bluuebunny/crossref_metadata_2025 huggingface dataset:
Interaction of Chlamydia psittaci with mouse peritoneal macrophages
Authors: Wyrick P B , Brownridge E A , Ivins B E
Published: January 2020

L-cell-grown Chlamydia psittaci elementary bodies (EB) were rapidly phagocytized by mouse peritoneal macrophages in vitro. However, the intracellular fate of chlamydiae in macrophages appeared to be dependent on the multiplicity of infection (MOI), i.e., the EB-to-macrophage ratio, and the treatment of the EB. At an MOI of 1:1 or less, survival is maximal, and growth and multiplication of live, untreated chlamydiae did occur. In contrast, at a high MOI (100:1), survival of chlamydiae is reduced, as confirmed by release of 3H-labeled nucleic acid into the supernatant. At the high MOI, macrophage damage occurred that resulted in significant release of the lactic dehydrogenase, beginning 2 h postinfection. This immediated macrophage cytotoxicity as abolished by pretreatment of EB with heat (5 min at 56 degrees C) and was reduced about 50% by coating EB with homologous antibody. Pretreatment of the chlamydia with heat or opsonizing antibody provides increased uptake of EB by macrophages but may contribute to increased destruction of these obligate intracellular pathogens in professional phagocytic cells.
Evidence for Quantitative Variability of Bacterial Opsonic Requirements
Authors: Guckian James C. , Christensen Wayne D. , Fine Douglas P.
Published: January 2020

We studied human serum opsonins by using combinations of heat inactivation and chelation to inhibit complement, adsorption to remove antibody, and trypan blue to inactivate the C3 receptor of human polymorphonuclear leukocytes.
Streptococcus pneumoniae
, serotype 25, required both complement and immunoglobulin for opsonization, even though that strain activated the alternative complement pathway. Both strains of
Escherichia coli
required antibody and complement, but varied in the degree of dependence on the C3 opsonin, since trypan blue moderately inhibited the killing of
E. coli
-1 and markedly inhibited the killing of
E. coli
-2.
Serratia marcescens
was opsonized in heat-inactivated serum (limited complement) or serum absorbed at 0°C with
S. marcescens
(limited antibody), but depended on the C3 receptor in absorbed serum.
S. marcescens
activated the alternative pathway. Thus, opsonic requirements varied with the availability of opsonins. Requirements for bacterial opsonization vary with species and strains within species, perhaps reflecting quantitative relationships among alternative and classical pathway activation of C3, efficiency of adsorption of C3 or immunoglobulin G to bacterial surfaces, and efficiency of attachment of these ligands to polymorphonuclear leukocyte receptors. Furthermore, although not always sufficient for opsonization, the C3 opsonin (activated through either the classical or alternative pathway) appears necessary for effective phagocytosis and killing of all strains studied.
Kinetics of the humoral and cellular immune response of guinea pigs after injection of the synthetic adjuvant N-acetylmuramyl L-alanyl-D-isoglutamine: comparison with Freund complete adjuvant
Authors: Souvannavong V , Adam A , Lederer E
Published: January 2020

We studied the time course of humoral and cellular immunity of guinea pigs injected with the synthetic adjuvant N-acetylmuramyl L-alanyl-D-isoglutamine (MDP Pasteur, 10 microgram) in Freund incomplete adjuvant; the kinetics were compared with those obtained with Freund complete adjuvant (50 microgram of whole Mycobacterium butyricum). The antibody response to ovalbumin was faster and higher with MDP, but dropped sooner to a low level; the secondary response was, however, again higher for MDP than for Freund complete adjuvant. Cellular immunity, as measured by delayed hypersensitivity, and migration inhibition factor production werepositive for both adjuvants. The same response was followed in animals injected with MDPA, the nonamidated analog of MDP; the same kinetics as for Freund incomplete adjuvant were obtained for the primary response, but the secondary response was stronger and gave a positive delayed hypersensitivity reaction.
Existence of multiple immunodeterminants in the type-specific capsular substance of group B type Ia streptococci
Authors: Kane J A , Karakawa W W
Published: January 2020

Rabbits immunized with group B, type Ia streptococci produce two distinct populations of protective antibodies. Evidence is presented indicating that these antibodies are directed against two major immunodeterminants which coexist in the specific type Ia antigen. Immunochemical data, using purified antibody preparations, indicate that the type substance, a sialic acid polymer consisting of galactose, glucose, glucosamine, and sialic acid, possesses two distinct immunodominant determinants, terminal sialic acid residues and a galactosyl oligosaccharide. Antibodies directed against either of these determinants were shown to possess in vivo and in vitro opsonic capabilities.
Biochemical examination of sera during systemic Candida infection in mice
Authors: Oblack D , Schwarz J , Holder I A
Published: January 2020

Candida pathogenesis was examined by intravenous challenge of mice with either C. albicans or C. guilliermondii. Animals were moribund 12 h postchallenge with C. albicans and were found to have the greatest number of organisms in the heart and kidney, severe interstitial myocarditis, and elevated serum levels of blood urea nitrogen, creatine phosphokinase, serum glutamic oxalacetic transaminase, serum glutamic pyruvic transaminase, and lactic dehydrogenase. In contrast, challenge with C. guilliermondii resulted in a significantly lower renal census, no myocarditis, and no significant change in the concentration of these serum constituents. Challenge with nonviable C. albicans did not produce the effects observed with viable organisms. Moreover, challenge with filamentous C. albicans resulted in biochemical alterations of lower magnitude and in lower mortality rates. These results indicated that altered serum biochemistries were correlated with the histopathology of fatal Candida infection and that there were distinct differences with C. guilliermondii and the dimorphic phases of C. albicans.
Separation of Soluble
Brucella
Antigens by Gel-Filtration Chromatography
Authors: McGhee Jerry R. , Freeman Bob A.
Published: January 2020

Soluble precipitating antigens of
Brucella suis
have been, in various degrees, purified by filtration on Sephadex gels. The most useful gels employed were Sephadex G-150, Sephadex G-200, and Sepharose 4B. Although not all fractions proved to be immunologically pure, some crude molecular-size estimates of most of the 13 soluble antigens of the
Brucella
cell could be given. In addition, monospecific antisera to three purified
Brucella
antigens have been prepared. By using purified preparations, physical and chemical data were obtained on two major antigens, E and
1
, and a minor antigen, f. Antigen E is not an agglutinogen and may be toxic. Antigen
1
is of low molecular weight and is neither toxic nor agglutinogenic. The minor antigen f is an agglutinogen as well as a precipitinogen and is found on the cell surface. Both major antigens, when purified, were immunogenic in rabbits.
Uptake of
3
H-Dihydrostreptomycin by Macrophages in Culture
Authors: Bonventre Peter F. , Imhoff John G.
Published: January 2020

Mouse peritoneal macrophages, in culture, concentrate significant amounts of
3
H-dihydrostreptomycin, provided that the incubation period is sufficiently extended. Macrophages cultured in vitro from both stimulated and unstimulated animals concentrate the antibiotic from growth or maintenance media. The increase in cell-associated radioactivity is linear for almost a week before a plateau is reached. Calculations based on intracellular volumes of the cells indicate that the intracellular concentration of dihydrostreptomycin may attain levels greater than five times that of the external milieu. No uptake is measurable at 4 C, suggesting an active mechanism of transport into the cell. Phagocytosis of killed bacteria during incubation did not increase uptake of the antibiotic nor did the addition of poly-
l
-ornithine to the medium augment uptake. A nonphagocytic cell line (BHK-21) concentrated
3
H-dihydrostreptomycin to a lesser extent than the macrophages. These observations suggest that a wide variety of mammalian cells may be permeable to the antibiotic, and thus potential bactericidal action on intracellular bacteria cannot be ignored.
Evidence on Possible Mycoplasma Etiology of Aster Yellows Disease I. Suppression of Symptom Development in Plants by Antibiotics
Authors: Davis R. E. , Whitcomb R. F.
Published: January 2020

Antibiotics suppressed development of aster yellows (AY) disease symptoms in plants of china aster [
Callistephus chinensis
(L.) Nees.] and annual chrysanthemum (
Chrysanthemum carinatum
, Schousb.). When inoculated chrysanthemum plants were treated by any of several techniques with tetracycline antibiotics or chloramphenicol, symptoms failed to appear during treatment but appeared 1 to 4 weeks after treatments were terminated. Under continuous administration of chlortetracycline, aster plants with AY symptoms developed symptomless axillary growth, including flowers. Streptomycin, oleandomycin, kanamycin, tylosin, carbomycin, polymyxin, bacitracin, neomycin, sulfanilamide, penicillin, vancomycin, or cycloserine had no discernible effect on development of AY symptoms. Treatment of plants with tetracycline antibiotics before exposure to inoculative (pathogen-transmitting) vectors delayed the appearance of symptoms or prevented AY infection. Remission of AY symptoms in inoculated plants treated with chlortetracycline was correlated with an inhibition of multiplication of AY agent, as measured by bioassay of extracts. The data give additional support to the hypothesis that aster yellows disease is caused by a mycoplasma-like microorganism.
Nonspecific immunostimulant activities of synthetic trehalose-6,6'-diesters (lower homologs of cord factor)
Authors: Parant M , Audibert F , Parant F , Chedid L , Soler E , Polonsky J , Lederer E
Published: January 2020

Mycobacterial cord factors (6,6'-diesters of trehalose with mycolic acids ranging from C80 to C90) have been shown to protect mice effectively against infection with Klebsiella pneumoniae or with Listeria monocytogenes. Our present findings indicate that the low-molecular-weight cord factor of Corynebacterium diphtheriae (with corynomycolic acids ranging from C28 PTO C36) is equally active. Moreover, its synthetic analog (with synthetic C32 mycolic acid) has the same activity. Two lower synthetic 6,6'-diesters of trehalose with C22 acids, which are described here for the first time, as well as dipalmitate and a dioleate of sucrose, were found inactive. The synthetic C76 trehalose diesters, which are capable of enhancing nonspecific resistance to infection, increase the immune response in mice, even when injected in metabolizable oil. They induce in the injected paws an inflammatory process weaker and more transient than the natural cord factor.
Molar Growth Yields and Enterotoxin B Production of
Staphylococcus aureus
S-6 with Amino Acids as Energy Sources
Authors: Keller Gabriela M. , Hanson Richard S. , Bergdoll Merlin S.
Published: January 2020

Staphylococcus aureus
S-6 was able to utilize glutamate, proline, histidine, aspartate, alanine, threonine, serine, or glycine as a major energy source, when these were added singly at a concentration of 2.5 g/liter to a base medium containing salts, vitamins, and 30 mg of each of 18 amino acids per liter. Molar growth yields (
Y
) for each of the eight amino acids were determined in aerobic batch cultures. Except with aspartate and serine, all molar growth yields decreased with increasing concentration of the energy source, whether or not it was present in excess. The average molar growth yields for both aspartate and serine were
Y
= 35 g/mol. The highest
Y
values obtained with the other amino acids were 79, 78, 56, 46, 58, and 21 g/mol for glutamate, proline, histidine, alanine, threonine, and glycine, respectively. Histidine was not utilized as an energy source at concentrations of less than 2 mM. Enterotoxin yields (mg of toxin per g [dry wt] of cells) higher than in the basal medium were obtained when proline, histidine, alanine, serine, and glycine were the major energy sources. When proline and alanine were the major energy sources, enterotoxin yields were not affected by the concentration, whereas toxin yields were reduced at high concentrations of the other six amino acids. The highest cell yields and
Y
values were obtained with glutamate, but its utilization as an energy source markedly reduced enterotoxin yields.
Platelet migration inhibtion as an indicator of immunologically mediated target cell injury in canine ehrlichiosis
Authors: Kakoma I , Carson C A , Ristic M , Stephenson E M , Hildebrandt P K , Huxsoll D L
Published: January 2020

A platelet migration inhibition test was devised to determine the presence of antiplatelet activity in serum collected from experimentally produced and natural cases of canine ehrlichiosis. The maximum platelet migration inhibition effect was observed during the acute phase of the disease and before the appearance of specific humoral antibody, measured by the indirect fluorescent-antibody test. Platelet migration inhibition may be one of the earliest events leading to pancytopenia. In most cases, sera positive for humoral antibodies also were positive for platelet migration inhibition, although no direct correlation was evident between the serological titer and the degree of platelet migration inhibition. Inoculation of dogs with uninfected canine blood did not induce the production of inhibition factor or antibody activity, which precluded a histocompatibility response to the cellular elements in the inoculum. Scanning electron microscopy indicated that the platelet inhibition factor interfered with platelet migration by inhibiting pseudopod formation. Affected platelets became rounded and showed evidence of clumping and leakage.
Experimental infection of the genital tract of female grivet monkeys by Mycoplasma hominis
Authors: Møller B R , Freundt E A , Black F T , Frederiksen P
Published: January 2020

Mycoplasma hominis, a common inhabitant of the mucosae of the genitourinary tract of human and nonhuman primates, was inoculated directly into the uterine tubes of five laparotomized grivet monkeys. A self-limiting acute salpingitis and parametritis developed within a few days in all animals. Although there were no clinical signs of overt disease, the gross pathology was characterized by pronounced oedematous swelling and hyperaemia of the tubes and parametria. Microscopically, cellular infiltrations of lymphocytes and some polymorphonuclear leukocytes were found in the acute phase in the subserosa and muscularis of the tubes and in the parametria. Granulation tissue and fat necrosis appeared at a later stage in the parametria. The infection was associated with a marked antibody response and a moderate rise of the erythrocyte sedimentation rate and leukocyte counts. The capability of M. hominis to produce salpingitis and parametritis in a nonhuman primate would seem to add rather significantly to the available evidence suggesting an etiological role of this organism in inflammatory diseases of the internal female genitals of humans.
Chromosomal locus for staphylococcal enterotoxin B
Authors: Shafer W M , Iandolo J J
Published: January 2020

The genetic locus of staphylococcal enterotoxin B (SEB) was investigated in the Staphylococcus aureus food-poisoning isolates, strains S6 and 277. Direct neutral sucrose gradient centrifugation analysis of sodium dodecyl sulfate-sodium chloride-mediated cleared lysates demonstrated that strain S6 contained a single 37S plasmid. Transductional analysis revealed that the 37S plasmid in S6 encoded for cadmium resistance (Cad) but not SEB. Additionally, elimination of cadmium resistance in S6 provided a plasmid-negative derivative that produced SEB at the same level as the parent. Examination of strain 277 showed two plasmids, a 37S species encoding for penicillin resistance (Penr) and a 21S species containing the gene(s) responsible for tetracycline resistance (Tetr). Elimination of the 37S, penr plasmid in 277 had no effect on SEB production, whereas introduction of the 21S tetr plasmid via transformation into strain 8325 (SEB--) did not confer enterotoxigenesis upon the transformants. The data obtained in this investigation suggest that the SEB gene(s) in these food-poisoning isolates of S. aureus is chromosomal.
Impaired host resistance to endotoxin and malaria in polychlorinated biphenyl- and hexachlorobenzene-treated mice
Authors: Loose L D , Silkworth J B , Pittman K A , Benitz K F , Mueller W
Published: January 2020

The in vivo effect of polychlorinated biphenyl (PCB) and hexachlorobenzene (HCB) on murine endotoxin sensitivity and resistance to malaria (Plasmodium berghei NYU-2) infection was studied. The dietary administration of 167 ppm (167 microgram/g) of PCB 1242 or HCB for 3 weeks resulted in an enhanced sensitivity to gram-negative endotoxin (Salmonella typhosa), which was further increased in animals maintained on the diets for 6 weeks. By 6 weeks, a 5.2- or 32-fold increase in endotoxin sensitivity was seen in mice fed PCB or HCB, respectively. A 20% decrease in mean survival time of mice fed PCB 1242 for 3 or 6 weeks and inoculated with malaria was demonstrated. Infected mice that received HCB for 3 or 6 weeks manifested a reduction in mean survival time of 24 or 31%, respectively. Histopathological examination revealed a normal thymus, spleen, mesenteric lymph nodes, and lungs. Centrilobular and pericentral hepatocyte hypertrophy, common to organochlorine exposure, was observed. Electron capture gas chromatographic analysis for PCB 1242 or HCB in the tissues examined histologically revealed a significant deposit of the xenobiotics. HCB concentration was approximately 16 to 25 times greater than that of PCB. The data indicate that environmental chemicals impair host resistance and that the alteration may be related to the presence of the chemicals in the lymphoreticular organs.
Siderophore production by Vibrio cholerae
Authors: Payne S M , Finkelstein R A
Published: January 2020

Vibrio cholerae produces a phenolate-type siderophore that stimulates growth of the organism in low-iron medium. This compound is similar, but not identical, to enterochelin, the siderophore produced by Salmonella and Escherichia coli.
Role for Macrophages and Thymus-Derived Lymphocytes in Cholera Toxin-Induced Immunosuppression
Authors: Lyons Susan F. , Friedman Herman
Published: January 2020

The exo-enterotoxin derived from
Vibrio cholerae
bacilli has marked immunomodulating activities, both in vivo and in vitro. In the present study, the mechanism whereby cholera toxin depresses the antibody-forming ability of murine splenocytes was investigated by in vitro reconstitution experiments. Spleen cells derived from mice treated with cholera toxin 2 days earlier were markedly deficient in their ability to respond to sheep erythrocytes upon challenge immunization in vitro. Addition of graded numbers of normal spleen cells to spleen cell cultures from toxin-treated mice partially restored the antibody response. Adherent splenocyte populations were even more effective in restoring antibody formation. Normal peritoneal exudate cells rich in macrophages were also capable of restoring the antibody-forming ability of toxin-pretreated splenocytes. Furthermore, thymus (T)-derived spleen cells from normal mice, as well as sheep erythrocyte “educated” T cells, were capable of restoring antibody formation to normal levels. The importance of T lymphocytes in restoring immune competence of spleen cell cultures from toxin-treated mice was shown by additional experiments in which T-depleted cell preparations were found to be ineffective in restoring antibody activity. These studies point to macrophages and T-derived lymphocytes as a major target for cholera toxin-induced immunosuppression.
Production and partial purification of Salmonella enterotoxin
Authors: Sedlock D M , Koupal L R , Deibel R H
Published: January 2020

By using a strain of Salmonella typhimurium, we detected the presence of an enterotoxin, as determined by the rabbit ileal loop assay, in various complex and defined media. The enterotoxin was concentrated by ultrafiltration of culture supernatant fluids and eluted in and adjacent to the void volume of a Sephadex G-100 column. This suggested that the enterotoxic factor was of a relatively high molecular weight, and additional evidence indicated it was heterogeneous in size. Further chromatography, using a diethylaminoethyl-cellulose anion exchanger, facilitated at least a 50-fold purification of the Salmonella enterotoxin.
Pathogenesis, Lethality, and Immunizing Effect of Experimental Cutaneous Cryptococcosis
Authors: Dykstra Mark A. , Friedman Lorraine
Published: January 2020

Mice were subcutaneously inoculated with small numbers of virulent
Cryptococcus neoformans
and divided into groups. Numbers of viable yeasts at the site were estimated at weekly intervals for 5 weeks on the basis of cultures of minced tissue excised from sacrificed animals. Organisms multiplied at the site for at least 4 weeks and were still detectable after the 5th week, although in reduced numbers. Agglutinins appeared within a week, but these antibodies were not detectable during the 2nd through the 5th week. Cryptococcal polysaccharide began to appear in the sera at 3 weeks, persisting through the duration of 5 weeks. All animals appeared healthy, but a few sickened after many months and died of systemic cryptococcosis. All of these events were observed in many separate experiments. The immunizing capacity of a cutaneous lesion was tested by challenging some of the above animals with viable
C. neoformans
after various intervals of time, either subcutaneously at a site distant from that of the vaccination or intravenously. Although we were unable to demonstrate reduced multiplication of yeasts in the brains, lungs, and spleens of intravenously challenged animals, it was possible to show that multiplication was inhibited at the site of subcutaneous challenge. It was noted also that vaccinated animals lived longer after lethal intravenous challenge than did nonvaccinated animals. The latter protection was observed, however, only when challenge followed vaccination by 3 weeks or longer, and it was effective only against a relatively low challenge dose. Mice were protected against a higher dose if they had previously received killed cryptococci, alternating subcutaneous and intraperitoneal inoculations, one of which contained a microbial adjuvant. No protection was observed in animals that were subcutaneously vaccinated with inert materials such as chitin, latex spheres, or even cryptococcal cell walls themselves.
Purification of cholera toxin and its subunits: new methods of preparation and the use of hypertoxinogenic mutants
Authors: Mekalanos J J , Collier R J , Romig W R
Published: January 2020

Cholera toxin was obtained in pure form by fractionation on two phosphocellulose columns successively. Cholera toxin and choleragenoid were quantitatively and selectively adsorbed to the first column in 10 mM phosphate buffer, pH 7.0, and were subsequently eluted with buffer of high ionic strength. The toxin was then separated from choleragenoid on the second column by chromatography at pH 8.3. The toxin obtained was highly active and pure as judged by electrophoresis, isoelectric focusing, and various immunological and chemical tests. Pure choleragenoid was by-product of the procedure. The A1 chain of the toxin was obtained in pure form by treating phosphocellulose-bound toxin with urea and a reducing agent. The anionic A1 peptide was thereby released, leaving a complex of the B and A2 chains (A25B) bound to the resin. The latter was then eluted and further purified to obtain nontoxic antigen. The overall yields of cholera toxin and choleragenoid were increased two- to threefold by the use of hypertoxinogenic mutants of Vibrio cholerae.
Characterization of plasmids that encode for the K88 colonization antigen
Authors: Shipley P L , Gyles C L , Falkow S
Published: January 2020

K88 antigen, and important virulence factor in porcine enteropathogenic Escherichia coli (EEC), can be transferred along with the ability to ferment the trisaccharide raffinose (Raf). The plasmids from a number of EEC strains that encode these two properties were isolated and characterized. In most strains the K88 and Raf genes were found on a single nonconjugative plasmid approximately 50 x 10(6) daltons in size. This plasmid core was conserved with only slight variation among the strains tested. In some transconjugants, larger conjugative plasmids were observed that were apparently recombinants between the Raf/K88 plasmid and a transfer fa(tor. Occasionally plasmids carrying only the raffinose fermentation genes arose by deletion of a deoxyribonucleic acid segment of about 20 x 10(6) daltons that included the K88 antigen gene(s).
Invasive aspergillosis: antiserum for circulating antigen produced after immunization with serum from infected rabbits
Authors: Lehmann P F , Reiss E
Published: January 2020

We describe a method for producing an antibody capable of detecting an antigen of Aspergillus fumigatus in serum and urine from infected rabbits and humans.
Adjuvant activity of synthetic 6-O-"mycoloyl"-N-acetylmuramyl-L-alanyl-D-isoglutamine and related compounds
Authors: Azuma I , Sugimura K , Yamawaki M , Uemiya M , Kusumoto S , Okada S , Shiba T , Yamamura Y
Published: January 2020

Adjuvant and antitumor activities of synthetic 6-O-"mycoloyl"-N-acetylmuramyl-L-alanyl-D-isoglutamine were examined. All the synthetic 6-O-corynomycoloyl-, 6-O-mocardomycoloyl-, and 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine were active as adjuvants for cell-mediated immune responses. However, 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine was less active as an adjuvant on circulating antibody formation. It was shown that pyrogenic activity of N-acetylmuramyldipeptide was reduced by 6-O-acylation with mycolic acid, but not with nocardomycolic or corynomycolic acid. Tumor-suppression activity was observed by the synthetic 6-O-mycoloyl-N-acetylmuramyl-L-alanyl-D-isoglutamine by using transplantable tumor in syngenic mice.
Lymphocyte cytotoxicity to influenza virus-infected cells: response to vaccination and virus infection
Authors: Greenberg S B , Criswell B S , Six H R , Couch R B
Published: January 2020

Peripheral blood leukocytes obtained from volunteers at various times following influenza vaccine or live influenza virus infection were assayed for cytotoxicity against influenza virus-infected cells. Cytotoxicity was highest on days 3 and 7 following vaccination with commercial A/Port Chalmers/1/73 inactivated influenza virus vaccine. Maximal cytotoxicity was found 9 days after infection induced by intranasal inoculation of a strain of A/Scotland/840/74 influenza virus. Individuals naturally infected with A/Victoria/3/75 were also found to have elevated cytotoxicity approximately 1 week after onset of illness. Cytotoxicity levels decreased toward base line approximately 30 days after the virus exposure. The effector mechanism(s) responsible for the early, transient elevation in specific immune release to influenza virus-infected cells may be different from the antibody-dependent cell cytotoxicity demonstrated in the peripheral blood leukocytes from volunteers who had a remote experience with influenza virus.
Proliferative and interferon responses by peripheral blood mononuclear cells after bone marrow transplantation in humans
Authors: Levin M J , Parkman R , Oxman M N , Rappeport J M , Simpson M , Leary P L
Published: January 2020

The capacity of peripheral blood mononuclear cells from bone marrow transplant recipients to proliferate and produce interferon in response to mitogens and specific antigens was tested. Proliferation in response to phytohemagglutinin or pokeweed mitogen occurred in cells from more than 90% of the recipients, and interferon was present in 60 to 70% of the supernatants from these cultures, even when tested as soon as 8 weeks after transplantation. Proliferation in response to bacterial antigens was infrequent, and interferon release was not detected. In the early post-transplantation period (less than 13 weeks), cells from only two of four cytomegalovirus (CMV) antibody-positive patients proliferated normally in response to CMV antigen and interferon release was detected only once. In the late post-transplantation period (more than 13 weeks), in only two of five instances did cells proliferating in response to CMV antigen release interferon. The response to CMV antigen of mononuclear cells from many transplant recipients differs from that of cells from normal controls.
Experimental
Haemophilus influenzae
Type b Meningitis: Immunological Investigation of the Infant Rat Model
Authors: Granoff Dan M. , Rockwell Richard
Published: January 2020

Infant rats inoculated intraperitoneally with
Haemophilus influenzae
type b develop bacteremia and meningitis. Rats were infected at 10 to 12 days of age and studied for the development of serum anticapsular antibody and bactericidal and opsonizing activity. Seven and 11 weeks after inoculation, convalescent animals showed a higher frequency of anticapsular antibody responses than uninfected controls, but 35 to 40% of the infected group had undetectable levels of anticapsular antibody (<0.10 μg/ml). In contrast, all of the convalescent animals, but none of the controls, showed moderate titers of serum bactericidal activity; and bactericidal activity persisted after absorption of the convalescent sera with type b capsule. Bactericidal activity was detected primarily in the eluted fraction corresponding to a molecular weight of 150,000 and was present in the offspring of convalescent females. Offspring of convalescent females were protected against challenge with
H. influenzae
type b, and control offspring could also be protected by passive immunization with convalescent serum which lacked detectable anticapsular antibody. Convalescent serum samples efficiently opsonized
H. influenzae
type b, and this activity persisted after absorption of the serum with capsular antigen. These data are consistent with the hypothesis that antibody to the noncapsular surface antigens of
H. influenzae
type b play an important role in host defenses.
Trypanosoma rhodesiense infection in congenitally athymic (nude) mice
Authors: Campbell G H , Esser K M , Phillips S M
Published: January 2020

Athymic nude mice (nu/nu), heterozygous litter mates (nu/+), and thymic reconstituted homozygous animals (nu/nu TxR) were infected with Trypanosoma rhodesiense. A reduced parasitemia and prolonged survival were observed in the nu/nu animals after infection and cure or after immunization with irradiated organisms. These studies indicate that resistance of mice to T. rhodesiense infection is relatively independent of thymic lymphocyte function.
Role of the surface coat in in vitro attachment and phagocytosis of Plasmodium berghei by peritoneal macrophages
Authors: Brooks C , Kreier J P
Published: January 2020

Evidence is presented to indicate that Plasmodium berghei merozoites, but not trophozotites, have an antiphagocytic capsule. The capsule appears to form around the developing merozoties of the schizont in the parasitophorous vacuole. Serum from animals immune to P. berghei reacts with this capsule. After reaction with immune serum, the antiphagocytic action of the capsule is lost. By the process of binding serum protein, the capsule becomes electron dense and can be readily visuallzed as the surface coat by electron microscopy. At physiological temperatures, phagocytosis by macrophages rapidly follows adhesion of antibody-coated parasites. Both tight and loose phagosomes are formed.
Studies on a canine intestinal spirochete: scanning electron microscopy of canine colonic mucosa
Authors: Turek J J , Meyer R C
Published: January 2020

A large homogeneous population of spirochetes was observed attached to the colonic mucosa of clinically normal dogs. Their association with the colonic mucosa and their mode of attachment resembled those of a spirochete reported in primates.
Toxicity of emulsified trehalose-6,6'-dimycolate (cord factor) in mice depends on size distribution of mineral oil droplets
Authors: Yarkoni E , Rapp H J
Published: January 2020

Toxicity of emulsified trehalose-6,6'-dimycolate administered intravenously into mice depended on the size distribution of the mineral oil droplets. Emulsions containing the greatest number of the largest oil droplets were the most toxic. Emulsions of trehalose-6,6'-dimycolate made with peanut oil or with olive oil were less toxic than those made with mineral oil.
Increased Tissue Conductance and Ion Transport in Guinea Pig Ileum After Exposure to
Staphylococcus aureus
Delta-Toxin In Vitro
Authors: O'Brien Alison D. , McClung H. Juhling , Kapral Frank A.
Published: January 2020

Prior studies had shown that
Staphylococcus aureus
delta-toxin was able to inhibit water absorption in guinea pig ileum and to elevate the cyclic AMP content of this tissue, but was unable to elicit certain cyclic AMP-mediated changes in Y-1 adrenal or Chinese hamster ovary cells. Because water movement passively follows the net movement of electrolytes in the gut, this study investigated the effect of delta-toxin on ion transport in guinea pig ileum maintained in vitro. The transmural potential difference (PD) of guinea pig ileum was measured and nullified with an automatic voltage clamp. The short circuit flowing under these conditions (
I

sc

) was measured, and the conductance was calculated (
I

sc

/PD). Unidirectional
22
Na
+
and
36
Cl
−
fluxes were measured. In a glucose-free Ringer solution, delta-toxin caused an immediate spike in PD and
I

sc

, and the extent and duration of the spike generally increased with increasing toxin concentration. The conductance of ileum was increased by delta-toxin, and this effect on conductance could be blocked by lecithin, a known inhibitor of delta-toxin. Tissue in the presence of glucose did not exhibit a spike in PD or
I

sc

when exposed to delta-toxin. In a glucose-free medium, delta-toxin caused a 1.5- to 2.5-fold increase in both the unidirectional absorption and secretion of Na
+
and Cl
−
, whereas the net secretion of Na
+
increased above basal levels. The observation that delta-toxin causes a prompt increase in intestinal ion flux lends credence to the concept that the elevation in cellular cyclic AMP, which occurs later, is a secondary response to the toxin. The rapid increase in ion flux may reflect the ability of delta-toxin to augment intercellular movement of ions across the mucosa rather than the stimulation of transcellular processes.
Immunoprophylaxis with BCG of experimental Echinococcus multilocularis infections
Authors: Reuben J M , Tanner C E , Rau M E
Published: January 2020

Previous studies have demonstrated that prophylactic treatment with BCG protects cotton rats (Sigmodon hispidus) against experimental infections with Echinococcus multilocularis; this treatment can, however, induce granulomatous reactions. In an attempt to identify a minimum prophylactic dose of BCG which would not induce granulomas, cotton rats were treated intraperitoneally with various doses of BCG (10(1) to 10(7) colony-forming units [CFU]) and then inoculated intraperitoneally with one brood capsule of the parasite. Consistent and complete protection was obtained by the inoculation of as few as 10(3) CFU of BCG. A dose of 10(1) CFU gave no protection whatsoever, and 10(2) CFU gave only partial protection. Doses larger than 10(3) (10(5), 10(7) CFU) also afforded complete protection but gave rise to granulomatous lesions. At the time of the inoculation of the parasite, protection coincided with a general elevation of leukocytes, especially cells of the monocyte/macrophage series. It is proposed that these results support evidence for the macrophage being the principal potential effector cell in hydatid disease.
Analysis of antigenic diversity among human cytomegaloviruses by kinetic neutralization tests with high-titered rabbit antisera
Authors: Waner J L , Weller T H
Published: January 2020

Neutralizing antisera to human cytomegalovirus were produced in rabbits with alkaline-buffered extracts of infected cell cultures. The antibody activity was complement dependent and associated primarily with the 7S immunoglobulin fraction. Antisera with homologous K values greater than 10.00 were shown to be suitable for neutralization kinetic studies and were so used to examine the antigenic relatedness of strains AD169, Davis, Esp, C-87, Kerr, and Towne. Based upon the degree of relationship as determined by normalized K values, antisera to the Davis or AD169 strains discriminated three antigenic groups, and an antiserum to strain Esp discriminated two groups.
Binding of Glucosyltransferase and Glucan Synthesis by
Streptococcus mutans
and Other Bacteria
Authors: Hamada Shigeyuki , Tai S. , Slade Hutton D.
Published: January 2020

Lyophilized and heat-treated cells from the seven serotypes of
Streptococcus mutans
were examined for their ability to bind added insoluble-product glucosyl-transferase (GTase) and to synthesize cell-associated glucan from [
14
C]sucrose. Lyophilized cells of serotypes
a
and
g
did not synthesize any more additional glucan than did the controls after exposure to GTase. These cells, however, synthesized four- to eightfold-greater quantities of glucan than did the cells of the remaining serotypes. Lyophilized cells of serotypes
b, c, d, e
, and
f
synthesized two- to threefold-greater quantities of glucan after exposure to GTase than did the controls without added enzyme. Lyophilized cells of serotypes
a
and
g
synthesized 6- to 10-fold-greater quantities of glucan than did heat-treated cells of the same strain after binding of GTase. Lyophilized cells of the remaining serotypes synthesized only 1.6- to 3.3-fold-greater quantities of glucan than did the heat-treated cells. These results demonstrate that heat treatment to inactivate cell-associated GTase does not create additional GTase binding sites in
S. mutans
and that serotypes
a
and
g
are considerably more active in cell-associated glucan synthesis than cells of the other five serotypes. Ten species of gram-positive and gram-negative bacteria from five genera which do not produce in vitro plaque synthesized 10- to 100-fold-less glucan than did the
S. mutans
strains after exposure to GTase. Of these species,
S. sanguis, Actinomyces viscosus
, and
A. naeslundii
synthesized the largest quantities of glucan. Three mutant strains of
S. mutans
which possess a reduced ability for in vitro adherence but do agglutinate with glucan or dextran synthesized only one-third as much glucan after binding of GTase as the control. These results are discussed in relation to in vitro and in vivo plaque development and the agglutination of
S. mutans.
The results support earlier findings which indicate that the presence of bacterial species other than
S. mutans
in smooth-surface dental plaque is due in part to contact of the cells with glucan in the developing plaque and not to the binding of cell-free GTase and the in situ synthesis of glucan. The results obtained with these representative strains of the seven serotypes of
S. mutans
may not apply to the same extent to other strains within the serotypes.
Scanning electron microscopy of experimental Trichophyton mentagrophytes infections in guinea pig skin
Authors: Hutton R D , Kerbs S , Yee K
Published: January 2020

Trichophyton mentagrophytes invasion of guinea pig skin was examined by scanning electron microscopy. Biopsies were obtained daily for 12 days from experimental infection sites. Dermatophyte invasion, examined in detail by scanning electron microscopy of cross-sectioned, prefixed skin was evidenced by: the appearance of hyphae within the stratum corneum; follicular invasion by hyphae, which remained initially within the follicle wall; emergence of the hyphae from the wall into the follicular canal; proliferation of the fungus down the follicle, with furrowing of the follicle wall and hair shaft cuticle; penetration of hyphae into the hair shaft by subcuticular and transcuticular routes; and massive peripilar hyphal proliferation with arthrosporogenesis. A three-dimensional perception of the invasion sequence of a dermatophyte in guinea pig skin was obtained by scanning electron microscopy.
Intraintestinal toxin in infant mice challenged intragastrically with Clostridium botulinum spores
Authors: Sugiyama H , Mills D C
Published: January 2020

Conventionally raised suckling mice were injected intragastrically with 10(5) spores of a Clostridium botulinum type A culture. Botulism was not observed, but 80% or more of mice challenged when 8 to 11 days old had botulinum toxin in the large intestine 3 days later. Mice younger than 7 days or older than 15 days were resistant to the challenge. When in vivo toxin production was started by spores given to 9-day-old mice, toxin was present in the intestine at 1 through 7 days postchallenge but with greatest consistency between 1 and 4 days. Total toxin in an intestine ranged up to 1,920 50% lethal doses as titrated intraperitoneally in adult mice. The dose infecting 50% of a group of 9-day-old mice was 700 (95% confidence limits of 170 to 3,000) spores per animal. Toxin was formed in the lumen of the large intestine; it was not associated with the ileum. Injection of 10(5) spores intraperitoneally into 9-day-old mice resulted in toxin production in the large intestines of 30% of the test animals.
Polymorphonuclear neutrophil chemotaxis under aerobic and anaerobic conditions
Authors: Casciato D A , Goldberg L S , Bluestone R
Published: January 2020

The motility of human polymorphonuclear neutrophils was studied in vitro under aerobic and anaerobic conditions. Chemotactic factors were generated from plasma with immune complexes or with whole bacteria (Staphylococcus aureus, Escherichia coli, and Bacteroides fragilis). Chemotaxis induced by chemotactic factors generated from immune complexes was identical under both conditions. However, chemotaxis utilizing chemotactic factors generated from bacteria was markedly depressed under anaerobic conditions. Mean random tubemoltility was not significantly different under aerobic and anaerobic conditions. These data indicate that different metabolic pathways may be involved in polymorphonuclear neutrophil movement. Some of these pathways require oxygen (chemotaxis in response to factors generated by bacteria in plasma), whereas others do not (random tube migration and chemotaxis in response to factors generated by immune complexes in plasma). These observations may be important in the induction of inflammatory responses within hypoxic tissues.
Role for Activated Macrophages in Resistance Against
Trichinella spiralis
Authors: Wing Edward J. , Remington Jack S.
Published: January 2020

To determine whether activated macrophages are important in resistance against the intestinal phase of nematode parasites, we studied
Trichinella spiralis
infections in mice with normal macrophages and in mice with macrophages activated by either chronic
Toxoplasma gondii
or acute
Listeria monocytogenes
infections. The peak
T. spiralis
adult worm burden in the intestines of normal C57BL/6 or Swiss Webster mice occurred from 6 to 14 days after infection. Subsequent expulsion of worms from the intestines occurred from 8 to 20 days after infection. C57BL/6 mice chronically infected with
T. gondii
and then challenged with
T. spiralis
larvae had significantly lower peak intestinal worm burdens (
P
< 0.05) than normal C57BL/6 mice similarly challenged. Swiss Webster mice infected 7 or 13 days earlier with
L. monocytogenes
and then challenged with
T. spiralis
larvae had significantly lower peak worm burdens (
P
< 0.01) than uninfected mice. The time of expulsion of adult worms was not affected by either infection. Swiss Webster mice infected 42 days earlier with
L. monocytogenes
(i.e., possessing lymphocytes sensitized to
L. monocytogenes
but not possessing activated macrophages) did not have a lower worm burden than uninfected mice. Serum factors (e.g., antibody) did not appear to play a role because normal mice injected with serum from
L. monocytogenes
-infected mice had worm burdens similar to those of mice injected with normal serum. The histopathology of intestines of mice infected with
T. gondii
or
L. monocytogenes
was the same as that of normal mice. When
T. spiralis
larvae were incubated with normal macrophages or macrophages from
T. spiralis
-infected mice in vitro for 24 h, the number of larvae with adherent
T. spiralis
macrophages was significantly (
P
< 0.005) greater than the number of larvae with adherent normal macrophages. These studies suggest a role for activated macrophages in resistance to
T. spiralis.
Comparative Immunological Analysis of Host Plasma Proteins Bound to Bloodstream Forms of
Trypanosoma brucei
Subspecies
Authors: Diffley Peter
Published: January 2020

The presence, location, host specificity, identity, and quantity of rat plasma proteins bound to bloodstream forms of
Trypanosoma brucei
subsp.
brucei, T. brucei
subsp.
rhodesiense
, and
T. brucei
subsp.
gambiense
were determined by a quantitative indirect fluorescent-antibody method and gel immunoassays. Fluorescence differences between trypanosomes obtained from rats and mice and treated with antiserum to normal rat plasma indicated that most, if not all, of the bound plasma proteins were host specific. Removal of plasma proteins by trypsinization of parasites provided evidence for their attachment to the surface of the parasite. The accreted proteins were found to be host albumin, immunoglobulin G (IgG), and complement (C3). The same quantities of these three plasma proteins were present on
T. brucei
subspecies collected from normal rats at 2 days postinfection, during low or peak parasitemias, or from cortisone-treated rats. IgM could only be detected on parasites collected from normal rats at peak parasitemia. With the aid of rocket immunoelectrophoresis, host albumin and IgG were found to account for 0.2 and 0.05%, respectively, of the total soluble proteins of the bloodstream forms. It was concluded from this study that: (i) host plasma proteins were bound to parasites early in the infection, suggesting a mechanism of adaptation to the mammalian host; (ii) the surface-bound IgG was not the result of a specific immune response against the parasites but might be the cause of C3 attachment; (iii) among the bloodstream forms of the three
T. brucei
subspecies, there were no differences in amounts of surface-bound albumin, IgG, or C3. A comparison between the present data dealing with
T. brucei
subspecies, on the one hand, and the previously published results concerning
T. congolense
, on the other, revealed significant differences in the amounts of the host plasma proteins attached to these hemoflagellates.
Significance of noncapsular antigens in protection against experimental Haemophilus influenzae type b disease: cross-reactivity
Authors: Myerowitz R L , Norden C W , Demchak T A
Published: January 2020

A bacterial strain, tentatively identified as an Actinobacillus species, was found to asymptomatically colonize the pharynx of some rats and to bear cell wall antigens which cross-react with noncapsular antigens of Haemophilus influenzae type b (HIb). The cross-reacting antigens appeared to be a heterogeneous mixture with varying molecular size and charge. The antigenic moieties are probably carbohydrate in nature. Antisera raised with this strain had both immunochemical and biological (bactericidal, opsonizing, and protective against experimental infection) activity against HIb. These findings lend further evidence to the idea that noncapsular antigens are important in the induction of resistance to HIb disease. The findings also raise the possibility of using bacteria which cross-react with noncapsular antigens for immunization against HIb disease in humans through nasopharyngeal or enteric colonization.