bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenomenon of hot-cold hemolysis: chelator-induced lysis of sphingomyelinase-treated erythrocytes
Authors: Smyth C J , Möllby R , Wadström T
Published: January 2020

Staphylococcus aureus produces a phospholipase C specific for sphingomyelin (beta-hemolysin). Erythrocytes with approximately 50% sphingomyelin in their membranes, e.g., from sheep, have been shown to have up to 60% of this phospholipid hydrolyzed by this enzyme at 37 C in isotonic buffered saline without hemolysis. Cooling of sphingomyelinase C-treated erythrocytes to 4 C causes complete lysis of the cells, a phenomenon known as hot-cold hemolysis. The addition of ethylenediaminetetraacetate (EDTA) to sheep erythrocytes preincubated with sphingomyelinase C was found to induce rapid hemolysis at 37 C. The treated cells became susceptible to chelator-induced hemolysis and to hot-cold hemolysis simultaneously, and the degree of lysis of both mechanisms increased equally with prolonged preincubation with sphingomyelinase C. Erythrocytes of species not readily susceptible to hot-cold hemolysis were equally insusceptible to chelator-induced lysis. Chelators of the EDTA series were the most effective, whereas chelators more specific for Ca2+, Zn2+, Fe2+, Cu2+, and Mg2+ were without effect. The rate of chelator-induced lysis was dependent on the preincubation period with beta-hemolysin and on the concentration of chelator added. The optimal concentration of EDTA was found to equal the amount of exogenously added Mg2+, a cation necessary for sphingomyelinase C activity. Hypotonicity increased the rate of chelator-induced hemolysis, whereas increasing the osmotic pressure to twice isotonic completely inhibited chelator-induced lysis. The data suggest that exogenously added and/or membrane-bound divalent cations are important for the stability of sphingomyelin-depleted membranes. The phenomenon of hot-cold hemolysis may be a consequence of the temperature dependence of divalent ion stabilization.
Induction of resistance with heat-killed compact-type strains of Staphylococcus aureus against challenge with the diffuse variant of the Smith strain of Staphylococcus aureus
Authors: Yoshida K , Ichiman Y , Otomo T
Published: January 2020

Active immunization of mice with high doses of heat-killed and autoclaved vaccine of unencapsulated strains of Staphylococcus aureus, which was grown in brain heart infusion media, protected against challenge with the Smith diffuse strain of Staphylococcus aureus. These organisms were capable of absorbing the protective antibody in rabbit hyperimmune sera prepared with the Smith diffuse strain. Also, mice treated with rabbit hyperimmune sera prepared with these strains (four out of six strains) protected against challenge with the Smith diffuse strain. Protective activities of these rabbit hyperimmune sera were assumed to be essentially identical to the protective antibody induced by the Smith diffuse strain.
Antigenic variation of Streptococcus mutans colonizing gnotobiotic rats
Authors: Bratthall D , Gibbons R J
Published: January 2020

Strains of Streptococcus mutans representative of serotypes b and d exhibited antigenic variation in both the oral cavity and in the intestinal canal of gnotobiotic rats. Laboratory-maintained cultures did not vary. The antigenic alterations observed were: (i) loss of detectable levels of both weakly reacting "strain" antigens and the type antigen; (ii) decreased production of the type antigen; (ii) production of altered type antigen; and (iv) production of an antigen not possessed by the parent strain. Immunization of animals before monoinfection with S. mutans strain Bob-1 (serotype d) appeared to increase the rate of emergence of antigenically altered mutants in the intestinal canal, and more diversely altered isolates were obtained. Antigenic variation may account in part for the variation noted by several investigators in attempting to immunize animals against S. mutans-induced dental caries.
Measurement of antibody to influenza virus neuraminidase by single radial hemolysis in agarose gels
Authors: Callow K A , Beare A S
Published: January 2020

A simple method of assaying anti-influenza neuraminidase antibodies in human sera was described. Suitable antigenic hybrid viruses were adsorbed to sheep erythrocytes, which were then incorporated into agarose gels. When sera were introduced into wells cut in the gels, zones of hemolysis were observed in the neighborhood of those containing neuraminidase antibodies. There was a direct relationship between zone size and antibody titer. No purification of adsorbed viruses was necessary. The test was rapid, required very simple reagents, gave results that agreed well with those given by conventional techniques, and appeared to be the most sensitive of four methods evaluated. Studies of cross-reactions by hyperimmune sera against homologous and heterologous neuraminidases and of absorption of neuraminidase antibodies from human sera indicated a high degree of specificity. The technique seems to be suitable for large-scale epidemiological investigations.
Effect of penicillin on the succession, attachment, and morphology of segmented, filamentous microbes in the murine small bowel
Authors: Davis C P , Savage D C
Published: January 2020

Indigenous segmented filamentous microbes attach to murine ileal epithelial cells. These microbes can be seen on the epithelial surface with a scanning electron microscope. They colonize preferentially the distal ileum in mice. Penicillin, placed in the animal's drinking water, eliminates the microbes from the mouse ileum, but recolinization of the ileum is observed 4 to 5 weeks after the penicillin treatment is stopped. Within 3 to 5 h after rats are given penicillin, the morphology of the microbes is changed. Their external surfaces are wrinkled or broken. Vacated and partially vacated attachment sites are observed. Almost all of the organisms disappear from murine ilea after the animals are exposed to penicillin for 10 h. These observations are discussed in relation to the microbe itself and in its interaction with ileal epithelial cells.
Specificity of secretory antibodies to bacterial immunogens
Authors: Ebersole J L , Molinari J A
Published: January 2020

The present investigation examined the specificity of the salivary immune response of axenic and conventional mice to topically administered Salmonella typhi, S. gallinarum, and S. typhimurium. Specific antibacterial antibodies were determined by passive hemagglutination and bacterial agglutination. Reciprocal antibody titers up to 320 were detected in saliva from mice immunized and assayed with homologous antigens. Antibodies to heterologous immunogens exhibited lower mean titers of 10 to 20 under identical conditions. High concentrations of specific antibodies to the somatic (O) antigen were detected in the saliva of mice administered these microorganisms; however, no significant differences in serum antibody levels were detectable after oral immunization. Only low levels of specific antiflagellar (H) antibodies were demonstrated in the saliva of immunized mice, whereas mean reciprocal titers of 20 were observed in the serum. Antibodies to the Vi antigen of S. typhi were detected in the saliva and serum of only those mice administered formalin-treated S. typhi. Examination of the classes of antibody elicited by these organisms indicated that immunoglobulin A (IgA) was the predominant class in saliva against the O antigens. The salivary response to the H antigens was comprised of both IgG and IgA, whereas the specific serum immunoglobulins were consistent with a primary humoral immune reaction. Local antibodies formed in response to the Vi antigen were exclusively IgG. Serum immunoglobulins produced after peroral administration of the somatic and virulence antigens were limited to the IgM class.
Interferon-producing capacity of germfree mice
Authors: Ito Y , Nisiyama Y , Shimokata K , Kimura Y , Nagata I
Published: January 2020

The general capacity of germfree mouse spleen cells to produce interferon in vitro in response to various stimuli was investigated. The interferon response of germfree mouse spleen cells in vitro, when compared with that of the conventionals, appears to be lower to some inducers. Interferon production in vitro stimulated by hemagglutinating virus of Japan (HVJ) or BHK-HVJ cells (BHK cells persistently infected with HVJ) was apparently suppressed in germfree mouse spleen cells as compared with the corresponding conventionals, whereas no difference of interferon production was observed between germfree and conventional mouse spleen cells in response to Newcastle disease virus, Escherichia coli endotoxin, poly(I:C), and phytohemagglutinin. Although monocontamination with HVJ had no enhancing effect on the interferon-producing ability of germfree mouse spleen cells in response to HVJ, conventionalization for 2 weeks greatly enhanced interferon-producing capacity.
Evidence for a one-hit theory in the immune bactericidal reaction and demonstration of a multi-hit response for hemolysis by streptolysin O and Clostridium perfringens theta-toxin
Authors: Inoue K , Akiyama Y , Kinoshita T , Higashi Y , Amano T
Published: January 2020

An analytical method was developed for estimating the number of hits necessary to lyse or kill cells in which various concentrations of the cells are treated with a constant amount of the lytic or killing agent in a constant reaction volume. The reaction may be due to a single-component agent or occur by a sequential chain of reactions due to a multi-component agent, even including side, abortive, or counter-reactions. It was clearly shown by this method that immune bactericidal reactions followed a one-hit theory. It was shown by this method that streptolysin O required four or five hits for hemolysis and Clostridium perfringens theta-toxin required two hits. These results were confirmed by both logarithmic dose-response and survival analyses. It was also shown that streptolysin O and theta-toxin can act complementarily on accumulation of the hits for hemolysis.
Role of plasma filtration in the intestinal fluid secretion mediated by infection with Salmonella typhimurium
Authors: Giannella R A , Rout W R , Formal S B , Collins H
Published: January 2020

The mechanisms whereby invasive enteropathogens, e.g., Salmonella typhimurium, induce intestinal secretion are largely unknown. Since these organisms penetrate the intestinal epithelium, disrupt the brush border, and evoke an acute inflammatory reaction, increased plasma filtration through a damaged, more permeable epithelium might contribute to the secretory process. To examine this possibility, the plasma-to-lumen clearance of two different sized molecules, [51Cr]albumin and [14C]mannitol, was measured in the in vivo rabbit ileal loop and in vivo rhesus monkey models of salmonellosis. In the rabbit ileal loop model, the clearance of neither molecule was increased when compared to cholera toxin-exposed loops. In the rhesus monkey, clearance of [14C]mannitol into the jejunum, ileum, and colon of Salmonella-infected animals did not differ from the observed in control animals. These data indicate that invasion of the intestinal mucosa by S. typhimurium has not substantially altered the permeability characteristics of the intestinal mucosa and that plasma filtration through a damaged, more permeable mucosa does not contribute to the Salmonella-induced intestinal secretory process.
Synthesis and release of polyribophosphate by Haemophilus influenzae type b in vitro
Authors: Anderson P , Pitt J , Smith D H
Published: January 2020

Polyribophosphate, the capsular polysaccharide antigen of Haemophilus influenzae type b, can be assayed in crude bacterial extracts by a combination of immunoprecipitation, hot-acid extraction, and colorimetry. In vitro, the antigen is synthesized during growth and, in well-buffered media, in early stationary phase as well. It is shed from the cell throughout the course of synthesis, and at 37 C almost all the extractable antigen has been released spontaneously within several hours after synthesis. Various strains are similar in extent of synthesis and gross composition of the antigen; however, strains differ in the rate of release during exponential phase. The antigen is released as (or as part of) a very large structure, although under certain culture conditions much smaller antigenic units are found.
Variability of the immunological state of germfree colostrum-deprived Minnesota miniature piglets
Authors: Setcavage T M , Kim Y B
Published: January 2020

Minnesota miniature piglets obtained by hysterectomy and deprived of colostrum were examined for the presence of immunoglobulin by immunoelectrophoresis, double-gel diffusion, and radial immunodiffusion techniques with specific anti-immunoglobulin chain sera. A large amount of variability existed between different litters of piglets and between different piglets within the same litter, ranging from no detectable immunoglobulin in the serum to very high immunoglobulin levels approaching that of the adult pig. All known classes of porcine immunoglobulin including immunoglobulin G, immunoglobulin M, and immunoglobulin A could be found in the sera from litters where there was extensive placental damage. This contaminating immunoglobulin was shown to have antibody activity to actinophage MSP-2 even when present in very low concentrations. The low level contamination with immunoglobulin G, which was the most frequently encountered type of contaminant, was demonstrated to be similar to sow immunoglobulin G both antigenically and in its molecular size. The data demonstrates that individual piglets must be tested for immunoglobulin content rather than being assumed to be immunologically "virgin" and emphasizes the need for an intact placenta barrier to obtain piglets free from maternal immunoglobulin and devoid of antigenic stimulation.
Analysis of the cell wall of five strains of Myocbacterium tuberculosis BCG and of an attenuated human strain, W 115
Authors: Phiet P H , Wietzerbin J , Zissman E , Petit J F , Lederer E
Published: January 2020

The chemical composition of the deproteinized, delipidated cell walls of five strains of BCG and of an attenuated human strain of Mycobacterium tuberculosis has been established, with special focus on their poly-L-glutamic acid content. All the cell walls have the same overall composition. Their poly-L-glutamic acid content varies from 0 to 4.6%. A correlation between the poly-L-glutamic acid content and the biological properties of the BCG strains reported in the literature could not be established.
Responses of mice immunized with influenza virus by serosol and parenteral routes
Authors: Scott G H , Sydiskis R J
Published: January 2020

Antibody levels in sera and respiratory secretions and resistance to respiratory infections were examined in mice given live infuenza virus in small-particle (2 mum) aerosols, large-particle (10 mum) aerosols, intraperitoneally, and subcutaneously. After parenteral administration antibody was found primarily in the serum, but small amounts were recovered in bronchoalveolar washings after 2 to 3 weeks. Specific antibody was present in both sera and bronchoalveolar washings from mice given virus in small-particle aerosols to achieve virus dissemination throughout the respiratory tract. Immunoglobulin A, immunoglobulin G, and trace amounts of immunoglobulin M, all specific for the infecting virus, were detected in bronchoalveolar washings of small-particle aerosol-infected mice. Virus administration in large-particle aerosols (for primary virus localization in upper respiratory tract) at doses greater than those required to initiate infection with small-particle aerosols failed to stimulate production of antibody in sera or bronchoalveolar washings. Small-particle aerosol-immunized mice were resistant to subsequent challenge with 10(2.0) respiratory median lethal doses of virulent virus, whereas large-particle aerosol-immunized mice were not protected. Parenteral immunization modified the course of the disease in challenged mice and reduce mortality rates but did not prevent reinfection of the respiratory tract.
Virulent Treponema pallidum: aerobe or anaerobe
Authors: Baseman J B , Nichols J C , Hayes N C
Published: January 2020

Substrate degradation and protein synthesis served as indicators of metabolism in virulent Treponema pallidum. Opitmal metabolic activity in these spirochetes was observed at 10 to 20% O2 concentrations, with markedly reduced activity at higher or lower O2 levels or under anaerobiosis; alternate functioning electron acceptors that might substitute for O2 were not found. Carbon monoxide and cyanide at concentrations that inactivate cytochrome oxidase were not effective metabolic poisons for T. pallidum, although Micrococcus lutea, a strict aerobe with cytochrome-dependent respiration, was inhibited under similar experimental conditions. Motility of virulent T. pallidum was vigorous in the presence of O2 and sluggish or inhibited in its absence, reinforcing the role of O2 in T. pallidum metabolism.
Host defenses in experimental scrub typhus: histopathological correlates
Authors: Catanzaro P J , Shirai A , Hilderbrandt P K , Osterman J V
Published: January 2020

Intraperitoneal (i.p.) infection of BALB/c mice with 1,000 50% mouse lethal doses of the Karp strain of Rickettsia tsutsugamushi was inevitably lethal, and associated pathological alterations were confined to the peritoneal cavity. These included: (i) continuous proliferation of rickettsial organisms in peritoneal macrophages until death; (ii) hepatic granulomas appearing 6 days after infection and increasing in size and number until death; (iii) splenomegaly, resulting principally from proliferation of lymphoid tissue, and (iv) terminal peritonitis. Under two circumstances, i.p. infections with R. tsutsugamushi were not lethal: (i) infection with 100 50% mouse infectious doses of the Gilliam strain, which, in fact, resulted in immune protection against otherwise lethal Karp challenge; and (ii) Karp infection of animals immunized with the Gilliam strain. In both cases, the associated pathological abnormalities were, as with primary Karp infection, restricted to the peritoneal cavity. Also similar was the striking splenomegaly due to lymphoid proliferation, which was particularly prominent in immunized animals. In contrast to primary and lethal Karp infection, however, these infections were characterized by: (i) minimal and transient proliferation of rickettsial organisms in peritoneal macrophages; (ii) disappearance of hepatic granulomas; and (iii) absence of peritonitis. It was concluded that the survival of an animal bearing an i.p. infection of scrub typhus depended on its ability to concentrate a sufficiently vigorous immune response in the peritoneal cavity, resulting in the evolution of rickettsiacidal macrophages capable of suppressing the infection.
Immune mechanism in Pasteurella multocida-infected mice
Authors: Woolcock J B , Collins F M
Published: January 2020

Various immunizing procedures were tested for their effect on the growth of Pasteurella multocida in specific pathogen-free CD-1 mice. Two injections with killed vaccine incorporated into Freund complete adjuvant gave excellent protection against parenteral challenge, but less to control the growth of an aerogenic inoculum. Protection with adjuvant-treated preparation was always superior to that seen with non-adjuvant-treated vaccines. A killed saline-suspended vaccine introduced aerogenically marginally increased resistance to small (2 to 5 mean lethal doses) aerogenic challenges. Live mycobacterium bovis (BCG) vaccine immunopotentiated a single dose of heat-killed P. multocida vaccine in terms of the protective immune response against a subsequent footpad challenge. Hyperimmune mouse serum, administered intraperitoneally, intramuscularly, or intravenously 1 to 7 days prior to subcutaneous challenge with 500 to 5,000 P. multocida, was highly protective. Mice injected in the thigh with 0.2 ml of hyperimmune serum prior to footpad challenge frequently developed a severe inflammatory response with local swelling within 48 h; these lesions often became severly abscessed with time. The passively protected, footpad-infected mice developed active immunity against a subsequent challenge given into the opposite footpad.
Effect of ambient temperatures on multiplication of attenuated transmissible gastroenteritis virus in the bodies of newborn piglets
Authors: Furuuchi S , Shimizu Y
Published: January 2020

Newborn piglets were found to be more resistant to infection with attenuated transmissible gastroenteritis virus when maintained at higher temperatures. This was attributed to a decreased rate of virus propagation and spreading in the bodies of the infected animals. The highest virus levels were detected in the tissues of piglets maintained at 8 to 12 C. In contrast, no virus was recovered from piglets maintained at 35 to 37.5 C. The virus was found only in the lymph nodes and respiratory organs in the piglets maintained at 20 to 23 C.
Role of Agrobacterium cell envelope lipopolysaccharide in infection site attachment
Authors: Whatley M H , Bodwin J S , Lippincott B B , Lippincott J A
Published: January 2020

Lipopolysaccharide (LPS) isolated from Agrobacterium tumefaciens inhibited tumor induction by virulent bacteria. LPS from site-binding strains was not effective if added to the plant wound shortly after the bacteria, and LPS from avirulent, non-site-binding strains of Agrobacterium was not inhibitory regardless of the order of addition. However, LPS and whole cells of avirulent strains NT1 and IIBNV6, which lack of Agrobacterim virulence plasmid, were inhibitory. Chromosomal deoxyribonucleic acid thus determines specificity of this essential component of the Agrobacterium infection process.
Genetic control of natural immunity to ecotropic mouse leukemia viruses: immune response genes
Authors: Nowinski R C
Published: January 2020

Humoral immune response to ectropic leukemia viruses in AKR and C57BL/6 mice was controlled by a gene that mapped in linkage group IX. Mice of the AKR strain had an immune nonresponsive allele of this gene, whereas mice of the C57BL/6 strain had an immune responsive allele. Antibody against murine leukemia virus (MuLV) reacted primarily with p15 protein of the viral envelope. It was concluded that the failure to find antibody production in AKR mice was the result of a genetic immunological defect, rather than the result of immunological tolerance that was induced by the persistent viremia of endogenous MuLV.
Multiplication of bovine parvovirus in two cell strains
Authors: Lubeck M D , Johnson F B
Published: January 2020

Bovine parvovirus has been found to infect two cell strains: buffalo lung fibroblasts and embryonic bovine tracheal cells. Infectivity titers were best determined by plaque assays. Immunofluorescence studies revealed intranuclear staining, whereas complement fixation tests confirmed bovine parvovirus antigen production. These cell strains represent economical and convenient cell types which can be used in studies on bovine parvovirus.
Experimental group B streptococcal infections in mice: hematogenous virulence and mucosal colonization
Authors: Furtado D
Published: January 2020

A group B streptococcus recovered from a blood specimen from a neonate with sepsis was used to evaluate the use of mice for studies characterizing the hematogenous virulence and the asymptomatic mucosal colonization of the vagina or of the respiratory tract by these bacteria. When injected intravenously, the 50% lethal dose for mice was 10(6); however, as few as 10(2) organisms produced septic deaths. In mice undergoing water diuresis, bacteriuria and pyelonephritis were not produced after direct bladder inoculation of the streptococci. Asymptomatic vaginal colonizations that persisted for 12 days were produced in both pregnant and virgin mice. Vaginal colonization before delivery did not result in transmission of infection to litters or in protection against subsequent oropharyngeal colonization in the suckling mice. In mice born of nonexposed mothers, oropharyngeal colonization was produced in both suckling and 3-week-old weaned mice. Whereas infection persisted for 14 days in all suckling mice, clearance occurred in over 50% of the weaned mice by day 14. The use of mice for studies on the virulence of the group B streptococci as well as for studies on the pathogenesis of disease by virulent strains is discussed.
Adjuvant effects of diethylaminoethyl-dextran
Authors: Houston W E , Crabbs C L , Kremer R J , Springer J W
Published: January 2020

Diethylaminoethyl-dextran exhibited a significant effect on the primary immune response of rhesus monkeys to formalin-inactivated Venezuelan equine encephalomyelitis virus vaccine (IVEE). Antibody formed to IVEE and adjuvant followed a classic immunoglobulin M-immunoglobulin G pattern; however, as compared with vaccine alone, use of this adjuvant with IVEE reduced the time required for onset of immunoglobulin G synthesis.
Immune interferon production by lymphoid cells: role in the inhibition of herpesviruses
Authors: Babiuk L A , Rouse B T
Published: January 2020

Bovine peripheral blood lymphocytes (PBL) obtained from infectious bovine rhinotracheitis (IBR) virus- and tuberculin-immunized animals produced large quantities of interferon within 24 h of in vitro stimulation by IBR and purified protein derivative antigens. Separation of PBL into populations enriched in T lymphocytes or B lymphocyte provided the antigen-specific step for immune interferon production. A 2- to 10-fold increase in interferon occurred when lymphocytes were combined with autologous macrophages. Although macrophages, even if treated with antilymphocyte serum to remove any contaminating lymphocytes, could produce some interferon, the augmented interferon produced by macrophage-lypmhocyte cultures was not dmpocytes. Direct physical contact between macrophages and lymphocytes was required for the production of enhanced levels of interferon. Antigen-antibody complexes of irradiated virus-infected cells in the presence of antibody were as efficient or better at stimulating interferon than was free antigen. Because IBR virus was inhibited by interferon levels stimulated in cultures by IBR antigen, it was suggested that the local production of interferon by immune cells might play a similar role in curtailing virus dissemination in vivo, thus leading to recovery from disease.
Proteins of typhus and spotted fever group rickettsiae
Authors: Eisemann C S , Osterman J V
Published: January 2020

Purified radioactive rickettsiae were obtained from irradiated and cycloheximide-inhibited L cells, and their proteins were analyzed by polyacrylamide gel electrophoresis. Rickettsial species could be distinguished by comparing the relative mobilities of constituent proteins after migration of two differentially labeled preparations in a single gel. Distinct differences were observed in gel patterns of rickettsiae from the typhus and spotted fever groups, as well as with different species within a group. Rickettsial organisms causing murine and epidemic typhus were clearly distinguished, as were the causative agentsof boutonneuse fever and rickettsialpox. The use of both internal and external molecular weight standards allowed molecular weight estimates for 19 proteins from both Rickettsia prowazekii and Rickettsia conorii. A flexible system for designating rickettsial proteins is proposed that lends itself to modification as more detailed analysis progresses.
Pseudomonas aeruginosa exotoxin: purification by preparative polyacrylamide gel electrophoresis and the development of a highly specific antitoxin serum
Authors: Callahan L T
Published: January 2020

Pseudomonas aeruginosa exotoxin has been purified to a specific activity of 12,000 to 16,000 mouse median lethal doses/mg of protein. Total recovery was about 25%, and the degree of purification was approximately 3,000-fold. Preparative polyacrylamide gel electrophoresis greatly facilitated purification. As judged by analytical disc gel electrophoresis, the purified toxin contained one major band of protein and only a negligible amount of contamination. Antiserum prepared against the purified toxin neutralized the lethal activity of crude toxin preprations and reacted by double immunodiffusion with a single component of concentrated broth cultures of P. aeruginosa isolates obtained from a clinical source.
Inhibition of phagocytosis by cryptococcal polysaccharide: dissociation of the attachment and ingestion phases of phagocytosis
Authors: Kozel T R , Mastroianni R P
Published: January 2020

The effects of cryptococcal polysaccharide and selected serum factors on (i) the attachment of Cryptococcus neoformans to macrophages and (ii) the subsequent ingestion of yeast cells by the macrophages were investigated. Percent attachment was measured after incubation of yeast cells with macrophages at 4 C. Percent engulfment was determined after incubation of yeast cells and macrophages at 37 C. Nonencapsulated yeast cells readily attached to macrophages at the low temperature and were engulfed at a high rate at 37 C, whereas encapsulated yeast cells attached to macrophages at low rates and were engulfed at low rates. Addition of varying doses of purified cryptococcal polysaccharide to nonencapsulated yeast cells inhibited attachment at approximately the same concentration of polysaccharide required for inhibition of engulfment. Nonencapsulated yeast cells that attached to macrophages at 4 C were eluted from the macrophages by addition of purified cryptococcal polysaccharide to the incubation medium. Heat-labile opsonins were not required for attachment of yeast cells to macrophages, but they were necessary for maximal initial rates of phagocytosis. Heat-stable components of serum facilitated attachment of cryptococci, but their most important function appeared to be triggering the ingestion of attached yeast. Specific antiserum had no effect on the attachment and engulfment of nonencapsulated cryptococci, and the antiserum produced only a small enhancement of the engulfment of encapsulated cryptococci.
Fluid and electrolyte transport in rhesus monkeys challenged intracecally with Shigella flexneri 2a
Authors: Kinsey M D , Formal S B , Dammin G J , Giannella R A
Published: January 2020

Shigella flexneri 2a is an invasive enteric pathogen that may produce diarrhea when ingested by human beings and subhuman primates. We have previously shown that shigella diarrhea correlates with water and electrolyte transport abnormalities in the jejunum and colon. Dysentery alone is associated only with colonic transport abnormalities. To define the relationship between invasion and inflammation of the colon and the occurrence of jejunal transport abnormalities, we studied water and electrolyte transport, histology, and bacteriology in rhesus monkeys that were infected by introducing S. flexneri 2a directly into the cecum. In contrast to the pattern of disease seen after oral administration, cecal inoculation resulted in clinical disease in 64% of animals, of which 94% manifested dysentery alone, rarely preceded by mild diarrhea. Histologically, invasion and inflammation was limited to the colon. Secretion of water and sodium occurred in the colon of infected monkeys when compared with controls, whereas transport was normal in the jejunum and ileum. These data further demonstrate that severe dysentery can result from cecal injection of shigellae, but also suggest that the occurrence of watery diarrhea requires and may result from an undefined interaction between the jejunal mucosa and the organisms during transit through the small intestine.
Influence of rodent malaria on the course of Leishmania enriettii infection of hamsters
Authors: Belehu A , Poulter L W , Turk J L
Published: January 2020

Plasmodium yoelii infection was established in hamsters, and the effect of this type of malaria on concurrent Leishmania enriettii infection was examined. It was found that the course of the L. enriettii infection was affected by P. yoelii and that this effect depended on the relative timing of the two infections. A chronic malarial infection with Plasmodium berghei was also established in hamsters, and this was found to affect the course of a concurrent L. enriettii infection in a similar manner to P. yoelii. These results are discussed in relation to current knowledge of the immunosuppressive effects of plasmodia.
Reaginic antibody production to protein antigens of Escherichia coli and Pseudomonas aeruginosa by mice
Authors: Danneman P J , Michael J G
Published: January 2020

Water-soluble antigens isolated from acetone-dried, gram-negative bacteria elicited reaginic antibody formation in mice. Antibodies specific for Escherichia coli antigens reacted with antigens isolated from several enterobacterial species tested, but not with antigens isolated from Pseudomonas aeruginosa. Reaginic antibodies induced by antigens isolated from a P. aeruginosa strain reacted with antigens isolated from several P.aeruginosa serotypes as well as with a purified protein component of the envelope of P. aeruginosa. The anti-Pseudomonas reagins did not cross-react with enterobacterial antigens. Antigenicity of the bacterial extracts was destroyed by trypsin treatment and reduced by heating, which suggested that the antigens were protein in nature. Whole bacterial cells adsorbed out reaginic antibodies, indicating that the antigens are located at or near the surface of the bacteria.
Neonatally thymectomized Lewis rats infected with Mycobacterium leprae: response to primary infection, secondary challenge, and large inocula
Authors: Fieldsteel A H , Levy L
Published: January 2020

Several experiments were carried out to measure the ability of neonatally thymectomized Lewis rats (NTLR) to limit multiplication of Mycobacterium leprae. NTLR inoculated in one hind footpad with 10(7) viable M. leprae and challenged in the other hind footpad with 5 x 10(3) organisms simultaneously or 120 or 180 days later permitted multiplication in both sites. By contrast, immunologically intact rats similarly inoculated did not permit multiplication from either inoculum. NTLR and immunologically normal BALB/c mice were equally susceptible to infection with M. leprae, in that multiplication occurred regularly in the footpads of both species when inoculated with a bacterial suspension diluted to provide five organisms per footpad. Finally, multiplication occurred when five viable M. leprae diluted with 10(7) heat-killed organisms were inoculated into the footpads of NTLR. Although there was some evidence that NTLR are not completely immunosuppressed, NTLR appear to be capable of detecting much smaller proportions of viable M. leprae than can be detected by immunologically normal mice.
Rhesus monkeys kidney cells persistently infected with Simian Virus 40: production of defective interfering virus and acquisition of the transformed phenotype
Authors: Norkin L C
Published: January 2020

Monolayer cultures of LLC-MK2 rhesus monkey kidney cells became persistently infected with simian virus 40 (SV40) when infected at a multiplicity of infection of 100 plaque-forming units/cell. A stable carrier state developed characterized by extensive viral proliferation without obvious cytopathic effect other than the slow growth of these cultures. By 11 weeks all cells produced the SV40 T antigen. In contrast, less than 5% of the cells produced V antigen. Virus-free clonal isolates were obtained by cloning in SV40 antiserum. Continuous cultivation in antiserum resulted in a temporary cure of unclone cultures. When virus did eventually reappear in the "cured" cultures the titers remained low. The virus produced by the carrier culture was defective at both 31 and 37% c, and it interfered with the growth of standard s40 during mixed infection of CV-1 green monkey kidney cells. All of the interfering activity in carrier culture homogenates could be sedimented by centrifugation at 109,000 x g for 3 h. These cultures were completely susceptible to vesicular stomatitis virus. Extensive viral deoxyribonucleic acid synthesis occurred in CV-1 cells infected with carrier culture virus. Carrier culture homogenates are only slightly less cytopathic to CV-1 cells than standard SV40. The carrier culture express several properties of SV40 transformation.
Mechanisms of immunity in typhus infections. VI. Differential opsonizing and neutralizing action of human typhus rickettsia-specific cytophilic antibodies in cultures of human macrophages
Authors: Beaman L , Wisseman C L
Published: January 2020

Human peripheral blood monocytes were incubated in vitro for 6 days to allow time for transformation into macrophage-like cells. Cytophilic antibodies in typhus convalescent human serum were demonstrated by addition of Rickettsia mooseri or Rickettsia prowazeki to passively sensitized human peripheral blood monocyte-derived macrophages that were held at 4 degrees C. Rosettes of rickettsiae were found around macrophages sensitized with immune serum but not around macrophages that had been incubated with normal serum. Inhibition of rosette formation occurred if the macrophages were maintained in normal human serum before addition of immune human serum. Rosettes of R. mooseri were also formed around monocytes obtained from an individual infected with R. mooseri. If the antibody-sensitized macrophages were maintained at 34 degrees C, enhanced phagocytosis of R. mooseri or R. prowazeki occurred as compared with macrophages exposed to normal human serum before infection. However, the cytophilic antibody did not significantly inhibit the subsequent growth of R. prowazeki within the macrophages. This is in contrast to results obtained when R. prowazeki was mixed with immune serum before addition to the macrophage. In the latter case, growth of R. prowazeki was largely inhibited. The significance of antibody cytophilic for macrophages in typhus infections is discussed.
Electron microscopic study on the interaction between normal guinea pig peritoneal macrophages and Coxiella burnetii
Authors: Kishimoto R A , Veltri B J , Canonico P G , Shirey F G , Walker J S
Published: January 2020

An electron microscopic study was conducted to explore the interaction between normal guinea pig peritoneal macrophages and phase I and II Coxeilla burnetii previously treated with either normal or immune serum. A comparison was made on the efficiency of phagocytosis and subsequent killing of rickettsiae by macrophages. Both phases of rickettsiae previously treated with normal serum multiplied within phagosomes after phagocytosis with resultant destruction of macrophages. In contrast, suspending rickettsiae in immune serum rendered them more susceptible to phagocytosis and potentiated their destruction within macrophages.
Association of viable and inactivated Salmonella typhimurium 395 MS and MR 10 with HeLa cells
Authors: Kihlström E , Edebo L
Published: January 2020

The mouse-virulent Salmonella typhimurium 395 MS, containing a complete lipopolysaccharide (LPS) structure with S-specific repeating units, and the nonvirulent, LPS-defective mutant 395 MR 10 (chemotype Rd), derived from it, were studied for their tendency to interact with HeLa cells. In the definition of interaction no distinction has been made between intracellular and cell membrane-attached bacteria. R10 bacteria were found to have a greater tendency to interact than MS bacteria. This difference was seen as early as 1 h after the start of incubation, but it became more pronounced beyond 3 h. Heat-killed and ultraviolet-killed R10 bacteria interacted with HeLa cells less than living ones. Killed MS bacteria interacted to an extent similar to that of living ones. These results are discussed in relation to the susceptibility of the bacteria to phagocytosis by professional phagocytic cells and to the physiochemical properties of the bacteria as measured by their distribution in a two-polymer, aqueous-phase system.
Selective adsorption of heterophile polyglycerophosphate antigen from antigen extracts of Streptococcus mutans and other gram-positive bacteria
Authors: Hamada S , Tai S , Slade H D
Published: January 2020

Hot saline extracts of Streptococcus mutans have been shown to contain antigenic substances which occasionally react nonspecifically with some antisera against whole cells of various serological groups and types of streptococci. Chromatography of the extract of S. mutans strain MT703 (serotype e) on a diethylaminoethyl-Sephadex A-25 column gave two principal antigens. One antigen was eluted without adsorption to the resin and was identified as the serotype-specific polysaccharide. The other antigen, which contained a large quantity of phosphorus, was absorbed to and released from the resin by gradient elution. It was reactive against the antisera specific for polyglycerophosphate (PGP) from group A Streptococcus pyogenes and/or S. mutans strain Ingbritt (type c). The PGP antigen was further purified by gel filtration with Sephadex G-75. Two peaks, PGP-1, and PGP-2, were obtained. Each possessed the same antigenic specificity to anti-PGP serum as shown by immunodiffusion. Chemical analyses revealed that the molar ratio of phosphorus to glycerol in both was about 1:1, although the protein content between the two was significantly different. PGP antigen was found to be widely distributed in hot saline extracts from various gram-positive bacteria, with a few exceptions. However, all gram-negative bacteria examined were free of PGP. The PGP in the hot saline extracts of various gram-positive bacteria possessed an essentially identical antigenic specificity. The addition of diethylaminoethyl-Sephadex A-25 resin to hot saline extracts successfully removed the cross-reacting PGP antigen. After adsorption of the extract from S. mutans, the supernatant contained only type-specific polysaccharide antigen, except type b, in which both type b-specific polysaccharide and PGP antigens were absorbed with the resin. This simple procedure should be useful for the removal of the PGP-type teichoic acid from antigen extracts of bacteria that contain uncharged polysaccharides.
Immunogenic and toxic properties of a purified lipopolysaccharide-protein complex from Pasteurella multocida
Authors: Ganfield D J , Rebers P A , Heddleston K L
Published: January 2020

An immunogenic fraction from Pasteurella multocida was found to consist chiefly of a high-molecular-weight protein-polysaccharide complex containing 25 to 27% protein and 10.7% carbohydrate. The starting material was obtained by differential centrifugation at 105,000 X g of saline extract of P. multocida cells and further purified by gel filtration on Sepharose 2B. Three peaks were usually obtained after gel filtraion.pharose 2B. Three peaks were usually obtained after gel filtration. The component in the first peak amounted to about 10% of the starting material and eluted in the void volume. It was predominately carbohydrate, although some protein was present. Two inoculations of 10 to 20 mug of the first component induced up to 80% protection in mice against a challenge inoculation with P. multocida that killed 100% of the controls. The second, or major, component amounted to about 75 to 95% of the starting material. This fraction contained 25 to 27% protein and 10.7% carbohydrate. Small amounts, 10 to 20 mug, induced active immunity in mice and turkeys, but large amounts could be lethal; the mean lethal dose was 195 mug for mice and 5.7 mug for 10-day-old chicken embryos. The components in the third peak were primarily proteins that gave reactions of nonidentity with the antigens of peak II in gel diffusion. The components present in the third fraction were definitely less effective in the induction of protective immunity than those present in the first or second. Analyses of the protective antigen(s) by the isoelectric focusing procedure in a pH 3 to 10 gradient showed that all of the precipitinogenic activity was found in the range of pH 3 to 4, with a peak at pH 3.7.
Establishment and distribution of Actinomyces viscosus and Actinomyces naeslundii in the human oral cavity
Authors: Ellen R P
Published: January 2020

The intraoral establishment and proportional distribution of suspected periodontal pathogens Actinomyces viscosus and Actinomyces naeslundii were studied using a recently developed differential plating medium, CNAC-20. Saliva and dental plaque samples were collected from 108 subjects ranging in age from infants to young adults; tongue and buccal mucosa samples were collected from only the adult subjects. Catalase-negative A. naeslundii was isolated from 40% of the predentate infants' and almost all other subjects' saliva samples. It predominated among CNAC-20 isolates in the saliva of subjects of all age groups, in the plaques of young children, and in the adult tongue samples. In contrast, catalase-positive A. viscosus was not isolated from predentate infant samples, and its frequency of isolation increased slowly with age (greater than 50% detection by age 7). A. viscosus was isolated in highest relative proportions from dental plaque and buccal mucosa samples. The two closely related species A. viscosus and A. naeslundii apparently differ in respect to factors determining the host age at which they colonize and their relative intraoral distribution in humans.
Isolation of a low-molecular-weight antibacterial system from human amniotic fluid
Authors: Schlievert P , Johnson W , Galask R P
Published: January 2020

A low-molecular-weight antibacterial system has been isolated from human amniotic fluid. The bacterial inhibitor requires the metal cation zinc and a peptide with a molecular weight of 630. The peptide component was purified using ultrafiltration, gel filtration, and ion-exchange chromatography. It can be inactivated by digestion with carboxypeptidase. The amino acid composition of the peptide is: 3 glutamine-glutamic acid, 2 glycine, and 1 lysine. Removal of zinc from the peptide has been shown to remove bacterial inhibitory activity.
Effect of warfarin on the induction and course of experimental endocarditis
Authors: Thompson J , Eulderink F , Lemkes H , van Furth R
Published: January 2020

The effect of warfarin treatment on an experimental endocarditis was studied in rabbits. Warfarin had no effect on the induction of a Streptococcus sanguis infection in catheter-induced endocardial vegetations, and the course of this infection was also unaltered. However, warfarin treatment resulted in rapidly progressive bacteremia, probably due to impaired circulation in clearing organs such as the lungs, liver, and spleen. Warfarin also reduced the survival time of the infected rabbits, in which pulmonary edema and extensive lung hemorrhages may have been a contributory factor.
Experimental herpes simplex virus carditis in mice
Authors: Grodums E I , Zbitnew A
Published: January 2020

Herpes simplex virus types 1 and 2 induced acute and chronic cardiac damage in suckling and weaning mice after intranasal inoculation. Signs of virus replication were detected by light, immunofluorescent, and electron microscopy techniques. Virtually all of the cardiac tissues appeared to be susceptible to herpes simplex virus. The myocardium, however, was most regularly affected. The viral lesions were discrete during the acute phase of infection. The cardiac damage, however, was more extensive in some of the chronically infected mice. Morphologically, these lesions either resembled the acute ones or were associated with inflammatory granulomatous and sclerotic changes.