigen provisionally termed K88ad. We propose to include this antigen into the international E. coli typing scheme. Ultrasonic extracts of field strains of E. coli possessing the K88ab, K88ac, or K88ad antigen and their E. coli K-12 K88+ transconjugants showed a specific K88 precipitation line in immunoelectrophoresis and double diffusion only when grown at 37 degrees C, but not when grown at 18 degrees C. By using agarose gels, K88ab, K88ac, and K88ad antigens showed anodic mobility in immunoelectrophoresis. When using Difco Noble agar gels, K88ad was not mobile or anodic, K88ab was cathodic; K88ac of 17 strains was cathodic and of 24 strains was anodic. The immunoelectrophoretic behavior of a K88 antigen (K88ab, K88ac, or K88ad) did not alter after transfer of the corresponding plasmid to E. coli K-12. Anodic and cathodic K88ac antigens could not be distinguished serologically. The differences between the results obtained in Noble agar gels and agarose gels are due to electro-endosmotic flow. We describe a procedure which increases the detection level of K88+ transconjugants in a mating mixture. It is based on the specific mannose-resistant attachment of K88+ cells to guinea pig erythrocytes.
Immunological and serological diversity of Neisseria gonorrhoeae: identification of new immunotypes and highly protective strains
Authors: Wong K H , Arko R J , Schalla W O , Steurer F J
Published: January 2020

Gonococci, irrespective of serotype or immunotype, varied significantly in their capacity to induce immunity in animal models, and in vitro serological relatedness did not always insure in vivo cross-protection. By using a serum bactericidal assay followed by in vivo cross-protection studies, new immunotypic strains which were highly protective were identified from cultures isolated in different geographical areas and from patients with various clinical manifestations. Beta-lactamase production and gonococcal immunotype did not appear as related characteristics in that certain penicillin-sensitive strains were highly effective in immunizing animals against infection with beta-lactamase producers. The findings of this study emphasize the importance of using appropriate biological tests and strains for the investigation of gonococcal immunity and vaccine development. Immunization with a combination of a few major gonococcal immunotypic immunogens may provide substantial protection against the majority of penicillin-sensitive and beta-lactamase-producing gonococci. Investigation of isolated immunotypic immunogens is in progress.
Immunity to Enterotoxigenic
Escherichia coli
Authors: Levine Myron M. , Nalin David R. , Hoover David L. , Bergquist Erick J. , Hornick Richard B. , Young Charles R.
Published: January 2020

Enterotoxigenic
Escherichia coli
strains represent the most frequent etiological agent of travelers diarrhea. Challenge studies with several of these strains were undertaken in volunteers to evaluate the mechanisms of disease-induced immunity. Seventeen students and other community volunteers were given 10
6
or 10
8
organisms of
E. coli
B7A (O148:H28), which produces heat-labile and heat-stable enterotoxins. Ten individuals developed diarrheal illness closely resembling natural travelers diarrhea; of these ten, rises in titer of serum antitoxin and anti-O antibody occurred in eight (80%). Eight of the volunteers who developed diarrhea in the first test agreed to undergo rechallenge 9 weeks later with 10
8
B7A organisms. Only one of these eight “veterans” developed diarrhea versus seven of twelve controls given the same challenge (
P
= 0.05). Despite clinical protection, all “veterans” excreted B7A after rechallenge. Four controls who developed diarrhea during the homologous B7A rechallenge test were rechallenged 9 weeks later with 10
9
organisms of
E. coli
strain E2528-C1 (O25:H-), which produces only heat-labile enterotoxin and possesses a different O, H, and pili antigen composition than B7A. Three of four “veterans” and two of six controls developed comparable diarrhea. These studies demonstrate that prior disease due to enterotoxigenic
E. coli
confers homologous immunity against subsequent challenge, and the operative mechanism apparently is not bactericidal and is not mediated by serum anti-O antibodies. Heterologous protection was not conferred where the only common antigen was heat-labile enterotoxin, indicating that serum infection-derived antitoxin to heat-labile enterotoxin also is not protective.
Chemical characterization and biological properties of lipopolysaccharides isolated from smooth and rough strains of Brucella abortus
Authors: Kreutzer D L , Buller C S , Robertson D C
Published: January 2020

The chemical composition and some of the biological activities of lipopolysaccharides (LPS) extracted from a smooth-intermediate strain (45/0) and a rough strain (45/20) of Brucella abortus have been examined. LPS were found in both the phenolic and aqueous extraction phases of strain 45/0, but only in the aqueous phase of 45/20. The phenolic LPS contained 9- to 16-fold-lower levels of heptose and reduced amounts of dideoxyaldoses compared with aqueous fractions. The major neutral sugars were glucose, galactose, and mannose. beta-Hydroxymyristic-acid, a common marker of enteric LPS, was not detected. Fatty acids present in highest amounts were hydroxylated and nonhydroxylated species with chain lengths of 16, 18, and 20 carbons. Only the phenolic LPS of strain 45/0 exhibited mouse lethality and a curable wasting disease; however, both phenolic and aqueous fractions caused carbohydrate depletion in mice. The toxicity of aqueous LPS could not be potentiated with Pb(OAc)4. These data, coupled with the lack of mitogenic activity for B-lymphocytes, are indicative of the unique structure-function relationships of Brucella LPS.
Isolation of mutants of Mycoplasma pneumoniae defective in hemadsorption
Authors: Hansen E J , Wilson R M , Baseman J B
Published: January 2020

Mutants of Mycoplasma pneumoniae incapable of hemadsorption were isolated by means of chemical mutagenesis. These hemadsorption-negative mutants did not attach to hamster tracheal rings in vitro at as high a frequency as that exhibited by the wild-type parent strain.
Suppression of In Vitro Antibody Response by Spleen Cells of Mice Infected with Friend-Associated Lymphatic Leukemia Virus
Authors: Bendinelli Mauro , Matteucci Donatella , Toniolo Antonio , Friedman Herman
Published: January 2020

The ability of spleen cells of mice infected with oncornaviruses to depress the in vitro antibody responsiveness of normal lymphoid cells was exploited in an attempt to clarify the role played by the lymphatic leukemia virus (LLV) component in the immunodepressive properties of the Friend leukemia complex. Spleen cells of mice infected with LLV or, for comparison, with the entire complex were added to cultures of sheep erythrocyte-primed uninfected spleen cells, and the antibody-forming cells produced by the latter, after antigen restimulation, were assayed. The addition within 2 days from culture initiation of low numbers of cells infected with either virus preparation suppressed all stages of the response affecting the production of both immunoglobulin M and immunoglobulin G antibody. The activity of infected cells resisted doses of ultraviolet radiation which inhibit cell multiplication but was abolished by disrupting the cells and was prevented by the presence of anti-LLV antibodies. The LLV-infected spleen cells responsible for suppression were not removed by treatments which selectively remove or kill macrophages and exhibited surface properties of B lymphocytes. These results were interpreted as indicating that the effect is due to virus (or viral products) released by B cells. The suppressing cells in the spleens of mice in the early days of Friend leukemia complex infection presented superimposable properties, supporting the concept that their activity is also due to the LLV they release in large quantities. However, in later stages of infection, the spleens of Friend leukemia complex-infected mice also contained non-B-suppressing cells possibly derived from the proliferation of nonlymphoid LLV-producing cells caused by the neoplastic process.
Response of sensitized and unsensitized human lymphocyte subpopulations to Plasmodium falciparum antigens
Authors: Wyler D J , Herrod H G , Weinbaum F I
Published: January 2020

Antigen preparations derived from Plasmodium falciparum-infected erythrocytes (but not from uninfected erythrocytes) can stimulate the in vitro proliferation of peripheral blood lymphocytes from malaria-sensitized as well as nonsensitized donors. The possibility that the nonspecific responses might be due to a parasite-derived B-cell mitogen has been previously suggested since polyclonal hypergammaglobulinemia is a frequent accompaniment of malaria infection. To test this hypothesis, we investigated the in vitro proliferative responses of purified T- and B-cell populations to malaria antigens. T but not B cells responded to the antigens. The addition of small numbers of T cells restored the ability of purified B cells to respond to lectin mitogens but not to malaria antigens. Falciparum malaria infection was associated with an increase in T-cell but not in B-cell proliferation in vivo, as assessed by the spontaneous tritiated thymidine incorporation of lymphocytes during a brief incubation in vitro. Our observations suggest that extracts of malaria parasites do not contain a B-cell mitogen but are antigenic as well as mitogenic for T cells.
Intramuscular and/or Intralumbar Postexposure Treatment of Rabies Virus-Infected Cynomolgus Monkeys with Human Interferon
Authors: Weinmann Ernst , Majer Mirko , Hilfenhaus Joachim
Published: January 2020

From 9 to 10 of 10 cynomolgus monkeys infected with rabies street virus died of rabies about 20 days postinfection (pI). Symptoms of illness appeared 1 to 4 days before death. In an attempt to protect infected animals from the disease, human leukocyte interferon (HIF) was administered intramuscularly (i.m.) near the site of infection or into the cerebrospinal fluid between the first and second lumbar vertebrae (i.e., intralumbarly [i.l.]). Multiple HIF doses given over a period of several days proved more effective than a single HIF dose. In every experiment, i.m. HIF treatment was started 1 day pI. The best result obtained was a survival rate of 7 of 10 monkeys. The i.l. HIF administration schedules, consisting of multiple doses given over a period of at least 8 days, were started on day 3, 7, or 11 pI. Here the best result noted was the protection of 5 of 10 treated monkeys. The latest successful postexposure i.l. HIF treatment began on day 11 pI. The highest protection rate, 8 survivors of 10 treated monkeys, was achieved by a combined i.m. and i.l. HIF treatment. From these results we conclude that human patients severely bitten by rabid animals should in addition to an active immunization be i.m. and i.l. treated with HIF. Particularly, i.l. HIF administration could be effective, even when given several days pI. Whether an HIF administration starting after the appearance of clinical symptoms of rabies can help cannot be decided upon from the studies made in this monkey model. The most obvious difference between rabies in humans and cynomolgus monkeys is the duration of illness between the outbreak of the disease and death (1 to 4 days only in this animal model). It might have been due to this short period of illness that i.l. and i.m. HIF treatment at the appearance of clinical symptoms failed to help any of the monkeys treated.
Immunobiology and species distribution of Mycobacterium tuberculosis antigen 5
Authors: Daniel T M , Ellner J J , Todd L S , McCoy D W , Payne V D , Anderson P A , Bhe F T
Published: January 2020

The immunobiology and mycobacterial species distribution of immunoabsorbent affinity chromatography-purified Mycobacterium tuberculosis antigen 5 have been studied. In delayed hypersensitivity skin tests, antigen 5 was nearly equipotent with tuberculin-purified protein derivative in sensitized guinea pigs. In vitro, antigen 5 was capable of stimulating the production of migration inhibitory factor by cultured lymphocytes from sensitized guinea pigs and humans. Antigen 5 stimulated thymidine incorporation by cultured guinea pig lymphocytes but did not stimulate thymidine incorporation by cultured human lymphocytes. Although erythrocytes were readily sensitized with antigen 5 for passive hemagglutination, their use did not offer any advantage over previous hemagglutination techniques for the serodiagnosis or evaluation of patients with tuberculosis. By immunoelectrophoresis and immunodiffusion, antigen 5 was readily identified in culture filtrates of 10 strains of M. tuberculosis and M. bovis but not in those of 30 strains of 12 other myobacterial species.
Adjuvant activity of 6-O-mycoloyl derivatives of N-acetylmuramyl-L-seryl-D-isoglutamine and related compounds in mice and guinea pigs
Authors: Uemiya M , Sugimura K , Kusama T , Saiki I , Yamawaki M , Azuma I , Yamamura Y
Published: January 2020

Adjuvant and antitumor activities of synthetic-6-O-mycoloyl-N-acetylmuramyl-L-seryl-D-isoglutamine and 6-O-mycoloyl-N-acetylmuramyl-glycyl-D-isoglutamine were examined in comparison with those of 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine. Synthetic 6-O-mycoloyl-N-acetylmuramyl-L-seryl-D-isoglutamine was active as an adjuvant for the induction of delayed-type hypersensitivity to m-[4-(4'-arsono-phenylazo)-phenyl]-N-acetyl-L-tyrosine in guinea pigs and for cell-mediated cytotoxicity in allogeneic mice. 6-O-mycoloyl-N-acetylmuramyl-glycyl-D-isoglutamine was inactive as an adjuvant for the induction of delayed-type hypersensitivity in guinea pigs; however, it was active for cell-mediated cytotoxicity in allogeneic mice. 6-O-mycoloyl-N-acetylmuramyl-L-seryl-D-isoglutamine and 6-O-mycoloyl-N-acetylmuramyl-glycyl-D-isoglutamine were not pyrogenic in rabbits. The antitumor activity of these 6-O-mycoloyl-N-acetylmuramyldipeptides was examined preliminarily by using transplantable syngeneic mouse tumors.
Adjuvant Activities in Production of Reaginic Antibody by Bacterial Cell Wall Peptidoglycan or Synthetic
N
-Acetylmuramyl Dipeptides in Mice
Authors: Ohkuni Hisashi , Norose Yoshihiko , Ohta Midori , Hayama Masaaki , Kimura Yoshitami , Tsujimoto Masaya , Kotani Shozo , Shiba Tetsuo , Kusumoto Shoichi , Yokogawa Kanae , Kawata Shigeo
Published: January 2020

This paper is concerned with the adjuvant activity in stimulatory immunoglobulin E production against ovalbumin (OA) by bacterial cell walls, cell wall peptidoglycan (PG), and their PG fragments and synthetic
N
-acetylmuramyl (MurNAc) dipeptides in A/J mice. A PG isolated from
Streptococcus pyogenes
, PG subunit polymer and dimer obtained from
Staphylococcus epidermidis
, and water-soluble fragments of cell walls or PG prepared from
Nocardia corynebacteriodes
and
Streptomyces gardneri
were found to enhance both the primary and secondary responses of anti-OA immunoglobulin E antibody production. It was suggested that the PG portion, either intact or highly degraded, was capable of enhancing the immunoglobulin E antibody production, and there was no need for the non-PG moiety or intactness of PG structure for the adjuvant activity. This finding was confirmed and extended by the use of synthetic MurNAc dipeptides. Among eight MurNAc dipeptides tested, MurNAc-
l
-Ala-
d
-isoGln, MurNAc-
l
-Ala-
d
-Gln, MurNAc-
l
-Ala-
d
-Glu, and MurNAc-
l
-Ser-
d
-isoGln were found active as an adjuvant in the stimulation of the primary and secondary reaginic anti-OA antibody production in a similar way to the cell wall PG and their fragments. None of the synthetic MurNAc-
l
-Ala-
l
-isoGln, MurNAc-
l
-Ala-
l
-Gln, MurNAc-
l
-Ala-
l
-Glu, and MurNAc-
l
-Ala-
d
-isoAsn, on the other hand, stimulated the anti-OA immunoglobulin E antibody production in either primary or secondary response, indicating the importance for the adjuvancy in immunoglobulin E production of the configuration of the glutamic acid residues adjacent to the
l
-Ala (or
l
-Ser) in muramyl dipeptides.
Resistance to Babesia spp. and Plasmodium sp. in mice pretreated with an extract of Coxiella burnetii
Authors: Clark I A
Published: January 2020

Mice injected intravenously with a commercially available extract of Coxiella burnetii prepared for use as the antigen in the complement fixation diagnostic test for Q fever were subsequently resistant to infection with Babesia microti, Babesia rodhaini, and Plasmodium vinckei petteri. The parasites appeared to die inside circulating erythrocytes. Protection was unaffected by exposing the pretreated mice to 900 rads on the day before they were infected. To explain these findings, it is postulated that pretreatment with Coxiella extract protects by potentiating the interferon-inducing capacity of the challenge dose of protozoa, which perhaps leads to enhanced of natural killer cells. Tumor necrosis factor also warrants investigation.
Abortion in laboratory animals induced by Moraxella bovis
Authors: Norman J O , Elissalde M H
Published: January 2020

Pregnant mice, guinea pigs, rats, and rabbits responded to injections of Moraxella bovis strain EPP-63(300) with abortion, death, embryo resorption, and production of small litters. The nature of response appeared to depend primarily on the number of viable cells injected and to some extent on the species of animal and stage of pregnancy. Intraperitoneal injection of mice with 3 X 10(5) viable cells induced 100% abortion and no deaths. Embryo resorption and smaller litters were induced with injection of lower doses of M. bovis. None of the rats used in this study aborted; however, resorption rates were higher in rats than mice. Bacteria-free filtrates prepared from aqueous extracts of blood agar on which M. bovis had been grown induced abortion in mice and guinea pigs, at a rate similar to that caused by viable cells. The lyophilized filtrate could be diluted to produce 100% abortion with no death of injected mice. Abortion-inducing, nonlethal doses of M. bovis and lyophilized filtrates appeared to have no effect on nonpregnant female or male mice.
Streptococcal infection of endocardial and other intravascular vegetations in rabbits: natural history and effect of dexamethasone
Authors: Francioli P B , Freedman L R
Published: January 2020

Experiments were designed to study the natural history of infection in different parts of the vascular system. Sterile vegetations were produced in rabbits by placing catheters in the inferior vena cava, tricuspid or aortic valves, and thoracic or abdominal aorta and then were infected by the intravenous inoculation of Streptococcus sanguis. At 1 day after bacterial challenge, all VEGS were infected, mean bacterial densities being highest in the VEGS of the aortic and tricuspid valves. By 14 days, there was a significant decrease in the mean bacterial density in all VEGS except for the aortic valve: the VEGS of the inferior vena cava and abdominal aorta were sterile, as were half of those of the thoracic aorta. There were no deaths except for animals with aortic valve infection. Dexamethasone inhibited the sterilization of the thoracic aorta VEGS, but was without effect on aortic valve VEGS, 5 mm distant. Sterilization of tricuspid valve VEGS after catheter removal was also inhibited by dexamethasone. Thus, there are host defense mechanisms which lead to the sterilization of infections everywhere in the vascular system except in the left side of the heart, and these mechanisms, as yet undefined, are inhibited by dexamethasone.