bluuebunny/crossref_metadata_2025 huggingface dataset:
Mycobacterium lepraemurium infection of nude athymic (nu/nu) mice
Authors: Lefford M J
Published: January 2020

Nude athymic (nu/nu) mice on a BALB/c background and their heterozygous euthymic litter mates (nu/+) were infected with either 10(8) or 10(6) Mycobacterium lepraemurium organisms intravenously or in the left hind footpad (LHF). After LHF infection with 10(8) M. lepraemurium organisms, nu/+ mice slowly developed a response that consisted of LHF swelling and local resistance to Listeria monocytogenes. The lower inoculum induced a proportionately lower response in nu/+ mice, but the nu/nu mice developed neither LHF swelling nor resistance to L. monocytogenes in response to either dose of M. lepraemurium. Counts of M. lepraemurium in the LHF revealed no difference between the nu/+ mice and nu/nu mice. After intravenous infection the nu/+ mice developed splenomegaly, but did not otherwise differ from nu/nu mice with respect to resistance to intravenous challenge with L. monocytogenes or growth of M. lepraemurium in the spleen. In light of the poor responsiveness of nu/+ mice in this experiment, they were then compared with CB6 and B6D2 mice, which are genetically susceptible and resistant to M. lepraemurium, respectively. These mice were infected with either 10(8) or 10(6) M. lepraemurium cells or 10(6) Mycobacterium bovis BCG cells in the LHF. Once again the nu/+ mice responded poorly to M. lepraemurium, the CB6 mice responded very strongly, and the B6D2 mice gave an intermediate response with respect to LHF swelling and resistance to L. monocytogenes. However, M. lepraemurium grew to higher numbers in the LHF of nu/+ and CB6 mice than in B6D2 mice, revealing, in CB6 mice, a dissociation between resistance to L. monocytogenes and M. lepraemurium. All three mouse strains responded strongly to M. bovis BCG, but there was a suggestion that nu/+ mice might be more susceptible to this agent than the other two strains. I concluded that the failure of nu/+ mice to restrict the growth of M. lepraemurium more than nu/nu mice was due to the intrinsic genetic susceptibility of both types of mice. In effect, the nu/+ mice behaved like nu/nu mice, as if they too were deficient in T lymphocytes that were responsive to M. lepraemurium.
Antimicrobial activities of dialysate-elicited and resident human peritoneal macrophages
Authors: Peterson P K , Gaziano E , Suh H J , Devalon M , Peterson L , Keane W F
Published: January 2020

Recent studies of the antimicrobial capacity of peritoneal macrophages (PM phi) isolated from patients undergoing chronic peritoneal dialysis have raised the question of whether these cells might be analogous to stimulated or activated murine PM phi. To explore this possibility, we compared PM phi from these patients (dialysate-elicited PM phi) with PM phi obtained from women undergoing laparoscopy (resident PM phi) in several in vitro assays of phagocyte function. Although bacterial phagocytosis by cells from both groups of donors was similar, significant differences were found in their chemiluminescence responses to opsonized zymosan. Although the mean peak luminol-enhanced chemiluminescence response of dialysate-elicited PM phi was 4.7 X 10(5) cpm, that of resident PM phi was only 1.3 X 10(5) cpm (P less than 0.05). In a lucigenin-enhanced chemiluminescence assay, dialysate-elicited PM phi again generated significantly greater chemiluminescence than did resident PM phi, suggesting that dialysate-elicited PM phi have a relatively increased capacity for O2- production. Using a fluorochrome microassay to assess the intracellular candidicidal activities of these cells, we found that dialysate-elicited PM phi killed 17% of cell-associated blastospores compared with only 1.5% killing by resident PM phi (P less than 0.05). These investigations led us to conclude that results of studies of the functional activity of dialysate-elicited PM phi cannot necessarily be extrapolated to resident PM phi and that dialysate-elicited PM phi do in some respects behave as stimulated or activated cells.
Synthesis of aerobactin and a 76,000-dalton iron-regulated outer membrane protein by Escherichia coli K-12-Shigella flexneri hybrids and by enteroinvasive strains of Escherichia coli
Authors: Griffiths E , Stevenson P , Hale T L , Formal S B
Published: January 2020

One of the chromosomal segments associated with the virulence of Shigella flexneri and transferred to Escherichia coli K-12 by conjugation has been shown to code for the production of aerobactin and for the synthesis of an iron-regulated 76,000-dalton (76K) outer membrane protein. Analysis of various E. coli K-12-S. flexneri transconjugants showed that the genes involved with the synthesis of aerobactin and with the production of the 76K protein were linked to the mtl region of the S. flexneri chromosome. S. flexneri itself synthesized a 76K protein in its outer membrane under iron restriction as well as traces of 81K and 74K proteins. An examination of four enteroinvasive strains of E. coli showed that each produced aerobactin and a 76K outer membrane protein during iron-restricted growth. The profile of the iron-regulated proteins expressed by the enteroinvasive strains of E. coli was virtually identical to that expressed by the laboratory-constructed E. coli K-12-S. flexneri hybrids under the same growth conditions.
Antibody specificity and antigen characterization of rat monoclonal antibodies against Streptococcus mutans cell wall-associated protein antigens
Authors: Ackermans F , Klein J P , Cormont F , Bazin H , Ogier J A , Frank R M , Vreven J
Published: January 2020

Monoclonal antibodies to Streptococcus mutans OMZ175 (serotype f) cell wall-associated antigens (wall-extracted antigens [WEA]) were derived from the fusion of Lou C plasmocytoma rat cells (IR 983 F) and spleen cells from Wistar R inbred rats immunized with WEA. Four cell lines producing monoclonal antibodies directed against a component of S. mutans WEA have been established. All four monoclonal antibodies reacted only with two antigens of WEA from S. mutans OMZ175 by Western blotting and immunoprecipitation techniques, enzyme-linked immunosorbent assay (ELISA), and competitive ELISA. Western blot analysis of WEA showed that the four monoclonal antibodies recognized two related cell wall-associated proteins with apparent molecular weights of 125,000 and 76,000. Immunoprecipitation of whole cells with the monoclonal antibodies confirmed the surface localization of the two antigens. The ELISA and competitive ELISA were used to analyze the distribution of the epitopes on seven S. mutans serotypes. All S. mutans serotypes were found to express the recognized epitopes; however, different reactivity patterns could be distinguished among the various strains tested, and the four monoclonal antibodies reacted only weakly with S. mutans serotypes d and g.
Characterization of the metabolism inhibition antigen of Mycoplasma arthritidis
Authors: Washburn L R , Ramsay J R , Roberts L K
Published: January 2020

The Mycoplasma arthritidis antigen(s) responsible for eliciting metabolism-inhibiting antibodies in rabbits has been partially characterized. Metabolism-inhibiting activity was absorbed from rabbit antisera by intact M. arthritidis cells and membranes but much less so by the soluble cytoplasmic fraction, indicating that the antigen is located on the outer membrane surface. It was stable to periodate and lipid extraction but labile to heat and proteolytic enzymes, indicating that it is protein in nature. Finally, it is most likely a tightly bound integral rather than a peripheral membrane protein, since it was not extracted by low-ionic-strength solutions or by the nonionic detergents Triton X-100, Nonidet P-40, and Tween 20. It was solubilized by both the anionic agent sodium deoxycholate and the zwitterionic detergent Zwittergent. Two two monoclonal antibodies with metabolism-inhibiting activity were produced. One recognized a 45,000-dalton surface protein; however, the other recognized an antigen which is probably of cytoplasmic origin, indicating that more than one cell component may be involved in the metabolism-inhibiting antibody response.
Cloning and expression of two Streptococcus mutans glucosyltransferases in Escherichia coli K-12
Authors: Gilpin M L , Russell R R , Morrissey P
Published: January 2020

Chromosomal DNA from Streptococcus mutans strain MFe28 (serotype h) was cloned in the bacteriophage vector lambda L47.1. Two classes of recombinants were found which expressed glucosyltransferase activity in phage plaques: (i) gtfS, which expressed a glucosyltransferase synthesizing a water-soluble, dextranase-sensitive glucan, and (ii) gtfI, which expressed a primer-dependent glucosyltransferase synthesizing an insoluble glucan.
Quantitative determination of bacterial replication in vivo
Authors: Morris Hooke A , Sordelli D O , Cerquetti M C , Vogt A J
Published: January 2020

A new methodology which permits the quantitative measurement of absolute bacterial replication in vivo is proposed. Mice were inoculated with mixtures of temperature-sensitive mutants and parental wild types, and the changes in the ratios of the two strains were measured. The number of wild-type generations was calculated from the declining ratios over time with the formula n = log (r0/rt)/log 2; n is the number of generations, and r0 and rt are the ratio of temperature-sensitive mutants to the parental wild type at time zero and at the times sampled throughout the experiment. The replication rate was determined by regression analysis. A mathematical argument for the formula is presented. Using this technique, we determined the mean generation times of Escherichia coli (33 min) and Pseudomonas aeruginosa (20 min) in the peritoneal cavities of mice, in the face of host clearance mechanisms during the first stages of infection.
Potentiation by sulfide of hydrogen peroxide-induced killing of Escherichia coli
Authors: Berglin E H , Carlsson J
Published: January 2020

L-Cysteine potentiates 100-fold the hydrogen peroxide-induced killing of a growing culture of Escherichia coli K-12 (Berglin et al., J. Bacteriol. 152:81-88). In the present study it is shown that hydrogen sulfide is formed from L-cysteine and that sodium sulfide could substitute for L-cysteine in the potentiation of hydrogen peroxide-induced killing of E. coli K-12. Addition of an amino acid, L-leucine, L-valine, or L-alanine, to an L-cysteine-containing medium with a growing culture of E. coli K-12 inhibited hydrogen sulfide formation and the potentiation of hydrogen peroxide-induced killing. These amino acids did not inhibit hydrogen sulfide formation from L-cysteine by a cell extract, and they did not inhibit the potentiation by sulfide of hydrogen peroxide-induced killing. This indicated that the amino acids protected the culture from L-cysteine-potentiated, hydrogen peroxide-induced killing by inhibiting the transport of L-cysteine into the cell. The potentiation by sodium sulfide of hydrogen peroxide-induced killing was abolished by the metal ion chelator 2,2'-bipyridyl. This indicated that metal ions, in addition to sulfide, were involved in the killing. Toxic effects of hydrogen peroxide are often presumed to be mediated by hydroxyl radicals formed in iron-catalyzed reactions. It was demonstrated that iron sulfide was more efficient than ferrous iron in catalyzing the formation of hydroxyl radicals from hydrogen peroxide. It was suggested that hydrogen sulfide formed in polymicrobial infections may play an important role in the host defense by potentiating the antimicrobial effect of hydrogen peroxide produced by phagocytic cells.
Correlation of plasmid type and disease caused by Coxiella burnetii
Authors: Samuel J E , Frazier M E , Mallavia L P
Published: January 2020

The obligate intracellular bacterium Coxiella burnetii is the etiological agent of acute Q-fever and chronic endocarditis in humans and of several zoonotic infections. The DNA from a variety of these disease isolates was compared for homology to the plasmid QpH1, found in the Nine Mile strain. Three patterns of homology were found in these isolates, i.e., one pattern identical to that of QpH1, one common to several endocarditis isolates and goat abortion isolates, and one common to the remaining group of endocarditis isolates. Plasmid DNA from the endocarditis-abortion isolate group, designated QpRS, was mapped by restriction enzyme analysis and compared with QpH1. These data show that QpRS was 2 to 3 kilobase pairs larger, contained DNA not found in QpH1, but was not generated from QpH1 by a single insertional event. Isolation of plasmid DNA from the second endocarditis group of isolates was not successful and may indicate that the plasmid has integrated into the chromosome. This analysis provides the first clear evidence that differences exist between C. burnetii isolates which cause various diseases, indicating that different C. burnetii strains may have unique virulence characteristics.
Immune Response to
Cryptococcus neoformans
Soluble Polysaccharide I. Serological Assay for Antigen and Antibody
Authors: Kozel Thomas R. , Cazin John
Published: January 2020

Chromium chloride was used as a coupling agent for the conjugation of purified cryptococcal polysaccharide to sheep erythrocytes. Sensitized erythrocytes were used in a passive hemagglutination (PHA) assay for antibody to cryptococcal polysaccharide and a passive hemagglutination inhibition (PHI) assay for antigen. The PHA assay was more sensitive than complement fixation, agglutination, or precipitation tests for antibody. The PHI assay could detect submicrogram quantities of soluble polysaccharide. Antigen or antibody could be detected in serum or spinal fluid from seven of eight patients with cryptococcosis. Tests for antigen or antibody were negative with sera from patients with histoplasmosis, blastomycosis, coccidioidomycosis, aspergillosis, or allescheriosis. A low frequency (3%) of positive reactors for antibody was found among sera from normal persons and from persons with unrelated diseases; whereas, all tests for antigen were negative. The assay showed a high degree of sensitivity for immunoglobulins of the immunoglobulin M class; however, cryptococcal antibody of the immunoglobulin G class was also detected. The immunological specificity of the polysaccharide preparation was due to carbohydrate rather than to protein associated with the polysaccharide.
Specific Hemagglutinin and a Modulator of Complement in Cockroach Hemolymph
Authors: Anderson Robert S. , Day Noorbibi K. B. , Good Robert A.
Published: January 2020

Natural hemagglutinin activity against vertebrate erythrocytes is present in the hemolymph of the cockroach
Blabarus craniifer
. The hemagglutinin titer against rabbit erythrocytes is high, whereas sheep and horse red cells agglutinate weakly. Hemagglutinin activity was depressed by the complement inhibitor, cobra venom factor. Cockroach hemagglutinin is heat-labile; all activity is destroyed by heating at 56 C for 1 hr. A humoral factor similar to the complement component 3 proactivator is also present in cockroach hemolymph. The formation of the cobra venom factor-hemolymph “complex” is dependent on the presence of divalent cations and will not proceed at 56 C. The hemolytic intermediate formed after treatment of cockroach hemolymph with cobra venom factor was active in the presence of serum treated with ethylenediaminetetraacetic acid to inactivate the early complement components.
Age-Associated Changes in Fecal Excretion Patterns of Strain 93 Chick Embryo Lethal Orphan Virus in Chicks
Authors: Clemmer D. I.
Published: January 2020

Oral incoulation of chickens with strain 93 (chick embryo lethal orphan) virus produced a subclinical infection of the gastrointestinal tract. The pattern of fecal virus excretion in birds infected at 4 or more weeks of age (adult pattern) differed from that in chicks infected when newly hatched (juvenile pattern). By comparison with the juvenile pattern, the adult pattern was characterized by lower peak titers of fecal virus, earlier decline in virus titer, and shorter duration of excretion. Quantitative studies on hatchmates infected at various ages showed that these characteristics are established sequentially with age: lower peak titers were observed in birds infected at 2 to 4 days of age; early decline in titer was first observed in birds infected at 14 days of age; and curtailed excretion was observed in birds infected at 21 days of age. Possible mechanisms operative in the autosterilization process are discussed.
Photodynamic Treatment of Herpes Keratitis
Authors: Moore Conard , Wallis Craig , Melnick Joseph L. , Kuns Michael D.
Published: January 2020

Photodynamic inactivation of virus was tested as a potential therapeutic tool for herpesvirus-induced keratitis, which is a leading cause of blindness and disability. Experimental keratitis produced in rabbits by infection with herpesvirus isolated from a human periorbital lesion was treated with proflavine and light. The herpetic keratitis could be rapidly resolved by the dye-light treatment, whereas improvement was not seen in untreated rabbits, nor in those treated with light alone or dye alone.
Salmonellosis in Orally Infected Specific Pathogen-Free C57B1 Mice
Authors: Collins F. M.
Published: January 2020

Specific pathogen-free C57B1 mice are 100 to 1,000 times as sensitive as CD-1 mice to intravenous or oral challenge by
Salmonella enteritidis
or
S. gallinarum.
Resistance to infection by
S. pullorum
was unaffected. Growth of
Listeria monocytogenes
and
Mycobacterium bovis
(BCG) in intravenously infected C57B1 mice was similar to that seen in CD-1 mice. Quantitative counts of viable
S. enteritidis
in the walls of the stomach, small intestine, cecum, and large intestine and in the corresponding intestinal contents showed that most of the oral challenge inoculum was rapidly inactivated so that, by 24 hr, less than 1% was still viable. Overnight starvation and pretreatment with bicarbonate solution increased the relative survival of the challenge approximately 10-fold. Despite the rapid and extensive inactivation of the oral inoculum within the normal intestine, significant numbers of salmonellae reached the liver and spleen by 48 hr, and this systemic infection was subsequently responsible for the death of a high proportion of the challenged animals.
Effect of an Interferon Stimulator, Polyinosinic: Polycytidylic Acid, on Experimental Fungus Infections
Authors: Worthington Michael , Hasenclever Herbert F.
Published: January 2020

In the present study, the administration of an interferon stimulator, polyinosinic: polycytidylic acid (In:Cn), increased the severity of both experimental
Candida albicans
and
Coccidioides immitis
infections in mice, as evidenced by earlier time of death and higher final mortality. In addition, kidney tissue levels of
C. albicans
at 3 and 5 days after infection were found to be about 40- to 300-fold higher in In:Cn treated mice than in control mice. Experimental
Cryptococcus neoformans
infection, however, was not significantly affected by In:Cn treatment.
Soluble Antigen of Blue-Tongue Virus
Authors: Wang C. S. , Lueker D. C. , Chow T. L.
Published: January 2020

A soluble viral antigen with possible diagnostic utility was obtained from blue-tongue virus-infected lamb kidney cells. It is a noninfectious proteinaceous substance without nucleic acid. The antigen is immunologically identical to one component of the viral particle, and in agar-gel diffusion tests it reacted with antiserum against several blue-tongue virus strains.
Characterization and Comparison of Mycobacterial Antigens by Discontinuous Pore Gradient Gel Electrophoresis
Authors: Affronti Lewis F. , Wright George L. , Reich Melvin
Published: January 2020

Concentrated culture filtrates and cell extracts were prepared from selected mycobacteria and analyzed by a multistage polyacrylamide electrophoretic procedure. Various staining procedures were used to detect protein, carbohydrate, nucleic acid, and lipid constituents. Visual examination of the more prominently stained components indicated differences and similarities in each species of
Mycobacteria
. The majority of the individual components were of small molecular weight; however, there appeared in all culture filtrates and extracts examined a slowly moving protein component of relatively large molecular size. Evidence based on immunodiffusion suggests that it is a common mycobacterial antigen.
Hemagglutinin Response of Human Subjects to Common Enterobacterial Antigen
Authors: Gorzynski E. A. , van Oss C. J. , Ambrus J. L. , Neter E.
Published: January 2020

Ten human volunteers were immunized intravenously with ethanol-soluble common enterobacterial antigen (CA) obtained from
Escherichia coli
O111. The immunogen contained 6 mg/ml, excluding salts. The antigen was given in amounts of 2 ml each in dilutions of 1:250 (48 μg) and 1:125 (96 μg), respectively, 1 week apart. Serum specimens were obtained prior to immunization and 1 week after the last injection. The CA hemagglutinins were titrated by means of the passive hemagglutination test, with CA from
E. coli
O14 as indicator antigen. The injections were well tolerated without side-effects by all subjects. A significant (> fourfold) increase in the titers of CA hemagglutinins was observed following immunization in 9 out of 10 volunteers. These results indicate the feasibility of immunization with CA of human subjects and suggest exploration of immunization against infection caused by
Enterobacteriaceae
producing this common enterobacterial antigen.
Failure of Cholera Enterotoxin to Alter Cyclic 3′, 5′- Adenosine Monophosphate-Mediated Responses in Toad Urinary Bladder
Authors: Lief Philip D. , Keusch Gerald T.
Published: January 2020

Cholera enterotoxin, which stimulates mammalian adenyl cyclase, failed to affect cyclic 3,5′-adenosine monophosphate-mediated functions in toad bladder. In addition, cyclase-mediated responsiveness to vasopressin was maintained.
Differential Susceptibility of Male and Female Mice to Encephalomyocarditis Virus: Effects of Castration, Adrenalectomy, and the Administration of Sex Hormones
Authors: Friedman Stanford B. , Grota Lee J. , Glasgow Lowell A.
Published: January 2020

Male mice have been noted to be more susceptible to encephalomyocarditis (EMC) virus than female animals. Castration of male mice abolished this difference and resulted in enhanced resistance similar to that of female mice. This increase in resistance in castrated males could be eliminated by adrenalectomy. Adrenalectomy alone increased susceptibility in male mice but not in female mice. The administration of a long-acting preparation of testosterone markedly increased susceptibility both in castrated male and female mice, as well as in intact females. Testosterone also had a similar effect in immature mice of both sexes. On the other hand, estradiol, but not progesterone, increased susceptibility to EMC virus in castrated mice of both sexes. These data indicate that testicular, ovarian, and adrenal hormones may influence resistance to EMC virus in the mouse and may be critical determinants of the clinical outcome of infection with this virus.
Time Relationships Between Injection of Antigen and Adjuvant III. Adjuvancy of
Bordetella pertussis
Given at Various Times After the Primary Antigenic Stimulus
Authors: Finger H. , Emmerling P. , Plager L.
Published: January 2020

The adjuvant activity of
Bordetella pertussis
was investigated, both at the cellular and humoral levels, when the bacterial adjuvant was given at various times after the primary antigenic stimulus of both 2 × 10
7
(suboptimal dose) and 4 × 10
8
(optimal dose) of sheep erythrocytes. In all experiments, both adjuvant and sheep erythrocytes were administered by the intraperitoneal route. Adjuvant activity was measured on the basis of the early and late phases of the primary response and on the degree of priming for the secondary immune reaction. A maximal adjuvant activity was found in mice which had received
B. pertussis
vaccine simultaneously with the antigen. Adjuvant effectiveness became less as the time interval between the injection of antigen and adjuvant increased. Adjuvancy also depended on the amount of antigen used as the primary antigenic stimulus. With 4 × 10
8
sheep erythrocytes, significantly increased priming for the secondary response was produced only when
B. pertussis
cells were administered within a period of 24 hr. When the bacterial adjuvant was administered either 48 or 72 hr after the primary antigenic stimulus, adjuvancy was found to be limited to the late phase of the primary response and to the prolonged development of antibody-forming cells during the secondary immune reaction. In contrast, significantly enhanced priming for the secondary response was detectable when the adjuvant was administered as late as 48 hr after primary immunization with 2 × 10
7
sheep erythrocytes. When the bacterial adjuvant was administered either 6, 24, 48, or 72 hr after the primary immunization with 2 × 10
7
sheep erythrocytes, the early phase of the primary 19
S
and 7
S
hemolysin response was found to be suppressed, and adjuvancy became detectable only thereafter.
Mycoplasma-Virus Interrelationships in Mouse Tracheal Organ Cultures
Authors: Westerberg Stephen C. , Smith Charles B. , Wiley Bill B. , Jensen Cynthia
Published: January 2020

The effects produced by single and mixed infections with
Mycoplasma pulmonis
and influenza A/PR-8 virus were studied in mouse tracheal organ cultures.
M. pulmonis
multiplied in the tracheal organ cultures, producing inhibition of ciliary activity and histologic tissue damage. The organism grew in close association with the cell membranes but did not appear to attach directly to the membranes or the cilia. Influenza A virus also replicated in tracheal organ cultures, producing ciliary inhibition and more extensive cytopathologic changes. Virus particles were seen by electron microscopy to attach to and cause clumping of the cilia. Simultaneous infection of the organ cultures with mycoplasma and virus resulted in more rapid inactivation of ciliary activity and greater tissue damage than occurred when the cultures were infected with only mycoplasma or virus. Presence of the virus appeared to have no effect on the growth of the mycoplasma; however, the mycoplasma partially interfered with virus replication.
Pathogenesis and Immunological Aspects of Experimental Histoplasmosis in Cynomolgus Monkeys (
Macaca fascicularis
)
Authors: Kaplan W. , Kaufman L. , McClure H. M.
Published: January 2020

Studies were carried out to obtain basic information on the pathogenesis of experimental histoplasmosis in Cynomolgus monkeys (
Macaca fascicularis
) and to determine whether such infected primates can be used as a source of positive reference sera in serological tests for histoplasmosis. Ten monkeys were inoculated intraperitoneally with approximately 9.1 × 10
7
viable
Histoplasma capsulatum
yeast-form cells or cell aggregates. At periodical intervals, their sera were tested for antibodies to
H. capsulatum
by the complement fixation (CF), immunodiffusion, and latex agglutination tests. Selected monkeys were also sacrificed at periodical intervals for cultural and pathological evaluation of their tissues. Infection with
H. capsulatum
elicited high-antibody responses, and the fungus disseminated to many organs. Initially, the infected monkeys developed CF titers as high as 1:256 to both histoplasmin and
H. capsulatum
yeast cell antigens. Subsequent challenges boosted the CF antibody titers to levels as high as 1:1,024. All of the monkeys developed M precipitins, and some also produced H precipitins. Latex agglutination titers as high as 1:1,024 were also demonstrated. Our findings show that Cynomolgus monkeys experimentally infected with
H. capsulatum
by the intraperitoneal route develop a mild form of histoplasmosis and that these animals can be used as a source of reference sera in serological tests for histoplasmosis.
Immunochemical Parameters of Some Commercial Conjugates for the Fluorescent Treponemal Antibody-Absorption Test
Authors: Hunter E. F. , Smith J. F. , Lewis J. S. , McGrew B. E. , Schmale J. D.
Published: January 2020

Fluorescein-labeled anti-human globulins were examined to determine the need for standardization of conjugates used in the fluorescent treponemal antibody-absorption (FTA-ABS) test. Twenty-one of 33 conjugates submitted by commercial manufacturers to the Reagents Control Activity, Venereal Disease Research Laboratory, for evaluation in the FTA-ABS test were available for study. Conjugates, after evaluation in FTA-ABS performance tests, were examined by immunoelectrophoresis, by titration against immunoglobulins G and M (IgG, IgM) with FTA-ABS techniques, and by the biuret protein and fluorescein diacetate methods for determining fluorescein to protein (F/P) ratios. The conjugates were predominately anti-IgG globulin with anti-light-chain activity. Differences were noted in the ability of some conjugates to detect IgM antibody. The F/P ratios of those conjugates that could be determined varied from 2.6 to 17.8 μg of fluorescein per mg of protein. The need to identify and standardize both the immunologic capabilities and the optimum F/P ratio for FTA-ABS test conjugates is presented.
Rabies Antibody Determination by Immunofluorescence in Tissue Culture
Authors: Debbie John G. , Andrulonis Jerome A. , Abelseth Melvin K.
Published: January 2020

A tissue culture (TC)-fluorescent antibody (FA) technique for the measurement of rabies-neutralizing antibody was found to be reliable and comparable to the standard mouse serum neutralization test. This test was performed with BHK-21 cells infected with the ERA vaccine virus strain on Lab-Tek TC chamber slides. A Flury high egg passage (HEP) rabies virus strain grown on continuous line of African green monkey (Vero) and on BHK-21 cells was investigated to determine its utilization in a TC-FA neutralization procedure. Although both the HEP and ERA viruses infected Vero and BHK-21 cells, the amount of fluorescent antigen observed was most consistent with ERA virus and BHK-21 cells.
Infection of Genital Tract and Transmission of Ocular Infection to Newborns by the Agent of Guinea Pig Inclusion Conjunctivitis
Authors: Mount David T. , Bigazzi Pierluigi E. , Barron Almen L.
Published: January 2020

Female guinea pigs were inoculated intravaginally with the agent of guinea pig inclusion conjunctivitis (Gp-ic). Evidence for infection was obtained by demonstration of Gp-ic inclusions in epithelial cells of genital tract smears, histopathology, recovery of Gp-ic during infection, and antibody response. Infection of the genital tract was produced with low doses (20-200 median egg lethal doses) of the agent. The infection lasted about 3 weeks, and there was no marked clinical response. Ocular infection that could be detected as early as 3 days after birth in offspring of infected mothers was also demonstrated.
Respiratory Diseases in Cyclophosphamide-Treated Mice II. Decreased Virulence of PR8 Influenza Virus
Authors: Singer S. H. , Noguchi P. , Kirschstein R. L.
Published: January 2020

Mice infected intranasally with the PR8 strain of influenza virus were treated with cyclophosphamide, a potent immunologic suppressor. During the first week of infection, mortality in the unmodified influenza infection averaged 65%, whereas in those animals also treated with cyclophosphamide it averaged 22.5%. After the first week, the mortality rate in the infected cyclophosphamide-treated animals rose to that seen during the first week in the animals only infected. This decreased mortality in the first week was found despite the fact that the cyclophosphamide-treated and infected animals had higher virus titers which persisted longer, decreased circulating antibody, and a decreased interferon response. This delayed mortality appeared to be related to the finding of decreased cellular infiltration in the lungs of infected cyclophosphamide-treated animals.
Adenylate cyclase activity of a 68,000-molecular-weight protein isolated from the outer membrane of Bordetella bronchiseptica
Authors: Novotny P , Chubb A P , Cownley K , Montaraz J A
Published: January 2020

A method was developed which is suitable for the isolation of substantial quantities of outer membrane proteins of Bordetella species in a water-soluble form. The extracted material may then be further fractionated in the absence of detergents by ion-exchange chromatography and preparative flat-bed isoelectrofocusing. These procedures facilitated the isolation of one of the proteins, of molecular weight 68,000, for which the antibody titer correlated with the degree of protection against nasal changes induced in specific-pathogen-free piglets by Bordetella bronchiseptica infection (P. Novotny, M. Kobisch, K. Cownley, A. P. Chubb, and J. A. Montaraz, Infect. Immun. 50:190-198). This protein, which banded between 7.0 and 7.6 pH in preparative isoelectrofocusing, may be further purified with a monoclonal immunosorbent. Immunopurified protein showed adenylate cyclase activity. The enzymatic activity was found to be unstable during processing; i.e., although the crude extract showed up to 150 nmol of cyclic AMP per mg/min, the immunopurified protein showed a maximum of only 200 nmol of cyclic AMP per mg/min. Two strains of B. bronchiseptica, isolated from herds of healthy pigs showing no signs of atrophic rhinitis, did not produce the 68,000-molecular-weight protein and were negative for adenylate cyclase. However, it is not known whether the 68,000-molecular-weight protein is a component of adenylate cyclase or whether it is an unrelated protein associated with this enzyme in some unknown way. Adenylate cyclase activity from culture supernatants of B. bronchiseptica, B. pertussis, and B. parapertussis can be absorbed equally to the same monoclonal immunosorbent.
Acute joint inflammation in mice after systemic injection of the cell wall, its peptidoglycan, and chemically defined peptidoglycan subunits from various bacteria
Authors: Koga T , Kakimoto K , Hirofuji T , Kotani S , Ohkuni H , Watanabe K , Okada N , Okada H , Sumiyoshi A , Saisho K
Published: January 2020

The systemic injection of an aqueous suspension of cell wall or its peptidoglycan (PG)-rich sonicate derived from group A streptococcus and Lactobacillus casei induced acute joint lesions in BALB/c, DBA/1J, (BALB/c X DBA/1J)F1, and C3H/He mouse strains, but not in C57BL/6, DBA/2, and AKR strains. Cell walls and their enzymatically degraded PG fragments from other bacteria as well as the synthetic disaccharide dipeptide and Lactobacillus plantarum cell wall-derived disaccharide tripeptide produced similar acute inflammation in susceptible BALB/c mice. Acute swelling and erythema of the ankles and wrists were observed as early as 3 h, reached maximum severity by day 2, and generally subsided by days 4 to 6 after injection. Histological studies showed synovial proliferation, marked infiltration of many mononuclear cells and a few polymorphonuclear leukocytes in the soft tissues, and extensive deposition of fibrinous exudate in the joint space. Antibody response was detectable against the PG fraction. However, anti-PG antibody does not seem to be responsible for the pathogenesis of this disease. On the other hand, experiments on decomplementation by cobra venom factor suggest that complement components are involved in the early phase of this arthritic model.
Demonstration of multiple antigenic determinants on Mycoplasma pneumoniae attachment protein by monoclonal antibodies
Authors: Hu P C , Huang C H , Huang Y S , Collier A M , Clyde W A
Published: January 2020

Distinct multiple antigenic determinants of the attachment protein of Mycoplasma pneumoniae have been identified by limited proteolytic cleavage, using specific monoclonal antibodies. Western blots prepared from the gels containing the cleaved fragments were probed with antiserum against M. pneumoniae or monoclonal antibodies. Five distinct bands with intact antigenic determinants were detected by the antiserum, of which two bands were each reactable with two monoclonal antibodies. A sequential binding assay suggested that these monoclonal antibodies recognized different antigenic sites of each band. These results demonstrate the existence of multiple antigenic sites on the attachment protein and describe procedures that should prove useful for identifying those antigenic sites critical to the specific attachment of M. pneumoniae.
Intracellular distribution of heat-labile enterotoxin in a clinical isolate of Escherichia coli
Authors: Clements J D , Lowe K L , Bonham L , el-Morshidy S
Published: January 2020

The intracellular distribution of heat-labile enterotoxin in a human isolate of enterotoxigenic Escherichia coli varied significantly as a result of changing incubation time, media, and degree of aeration. Direct comparison with a K-12 plasmid recipient revealed a similar but less dramatic response to environmental factors.
Monoclonal antibodies produced against sporozoites of the human parasite Plasmodium malariae abolish infectivity of sporozoites of the simian parasite Plasmodium brasilianum
Authors: Cochrane A H , Barnwell J W , Collins W E , Nussenzweig R S
Published: January 2020

We have used a sporozoite neutralization assay to define the biological relevance of the cross-reactivity of two monoclonal antibodies, raised against sporozoites of the human parasite Plasmodium malariae (Uganda 1/CDC), with sporozoites of the simian parasite Plasmodium brasilianum (Colombian). In vitro incubation of each of these two monoclonal antibodies with sporozoites of P. brasilianum totally abolished the infectivity of these parasites for Saimiri sciureus. Using Western blot analysis and one of the P. malariae monoclonal antibodies, we identified two sporozoite proteins characteristic of the Colombian isolate of P. brasilianum with apparent molecular weights of 56,000 and 66,000. The same monoclonal antibody identified two proteins in an extract of the Peruvian isolate of P. brasilianum with apparent molecular weights of 59,000 and 69,000.
Unimpaired function of human phagocytes in the presence of phagocytosis-resistant group A streptococci
Authors: Manjula B N , Schmidt M L , Fischetti V A
Published: January 2020

The resistance to phagocytosis of the group A streptococci has been attributed mainly to the presence of the surface antigen, M protein. In the present study, we addressed the question of whether the phagocytosis resistance of the group A streptococci is due to their ability to impair the function of the phagocytic cells. The results of these studies demonstrate that the presence of a large excess of a phagocytosis-resistant strain of streptococci does not significantly interfere with either the antibody-independent or the antibody-dependent phagocytosis of streptococci. Apparently, a phagocytosis-resistant strain of streptococci does not bring about a generalized deactivation of the phagocytic plasma membrane. This suggests that if the resistance of the group A streptococci is due to any deactivating influence at all on the phagocytic plasma membrane, it is likely to be confined to the contact area of the cocci with the phagocyte.
Dissection of macrophage tumoricidal and protozoacidal activities using T-cell hybridomas and recombinant lymphokines
Authors: Futch W S , Schook L B
Published: January 2020

Macrophage (M phi) phenotype and function can be modulated by various T-cell lymphokines (LK). The alteration of M phi phenotype is a result of LK concentration, duration of exposure, and the level of M phi activation when obtained from in vivo sources through elicitation by either sterile irritants or cellular immune mechanisms. To dissect M phi activation into discrete signals, we constructed T-cell hybridomas by fusing hypoxanthine-aminopterin-thymidine-sensitive BW5147 cells with nylon wool-purified, concanavalin A-stimulated T cells. The resulting T-cell hybrids were screened for their ability to (i) protect M phi from the cytopathic effect of Naegleria lysates, (ii) induce class II major histocompatibility complex gene product (Ia antigen) expression, (iii) increase tumoricidal and cytostatic activity, and (iv) alter ectoenzyme profiles on either resident or thioglycolate-elicited M phi. Two hybridomas (T-3 and T-9) were selected for further evaluation because of their activity patterns. Supernatants from T-3 and T-9 were compared with cloned gamma-interferon (IFN-gamma) for alterations of biological activities. Both T-3 and T-9 were able to protect resident-M phi cells from Naegleria lysate but had no protective effect on thioglycolate-induced M phi. T-9 supernatant had patterns of activity similar to IFN-gamma, whereas T-3 patterns were different. The addition of anti- IFN-gamma removed T-9 cytostatic activity while not affecting T-3-induced activity. The LK inducing protection from the cytopathic effect of Naegleria lysate is not IFN-gamma but another molecular moiety. We conclude that the activation of M phi for the destruction of tumor cells and amoebae may occur via different mechanisms.
Migration patterns of pathogenic and nonpathogenic Naegleria spp
Authors: Thong Y H , Ferrante A
Published: January 2020

Four species of Naegleria were tested for their ability to migrate under agarose. Pathogenic N. fowleri strains exhibited rapid locomotion at 37 degrees C. Environmental isolates of N. fowleri moved faster than clinical isolates which had been kept in axenic culture for longer periods, and this result was confirmed by using the 84-2205-7 strain kept in axenic culture for 1 or 5 months. Nonpathogenic N. gruberi strains migrated actively at 28 degrees C but not at 37 degrees C; moreover, even at 28 degrees C, active amoebae constituted only a small proportion of the whole. The temperature-tolerant, nonpathogenic species N. lovaniensis moved more slowly than N. fowleri at 37 degrees C. In contrast, N. australiensis, which is temperature tolerant as well as pathogenic for mice, migrated at a rate comparable to that of N. fowleri. There appears to be a direct correlation between the locomotive ability of free-living amoebae and their pathogenic potential.
Bactericidal effect of normal swine sera on Treponema hyodysenteriae
Authors: Joens L A , Nuessen M E
Published: January 2020

Treponema hyodysenteriae was incubated in 20% normal swine sera (NSS) at 37 degrees C for 4 h, and viability was determined by a plate dilution method. NSS was bactericidal for nonpathogenic T. innocens and avirulent T. hyodysenteriae, but not for virulent T. hyodysenteriae isolates. Heat inactivation at 56 degrees C for 30 min, treatment with EDTA or EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid], or removal of immunoglobulin M eliminated the bactericidal activity of NSS. However, removal of the alternate complement pathway with 10 mg of bentonite per ml did not remove bactericidal activity of NSS. Incubation of virulent isolates of T. hyodysenteriae in the presence of specific antisera plus NSS resulted in bactericidal activity. These data suggest that complement and natural antibody may be involved in protecting the host from T. innocens or avirulent T. hyodysenteriae and that T. hyodysenteriae antibody plus complement are involved in protecting convalescent pigs from re-exposure to swine dysentery.
Cell surface characteristics of coagulase-negative staphylococci and their adherence to fluorinated poly(ethylenepropylene)
Authors: Hogt A H , Dankert J , Hulstaert C E , Feijen J
Published: January 2020

The ability of 21 nonencapsulated and 15 encapsulated coagulase-negative staphylococci (CNS) to adhere to xylene in xylene-water emulsions and to fluorinated poly(ethylenepropylene) (FEP) films revealed remarkable differences. Nonencapsulated CNS strains adhered well to FEP, whereas their adherence to xylene ranged widely. Encapsulated strains with low adherence to xylene showed slight adherence to FEP. Encapsulated strains which adhered well to xylene ranged widely in their adherence to FEP. It was concluded that results obtained from the xylene adherence test were not predictive of the adherence of CNS to the hydrophobic FEP surface. The number of nonwashed, slime-producing CNS strains adhering to FEP was similar to that of washed bacteria of the same strains. Bacterial adherence to FEP was decreased when FEP films were exposed to a solution containing extracellular products (EP) obtained from a slime-producing CNS strain. Bacterial adherence to xylene also decreased when the bacterial suspensions contained EP. Apparently, initial adherence of CNS to FEP and xylene is hampered by EP. Nonencapsulated and encapsulated CNS pretreated with proteolytic enzymes failed to adhere to xylene and FEP, indicating that intact surface proteins or constituents associated with surface proteins mediated their adherence to xylene and FEP. Freeze-etch replicas of a CNS strain adhering to FEP showed a smooth, flattened area on the bacterial surface at the contact site of the bacteria with the FEP, indicating that an external layer was present at the bacterial surface.
Analysis of damage to human ciliated nasopharyngeal epithelium by Neisseria meningitidis
Authors: Stephens D S , Whitney A M , Melly M A , Hoffman L H , Farley M M , Frasch C E
Published: January 2020

We used an in vitro model of human nasopharyngeal tissue in organ culture to evaluate the effects of Neisseria meningitidis on human cilia and ciliary function. Encapsulated, viable meningococci damaged ciliated epithelium of nasopharyngeal organ cultures, whereas Neisseria subflava, a commensal species, did not. Meningococcus-induced ciliary damage was due to loss of ciliated cells to which meningococci were not attached. Damage was seen with piliated and nonpiliated meningococci and did not appear to require the presence of other specific meningococcal surface proteins. Meningococcal viability was a requirement for both ciliary damage and interactions of meningococci with microvilli of nonciliated epithelial cells. That is, filter-sterilized supernatants from meningococcus-infected organ cultures, heat-killed meningococci at high inoculum, and purified meningococcal or gonococcal lipopolysaccharide at concentrations of 100 micrograms/ml did not damage ciliary activity of nasopharyngeal organ cultures. In contrast, meningococcal lipopolysaccharide at 10 micrograms/ml markedly damaged ciliary activity of human fallopian tube organ cultures, suggesting a selective toxicity of lipopolysaccharide for specific human ciliated cells. Damage to nasopharyngeal ciliated epithelium by N. meningitidis may be an important first step in meningococcal colonization of the human nasopharynx, but meningococcal lipopolysaccharide does not appear to be directly responsible for this toxicity.
Environmental modulation of lipopolysaccharide chain length alters the sensitivity of Escherichia coli to the neutrophil bactericidal/permeability-increasing protein
Authors: Weiss J , Hutzler M , Kao L
Published: January 2020

We have shown previously that the sensitivity of Escherichia coli to the neutrophil bactericidal/permeability-increasing protein (BPI) depends mainly on the polysaccharide chain length of outer membrane lipopolysaccharides (LPS) (J. Weiss, S. Beckerdite-Quagliata, and P. Elsbach, J. Clin. Invest. 65:619-628, 1980). Thus, rough strains of E. coli producing only short-chain LPS are more sensitive to BPI than smooth strains that produce LPS with varied chain lengths. We now show that changes in the bacterial growth environment can modify BPI sensitivity of smooth E. coli as much as 30-fold depending on the bacterial strain and the growth conditions examined. Changes in BPI sensitivity paralleled differences in binding affinity of E. coli for BPI and closely correlated with changes in the chain length of LPS produced under different growth conditions, as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. No concomitent changes in either the number of LPS molecules per cell or the bacterial protein profile were detected. Rough strains showed little or no growth-dependent variation in BPI sensitivity, further indicating that subtle alterations in bacterial constituents other than LPS do not significantly affect bacterial sensitivity to BPI. Thus, the BPI sensitivity of E. coli can be modulated not only by the genotypic conversion of the LPS phenotype, but also by environmental effects on LPS-polysaccharide formation in wild-type strains.