bluuebunny/crossref_metadata_2025 huggingface dataset:
Class-specific antibody response to group B Neisseria meningitidis capsular polysaccharide: use of polylysine precoating in an enzyme-linked immunosorbent assay
Authors: Leinonen M , Frasch C E
Published: January 2020

An enzyme-linked immunosorbent assay was developed for measuring immunoglobulin class-specific antibodies to Neisseria meningitidis capsular polysaccharides. The polysaccharides were fixed to the solid phase by precoating the polystyrene surface with basic polyamino acids. Polylysine precoating was found to give optimal antibody values with the meningococcal polysaccharides studied. The enzyme-linked immunosorbent assay was used to measure antibodies against group B meningococcal polysaccharide in healthy adults and in patients with group B meningitis. Antibodies to group B polysaccharide in sera obtained from healthy adults were primarily immunoglobulin M (IgM). Although the antibody response to disease was mostly IgM, both IgG and IgM antibodies increased. Infection with group B organisms stimulated anti-group B polysaccharide antibodies even in young patients, aged 6 to 7 months, and the magnitude of the B polysaccharide response increased with the age of the patient.
In vitro effect of hydrocortisone on the attachment and ingestion phases of immunoglobulin G- and complement component 3b-mediated phagocytosis by human neutrophils
Authors: Forslid J , Hed J
Published: January 2020

The fluorescence quenching method (FQ method) was used to investigate the effect of hydrocortisone on the attachment and ingestion phases of immunoglobulin G (IgG)- and complement component 3b (C3b)-mediated phagocytosis by human neutrophils (PMNs). The results were compared with metabolic activity (O2- release) of the phagocytes. When the PMNs were treated with 5 X 10(-5) M hydrocortisone or more, both IgG-mediated and C3b-mediated interactions decreased. The number of intracellular particles decreased as the total number of PMN-associated particles decreased, indicating an effect mainly on particle attachment. This was substantiated by the fact that pretreatment of the PMN with cytochalasin B resulted in a hydrocortisone dose-related decrease of interacting particles. The FQ method made it possible to quantify the stimulus-phagocyte interaction in relation to the metabolic response. Superoxide anion release decreased at the highest concentration of hydrocortisone used (5 X 10(-3) M), which merely reflected the decreased number of interacting particles. No reduction in metabolic activation was obtained when the superoxide anion release was correlated with the number of interacting yeast-IgG particles. The results indicate that hydrocortisone primarily affects the binding capacity of Fc and C3b receptors, resulting in decreased metabolic activation. The effector functions, e.g., ingestion and metabolic activation, were not affected by hydrocortisone in this study.
Reversible increase in the saturation of C18 fatty acids induced by diphtheria toxin in tissue culture cells
Authors: Apostolov K , Barker W
Published: January 2020

Diptheria toxin (DT) reversibly increases the saturation of the long-chain C18 fatty acids (C18FA) in concentrations which are at least 100-fold smaller than the concentrations which completely inhibit host cell protein synthesis. The concentrations required for induction of the increase in the saturation of the C18FA in fibroblasts are 100-fold smaller than the concentrations for the epithelial cells. The increase in saturation of the C18FA was proportional to the concentration of DT. A cytopathic effect appeared with DT concentrations which induced a near-maximal increase in the saturation of the C18FA, but the cells remained viable. However, at very high concentrations of DT and depending on the type of cell, there was no change in the fatty acids, and the treated cells disintegrated. Interferon, which also induces a reversible increase in the saturation of the C18FA (K. Apostolov and W. Barker, FEBS Lett. 126:261-264, 1981), produces a cumulative effect when used in conjunction with DT. This additive effect of DT and interferon is discussed in the light of other similar biological activities of these agents.
Synergistic effect of Candida albicans and Staphylococcus aureus on mouse mortality
Authors: Carlson E
Published: January 2020

A synergistic effect on mouse mortality was demonstrated in combined infection of mice with Candida albicans and a Staphylococcus aureus strain isolated from a patient with toxic shock syndrome. Mice exhibited high resistance when inoculated intraperitoneally by either pathogen alone. Dual infection with the two organisms together, however, at doses which separately caused no animal deaths, resulted in 100% mortality. This synergistic effect could not be reproduced when either of the agents was heat inactivated.
Suppression of tuberculoimmunity in mice by Evans blue dye
Authors: Crowle A J , May M H
Published: January 2020

Evans blue dye, which acts as an adjuvant in mice to promote induction of cell-mediated immunity, was found earlier to apparently act contrarily and decrease tuberculoimmunity. This phenomenon was investigated here in a long-term experiment examining the persistence of the suppressed immunity. Systemic suppression after injection of only 1 mg of dye was confirmed and was found to last for more than 2 weeks but less than 11 weeks. Localized evidence for dye-caused depression of tuberculoimmunity also was seen by the development in dye-injected mice immunized with emulsified whole bacilli of caseating tubercles at sites of pulmonary infection, a very rare type of pathology for tuberculosis in mice. The experiment, in addition, detected an aging-related decline in mice of resistance to tuberculosis evident by shortened survival, enlarged pulmonary tubercles, and increased susceptibility to systemic tuberculin reaction.
Effect of staphylococcal alpha-toxin on phagocytosis of staphylococci by human polymorphonuclear leukocytes
Authors: Gemmell C G , Peterson P K , Schmeling D J , Quie P G
Published: January 2020

Evidence is presented which shows that alpha-toxin elaborated by Staphylococcus aureus can affect certain of the biological functions of human polymorphonuclear leukocytes (PMN). The pretreatment of PMNs with low doses (less than 10 hemolytic units) of purified toxin enhanced their ability to phagocytose and to kill serum-opsonized staphylococci. At higher doses (greater than or equal to 10 hemolytic units), a certain amount of cell damage was caused (detectable by trypan blue uptake), which resulted in a reduction of their phagocytic capacity. These effects were not seen either with heat-inactivated (60 degrees C, 30 min) or with antibody-neutralized toxin. It is thought that the toxin can bind onto the membrane of the PMN, either exposing additional receptor sites for opsonized bacteria or speeding up the transport of staphylococci across the membrane. Once ingested, the bacteria were more readily killed by the PMNs.
Kinetics and genetics of herpes simplex virus-induced antibody formation in mice
Authors: Knoblich A , Görtz J , Härle-Grupp V , Falke D
Published: January 2020

The kinetics of antibody synthesis was investigated after intraperitoneal, subcutaneous, and footpad infection of various strains of mice with herpes simplex virus. Immunoglobulin M antibodies appeared 5 days after and immunoglobulin G antibodies appeared 10 to 12 days after intraperitoneal infection with herpes simplex virus type 1. The major histocompatibility complex and the background genome of inbred mice were not found to have a systematical influence on antibody synthesis. Female mice, however, consistently produced more antibodies than did male if the infection was done intraperitoneally, but not if it was done subcutaneously or into footpads. Castration considerably increased the amount of antibodies produced by male mice. The difference in antibody formation between females and males could be abolished by injection of silica; moreover, antibody titers were enhanced by this treatment. This has also been found by immunization with a Formalin-inactivated herpes simplex virus vaccine. The effect of silica in enhancing antibody formation could be observed up to 12 days after infection. Infectious virus could be detected up to 2 days after infection, and herpes simplex virus type 1 antibody-stimulating antigens could be detected up to 4 days in ultrasonicates of macrophages. The assumption is made that androgen-sensitive cell populations, including macrophages and their soluble products, are involved in antibody-depressing mechanisms.
De novo purine synthesis, purine salvage, and DNA synthesis in normal and Lesch-Nyhan fibroblasts infected with Mycoplasma pneumoniae
Authors: Upchurch S , Gabridge M G
Published: January 2020

The effects of Mycoplasma pneumoniae on host cell metabolism were studied by using two types of host cells, MRC-5 human lung fibroblasts, a normal cell line, and Lesch-Nyhan fibroblasts, a cell line deficient in hypoxanthine-guanine phosphoribosyl transferase (EC 2.4.2.8). The susceptibilities of the two cell types were determined by infecting the cells with M. pneumoniae at different multiplicities of infection (MOI). Our data indicate that the Lesch-Nyhan cells were four times more susceptible to damage by M. pneumoniae than the MRC-5 cells. The effects of different MOIs (10 and 50) on de novo purine synthesis. DNA synthesis, and the development of a cytopathic effect were determined. In both cell types, the higher MOI inhibited de novo purine synthesis to a greater extent than the lower MOI. This correlated closely with the cytopathic effect which developed in the monolayers (i.e., the more the inhibition of de novo purine synthesis, the greater the cytopathic effect which developed). In the Lesch-Nyhan cells, DNA synthesis was completely inhibited by the high MOI, whereas in the MRC-5 cells, DNA synthesis was stimulated by the high MOI. In the MRC-5 cells infected with M. pneumoniae, purine salvage activity increased, as indicated by an increase in adenosine deaminase (EC 3.5.4.4) activity. These data indicate that M. pneumoniae alters host cell metabolism, particularly the nucleic acid metabolic pathways. This may explain in part the mechanism of pathogenesis of M. pneumoniae infection.
Protective effect of a muramyl dipeptide analog encapsulated in or mixed with liposomes against Candida albicans infection
Authors: Fraser-Smith E B , Eppstein D A , Larsen M A , Matthews T R
Published: January 2020

Encapsulation of N-acetylmuramyl-L-alpha-aminobutyryl-D-isoglutamine in multilamellar vesicles composed of phosphatidylcholine, cholesterol, and phosphatidylserine (7:6.7:3) or phosphatidylcholine and phosphatidylserine (7:3) reduced the amount of drug needed to protect against a Candida albicans intravenous infection. The 50% effective doses for encapsulated and free drug were 5.5 and greater than 80 mg/kg, respectively. The optimum treatment was twice (at days 4 and 2 preinfection) by the intravenous route. Intraperitoneal, subcutaneous, and oral routes of administration were ineffective. The same potentiation of anti-Candida activity was observed whether the lower dose of drug was encapsulated in multilamellar vesicles, mixed with multilamellar vesicles, or given either 1 h before or 1 h after multilamellar vesicles. It was postulated that the mechanism of action involved the retention of the liposomes by organs of the reticuloendothelial system, resulting in an enhanced response of the macrophages to the immunostimulating activity of the N-acetylmuramyl-L-alpha-aminobutyryl-D-isoglutamine given in conjunction with the vesicles.
Characterization of plasmids in bacterial fish pathogen
Authors: Toranzo A E , Barja J L , Colwell R R , Hetrick F M
Published: January 2020

Plasmid profiles of representative fish pathogens, Aeromonas salmonicida, Aeromonas hydrophila, Vibrio anguillarum, Pasteurella piscicida, Yersinia ruckeri, Edwardsiella tarda, and Renibacterium salmoninarum, were determined by agarose gel electrophoresis with four different plasmid detection methods. A combination of two methods was required to detect the plasmids present in these strains and to calculate precisely the molecular weights of the plasmids. Of 38 strains, 28 harbored one or more plasmids, with the majority of strains demonstrating multiplasmid banding. Similarity in plasmid banding between strains was noted and related to geographic source. Five strains of A. salmonicida possessed six plasmid bands having molecular weights of 8.6 X 10(6), 8.4 X 10(6), 8.1 X 10(6), 3.6 X 10(6), 3.5 X 10(6), and 3.4 X 10(6). Four P. piscicida isolates shared three plasmid bands having molecular weights of 37 X 10(6), 15 X 10(6), and 5 X 10(6), and five A. hydrophila strains harbored a common plasmid having a molecular weight of ca. 20 X 10(6) to 30 X 10(6). The highest-molecular-weight plasmids (145 X 10(6) and 130 X 10(6) were detected in V. anguillarum. From curing experiments, it was found that in A. hydrophila strain 79-62, a loss of resistance to tetracycline was associated with loss of plasmid content in all susceptible derivatives, suggesting plasmid-mediated tetracycline resistance. Cell surface characteristics and metabolic properties were also modified in cured derivatives of A. hydrophila strain 79-62.
Transformed and normal cell surface glycoproteins that induce interferon production by nonsensitized lymphocytes
Authors: Smith E M , Hughes T K , Blalock J E
Published: January 2020

Nonsensitized human and mouse lymphocytes produce interferon (IFN)-alpha and IFN-alpha/beta, respectively, in response to most transformed cells, as well as normal xenogeneic cells. The treatment of transformed human (WISH) and mouse (L) or normal mouse embryo (ME) cells with trypsin, pepsin, or neuraminidase resulted in a loss of the cell's ability to induce IFN-alpha or alpha/beta. These findings suggested that a cell surface glycoprotein is responsible for the induction of IFN-alpha and alpha/beta. The sonication of WISH, L, and ME cells released glycoproteins which retained IFN-inducing activity in the soluble form. The inducing molecules from WISH, L, and ME cells had molecular weights of 130,000, 90,000, and 68,000, respectively. Further purification, plus a confirmation of the glycoprotein nature of the inducer molecule, was shown by specifically binding and eluting L cell inducer bioactivity from a concanavalin A-Sepharose affinity column.
Staphylococcal peptidoglycan: T-cell-dependent mitogen and relatively T-cell-independent polyclonal B-cell activator of human lymphocytes
Authors: Levinson A I , Dziarski A , Zweiman B , Dziarski R
Published: January 2020

Staphylococcal cell wall products have been widely examined as probes for dissection of in vitro human immune responses. Mitogenic and polyclonal B-cell-activating properties have been attributed to intact cell walls or the protein A constituent thereof. We now report that staphylococcal peptidoglycan (PG), the major cell wall constituent, is not only a potent mitogen but also a polyclonal B-cell activator for human peripheral blood mononuclear cells (PBM). PG-induced proliferative responses of human PBM were comparable to that observed in pokeweed mitogen-stimulated cultures. As was true for pokeweed mitogen, PG-induced proliferation required the presence of T-cell help. Cultures of human PBM with PG also resulted in B-cell differentiation as reflected by an increase in numbers of immunoglobulin-secreting cells in stimulated cultures. In contrast to the proliferative response, PG-induced B-cell differentiation was relatively T-cell independent. This point became apparent when B-cell fractions were partially depleted of excessive numbers of monocytes before culture. Also, B-cell proliferation did not appear to be a major prerequisite for PG-induced B-cell differentiation responses. These data indicate that PG is a potent T-cell-dependent mitogen and relatively T-cell-independent polyclonal B-cell activator of human lymphocytes.
Non-cariogenicity of the disaccharide palatinose in experimental dental caries of rats
Authors: Ooshima T , Izumitani A , Sobue S , Okahashi N , Hamada S
Published: January 2020

The caries-inducing activity of palatinose (isomaltulose, alpha-D-glucopyranosyl-1,6-fructose) was examined in in vitro and in vivo experiments, comparing it with other carbohydrates. When Streptococcus mutans was successively subcultured in a broth medium containing 1% palatinose, the strains belonging to serotype a, d, or g did not ferment palatinose, whereas the strains belonging to serotype b, c, e, or f did ferment palatinose. Furthermore, palatinose significantly inhibited the synthesis of insoluble glucan from sucrose by S. mutans. Specific-pathogen-free rats which had been infected with S. mutans 6715 and fed a diet containing 56% palatinose did not develop significant dental caries. However, rats infected similarly, but fed a diet containing sucrose, glucose, fructose, or a glucose-fructose mixture manifested significant caries when compared with the noninfected, sucrose-fed control rats. Furthermore, it was found that replacement of half of the sucrose content with palatinose resulted in decreased caries development compared with caries development in rats fed the sucrose diet.
Suppression of Murine Lymphocyte Mitogen Responses by Exopolysaccharide from
Capnocytophaga ochracea
Authors: Bolton Ronald W. , Dyer John K.
Published: January 2020

An extracellular polysaccharide was purified from culture supernatants of
Capnocytophaga ochracea
25, a gram-negative bacillus associated with human periodontal disease. The extracellular polysaccharide suppressed in vitro mitogenic responses of murine splenic lymphocytes to concanavalin A and lipopolysaccharide. This suppression wad dose dependent, persisted up to 120 h, and was not caused by direct toxicity of the extracellular polysaccharide.
Correlation of Virulence and Collagenolytic Activity in
Entamoeba histolytica
Authors: Gadasi Hana , Kessler Efrat
Published: January 2020

The levels of collagenolytic activity of strains HM-1:1 MSS (HM-1), (HM-1), 200-NIH, and HK-9 of
Entamoeba histolytica
were compared. Collagen degradation was evaluated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Conditioned media as well as extracts of the highly virulent strain HM-1 effectively degraded native type I collagen. Significantly lower activity was found in the analogous fractions of strains 200-NIH and HK-9, which are not as virulent. The collagenolytic activity of strain HM-1 was associated with the isolated plasma membrane fraction and could be eluted from the membranes by buffers of high ionic strength, indicating that it is not an integral membrane protein. Unlike the vertebrate and the clostridial collagenases, the collagenolytic activity of
E. histolytica
HM-1 was enhanced in presence of dithiothreitol and was inhibited by
N
-ethylmaleimide. The correlation between virulence of the individual strains and their collagenolytic activity suggests that collagenase might play a role in pathogenesis of amoebiasis. The localization of the enzyme on the plasma membrane and its presence in the extracellular medium favor this view.
Temperature-sensitive events during the replication of the attenuated S-1 clone of dengue type 2 virus
Authors: Eckels K H , Summers P L , Russell P K
Published: January 2020

Temperature-sensitive events occurring during the replication of the attenuated S-1 clone of dengue type 2 virus were examined. The S-1 clone was more thermolabile than the parent virus at the nonpermissive temperature of 38.5 degrees C. Adsorption experiments in fetal rhesus monkey lung cells revealed an inefficient adsorption of S-1 at 38.5 degrees C compared with the parent virus, suggesting an alteration in a thermolabile virion protein important in adsorption. The production of S-1 viral RNA and antigen occurred at the nonpermissive temperature, which indicated that early events in the replication cycle of S-1 were not affected. Release of infectious virus at 38.5 degrees C was not impaired; however, lower amounts of infectious virus in infected cells at the nonpermissive temperature indicated that maturation of the S-1 clone was suppressed.
Frequency of Cytotoxic T Lymphocyte Precursors to Herpes Simplex Virus Type 1 as Determined by Limiting Dilution Analysis
Authors: Rouse Barry T. , Larsen Hal S. , Wagner Hermann
Published: January 2020

The conditions for establishing a limiting dilution assay to measure cytotoxic T lymphocyte precursors (CTL-P) against herpes simplex virus type 1 (HSV-1) were determined. Analysis by Poisson statistics demonstrated that the estimated frequency of HSV-1-reactive cells in the spleens of normal mice was less than 1/250,000. In contrast, mice immunized previously with infectious HSV-1 demonstrated a CTL-P frequency between 1/3,500 and 1/15,670. The generation of a maximum cytotoxic T lymphocyte response required that mice be primed in vivo with infectious virus. Immunization with inactivated virus either failed to elicit detectable CTL-P frequencies or gave frequencies markedly less than those induced with infectious virus. To obtain positive cultures, the responder cell population had to be exposed to stimulator splenocytes expressing viral antigens. Normal splenocytes without virus or normal splenocytes with T cell growth factor did not result in significant cytotoxicity. Split culture analysis comparing cytotoxicity against syngeneic and allogeneic virus-infected targets provided evidence for specificity,
H-2
restriction, and the T cell nature of the CTL-P. It was determined that precursors were eliminated by treatment with anti-Thy 1, Lyt 2.1, or Lyt 1.1, indicating the CTL-P were Lyt 1
+
2
+
cells. Cytotoxicity was reduced after treatment of the responders with anti-Lyt 2 plus complement, which gave further evidence of the T cell nature of the cytotoxic T lymphocytes. These experiments demonstrated the feasibility of using the limiting dilution approach as a highly sensitive and quantitative means to measure the cell-mediated immune response to HSV-1 antigens.
Agglutination of Mouse Erythrocytes by
Eperythrozoon coccoides
Authors: Iralu Vichazelhu , Ganong Kevin D.
Published: January 2020

Erythrocytes from blood of mice infected with
Eperythrozoon coccoides
for 3 or 4 days agglutinated spontaneously. Washed
E. coccoides
particles agglutinated washed erythrocytes of uninfected mice.
E. coccoides
-mediated agglutination of normal mouse erythrocytes would be an excellent system for studies of bacterial adhesion.
Antigen analysis of several pathogenic strains of Trichomonas vaginalis
Authors: Alderete J F
Published: January 2020

Analysis of several human strains of Trichomonas vaginalis and one bovine strain of Tritrichomonas foetus was accomplished with standard sodium dodecyl sulfate-gel electrophoresis and fluorography technology. Highly motile, live trichomonads were radiolabeled, and total trichloroacetic acid-precipitated proteins were electrophoresed. Complex protein profiles of the various human strains of T. vaginalis were obtained with proteins ranging in molecular weight from 20,000 to greater than 200,000. The parasite biosynthesis of the Coomassie brilliant blue-stained protein bands was demonstrated by efficient radiolabeling of trichomonads with [35S]methionine or a 3H-amino acid digest before electrophoresis and fluorography. Immunogenic trichomonal proteins were then identified by a radioimmunoprecipitation method. A detergent extract of [35S] methionine-labeled T. vaginalis proteins was mixed with serum from an immunized rabbit or pooled sera from subcutaneously infected mice and soluble antibody-antigen complexes isolated by adsorption to protein A-bearing Staphylococcus aureus. The radiolabeled protein antigens were then identified by gel electrophoresis and fluorography. Immunized rabbit serum and pooled sera from challenged mice contained high-titered antibody which reacted with numerous high- and low-molecular-weight proteins. Individual subcutaneously infected mice were found to possess identical antibody responses to these immunogenic trichomonal proteins. A high degree of serological cross-reactivity among the various trichomonads was demonstrated. No differences in the composition of immunogenic proteins were observed among cultures grown in vitro for various lengths of time under the experimental conditions employed. Finally, electrophoretic analysis of cloned colonies of T. vaginalis organisms revealed no differences in their protein composition. The biological relevance of these observations is discussed.
Evaluation of the protective efficacy of Haemophilus influenzae type b vaccines in an animal model
Authors: Halsey N A , Johansen T L , Bowman L C , Glode M P
Published: January 2020

The infant rat model for Haemophilus influenzae type b (Hib) disease was modified to test the protective efficacy of candidate vaccines. The administration of 0.4 micrograms of polyribosylribitol phosphate (PRP) combined with pertussis vaccine or with diphtheria toxoid-tetanus toxoid-pertussis vaccine (DTP) to infant rats at 5, 10, and 15 days of age resulted in significant (P less than 0.001) protection against Hib bacteremia after challenge at 20 days of age. A dose-response effect was demonstrated, and suppression of the protective effect was noted with high doses of PRP. The administration of pertussis vaccine alone or DTP alone did not protect against Hib bacteremia. The degree of protection against Hib bacteremia correlated (P less than 0.005) with the serum anti-PRP antibody response measured before bacterial challenge. High-molecular-weight PRP preparations administered alone or in combination with pertussis vaccine were less effective than standard PRP combined with pertussis vaccine. These data compare favorably with the antibody response in human infants receiving the same vaccines. The observations in this animal model should facilitate the development and testing of other vaccine preparations for human infants.
Chemical and immunological comparison of surface fibrils of strains representing six taxonomic groups of Actinomyces viscosus and Actinomyces naeslundii
Authors: Masuda N , Ellen R P , Fillery E D , Grove D A
Published: January 2020

Human isolates of Actinomyces viscosus and Actinomyces naeslundii have been divided into six clusters in a numerical taxonomy study. Surface fibrils of strains representing these clusters were isolated and purified. Chemical analyses revealed that the major component of all fibrils was protein and that although differences in percentages of specific amino acid residues were found, the relative proportions of basic, acidic, polar uncharged, and nonpolar amino acids were rather similar among clusters. All of the fibrils except those from strain B236 (cluster 2) either failed to migrate or penetrated only slightly into gels during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, even after boiling, reduction, or alkylation. Immunological studies by electron microscopic examination of fibril-antibody immunocomplexes, whole bacterial cell agglutination, inhibition of hemagglutination, and immunofluorescence by using antifibril antisera and antibodies demonstrated that strains of typical A. naeslundii (cluster 5) have a specific fibril-associated antigen(s) distinct from those of strains of other clusters. Cross-reactions for atypical A. naeslundii (cluster 3) were few. The fibrils from A. viscosus clusters 1, 2, 4, and 6 demonstrated several cross-reactions. By absorbing antifibril antibodies with cross-reactive strains it was possible to obtain cluster-specific antibodies, as determined by whole cell agglutination, only for cluster 5. Absorbed antifibril antisera for both A. naeslundii clusters 3 and 5 were specific by indirect immunofluorescence, whereas anti-cluster 1 fibril antisera cross-reacted only with other A. viscosus cluster representatives. Purification of Actinomyces fibrils by methods used for appendages of other species yields preparations containing common antigens among taxonomic groups. However, absorbing antifibril antisera, gamma globulin, or both has promise for producing cluster-specific reagents useful in identification.
Chronic colonization of rat airways with Pseudomonas aeruginosa
Authors: Boyd R L , Ramphal R , Rice R , Mangos J A
Published: January 2020

Colonization of the airways of rats by Pseudomonas aeruginosa was established by treating the animals with hexamethylphosphoramide (HMPA) and inoculating with P. aeruginosa. Male Sprague-Dawley rats were given tap water (controls) or HMPA in the drinking water at 2 or 4 mg/ml. The ciliated cells of the airway epithelium were denuded, and microulcerative lesions in the epithelium were induced in the HMPA-treated rats. After 2 weeks of treatment, the rats were inoculated by transoral intratracheal instillation with 5 X 10(7) CFU of P. aeruginosa obtained from a cystic fibrosis patient. Two weeks after inoculation, P. aeruginosa was cultured from the airways, and scanning and transmission electron microscopy showed bacilli adhering to or invading the injured airway epithelium. P. aeruginosa was present in tracheal and intrapulmonary tissue homogenates of 9% of the P. aeruginosa-inoculated control rats (n = 22) as compared with 61% of the 2-mg/ml (n = 18) and 65% of the 4-mg/ml (n = 20) HMPA-treated rats (P less than 0.05). No dose-response relationship was found between 2 and 4 mg of HMPA per ml and colonization. Contamination of 47% of all of the rats with Mycoplasma pulmonis, as indicated by a positive enzyme-linked immunosorbent assay for immunoglobulin G, had no discernible significant effect on colonization by P. aeruginosa. These results indicate that colonization of the rat airway by P. aeruginosa can be achieved experimentally by treating the animals with HMPA. This research supports the hypothesis that colonization by P. aeruginosa may occur in airways where the ciliated epithelium has been injured and epithelial lesions exist.
Pathogenicity of
Dermatophilus
and
Geodermatophilus
Authors: Gordon Morris A. , Perrin Ula
Published: January 2020

Cutaneous infection in laboratory animals could not be induced with any of several strains of
Geodermatophilus
. A model for consistent production of streptotrichosis in rabbits, with cultures of
Dermatophilus congolensis
, is presented.
Influence of Glucose Levels on the In Vitro Phagocytosis of Bacteria by Human Neutrophils
Authors: van Oss Carel J.
Published: January 2020

A previously developed in vitro method for studying the phagocytosis of bacteria and particles by human neutrophils was used to investigate the influence of different glucose levels on phagocytosis. It was found that high glucose levels (200, 400, and 800 mg of glucose/100 ml) significantly depressed the phagocytosis of
Staphylococcus epidermidis, S. aureus
, and
Escherichia coli
. At very low glucose levels, a somewhat decreased phagocytic activity was noted. The strongest phagocytic activity occurred at glucose concentrations of 50 and 100 mg/100 ml. A second effect noted at the higher (200, 400 and 800 mg/100 ml) glucose concentrations was a decreased adhesiveness of the neutrophils to solid surfaces. The mechanism of the decrease in phagocytosis and in neutrophil adhesiveness at higher glucose levels is unknown, but it is not linked to increased osmotic pressures due to the presence of glucose, as ethanol, at the same and even higher osmolal concentrations, had no effect on the phagocytosis. These results show that not only the phagocytic activity of those neutrophils that do adhere to a solid surface is diminished at higher glucose concentrations but also that fewer neutrophils adhere to solid surfaces at higher glucose levels. These two phenomena combined may provide at least part of the explanation for the well-known decrease in resistance to bacterial infections of diabetics.
Characterization of Antigens from the Yeast Phase of
Histoplasma capsulatum
Authors: Garcia John P. , Howard Dexter H.
Published: January 2020

Preliminary studies established methods for obtaining maximum yields of viable cells. Liquid shaken cultures of
Histoplasma capsulatum
provided a maximum of 4 × 10
8
to 5 × 10
8
cells/ml regardless of the inoculum size. Under optimum conditions, cells were viable (90 to 95%) for 168 to 240 hr. Generation times ranged from 7.52 to 8.36 hr. Immunodiffusion, immunoelectrophoresis, and ultracentrifugation studies on phenol and ethylenediamine extracts of intact cells and cell walls revealed the presence of two components in the ethylenediamine extracts and three in the phenol preparations. The ethylenediamine extracts from intact cells and cell walls seemed to be identical although one of the components was more abundant in cell walls. Mice injected intraperitoneally with intact cells or cell walls were protected against intravenous challenge with
H. capsulatum
. Among the extracts, the water-soluble ethylenediamine extract from cell walls was most immunogenic. The other extracts gave only a light protection or none at all. Intact cells and cell walls were slightly toxic to mice. Two of the extracts were toxic when incorporated into Freund's complete adjuvant.
Age-Related Susceptibility to
Haemophilus influenzae
Type b Disease in Rabbits
Authors: Schneerson Rachel , Robbins John B.
Published: January 2020

Anti-type b antibodies were detected in newborns and in most 6- to 8-week-old rabbits, similar to those observed in humans. Culture of the pharynx and rectum of rabbits at varied ages failed to yield
Haemophilus influenzae
type b. Bacteria with cross-reacting antigens were observed in both the pharyngeal and rectal cultures from rabbits of varied ages. Rabbits without preexisting serum anti-type b antibodies were highly susceptible to septicemia and death after injection of small numbers of
H. influenzae
type b. Passively acquired antibody, whether observed in newborns or injected into animals without preexisting antibodies, conferred a high degree of protection against
H. influenzae
type b.
Separation of Two Hemolysins from
Aeromonas hydrophila
by Isoelectric Focusing
Authors: Wretlind Bengt , Möllby Roland , Wadström Torkel
Published: January 2020

Two hemolytic proteins (hemolysin I and II) with isoelectric points of 5.5 and 4.3 were purified by isoelectric focusing from the culture supernatant fluid of
Aeromonas hydrophila
. The purified hemolysins were unstable after dialysis and upon heating at 37 C and were inactivated by proteolytic enzymes. The two hemolysins were lethal for mice in a dose of 10 μg (hemolysin I) and 3 μg (hemolysin II) and showed similar hemolytic spectra on red cells of different species. Both were also cytotoxic for HeLa cells and human fibroblasts and gave dermonecrosis in rabbits.
Identification of Enterotoxin E
Authors: Bergdoll Merlin S. , Borja Concordia R. , Robbins Ruth N. , Weiss Karl F.
Published: January 2020

Identification of a new enterotoxin was accomplished by purification of the enterotoxin produced by staphylococcal strain FRI-326 and by preparation of specific antitoxin to the enterotoxin. Toxicity of the preparations was determined in rhesus monkeys, and specificity of the enterotoxin-antitoxin reaction was determined in gel diffusion plates. The enterotoxin was designated enterotoxin E.
Type-Specific Antigens of Group B Type Ic Streptococci
Authors: Wilkinson Hazel W. , Eagon R. G.
Published: January 2020

The type-specific antigens of group B type Ic (old designation type Ii) streptococci were extracted, purified, and characterized by serological and chemical methods. The Ia antigen, shared by types Ia and Ic, is a polysaccharide composed of 69% galactose and 25% glucosamine (i.e., 31%
N
-acetyl-glucosamine). However, these monosaccharides failed to inhibit significantly the quantitative precipitin reactions between purified antigen and type Ia antiserum. Indications are that the immunodominant group of this antigen consists of more than a simple monosaccharide. The Ic antigen, shared by types Ib and Ic, is a protein unrelated to the X and R protein antigens. Ic antigen consists of two serologically active determinants, one of which is susceptible to both trypsin and pepsin digestion and the other to pepsin but not to trypsin digestion. Acrylamide gel electrophoresis of the partially purified Ic antigen resulted in the occurrence of both determinants throughout the length of the gel, as shown by double gel diffusion slides.
Electron Microscopic Localization of Alpha Toxin Within the Staphylococcal Cell by Ferritin-Labeled Antibody
Authors: Coulter J. R. , Mukherjee T. M.
Published: January 2020

Highly purified staphylococcal alpha toxin has been used to produce monospecific anti-alpha antibody in rabbits. Gamma globulin prepared from the serum of these rabbits was coupled with ferritin by using toluene diisocyanate. Staphylococcal cells which had been disrupted by two passages through an LKB X-press were treated with this conjugate. Electron microscopic examination of this material showed alpha toxin or an antigenically mature precursor located on the cytoplasmic membrane. The possible function of alpha toxin in this situation is discussed.
Characteristics of
Escherichia coli
Serotypes in the Yanomama, a Primitive Indian Tribe of South America
Authors: Eveland W. C. , Oliver W. J. , Neel J. V.
Published: January 2020

From stool samples of isolated subjects from members of the Yanomama tribe of South America, 432 isolates of
Escherichia coli
were obtained from 72 individuals. Two hundred and four of these strains were typable with a standard panel of 147 O antisera; included in the above were eight enteropathogenic strains. From the untypable strains, antisera were produced, and 13 serologically distinct O serotypes were identified. These data substantiate the ubiquity of known strains of
E. coli
as microhabitants of man's internal environment. The finding of 13 new O serotypes suggests that, in efforts to understand the ecosystem of primitive man, the internal milieu must also be investigated.
Detection of type-specific antibody to herpes simplex virus type 1 by radioimmunoassay with herpes simplex virus type 1 glycoprotein C purified with monoclonal antibody
Authors: Arvin A M , Koropchak C M , Yeager A S , Pereira L
Published: January 2020

Herpes simplex virus type 1 (HSV-1) and HSV-2 specify at least four glycoproteins designated gA/gB, gC, gD, and gE. Previous studies have shown that gC produced by HSV-1 is antigenically distinct from the corresponding HSV-2 glycoprotein. With the exception of gC, the glycoproteins of both serotypes share antigenic sites. Standard serological assays fail to differentiate the antibody to the shared antigenic determinants from the type-specific antibody. In this paper, we describe a procedure for purifying gC from HSV-1-infected cell extracts with an immunoadsorbent prepared with an HCL monoclonal antibody. When used in a solid-phase radioimmunoassay, gC proved to be a type-specific antigen for quantitation of antibody to HSV-1. Among individuals who had no antibody to HSV at the onset of infection, all of those with primary HSV-1 infection developed antibody to gC. Subjects with primary HSV-2 infection failed to develop antibody reactive with gC of HSV-1 (P less than 0.01). Both immunoglobulin G and M antibodies against gC were detected in sera from subjects with either primary or recurrent HSV-1 infection. Higher antibody titers to gC were found in sera from individuals with recurrent infection than in sera from those with primary HSV-1 infection.
HLA-restricted T lymphocyte-mediated cytotoxicity against herpes simplex virus-infected cells in humans
Authors: Yasukawa M , Shiroguchi T , Kobayashi Y
Published: January 2020

Cytotoxic T lymphocytes (CTL) against herpes simplex virus (HSV) were induced in vitro from human peripheral blood lymphocytes by stimulation with HSV antigen. CTL generated by HSV type 1 (HSV-1) antigen stimulation killed not only HSV-1-infected target cells but also HSV type 2 (HSV-2)-infected target cells, though at a lower level. This evidence suggests that CTL against HSV recognize the HSV type-specific and type-common determinants on HSV-infected target cells. These CTL were generated from high responders against HSV-1 antigen as measured by antigen-specific T lymphocyte proliferation in vitro, but not to such an efficient degree from low responders. The cytotoxic activities of CTL against the allogeneic HSV-infected target cells were high when at least one of the HLA-A or -B antigens was shared. However, the HLA-A and -B nonidentical target cells were not killed effectively. The data presented here suggest the possibility of HLA restriction of HSV-specific CTL in humans.
Granulocyte phagocytosis of Streptococcus pneumoniae in properdin-deficient serum
Authors: Braconier J H , Odeberg H , Sjöholm A G
Published: January 2020

Properdin (P)-deficient human serum containing type-specific anticapsular antibodies and having an intact classical pathway of complement did not support efficient granulocyte phagocytosis of Streptococcus pneumoniae serotypes 6A, 14, 19F, 23F, or 35 in an opsonophagocytic assay. Compared with pooled control serum, the difference was most pronounced for serotypes 14 and 23F and at a final serum concentration of 16%. The reduced phagocytic killing of S. pneumoniae serotype 23F in P-deficient serum was due rather to defective opsonization than to impaired intracellular killing. The uptake of C3 by the serotype 23F strain was found to be low in P-deficient serum. The addition of native P promoted C3 fixation as well as opsonization. The activity of P was abolished by heat treatment (56 degrees C, 30 min). Experiments with Mg EGTA to block C1 activation suggested that serotype 23F pneumococci were more dependent on the classical pathway for opsonization than were serotype 35 pneumococci, which appear to be partly opsonized through the alternative pathway alone.
Further Studies on the Immunogenicity of
Haemophilus influenzae
Type b and Pneumococcal Type 6A Polysaccharide-Protein Conjugates
Authors: Chu Chiayung , Schneerson Rachel , Robbins John B. , Rastogi Suresh C.
Published: January 2020

Conjugates were prepared by carbodiimide-mediated coupling of adipic acid hydrazide derivatives of
Haemophilus influenzae
type b (Hib),
Escherichia coli
K100, and pneumococcal 6A (Pn6A) polysaccharides with tetanus toxoid (TT), as an example of a “useful” carrier, and horseshoe crab hemocyanin (HCH), as an example of a “nonsense” carrier. These conjugates were injected into NIH mice, and their serum antibody responses to the polysaccharides and proteins were characterized. As originally reported, Hib conjugates increased the immunogenicity of the capsular polysaccharide and elicited greater than the estimated protective levels of anti-Hib antibodies in most recipients after one injection and in all after the third injection (Schneerson et al., J. Exp. Med.
152
:361-376, 1980). Both Hib conjugates induced similar anti-Hib responses. The K100-HCH conjugate was more immunogenic than the K100-TT conjugate and elicited anti-Hib responses similar to the Hib conjugates after the third injection. Simultaneous injection of the K100 and the Hib conjugates did not enhance the anti-Hib response. The Pn6A-TT conjugate induced low levels of anti-Hib antibodies; when injected simultaneously with the Hib conjugates, the anti-Hib response was enhanced, as all mice responded after the first injection and with higher levels of anti-Hib than observed with the Hib conjugates alone (
P
< 0.05). The Pn6A conjugates were not as immunogenic as the Hib conjugates. Pn6A-TT was more effective than was Pn6A-HCH; it elicited anti-Pn6A (>100 ng of antibody nitrogen per ml) in 6 of 10 mice after the third injection. The addition of the Hib-HCH conjugate to the Pn6A-TT conjugate increased the anti-Pn6A response with a higher geometric mean antibody titer, and 9 of 10 mice responded after the third injection. A preparation of diphtheria toxoid, TT, and pertussis vaccine increased the anti-Hib antibody levels after the first injection only in mice receiving Hib-TT, but not in mice receiving Hib-HCH, suggesting that additional carrier protein (TT) enhanced the anti-polysaccharide response. Simultaneous injection of Hib and Pn6A conjugates with the same or different carriers resulted in an enhanced serum antibody response to each polysaccharide. The anti-tetanus toxin response reached protective levels (>0.01 U/ml) in most mice after the first injection and in all mice after the second and third injections of TT conjugates. A progressive increase in the anti-HCH response with each additional injection was noted in animals receiving HCH conjugates. Animals receiving the diphtheria toxoid-TT-pertussis vaccine preparation responded with a greater increase in anti-carrier antibody than those receiving the conjugates alone. This method of synthesis provided conjugates capable of inducing protective levels of antibodies to both the polysaccharides and carrier proteins.
Tolerance and suppression of immunity to herpes simplex virus: different presentations of antigens induce different types of suppressor cells
Authors: Schrier R D , Pizer L I , Moorhead J W
Published: January 2020

In this report, we examine tolerance (hyporesponsiveness) and suppression of delayed hypersensitivity (DH) to herpes simplex virus (HSV) in mice, using two different forms of tolerogen: HSV particles and HSV-infected spleen cells. The intravenous injection of mice with either HSV particles or spleen cells 7 days before subcutaneous immunization with virus induced a profound state of unresponsiveness. This unresponsive state was mediated, at least in part, by suppressor T cells (Ts), which were demonstrated by passive transfer to naive recipients. However, different types of Ts were induced depending on the form of the tolerogen. The injection of HSV particles induced Ts which suppressed the induction but not the expression of DH. On the other hand, the injection of HSV spleen cells induced two types of Ts: one which inhibited the induction of the DH response and one which inhibited the expression of DH to HSV. Both tolerance and Ts are virus specific (i.e., the DH response to an unrelated virus was not inhibited) but not type specific for HSV type 1 and HSV type 2. Since both virus particles and virus-infected cells may be present in the blood during HSV infection, the induction of this type of immune regulation may influence the outcome of both acute and latent HSV infections.
Identification of the major antigenic determinants on lipoglycans from Acholeplasma granularum and Acholeplasma axanthum
Authors: Al-Samarrai T H , Smith P F , Lynn R J
Published: January 2020

The major antigenic determinants of the lipoglycans from Acholeplasma granularum and Acholeplasma axanthum were found to be the oligosaccharide sequences Glcp(beta 1 leads to 2)-Glcp(alpha 1 leads to 4)-Glcp and Glcp(beta 1 leads to 2)-Glcp(beta 1 leads to 2)-Glcp, respectively. The disaccharides sophorose and maltose inhibited hemagglutination by specific antiserum of sheep erythrocytes coated with lipoglycan from A. granularum. Only sophorose possessed this capacity with the lipoglycan from A. axanthum. Periodate oxidation destroyed the ability of both lipoglycans to interact with specific antibody, a result compatible with structural data on the lipoglycans.
Regulation of expression of Escherichia coli pilus K99
Authors: Isaacson R E
Published: January 2020

An immunoassay demonstrated that the assembled K99 pilus on the surface of Escherichia coli grown in minimal medium appeared during the logarithmic phase of growth, but the synthesis of K99 subunits, as measured by nonequilibrium two-dimensional gel electrophoresis, occurred throughout the life cycle of the cell. Contrary to other reports, the addition of glucose to the growth medium did not affect K99 pilus assembly or subunit synthesis, although in a K99+ adenyl cyclase (cya) mutant, subunit synthesis was reduced. There was no reduction in the amount of assembled K99 on the cell surface of the cya mutant compared with the wild-type parent. The addition of L-alanine to minimal medium repressed K99 synthesis. However, if L-threonine or L-isoleucine was also included in the growth medium, the effect of L-alanine was reduced. Chloramphenicol caused a complete inhibition of K99 subunit synthesis, but assembly proceeded normally. Growth at 18 degrees C inhibited both subunit synthesis and pilus assembly. Approximately 92% of all cellular K99 was associated with the outer membrane, and 4% was associated with the inner membrane. No K99 was detected in the cytoplasm.