bluuebunny/crossref_metadata_2025 huggingface dataset:
Pathogenesis of Tularemia in Monkeys Aerogenically Exposed to
Francisella tularensis
425
Authors: Schricker Robert L. , Eigelsbach Henry T. , Mitten John Q. , Hall William C.
Published: January 2020

The pathogenesis of tularemia was studied in groups of rhesus monkeys (
Macaca mulatta
) that inhaled graded 10-fold doses ranging from 10 through 10
6
organisms of
Francisella tularensis
425, a strain highly virulent for the white mouse but of reduced virulence for the domestic rabbit. Mean incubation periods ranged from 3 to 6 days followed by acute illness lasting 5 to 11 days with subsequent recovery of most animals. The higher inhaled doses resulted in shorter incubation periods, longer and more severe acute illnesses, and 18% mortality at the highest dose. Strain 425 multiplied in the lungs, disseminated to the regional lymph nodes, and became systemic. Maximal bacterial populations in tissues were reached by the 7th day after exposure of the animals regardless of the number of organisms inhaled.
F. tularensis
was no longer recoverable from any of six tissues examined 2 months after exposure. The most significant tissue changes occurred in the lungs; these consisted of foci of liquefaction necrosis, lobular consolidation, and pleural effusion and adhesions. The data indicate that the inhaled dose of strain 425 determined the maximal growth of the organism in the lungs which in turn influenced the severity of the usually self-limiting pneumonia and systemic infection. Although the monkey is less resistant to tularemia than is man, this laboratory animal when infected with
F. tularensis
425 provides a useful model for the self-limiting type of human pulmonary tularemia usually observed in Europe and Asia but to a lesser extent in North America.
Activity of Macrophage and Neutrophil Cellular Fractions from Normal and Immune Sheep Against
Listeria monocytogenes
Authors: Outteridge P. M. , Osebold J. W. , Zee Y. C.
Published: January 2020

Cellular immunity to
Listeria monocytogenes
infection was studied by assaying for antibacterial activity in fractions of leukocytes collected from the peritoneal cavity, lungs, and mammary glands of immunized sheep. The cells were collected in populations that were largely either macrophages or neutrophils. Mechanically disrupted cells were divided into nuclear, lysosomal, and supernatant fluid fractions and then subjected to freezing and thawing. Comparison with similarly treated rabbit cells showed that greater fragility exists in the lysosomes of sheep cells, as indicated by the amount of acid phosphatase activity released. Inhibition of bacterial growth was assayed in a broth medium at
p
H 4.6. As expected, nuclear and lysosomal fractions from neutrophils were inhibitory. Some antibacterial activity was found in nuclear fractions of macrophages. The lysosomes of macrophages collected from the peritoneal cavity and the mammary gland did not inhibit the growth of
L. monocytogenes.
Peritoneal macrophages were allowed to interact with sensitized lymphocytes and an avirulent strain of
L. monocytogenes
for 4 hr prior to disruption and fractionation, but antibacterial activity was not detected. Pulmonary alveolar macrophages from 5 out of 16 sheep contained
Listeria
inhibitory activity in their lysosomes. The mechanism was inhibitory but not bactericidal.
Immunogenicity of Egg-Grown Influenza Vaccine: Inhibition by Immunization of Recipients with Host Antigen
Authors: Lindenmann J.
Published: January 2020

Rabbit antibody directed against normal allantoic fluid reduced the protective power of influenza vaccine prepared from egg-grown PR8 virus. A similar vaccine prepared from mouse lung virus was not inhibited. Preimmunization of mice with normal allantoic fluid or with egg-grown influenza B/Lee vaccine (crude or purified) inhibited the protective power of PR8 egg vaccine but did not affect a similar lung vaccine.
Serological Comparisons Among Hart Park Virus and Strains of Flanders Virus
Authors: Boyd Kenneth R.
Published: January 2020

The antigenic relationships among Hart Park (HP) virus strain AR 70, Flanders virus strain 61-7484, and other related strains were investigated. Strains AR 70 and 61-7484 were shown to be antigenically related but different from each other in neutralization, complement fixation, and double-diffusion tests. Two strains isolated in Texas and one strain each, isolated in Illinois and Florida, appeared to be closely related if not identical to Flanders virus (61-7484) and antigenically related but different from HP virus AR 70. Ten additional strains isolated in 10 different states were compared to the two prototype strains. Seven of the 10 could not be distinguished from Flanders virus, and all 10 strains were different from HP virus. The results indicate that a virus-type consisting of viruses antigenically related if not identical to Flanders virus is widely distributed within the United States, whereas HP virus is antigenically related but different from this type.
Characteristics of a Swine Papovavirus
Authors: Newman Joseph T. , Smith Kendall O.
Published: January 2020

A new member of the papovavirus group has been isolated and appears to infect swine. The new agent, tentatively named swine papovavirus, appears to be very defective and replicates only within a very narrow host cell range. The original source of the isolate is under investigation. Preliminary evidence suggests that the origin of swine papovavirus is either a stable pig kidney cell line or pancreas-derived trypsin
Mapping of a cloned glucosyltransferase gene in Streptococcus mutans
Authors: Perry D , Nilsen L J , Kuramitsu H K
Published: January 2020

A cloned glucosyltransferase (gtfA) fragment, inserted adjacent to an erythromycin resistance (Eryr) marker in plasmid pVA891, was used in transformation experiments to determine the genetic location of gftA on the Streptococcus mutans chromosome. Eryr (gftA) cotransformed with a methionine (Met+) marker at a frequency of approximately 23%, whereas cotransfer with a number of other markers was not observed. The number of Met+ transformants was approximately 50-fold greater than the number of Eryr transformants. Furthermore, over 20% of the Eryr transformants were always Met+, whereas less than 1% of the Met+ transformants were Eryr, indicating the extreme asymmetrical cotransfer of these markers. The results indicate that S. mutans genes can be mapped by this procedure.
Correlation of serum opsonins with in vitro phagocytosis of Streptococcus pneumoniae
Authors: Chudwin D S , Artrip S G , Korenblit A , Schiffman G , Rao S
Published: January 2020

C-reactive protein (CRP), an acute-phase reactant which binds to phosphocholine (PC) in the pneumococcal cell wall, and anti-PC antibodies are protective against experimental pneumococcal bacteremia in mice. To determine the relative opsonic capacities of CRP and anti-PC compared with those of antibodies against pneumococcal capsular polysaccharides (anti-PCP), we correlated in vitro opsonic activity for serotype 7F Streptococcus pneumoniae with concentrations of CRP, anti-PC, and anti-type 7F PCP in human sera from 10 normal subjects and 38 patients with sickle cell (SS) disease, a high-risk group for pneumococcal infection. Opsonic activity, measured by a radiolabeled bacterial uptake assay, correlated with anti-PCP levels but not with CRP or anti-PC in both the normal subjects and patients with SS disease. Addition of CRP to normal sera did not increase opsonic activity for serotypes 4 and 7F S. pneumoniae, although it did so for serotype 27, a nonpathogenic strain unique for having PC in its capsule. CRP and anti-PC were not effective opsonins when they bound to the pneumococcal cell wall rather than the capsule. The protective effects of CRP or anti-PC against these serotypes may be produced by means other than complement-dependent opsonization.
Bacterial adherence to human endothelial cells in vitro
Authors: Ogawa S K , Yurberg E R , Hatcher V B , Levitt M A , Lowy F D
Published: January 2020

Differences in the ability of bacteria to adhere to normal valvular endothelium may account for the predominance of particular species as pathogens in acute endocarditis. An in vitro adherence assay was developed to simulate the host surface encountered in acute bacterial endocarditis by using confluent monolayers of human endothelial cells. Adherence of 32 gram-positive and -negative blood culture isolates to this surface was compared. All five Staphylococcus aureus strains tested were highly adherent to endothelial cells, as was one gram-negative strain (Serratia marcescens). The remaining gram-positive and -negative isolates, including four viridans streptococci, were relatively nonadherent. Transmission electron microscopy demonstrated attachment of Staphylococcus aureus and invagination of the underlying endothelial cell membrane at 1 h followed by engulfment of large numbers of bacteria after 3 h. The intracellular bacteria appeared to be contained within vacuoles. Preferential attachment of some strains of bacteria, in particular Staphylococcus aureus, to human endothelial cells occurred in vitro, suggesting that adherence is an important determinant of bacterial pathogenicity in acute endocarditis. Active uptake of bacteria by endothelial cells may help account for the virulence of Staphylococcus aureus in endovascular infections and for the ability of this organism to establish multiple metastatic foci of infection.
Purification and vaccine potential of Klebsiella capsular polysaccharides
Authors: Cryz S J , Fürer E , Germanier R
Published: January 2020

Capsular polysaccharide (CPS) from 18 Klebsiella strains of different capsular types was isolated and characterized. Purified CPSs were composed primarily of carbohydrate with trace quantities of protein, nucleic acids, and lipopolysaccharide. All CPSs were of a high molecular weight, possessing a Kd of 0.01 to 0.11 as determined by gel filtration over Sepharose CL-4B. Low levels of lipopolysaccharide present in all preparations were responsible for the highly pyrogenic nature of one-half of the CPS preparations. Treatment of capsular material with dilute NaOH in 95% ethanol markedly reduced the pyrogenicity of all preparations and had a negligible effect on their molecular weight. The immunogenicity of the various native CPSs for mice varied considerably from serotype to serotype, but all evoked an anticapsular immunoglobulin G response. Five of 18 NaOH-treated polysaccharides were significantly (P less than 0.05) less immunogenic than their native counterparts. Human immunoglobulin G prepared from volunteers immunized with either native or NaOH-treated KP1-0 capsular polysaccharide was equally effective at preventing experimental fatal Klebsiella pneumoniae burn wound sepsis in mice.
Bacterial adherence and hemolysin production from Escherichia coli induces histamine and leukotriene release from various cells
Authors: Scheffer J , König W , Hacker J , Goebel W
Published: January 2020

We investigated the role of bacterial adherence and hemolysin production from Escherichia coli parent and genetically cloned strains as to their effects on histamine release from rat mast cells and leukotriene generation from human polymorphonuclear granulocytes. These mediators were involved in the induction of inflammatory disease processes and led, for example, to enhancement of vascular permeability, chemotaxis (leukotriene B4 [LTB4]), chemoaggregation, lysosomal enzyme release, and smooth muscle contraction, (LTC4, LTD4, and LTE4). Washed bacteria (E. coli K-12 MS+ Hly +/-; E. coli 536 MS+ MR +/-) as well as their culture supernatants were analyzed. Washed E. coli K-12 (Hly+), unlike Hly- strains, induced high amounts of histamine release from rat mast cells and chemotactic activity from human polymorphonuclear granulocytes. Significant leukotriene release was obtained with washed E. coli K-12 Hly+ strains and their bacterial culture supernatants. Leukotriene induction was dependent on the amount of hemolysin activity present in the supernatant. However, additional soluble factors should also be considered. The presence of hemolysin appeared to accelerate and enhance the rate of phagocytosis of bacteria by neutrophils. When E. coli 536 (MS+ MR +/- Hly +/-) strains were analyzed, the simultaneous presence of MR+ pili and hemolysin production led to an increase in histamine release as compared with MR- Hly+ strains. The genetically cloned MR+ Hly+ E. coli 536 strain induced higher amounts of leukotrienes as compared with the wild-type strain. Our data suggest a potent role for adhesins and hemolysin as virulence factors in inducing the release of inflammatory mediators.
Influence of potential virulence determinants on Bordetella bronchiseptica-induced ciliostasis
Authors: Bemis D A , Wilson S A
Published: January 2020

Eighteen strains of Bordetella bronchiseptica, selected on the basis of previously determined phenotypic characteristics, were examined for their ability to induce ciliostasis in canine tracheal outgrowth cultures. Fifteen strains grown on brucella agar caused ciliostasis. Strains that did not cause ciliostasis were stable, nonpiliated, and nonhemolytic, they did not produce extracellular adenylate cyclase, and were morphologically indistinguishable from rough-phase variants on brucella agar. Plasmids were detected in only five of the strains which induced ciliostasis, transfer of plasmids from four of these strains to one which did not induce ciliostasis did not alter its virulence or colony morphology. All strains which were hemolytic on Bordet-Gengou agar produced extracellular adenylate cyclase. Two nonhemolytic strains, one which produced only rough colonies on brucella agar, also induced ciliostasis. Two types of colony (phase) variation were observed: one recognizable on both brucella agar and Bordet-Gengou agar at frequencies of less than or equal to 10(-2), associated with multiple loss of virulence determinants, and the other recognizable only on Bordet-Gengou agar at frequencies of greater than or equal to 10(-2), associated with flagellum expression. The possession of readily detectable somatic pili was the only phenotypic characteristic consistently associated with the ability to induce ciliostasis. Formalin-killed and chloramphenicol-inhibited B. bronchiseptica strain 110H organisms had detectable pili and attached to cilia, but did not cause ciliostasis. Protease-treated B. bronchiseptica strain 110H organisms did not have detectable pili and in the presence of chloramphenicol did not attach to cilia. Attachment to cilia, although not in itself sufficient to cause ciliostasis, is intimately associated with and may be required for the induction of ciliostasis by B. bronchiseptica strains.
Influence of estrogen on host resistance: increased susceptibility of mice to Listeria monocytogenes correlates with depressed production of interleukin 2
Authors: Pung O J , Tucker A N , Vore S J , Luster M I
Published: January 2020

Mice given pharmacological levels of the synthetic estrogen diethylstilbestrol demonstrated a marked increase in susceptibility to infection with Listeria monocytogenes. Experiments were performed in an effort to determine the mechanism(s) by which estrogen treatment increases the susceptibility of mice to L. monocytogenes infection. Estrogen exposure depressed the in vivo proliferative response of splenic lymphocytes to L. monocytogenes, which correlated with the decreased in vitro response of these cells to phytohemagglutinin. Interleukin 2 (IL 2) production by splenic lymphocytes from estrogen-treated mice was decreased, although these cells were capable of proliferating normally in response to exogenous IL 2. Interleukin 1 production by peritoneal macrophages was not depressed by estrogen exposure. The number of bacteria observed in the spleens of estrogen-exposed mice challenged with L. monocytogenes was reduced by IL 2 administration. Thus, estrogens may decrease host resistance to L. monocytogenes by inhibiting IL 2 production and the subsequent proliferation of antigen-sensitized T lymphocytes required for recovery.
Factors governing adherence of Candida species to plastic surfaces
Authors: Klotz S A , Drutz D J , Zajic J E
Published: January 2020

The ability of Candida albicans and Candida spp. to adhere to inert polymeric surfaces may allow these organisms direct ingress into the human host. Biophysical characterization of this adherence shows that the forces responsible for such adherence are attractive London-van der Waals forces (or hydrophobic forces) and electrostatic forces. The hydrophobic affinity of yeasts was determined by (i) a water-hydrocarbon two-phase assay and by (ii) measurement of the contact angle (theta) of a liquid droplet on a monolayer of yeast cells. The hydrophobicity of the yeasts correlated with the tendency of yeasts to adhere to polystyrene and was reduced in the presence of Tween 20. The adherence of yeasts to polymers of increasing hydrophobicity (determined by the contact angle method) was directly proportional to theta. Yeast surface charges were altered by selectively blocking amino and carboxyl groups. The more positively charged yeasts adhered in greater numbers. Increasing the molarity of NaCl increased yeast adherence. These forces probably contribute to the negative cooperativity (determined by Scatchard and Hill plot) that characterizes the adherence of yeasts to polymers.
The ability of Salmonella typhimurium to produce the siderophore enterobactin is not a virulence factor in mouse typhoid
Authors: Benjamin W H , Turnbough C L , Posey B S , Briles D E
Published: January 2020

One of the nonspecific defense mechanisms of higher animals is their ability to limit iron availability to infecting bacteria. Thus it has been argued that all pathogenic bacteria must have special mechanisms to obtain iron in the host environment. Salmonella typhimurium is known to produce a siderophore, enterobactin, with which it can obtain iron from host transferrin. Previous studies have indicated that the production of this molecule is necessary for the ability of intraperitoneally injected. S. typhimurium cells to cause mouse typhoid, a largely intracellular infection. We have reexamined this finding with wild-type S. typhimurium and isogenic strains carrying the nonenterobactin-producing mutation ent-1 or ent-7. Our findings demonstrate that, although enterobactin production is necessary for growth in normal mouse serum, it does not affect the ability of S. typhimurium to cause mouse typhoid. Based on these findings and published results of other investigators on the role of siderophores in intracellular pathogens, a more comprehensive investigation of the importance of siderophores in intracellular infections may be warranted.
Identification of precursor of phagocytosis-stimulating factor in guinea pig polymorphonuclear neutrophils
Authors: Ishibashi Y , Yamashita T
Published: January 2020

A precursor of phagocytosis-stimulating factor (PSF) was identified by immunoblot assay with purified anti-PSF antibodies. The purified anti-PSF antibodies recognized not only the PSF with a molecular weight of 16,000 (16K) but also the 36K protein with a pI of 6.5 in the granule fraction of polymorphonuclear neutrophils, indicating that this 36K protein has an antigenic determinant common to PSF. In addition, the appearance of the PSF during phagocytosis by polymorphonuclear neutrophils was closely correlated to the decrease of the 36K protein. These results suggest that the 36K protein is the precursor of PSF which is converted to biologically active PSF in the granules during phagocytosis.
In vitro adherence of Escherichia coli to endometrial epithelial cells of rats and influence of estradiol
Authors: Nishikawa Y , Baba T
Published: January 2020

The influence of ovarian hormones on the adhesion of Escherichia coli to endometrial epithelial cells was investigated in an in vitro system. Endometrial cells liberated by collagenase from rat uteri were used. Optimal test conditions were obtained when 5 X 10(8) E. coli bacteria were added to 10(5) epithelial cells and incubated for 60 min. The adhesion of the organisms was inhibited by the addition of either mannose or alpha-methyl-D-mannopyranoside. When epithelial cells collected from uteri of estradiol-treated rats were used, the number of E. coli adhering to the cells was markedly lower than that adhering to epithelial cells collected from control rats. These results suggest that E. coli adheres to endometrial epithelial cells with so-called type 1 pili and that estradiol alters the nature of the endometrial epithelium and prevents the adherence of the organisms to the cells.
Protection against infection with Neisseria meningitidis group B serotype 2b by passive immunization with serotype-specific monoclonal antibody
Authors: Brodeur B R , Larose Y , Tsang P , Hamel J , Ashton F , Ryan A
Published: January 2020

Hybridomas derived from mice immunized with Neisseria meningitidis serogroup B serotype 2b (B,2b) outer membrane preparations produced monoclonal antibodies (MAbs) specific for major outer membrane proteins of classes 1, 2, and 5. The MAbs were examined by enzyme-linked immunosorbent assay against a selected panel of seven strains of N. meningitidis (B,2b) of different sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns, a serotype 2a, and a nontypable strain. The five MAbs selected were all bactericidal and of different immunoglobulin subclasses. None of the MAbs reacted with other bacterial strains in a dot-enzyme immunoassay. The corresponding antigenic determinant for each MAb was localized on a specific outer membrane protein by immunoblotting of sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of major outer membrane proteins. MAbs M5-11 and M5-30 bound to the class 2 protein and were serotype 2b specific. MAb M2-20 bound to the class 1 protein, and MAbs M5-16 and M5-19 bound to the class 5 protein. A mouse model of infection was established whereby a local infection progressed to lethal bacteremia over 3 days, and 50% of the animals were killed with an intraperitoneal injection of 10 meningococci plus 4% mucin and 1.6% hemoglobin. The ability of the MAbs to provide passive protection against experimental infection with N. meningitidis (B,2b) was examined. Both serotype-specific MAbs M5-11 and M5-30 were highly protective even though they were of different immunoglobulin subclasses. The class 5-specific MAb offered no protection, while the class 1-specific MAb gave limited protection. It may therefore be possible to provide protection against serotype 2b infection by using as vaccine the class 2 serotype-specific surface-exposed outer membrane protein epitopes defined by MAb M5-11 or M5-30.
Growth inhibition of Babesia bovis in culture by secretions from bovine mononuclear phagocytes
Authors: Montealegre F , Levy M G , Ristic M , James M A
Published: January 2020

Bovine blood mononuclear phagocytes from babesia-free cattle were cultured in vitro. Cell monolayers were treated with culture-derived soluble Babesia bovis exoantigens, immune complexes, and bovine anti-B. bovis immune serum. Subsequently, the monolayers were washed free of the reagents and allowed to develop further in the presence of standard culture medium. Transfer of supernatant media from these cultures to those of B. bovis revealed the presence of growth-inhibiting factors. These factors were thermostable, nondialyzable, and were degraded by freeze-thawing, and their action was concentration dependent. Supernatants from antigen- and immune complex-treated monolayers demonstrated greater inhibitory effects than did supernatants from antibody-treated or untreated monolayers. Erythrocytes incubated with supernatant medium from antigen-treated monolayers did not support growth of B. bovis as well as did erythrocytes incubated with supernatants from untreated monocyte monolayers. This result suggests that the mechanism of action of soluble factors could be through some modification of the erythrocyte such as the blockage of active transport of essential nutrients.
Genetic control of systemic Leishmania major infections: dissociation of intrahepatic amastigote replication from control by the Lsh gene
Authors: Mock B A , Fortier A H , Potter M , Nacy C A
Published: January 2020

Systemic disease induced by Leishmania major was estimated by microscopic examination of liver impression smears and determination of numbers of intrahepatic amastigotes in intravenously or subcutaneously infected inbred, hybrid, and congenic mice. The distribution of susceptible phenotypes among these mice, particularly the susceptibility of a strain congenic for Lshr, strongly suggested that Lsh, a gene which controls intrahepatic replication of Leishmania donovani, does not influence systemic disease by L. major.
Strain differences in susceptibility to murine respiratory mycoplasmosis in C57BL/6 and C3H/HeN mice
Authors: Davis J K , Parker R F , White H , Dziedzic D , Taylor G , Davidson M K , Cox N R , Cassell G H
Published: January 2020

Not only is murine respiratory mycoplasmosis, due to Mycoplasma pulmonis, a complication of biomedical research, it provides excellent animal models to study the development of a naturally occurring respiratory disease induced by an infectious agent. The understanding of pathogenic mechanisms of disease can be greatly facilitated by studying genetic differences in susceptibility. Five strains of mice with various H-2K haplotypes were examined for their susceptibility to murine respiratory mycoplasmosis; of these, C57BL/6 and C3H/HeN mice were chosen for additional study. There were no significant differences in the incidence of infection in either the upper or lower respiratory tract or in the severity of upper respiratory tract lesions in the two strains as determined at 14 days postinfection. In striking contrast, the C57BL/6 mice were significantly more resistant to the development of gross and microscopic lung lesions and to death due to pneumonia as shown by an almost 100-fold difference in the 50% lethal dose, 50% gross pneumonia dose, and 50% microscopic lesion dose. The most apparent differences in lung lesions between the two strains were in the severity of acute lesions of the bronchial epithelium, the amount of mixed inflammatory response in the alveoli, and the amount of lymphoid infiltrates. All were significantly more severe in C3H/HeN mice. In addition, more C3H/HeN mice developed antibody responses to M. pulmonis. The amount of antibody correlated with lesion severity in both strains.
Lymphoid procoagulant response to bacterial endotoxin in the rat
Authors: Lando P A , Edgington T S
Published: January 2020

A number of species respond to bacterial endotoxin (lipopolysaccharide [LPS]) wherein cells of the monocyte-macrophage lineage are rapidly induced either directly or via T-cell collaboration to initiate the extrinsic coagulation protease pathway. This results in fibrin formation and deposition as well as consumption of plasma coagulation proteins. It has been claimed that this cellular response, basic to the Shwartzman reaction, is lacking in rats and may account for the more limited severity of the Shwartzman reaction in this species. We examined the in vitro lymphoid procoagulant response in Fischer 344, Brown Norway, and Lewis rats. When peripheral blood mononuclear cells (PBM) were stimulated in vitro with LPS, a procoagulant activity (PCA) response was observed when assayed by acceleration of clotting of recalcified human or rat platelet-poor plasma. The response was rapid, with a maximum achieved at 4 h. PCA was not physically dissociated from viable PBM by 5 mM EDTA, which is consistent with the presence of an intrinsic plasma membrane initiator molecule rather than calcium-bound gamma-carboxylated glutamic acid-containing proteases. The induction of monocyte PCA was prevented by incubation of cells with cycloheximide or actinomycin D, implicating a new biosynthetic requirement. Cultivation of PBM with warfarin did not diminish the function of the effector PCA, nor did vitamin K augment the function of the endotoxin-induced PCA, indicating that the functional activity was not attributable to gamma-carboxylated glutamic acid-containing proteins. No inhibition of the cellular PCA molecule was produced by serine protease inhibitors. The LPS-induced PCA appeared to involve a tissue factor-like molecule since both factors X and VII were required in mediating PCA. Isolation of monocytes and T lymphocytes from LPS-stimulated PBM demonstrated that PCA was present in the monocyte-rich fraction. When isolated rat T lymphocytes and monocytes were separately exposed to LPS, PCA was not induced. In contrast, when the cells were combined, LPS induced PCA, indicating that the PCA response involved cellular collaboration between cells present in T lymphocyte and monocyte populations.
Prevention and therapy of experimental Escherichia coli infection with monoclonal antibody
Authors: Kim K S , Cross A S , Zollinger W , Sadoff J
Published: January 2020

Mouse hybridoma antibody of immunoglobulin class M prepared with live group B meningococci was evaluated for its ability to protect against and treat Escherichia coli infections in a newborn-rat model. In these studies, antibody was administered intraperitoneally and bacteria were administered subcutaneously to avoid introducing the antibody and bacteria to the same site. The activity of this hybridoma antibody was specific; the antibody provided protection against the K-1 strain, but not against the K-92 strain. In addition, the amount of the antibody required for protection was dependent upon the size of bacterial challenge. With an increase of the bacterial inocula from the 100% lethal dose to 10 times the 100% lethal dose there was a threefold increase in the amount of the antibody required for 50% protection. Similarly, therapeutic efficacy of the antibody was also dependent upon the magnitude of bacteremia before therapy. The antibody successfully cleared the bacteremia only when the pretherapy bacterial counts in blood were less than 10(4) CFU/ml. These findings suggest that the monoclonal immunoglobulin M antibody against the capsular polysaccharide of the group B meningococcus may be useful in the prevention and treatment of K-1 E. coli infections.
Contact-dependent cytopathogenic mechanisms of Trichomonas vaginalis
Authors: Krieger J N , Ravdin J I , Rein M F
Published: January 2020

The cytopathogenic mechanisms of Trichomonas vaginalis have been debated since the 1940s. We examined the following three proposed pathogenic mechanisms: contact-dependent extracellular killing, cytophagocytosis, and extracellular cytotoxins. Serial observations of Chinese hamster ovary (CHO) cell monolayers exposed to trichomonads revealed that (i) trichomonads form clumps, (ii) the clumps adhere to cells in culture, and (iii) monolayer destruction occurs only in areas of contact with T. vaginalis. Kinetic analysis of target cell killing by trichomonads revealed that the probability of CHO cell death was related to the probability of contact with T. vaginalis, supporting the observation by microscopy that trichomonads kill cells only by direct contact. Simultaneous studies of 111indium oxine label release from CHO cells and trypan blue dye exclusion demonstrated that T. vaginalis kills target cells without phagocytosis. Filtrates of trichomonad cultures or from media in which trichomonads were killing CHO cells had no effect on CHO cell monolayers, indicating that trichomonads do not kill cells by a cell-free or secreted cytotoxin. The microfilament inhibitor cytochalasin D (10 micrograms/ml) inhibited trichomonad killing of CHO cell monolayers by 80% (P less than 0.0001). In contrast, the microtubule inhibitor vinblastine (10(-6) M) caused only 17% inhibition of trichomonad destruction of CHO cell monolayers (P less than 0.020), whereas colchicine (10(-6) M) had no effect. T. vaginalis kills target cells by direct contact without phagocytosis. This event requires intact trichomonad microfilament function; microtubule function appears not to be essential.
Cloning of Mycobacterium bovis BCG DNA and expression of antigens in Escherichia coli
Authors: Thole J E , Dauwerse H G , Das P K , Groothuis D G , Schouls L M , van Embden J D
Published: January 2020

A gene bank of Mycobacterium bovis BCG DNA in Escherichia coli was constructed by cloning Sau3A-cleaved mycobacterium DNA fragments into the lambda vector EMBL3. The expression of mycobacterial antigens was analyzed by Western blotting with hyperimmune rabbit sera. Among 770 clones tested, several were found that produced various mycobacterial antigens in low amounts, with concentrations generally close to the detection limit. One particular clone was chosen for further investigation. This clone produced a 64-kilodalton (kDa) antigen. By placing the lambda promoter PL in front of the structural gene of this antigen, an overproducing E. coli strain was obtained. Rocket-line immunoelectrophoresis experiments showed that antigens cross-reacting with the 64-kDa protein are present in a wide variety of mycobacteria and also in so-called purified protein derivatives which are routinely used for skin tests. Preliminary experiments indicate the presence of antibodies against the 64-kDa antigen in sera from tuberculosis patients.
Phosphoenolpyruvate-sugar phosphotransferase transport system of Streptococcus mutans: purification of HPr and enzyme I and determination of their intracellular concentrations by rocket immunoelectrophoresis
Authors: Thibault L , Vadeboncoeur C
Published: January 2020

Enzyme I and HPr, the general proteins of the phosphoenolpyruvate-sugar phosphotransferase system, play a pivotal role in the control of sugar utilization in gram-negative and gram-positive bacteria. To determine whether growth conditions could modify the rate of biosynthesis of these proteins in Streptococcus mutans, we first purified to homogeneity enzyme I and HPr from S. mutans ATCC 27352. Using specific antibodies obtained against these proteins, we determined by rocket electrophoresis the intracellular levels of enzyme I and HPr in cells of S. mutans 27352 grown under various batch culture conditions and in a number of glucose-grown cells of other strains of S. mutans. HPr was purified by the procedure reported by Gauthier et al. (L. Gauthier, D. Mayrand, and C. Vadeboncoeur, J. Bacteriol. 160:755-763, 1984) and displayed a single band with a molecular weight of 6,650 when analyzed by sodium dodecyl sulfate-urea gel electrophoresis. Enzyme I was purified by DEAE-cellulose chromatography, affinity chromatography on an anti-Streptococcus salivarius column, and preparative electrophoresis. The protein migrated as a single band in native and denaturating gel electrophoresis. The subunit molecular weight of enzyme I determined by electrophoresis under denaturating conditions was 68,000. In gel filtration chromatography at 4 degrees C, the enzyme migrated as a 135,000- to 160,000-molecular-weight species, suggesting that enzyme I is a dimer. In double immunodiffusion experiments, antibodies against HPr reacted with several oral streptococci, Streptococcus lactis, Streptococcus faecium, and Lactobacillus casei, but not with Bacillus subtilis, Staphylococcus aureus, and Escherichia coli. Antibodies against enzyme I of S. mutans 27352 cross-reacted with enzyme I from all the other oral streptococci tested. No cross-reaction was observed with other gram-positive and gram-negative bacteria. The levels of enzyme I and HPr determined by rocket electrophoresis in S. mutans 27352 varied at the most by twofold, depending on the growth conditions. Glucose-grown cells of other S. mutans strains contained levels of enzyme I and HPr which were similar to those found in S. mutans 27352.
Induction of a macrophage migration enhancement factor after desensitization of tuberculin-positive rabbits with purified protein derivative
Authors: Gordon M R , Takata I , Myrvik Q N
Published: January 2020

The production of a macrophage migration enhancement factor (MEF) has been achieved as a consequence of administering a desensitizing dose of purified protein derivative (PPD) to Mycobacterium bovis BCG-sensitized rabbits. The migration-enhancing effect was first demonstrated when alveolar macrophages (AM) harvested from desensitized rabbits exhibited marked migration stimulation; whereas maximum migration enhancement was observed 8 days after the administration of PPD, migration enhancement of the AM from these rabbits persisted for up to 12 days. Sera from BCG-sensitized, PPD-desensitized animals exhibited a peak of MEF activity 4 days after desensitization. Maximal MEF activity was demonstrated in culture supernatants of nonadherent spleen cells harvested 8 days after the intravenous desensitizing dose of PPD was given. Control spleen cell culture supernatants did not produce detectable MEF. The route of desensitization with PPD was critical. When PPD was administrated intratracheally, MEF activity was not induced. The intravenous administration of BCG after PPD desensitization reversed migration enhancement to strong migration inhibition. Ammonium sulfate fractionation indicated that two fractions contained MEF activity. MEF activity was retained by dialysis membranes with a 15,000-molecular-weight cutoff but passed through dialysis membranes with a 25,000-molecular-weight cutoff. The mixture of migration inhibition factor with MEF-containing supernatants resulted in the mutual cancellation of both activities. These observations suggest that MEF may be a modulator of macrophage effector responses mediated by migration inhibition factor.
In vitro cultivation of Toxoplasma gondii cysts in astrocytes in the presence of gamma interferon
Authors: Jones T C , Bienz K A , Erb P
Published: January 2020

Long-term culturing of Toxoplasma gondii cysts was accomplished in vitro in association with murine astrocytes and intermittent additions of gamma interferon to the media. Phase-contrast microscopy was used to follow the stages of cyst development, and electron microscopy confirmed the presence of morphologic characteristics of T. gondii cysts. T. gondii cysts formed in vitro had a single trilaminar membrane during both intracellular and extracellular existence and contained amorphous electron-dense material either throughout the cyst or in a uniform layer under the trilaminar membrane. The bradyzoites were similar to previous descriptions of tachyzoites in vitro except that they were smaller and contained numerous electron-lucent vacuoles. Gamma interferon is not necessary for cyst formation, but it controls the division of tachyzoites and may allow cysts to remain for prolonged periods without rupturing. In vitro-cultivated T. gondii cysts will be useful for producing bradyzoite and cyst antigens and for measuring the effects of antimicrobial agents and immune modulators on the viability of intracystic T. gondii.
Experimental infection of infant rabbits with verotoxin-producing Escherichia coli
Authors: Pai C H , Kelly J K , Meyers G L
Published: January 2020

To study the pathogenesis of diarrheal disease due to verotoxin (VT)-producing Escherichia coli, 3-day-old rabbits were inoculated intragastrically with live E. coli O157:H7 (high VT producer), E. coli O113:K75:H21 (low VT producer), or O157:H45 (VT negative) and were examined for clinical symptoms, bacterial colonization, presence of detectable free VT in the intestines, and histological changes. Diarrhea developed consistently with 10(8) bacteria of E. coli O157:H7 but was observed only infrequently with even a higher dose of E. coli O113:K75:H21. VT-negative strains failed to cause diarrhea under the same experimental conditions. E. coli O157:H7 was recovered from the colon of infected animals in a significantly higher concentration than from the small intestine, and the clinical symptoms correlated with the presence of detectable free VT in the colon. Histological changes were seen mainly in the mid- and distal colon; these changes were characterized by a vast increase in apoptosis in the surface epithelium, increased mitotic activity in the crypts, mucin depletion, and a mild to moderate infiltration of neutrophils in the lamina propria and epithelium. Multiple foci of attached bacteria were seen on the surface epithelium of the gut-associated lymphoid tissue, cecum, and colon. Bacteria were never seen in epithelial cells or the lamina propria. These mucosal abnormalities as well as clinical symptoms were reproduced in infant rabbits by the intragastric administration of VT alone. These results are consistent with the hypothesis that VT plays a major role in the pathogenesis of diarrhea caused by E. coli O157:H7 and other VT-producing E. coli.
Encapsulation and melanin formation as indicators of virulence in Cryptococcus neoformans
Authors: Kwon-Chung K J , Rhodes J C
Published: January 2020

Acapsular (Cap-) mutants of Cryptococcus neoformans var. neoformans that produce melanin (Mel+) on diphenol media at 30 degrees C but not at 37 degrees C were found to be avirulent for mice. Compared with wild-type isolates, the mutants had a lower rate of L-3,4-dihydroxyphenylalanine uptake at 37 degrees C and showed an insignificant level of phenoloxidase activity at both temperatures. To study the relationship of Cap and Mel phenotypes to virulence in mice, we crossed one of the mutants (Cap- Mel-) with a wild type (Cap+ Mel+) to obtain four classes of progeny (Cap+ Mel+, Cap+ Mel-, Cap- Mel+, and Cap- Mel-). The progeny with the Cap+ Mel+ phenotype and the wild-type parent (Cap+ Mel+) were inoculated into mice (10(6) cells per mouse) and, within 40 days, produced fatal infection in 90 to 100% of the animals. None of the other three phenotypes produced fatal infection within the same period. While progeny with the Cap+ Mel- phenotype did produce fatal infection after 40 days, 70 to 90% of the mice survived at least until day 70. However, in the isolates recovered from the brain tissue of a mouse that died on day 68, nearly 40% of the CFU had reverted to the Cap+ Mel+ type. The virulence of one of these revertant Cap+ Mel+ isolates was compared with that of a Cap+ Mel- isolate recovered from the same tissue. One hundred percent of the mice inoculated with the revertant died within 35 days, while no fatal infection was produced in the mice inoculated with the Cap+ Mel- isolate within the same period. The isolates with the Cap- Mel+ or Cap- Mel- phenotype not only failed to produce fatal infection but failed to revert to the Cap+ Mel+ type in the mouse brain during the experimental period. These results indicate that both the Cap+ phenotype and the Mel+ phenotype are important indicators of virulence in C. neoformans.
Modulation of bone metabolism by two chemically distinct lipopolysaccharide fractions from Bacteroides gingivalis
Authors: Millar S J , Goldstein E G , Levine M J , Hausmann E
Published: January 2020

Two separate species of lipopolysaccharide (LPS) from Bacteroides gingivalis 381 have been isolated. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated not only the heterogeneity of each species, but also that they represented high- and low-molecular-weight LPS entities. Although both contained the same carbohydrate and fatty acid components, the proportions of these differed between the LPS species. The direct effects of these two species in modulation of bone resorption and bone collagen and noncollagen protein synthesis have been examined. In a bone resorption assay, these two species stimulated 45Ca release from prelabeled fetal rat bones in a concentration range of 0.5 to 3.0 micrograms/ml. The two LPS species also elicited a 30 to 40% reduction in collagen protein formation at 10 micrograms/ml. Responses of the same order of magnitude were observed with LPS from Salmonella minnesota at 10 micrograms/ml. The higher-molecular-weight LPS species also significantly inhibited noncollagen protein formation. This is the first report that LPS from B. gingivalis 381, a suspected periodontal pathogen, inhibits bone collagen formation and, in conjunction with the bone resorption potency, further implicates LPS in alveolar bone loss associated with periodontal disease.
Treatment of alveolar macrophages with cytochalasin D inhibits uptake and subsequent growth of Legionella pneumophila
Authors: Elliott J A , Winn W C
Published: January 2020

Legionella pneumophila multiplied rapidly in guinea pig and rat alveolar macrophages but failed to grow when phagocytic activity was inhibited by pretreatment with 0.5 or 1.0 microgram of cytochalasin D per ml. Attachment was not inhibited by cytochalasin D. No extracellular multiplication occurred when L. pneumophila were in close proximity to viable functional macrophages or even when the bacteria were attached to plasma membranes of the macrophages. Nonopsonized L. pneumophila were avidly phagocytized by alveolar macrophages. When bacteria were centrifuged onto a cell pellet, more than 85% of the phagocytes contained one or more bacteria within 15 min. In contrast, under the same conditions only approximately 15% of the macrophages contained nonopsonized Escherichia coli or Staphylococcus aureus. Phagocytosis of L. pneumophila by untreated guinea pig macrophages occurred by extension of pseudopodia around the bacteria in a classical manner. The failure of the bacteria to actively penetrate the phagocyte suggests that their intracellular survival must not depend on avoidance of a phagosome but rather on an inhibition of or resistance to subsequent microbicidal functions of the macrophage.
Transfer of resistance to syphilitic infection from maternal to newborn guinea pigs
Authors: Pavia C S
Published: January 2020

Partial protection against cutaneous syphilis and dissemination of treponemes to the regional lymph nodes was achievable in Treponema pallidum-infected neonatal strain 2 guinea pigs having syphilis-immune mothers. This evidence for enhanced resistance was associated with the natural transfer of antitreponemal, maternally derived immunoglobulins before parturition.
Mucus colonization as a determinant of pathogenicity in intestinal infection by Campylobacter jejuni: a mouse cecal model
Authors: Lee A , O'Rourke J L , Barrington P J , Trust T J
Published: January 2020

Human isolates of the intestinal pathogen Campylobacter jejuni have been shown to colonize mucus on the outer surface and deep within the intestinal crypts of gnotobiotic or germfree mice. The cecal crypts are preferentially colonized. A model of mucus colonization by C. jejuni in the mouse cecum has been developed, using antibiotic- and magnesium sulfate-treated specific-pathogen-free animals. These spiral-shaped bacteria colonize the mucus in a similar manner to the normal spiral-shaped microbiota. No evidence of adhesion to the intestinal surface was found with a wide variety of microscopic techniques. The campylobacters were seen to be highly motile in living preparations of gut tissue and rapidly tracked along intestinal mucus. Just as many of the normal spiral-shaped bacteria of intestinal surfaces can achieve close association with the epithelium through mucus association and do not adhere to the surface, C. jejuni colonizes the intestinal mucosa via mucus colonization. Thus, a major determinant of pathogenicity in intestinal infection with C. jejuni is proposed to be an ability to colonize intestinal mucus. The possession of specific adhesins is unlikely to be a significant determinant of pathogenicity. Better understanding of the mechanism of mucus association and the properties of the bacterium that are responsible will provide a basis for the rational selection of preventative measures. The model of mucus association in adult antibiotic-treated mice provides an opportunity for colonization studies with variant organisms and immunization studies.
Chromosomal map position of genes encoding P adhesins in uropathogenic Escherichia coli
Authors: Hull R , Bieler S , Falkow S , Hull S
Published: January 2020

Escherichia coli isolates from upper urinary tract infections frequently express adherence to human uroepithelium and D-mannose-resistant hemagglutination of human erythrocytes. Such adherence is usually associated with P pili encoded by the pap operon(s). In this paper, we report approximate chromosomal map positions for two copies of the pap operon. Only one copy expressed an adhesin capable of D-mannose-resistant hemagglutination, although both expressed P-related antigen.