bluuebunny/crossref_metadata_2025 huggingface dataset:
Rabbit polymorphonuclear leukocytes do not secrete endogenous pyrogens or interleukin 1 when stimulated by endotoxin, polyinosine:polycytosine, or muramyl dipeptide
Authors: Windle B E , Murphy P A , Cooperman S
Published: January 2020

Rabbit polymorphonuclear leukocytes were purified from rabbit blood by centrifugation on colloidal silica gradients followed by sedimentation in 4% Ficoll. The purified neutrophils had normal random motility, responded to chemotactic stimuli, phagocytosed zymosan particles, made superoxide, and phagocytosed and killed bacteria. However, they did not secret endogenous pyrogens either spontaneously or in response to stimulation with endotoxin, polyinosine:polycytosine, or muramyl dipeptide. Macrophages isolated on the same gradients secreted some pyrogen spontaneously and secreted considerably more in response to the same three stimuli. This evidence reinforces the idea that macrophages are the only source of endogenous pyrogens, and that pyrogens secreted by cell populations that are rich in neutrophils are to be attributed to the monocytes or macrophages that the cell populations contain.
Activation of macrophages by lymphokines: enhancement of phagosome-lysosome fusion and killing of Coccidioides immitis
Authors: Beaman L , Benjamini E , Pappagianis D
Published: January 2020

Previously, it was shown that arthroconidia of Coccidioides immitis appear to inhibit phagosome-lysosome fusion and survive within normal mouse peritoneal macrophages. However, when these macrophages are exposed to antigen-stimulated T lymphocytes from immune mice, activation occurs, leading to enhanced phagosome-lysosome fusion and killing of C. immitis. Results indicate that the activation of macrophages can be effected after incubation with soluble lymphocyte product(s) (lymphokines). The activation of macrophages results if the macrophages are exposed to the lymphokine before, but not after, infection. The results indicate that the lymphocyte population responsible for the elaboration of the lymphokine is phenotypically Lyt1+2- and that activation of macrophages by the lymphokine can occur across H-2 histocompatibility barriers.
Relationship between cecal population levels of indigenous bacteria and translocation to the mesenteric lymph nodes
Authors: Steffen E K , Berg R D
Published: January 2020

Translocation is defined as the passage of viable bacteria from the gastrointestinal tract to the mesenteric lymph nodes (MLN) and other organs. The extent of translocation of certain indigenous, oxygen-tolerant bacteria from the cecum to the MLN, spleen, liver, kidney, and peritoneal cavity were determined in diassociated or triassociated gnotobiotic mice. Minimal bacterial translocation occurred to the spleen, liver, kidney, or peritoneal cavity. However, most bacterial strains readily translocated to the MLN. The percentage of the total population of each bacterial strain in the ceca was compared with the percentage of the total population of that strain in the MLN. There was a direct relationship between the numbers of a particular bacterial strain populating the ceca of diassociated or triassociated mice and the numbers of viable bacteria of this strain present in the MLN. Thus, the cecal population level of a particular bacterial strain determined the numbers of viable bacteria of this strain translocating to the MLN. The translocation of these bacterial strains from the gastrointestinal tract is an important first step in the pathogenesis of infection caused by members of the normal intestinal microflora.
Enhancement of mannose-mediated stimulation of human granulocytes by type 1 fimbriae aggregated with antibodies on Escherichia coli surfaces
Authors: Perry A , Ofek I , Silverblatt F J
Published: January 2020

In the present study, we assayed protein iodination in human granulocytes after interaction of the cells with mannose-specific (MS) type 1 fimbriated (MS+) and nonfimbriated (MS-) phenotypes of Escherichia coli pretreated with various amounts of anti-E. coli and antifimbrial antibodies. The MS+ phenotype stimulated protein iodination in granulocytes and possessed potent MS activity as measured by yeast aggregometry. In contrast, the MS- phenotype lacked all these activities. MS+ pretreated with moderate concentrations of antibodies, however, showed up to a 15-fold increase in granulocyte stimulation as compared to granulocyte stimulation induced by the non-antibody-treated MS+ phenotype or by the antibody-treated MS- phenotype. This marked antibody-mediated increase in stimulation of granulocytes was (i) dependent on the antibody concentrations, (ii) markedly reduced by methyl-alpha-D-mannoside, (iii) caused by immunoglobulin G as well as by F(ab')2, (iv) caused only by antifimbrial antibodies, (v) associated with cross-linked fimbriae seen as "bundles" under an electron microscope, and (vi) mimicked by treating MS+ bacteria with a certain range of glutaraldehyde. The data taken together support the hypothesis that, although cross-linking of fimbriae reduced the density of functional MS fimbriae over the surface of antibody-treated bacteria and consequently reduced the ability of these organisms to agglutinate yeast cells, the resulting bundles of MS fimbriae were far more effective at stimulating granulocytes because, bound together, they were better equipped to aggregate the mannose-containing receptors on the granulocyte surface.
Characterization of serological cross-reactivity between polysaccharide antigens of Streptococcus mutans serotypes c and d
Authors: Grossi S , Prakobphol A , Linzer R , Campbell L K , Knox K W
Published: January 2020

Immunological assays with antisera prepared against purified Streptococcus mutans serotype c polysaccharide demonstrated that a cross-reacting determinant on c polysaccharide reacted with the wall-associated rhamnose-glucose polysaccharide from S. mutans serotype d. Studies with 60 antisera prepared against chemostat cultures of S. mutans Ingbritt (c) demonstrated that the rhamnose-glucose polysaccharide cross-reactive determinant was consistently expressed on c antigen under a variety of growth conditions.
Polyclonal activation of B-lymphocytes in vivo by Salmonella typhimurium lipoprotein
Authors: Johnson R B , Köhl S , Bessler W G
Published: January 2020

Lipoprotein prepared from the outer membrane of Salmonella typhimurium is a polyclonal activator of murine B-lymphocytes. It was shown to be mitogenic for splenic cultures, stimulating increased incorporation of [3H]thymidine into DNA. When injected intravenously into mice, the lipoprotein induced splenomegaly and polyclonal B-cell activation. The latter was evident from an increase in the number of plaque-forming cells against trinitrophenylated sheep erythrocytes. Similar results were obtained with Escherichia coli lipoprotein.
Effect of the Pesticide Carbaryl on Replication of Human and Simian Varicella Viruses
Authors: Schmidt Nathalie J.
Published: January 2020

Pretreatment of host cells with carbaryl delayed the early spread of simian and human varicella virus infections. Toward the end of the growth cycle there was an apparent enhancement of infection, since treated cultures showed more infectious centers than did untreated ones in which infectivity had reached maximum levels and then declined.
Serological Cross-Reaction Between Lipids of
Mycoplasma pneumoniae
and
Mycoplasma neurolyticum
Authors: Kenny George E.
Published: January 2020

The complement-fixing activity of crude lipid extracts of 10
Mycoplasma
species was compared with that of whole organism antigens employing immune rabbit serum. Five species (
M. pneumoniae, M. neurolyticum, M. granularum, M. laidlawii
, and
M. fermentans
) showed serological activity, whereas the remaining five species (
M. canis, M. felis, M. gallisepticum, M. hyorhinis
, and
M. pulmonis
) did not show significant activity in their lipid fractions. The lipid fractions of the five species which had serological activity in their lipid fractions showed three groups of serological specificity.
M. pneumoniae
cross-reacted with
M. neurolyticum, M. granularum
cross-reacted with
M. laidlawii
, and
M. fermentans
showed specific activity. Acute and convalescent sera from human pneumonia patients from whom
M. pneumoniae
had been isolated showed antibody increases which could be measured nearly as well by lipids of
M. neurolyticum
as by those of
M. pneumoniae
. Only a few human convalescent sera showed antibody measureable by lipids of
M. granularum, M. pneumoniae
did not cross-react with
M. neurolyticum
by other serological parameters such as growth inhibition on agar or double immunodiffusion, indicating that only the lipid antigens of these two species cross-react.
Reduced Germination of
Clostridium botulinum
Type A Spores In Vitro by Polymorphonuclear Leukocytes from Chronic Granulomatous Disease
Authors: Suzuki Jon B. , Grecz Nicholas , Windhorst Dorothy
Published: January 2020

Leukocytes from two patients with chronic granulomatous disease of childhood (CGD) and from one female carrier were examined for their capacity to induce germination of
Clostridium botulinum
type A spores in vitro. Normal human leukocytes induced germination of
C. botulinum
spores to the same extent as guinea pig neutrophils. Germination was depressed by more than half when cells from CGD patients were used. A noticeable, but less severe, abnormality was present in leukocytes from a female carrier for this X-linked trait. CGD leukocytes are defective in cellular production of H
2
O
2
, and the latter is known to be effective in inducing germination of clostridial spores. Lysozyme is also known to be effective in spore germination and is present in CGD leukocytes in normal amounts. The posibility that
C. botulinum
spores require a mechanism which sensitizes them to the action of H
2
O
2
and lysozyme is raised, and it is suggested that the methodology used in this work measures a defect of such a mechanism in CGD leukocytes.
Epidemiology of a Herpesvirus of Turkeys: Possible Sources and Spread of Infection in Turkey Flocks
Authors: Witter R. L. , Solomon J. J.
Published: January 2020

Parameters of turkey herpesvirus (HVT) infection were followed sequentially from hatching through 20 weeks in two commercial turkey flocks. Maternal antibody was uniformly present at hatching but decreased below detectable levels by the 3rd week. Virus was first isolated at the 5th and 6th weeks and thereafter spread rapidly within the flocks. Development of active antibody followed that of viremia by 1 week. Both virus and antibody were present in virtually all turkeys after the 7th or 10th week; however, the incidence of tumors was negligible. Dust and litter from infected flocks were negative for HVT by poult bioassay, but the infection was easily transmitted by exposure of poults to dirty isolation cages or air from HVT-infected turkeys. Attempts to demonstrate congenital transmission of HVT by direct virus isolation from embryos of infected breeders or by isolation rearing procedures were unsuccessful.
Experimental Salmonellosis: Hypersensitivity to Cell Wall Lipopolysaccharide and Anti-Infectious Resistance of Mice Infected with
Salmonella
Authors: Kawakami Masaya , Hara Yoshihiko , Osawa Nobutaka
Published: January 2020

Mice infected with various strains of
Salmonella enteritidis
and
S. typhimurium
were found to be more sensitive to the cell wall lipopolysaccharide (LPS) extracted from certain strains of
Salmonella
than noninfected mice. This hypersensitivity was induced by those smooth or rough strains which possessed a polysaccharide chain longer than that of a glucoseless mutant. The major antigenic group participating in this hypersensitivity was presumed to be in rough core polysaccharide sequence because fractions containing either O side chain or LPS of a heptoseless mutant were ineffective in provoking a hypersensitivity reaction. Conditions for the induction and the phases of development of this hypersensitivity to LPS and for anti-infectious resistance were shown to be different. Present and previously obtained results suggest that the antigens participating in each of these two conditions were different.
Characterization of an Equine Infectious Anemia Antigen Extracted from Infected Horse Spleen Tissue
Authors: Norcross N. L. , Coggins L.
Published: January 2020

The spleens of horses infected with equine infectious anemia contain an antigen that is useful for a diagnostic immunodiffusion test. This antigen was extracted from the spleen by homogenization of the tissue, centrifugation, and precipitation from the supernatant fluid at 50% saturation with (NH
4
)
2
SO
4
. The antigen was purified by subjecting it to two cycles of electrophoresis in a continuous free-flow electrophoresis cell and finally filtering through a column of Sephadex G-200 gel. The antigen was found to be a small protein with a molecular weight of 27,500 and sedimentation coefficient of 2.1
S
. Its density was about 1.18 and its isoelectric point 5.8. At 45 to 50 C, it coagulated, losing its antigenicity. The antigen was useful for assaying antibody in the serum from infected horses by using the complement fixation test or the immunodiffusion test. Complement-fixing antibodies were found to be more transient than the precipitating antibodies.
Product of
Staphylococcus aureus
Responsible for the Scalded-Skin Syndrome
Authors: Kapral F. A. , Miller Margaret M.
Published: January 2020

Certain
Staphylococcus aureus
strains of phage group 2 produced a protein distinct from the alpha and delta toxins which was capable of causing generalized exfoliation in neonatal mice and presumably is responsible for the scalded-skin syndrome in humans. This protein, named “exfoliatin,” was purified and found to have a molecular weight of approximately 24,000. Exfoliatin was acid-labile, rather heat-stabile, and antigenic.
Compact-Type Growth of
Staphylococcus epidermidis
Strains in Serum-Soft Agar
Authors: Yoshida Kosaku , Smith Melvin R. , Naito Yoshiko
Published: January 2020

Of 120
Staphylococcus epidermidis
strains tested, 25.8% were of the compact type in serum-soft agar. Compact variants showed higher production of metabolites than counterpart diffuse variants.
Growth Advantage of Fast Lactose-Fermenting Mutants of Slow Lactose-Fermenting Coliform Bacteria in Gnotobiotic Rats and Mice
Authors: Wilkins Tracy D. , Franzese Jacqueline A.
Published: January 2020

When germ-free rats or mice were mono-associated with either
Escherichia coli lac
−
(ATCC 15939) or a slow lactose-fermenting coliform (
Paracolobactrum coliforme
ATCC 11605), fast lactose-fermenting mutants overgrew the parent organisms.
Development of specific and cross-reactive lymphocyte proliferative responses during chronic immunizing infections with Rickettsia tsutsugamushi
Authors: Jerrells T R , Osterman J V
Published: January 2020

The development of antigen-responsive lymphocytes was followed in mice immunized with the Gilliam, Karp, or Kato strains of Rickettsia tsutsugamushi by utilizing an in vitro lymphocyte proliferation assay. Subcutaneous immunization with viable rickettsiae of all three strains resulted in the appearance of lymphocytes in the spleen responding to irradiated tissue culture-grown rickettsiae used as stimulating antigens. Although all animals demonstrated antigen-induced proliferation elicited by homologous antigen by 14 days after immunization, the time of peak responsiveness varied, depending on the strain of rickettsiae used for immunization. In all cases, peak proliferative responses occurred at a time after immunization that was after the previously reported time after immunization at which resistance to rechallenge was observed. Reactivity to heterologous strains of R tsutsugamushi developed roughly in parallel with homologous reactivity in Karp- and Gilliam-immunized mice, with a marked degree of heterologous reactivity evident. Kato-immunized mice demonstrated greater reactivity to heterologous antigens early in the development of antigen reactivity and demonstrated a somewhat greater degree of cross-reactivity, relative to homologous responses, than the other groups. It was found that nylon wool-nonadherent immune cells, if cultured with antigen and adherent cells obtained from normal spleens or peritoneal exudates, responded in culture. The thymus-derived lymphocyte nature of the responding cell was further suggested when treatment of immune spleen cells with anti-Thy 1.2 serum and complement eliminated antigen response.
Isolation of Rhinovirus Intertypes Related to Either Rhinoviruses 12 and 78 or 36 and 58
Authors: Halfpap Laurel M. , Cooney Marion K.
Published: January 2020

Many antigenic relationships have been demonstrated among the 90 rhinovirus serotypes. Among these are reciprocal cross-reactions between serotypes 12 and 78 and between serotypes 36 and 58. Neutralizing-antibody titers to homologous virus of the related pairs are generally 16- to 64-fold higher than to the heterologous member, and neutralization by heterologous antiserum in the pools is not seen with prototype viruses. However, a number of isolates were encountered which gave anomolous results when tested with the antiserum pools in fetal tonsil cells. When these strains were tested in fetal tonsil cells against the monospecific antisera composing the pools, it was shown that several isolates were apparently intertypes, neutralized equally by antisera to related types 12 and 78 or 36 and 58. Isolate 1104, an apparent intertype between serotypes 36 and 58, and isolate 9433, intermediate between serotypes 12 and 78, were selected to use as immunogens in rabbits. When tested in HeLa cells, antiserum prepared against isolate 1104 neutralized isolates 1104, 58, and 36 at titers of 1280, 640, and 40, respectively. The
k
values against isolates 1104, 58, and 36 were 356, 145, and 4, respectively, indicating a much closer relationship of isolate 1104 to type 58 than to type 36. Similar results were obtained with isolate 9433. The neutralizing-antibody titer of anti-9433 serum was 160 against both 9433 and type 78 and was 20 against type 12. The
k
values of anti-9433 serum against 9433, 78, and 12 were 161, 111, and 2, respectively, indicating that 9433 and 78 were nearly identical. However, the respective neutralizing-antibody titers of anti-78 serum to type 78 and isolate 9433 were 640 and 80, and the respective
k
values were 172 and 85, demonstrating some antigenic differences. The discovery of intertypes confirms the antigenic variation among rhinoviruses, and the intertypes may represent links in the evolution of types. These observations also demonstrate that isolates in first or second passage in diploid cells may display an antigenic profile different from that seen in HeLa cells at high HeLa cell passage level.
Receptor-mediated entry of Pseudomonas toxin: methylamine blocks clustering step
Authors: Morris R E , Manhart M D , Saelinger C B
Published: January 2020

Clustering of ligands into coated regions of the plasma membrane is an early step in receptor-mediated endocytosis. The association of Pseudomonas exotoxin A (PE) with mouse LM fibroblasts was visualized by using biotinyl-PE and avidingold. Movement of PE into coated regions occurred within 30 s of warming monolayers to 37 degrees C. This clustering was stopped by the primary amines methylamine and ammonium chloride but was not altered by the tertiary amine chloroquine. Toxin internalization was rapid, with a half-time of approximately 5 min. Although primary amines stopped clustering, they did not alter the rate of toxin internalization; they did alter the route followed after entry. We have shown previously that methylamine protects cells from the lethal action of PE. Here we suggest that methylamine protects, at least in part, by blocking clustering, and that receptor-mediated endocytosis is required for efficient expression of PE toxicity.
Genetic Mapping of the
tox-1000
Locus of
Vibrio cholerae
El Tor Strain RJ1
Authors: Saunders David W. , Bramucci Michael G.
Published: January 2020

The results of a genetic cross between a
Vibrio cholerae
RJ1 donor and a
V. cholerae
3083-2 recipient suggest that the map position of
tox-1000
is between
his
and
trp
.
Comparison of the immunogenicity of vaccines prepared from viable Mycobacterium bovis BCG, heat-killed Mycobacterium leprae, and a mixture of the two for normal and M. leprae-tolerant mice
Authors: Shepard C C , van Landingham R M , Walker L L , Ye S Z
Published: January 2020

Intradermal vaccines consisting of viable Mycobacterium bovis BCG, heat-killed Mycobacterium leprae, or mixtures of the two were titrated in mice in doses of 10(5.2), 10(5.8), 10(6.4), 10(7.0), and 10(7.6) acid-fast bacilli. The immune response was measured by sensitization (48 to 72 h foot pad enlargement on challenge with 10(7.0) heat-killed M. leprae) and by protection against infection with a viable M. leprae challenge. There was increasing response with increasing dose of vaccine, and overall the responses to the three vaccines were similar. At the lowest dose, however, the combination of BCG and M. leprae gave superior protection. The local reaction to the vaccines in the lower dose range was less severe with the M. leprae vaccine. In another experiment, the three vaccines were compared in normal mice and in mice that had been rendered tolerant by intravenous injection of M. leprae. The tolerant mice developed no measurable sensitization on vaccination with M. leprae, but they developed partial but distinct sensitization on vaccination with BCG, alone or in combination with M. leprae. The tolerant mice developed little or no protection with any of the vaccines, however.
Polyclonal activation of human peripheral blood B lymphocytes by Fusobacterium nucleatum
Authors: Mangan D F , Lopatin D E
Published: January 2020

The present study examined in vitro polyclonal human B-lymphocyte (B-cell) activation (PBA) by Fusobacterium nucleatum. Pokeweed mitogen, a well-studied PBA activator, was included in some experiments for comparison. PBA was determined by the total immunoglobulin A, G, and M concentrations in the culture supernatants as measured by micro-enzyme-linked immunosorbent assay. F. nucleatum, at concentrations between 1 and 10 micrograms/ml, stimulated optimal PBA in monocyte-depleted cultures, whereas the pokeweed mitogen response was optimal in unfractionated, monocyte-containing cultures. Immunoglobulin synthesis occurred primarily between days 6 and 8 after stimulation with F. nucleatum. T lymphocytes enhanced the PBA response to F. nucleatum, particularly at a T- to B-cell ratio of 1:1. Immunoglobulin production was greater in round-bottomed wells than in flat-bottomed wells at lymphocyte concentrations of 200,000 cells per well. The PBA response, however, increased dramatically in flat-bottomed wells containing higher lymphocyte concentrations, suggesting that PBA is enhanced by cell-to-cell contact. A delay in stimulation of the lymphocytes with F. nucleatum resulted in diminished immunoglobulin production. The results provide information on the regulation of in vitro PBA induced by F. nucleatum. The data also suggest that there may be differences in the mechanisms by which F. nucleatum and pokeweed mitogen stimulate PBA.
Effect of splenectomy on Trypanosoma lewisi infection in young rats
Authors: Llende M , Santiago-Delpín E A , Lavergne J A
Published: January 2020

The effect of splenectomy on animals infected with Trypanosoma lewisi is unclear, and previous reports are inconclusive or conflictive. We splenectomized rats of different ages after they had been infected with T. lewisi. Female Sprague-Dawley rats, Hemobartonella-free, were assigned to four groups according to weight: 80, 108, 140, and 170 g. Each group had splenectomized, sham-operated, and nonoperated control subgroups, all infected with T. lewisi (0.5 ml of 10(6) parasites per ml) 90 h before surgery. Before surgery, parasite levels in host blood were similar. At 24 h after splenectomy in all groups, regardless of weight, blood parasite levels were much higher than they were in sham-operated or control animals (P less than 0.001 to P less than 0.0001; analysis of variance). Younger rats (80 and 108 g) had a higher mortality rate after splenectomy than sham-operated and control animals. Older rats (150 and 170 g) had no mortality. These results show the impact of age and the importance of the spleen on parasite-host interactions in rats infected with T. lewisi.
Protection against human and porcine enterotoxigenic strains of Escherichia coli in rats immunized with a cross-linked toxoid vaccine
Authors: Klipstein F A , Engert R F , Clements J D , Houghten R A
Published: January 2020

To compare their relative immunogenicities, we used synthetically produced Escherichia coli heat-stable toxin coupled to a protein carrier and the B subunit of porcine heat-labile toxin separately in graded dosages to immunize rats. Equivalent antigen unit dosages of each toxin raised approximately the same level of mucosal immunoglobulin A (IgA) antitoxin response and degree of protection against a challenge with respective heat-stable- or heat-labile-toxin-producing viable bacteria. Conjugation conditions were identified, therefore, which yielded a vaccine of these toxins, cross-linked by the carbodiimide reaction, that consisted of equal antigenic proportions of each toxin component as determined by enzyme-linked immunosorbent assay and expressed in antigen units. The dose-related response to immunization with this vaccine was the same as the response to its components given separately. The toxicity of the heat-stable toxin component was reduced greater than 600-fold. Immunization with optimal antigen unit dosages of the vaccine gave greater than or equal to sixfold increases in mucosal IgA antitoxin titers and provided significant (P less than 0.001) protection against challenge with heterologous serotypes of viable strains, of either human or porcine origin, that produce heat-stable or heat-labile toxin or both.
Serological comparison of canine rotavirus with various simian and human rotaviruses by plaque reduction neutralization and hemagglutination inhibition tests
Authors: Hoshino Y , Wyatt R G , Greenberg H B , Kalica A R , Flores J , Kapikian A Z
Published: January 2020

By the plaque reduction neutralization test, the CU-1 strain of canine rotavirus was similar, if not identical, to three strains (no. 14, no. 15, and P) of the tentatively designated third human rotavirus serotype. In addition, strain CU-1 demonstrated a one-way antigenic relationship with two other strains (M and B) of the third human rotavirus serotype. The CU-1 strain of canine rotavirus hemagglutinated human group O, rhesus monkey, dog, sheep, and guinea pig erythrocytes. A two-way antigenic relationship between canine (CU-1) and simian (MMU 18006 and SA11) rotaviruses demonstrated previously by the plaque reduction neutralization test was confirmed further with two additional isolates (A79-10 and LSU 79C-36) of canine rotavirus by the plaque reduction neutralization test and the hemagglutination inhibition test. The CU-1 strain of canine rotavirus, which is known to be distinct from two well-characterized human rotavirus serotypes (Wa and DS-1), was also found to be distinct from the St. Thomas no. 4 strain, which is a newly defined fourth human rotavirus serotype. Thus, this canine strain, which is related antigenically to one of four human rotavirus serotypes, is another example of an animal rotavirus which shares serotype specificity with a human rotavirus.
Adherence of mucoid and nonmucoid Pseudomonas aeruginosa to acid-injured tracheal epithelium
Authors: Ramphal R , Pyle M
Published: January 2020

A model of the adherence of Pseudomonas aeruginosa to injured lower respiratory tract cells is described. Mouse tracheas were injured by exposure to 0.1 N HCl for 15 min, cut into pieces of 2 to 4 rings, and placed in petri dishes for adherence studies. Adherence was quantitated by direct count of the number of bacteria attached to a fixed surface area as viewed by scanning electron microscopy. Optimal conditions for study were 1 h of incubation with an inoculum of 10(7) to 10(8) CFU/ml. Both mucoid and nonmucoid P. aeruginosa adhered to the injured cells in this system, and no major differences in adherence between these strains was apparent. Adherence to injured tracheal cells was a phenomenon limited to P. aeruginosa and did not occur with Escherichia coli and Klebsiella pneumoniae. Besides adherence to injured cells, this model allowed the demonstration of adherence of mucoid P. aeruginosa and K. pneumoniae to mucin strands of uninjured control tracheas. This model is an alternative to the buccal cell model and has the advantage of allowing the study of the adherence of both mucoid and nonmucoid strains of P. aeruginosa.
Angiotensin-1-converting enzyme activity of murine macrophages isolated from granulomas elicited by eggs of Schistosoma mansoni
Authors: Krulewitz A H , Stadecker M J , Wright J A , Fanburg B L
Published: January 2020

Angiotensin-1-converting enzyme (A1CE) activity was assessed in macrophages isolated from hepatic egg granulomas at 7 and 20 weeks after the infection of mice with Schistosoma mansoni. At 2 h after isolation, granuloma macrophages (GM) from chronically infected mice (20 weeks) displayed significantly higher levels of A1CE than did GM from mice 7 weeks after infection; both of the values were significantly higher than control peritoneal resident macrophages from normal mice. A1CE activity in GM and their respective culture supernatants declined over a 72-h period of in vitro cultivation; however, the decline was abrogated with the addition of dexamethasone to the culture medium. GM from athymic mice displayed significantly higher A1CE activity than did those from normal controls. Our data confirm and extend previous findings of others that GM are a major source of A1CE in murine schistosomiasis and that lymphocyte function may regulate GM A1CE production.
Correlation between the presence of a fifty-megadalton plasmid in Salmonella dublin and virulence for mice
Authors: Terakado N , Sekizaki T , Hashimoto K , Naitoh S
Published: January 2020

All of the Salmonella dublin organisms isolated from cattle in Japan (20 strains) and in Europe (2 strains) harbored one plasmid of approximately 50 megadaltons (Mdal). The biochemical reactions of a derivative strain (5230P-) cured of the 50-Mdal plasmid were those of its parental strain 5230P+. However, the strain 5230P- proved to be 100- to 1,000-fold less virulent for mice compared with that of the strain 5230P+, suggesting that the virulent phenotypes of S. dublin may be mediated by the 50-Mdal plasmid.
Hepatitis B vaccine: further studies in children with previously acquired hepatitis B surface antigenemia
Authors: Barin F , Yvonnet B , Goudeau A , Coursaget P , Chiron J P , Denis F , Mar I D
Published: January 2020

Three doses of inactivated hepatitis B vaccine were given at 1-month intervals to 31 hepatitis B surface antigen (HBsAg)-positive Senegalese children aged between 3 and 24 months. A control group of 18 HBsAg-positive Senegalese children received diphtheria-tetanus-polio vaccine. Immunization of HBsAg-positive infants with hepatitis B vaccine was safe but inefficient. After a 12-month follow-up, the prevalence of HBsAg chronic carriers was not significantly reduced in the hepatitis B vaccine group as compared with the control group: 48.4 and 66.7%, respectively. The presence of hepatitis B antigen was found to be a major risk factor for HBsAg-positive children to develop a chronic carrier state. The risk of developing an HBsAg chronic carrier state was also related to advancing age at time of enrollment in the study.
Isolation and characterization of a plasmid from phase I Coxiella burnetii
Authors: Samuel J E , Frazier M E , Kahn M L , Thomashow L S , Mallavia L P
Published: January 2020

The DNA from the Nine Mile phase I strain of Coxiella burnetti, the etiological agent of Q fever, has been isolated and purified by cesium chloride-ethidium bromide density gradient centrifugation. A fraction of this DNA has a density characteristic of plasmid DNA. The plasmid DNA was cut with 20 different restriction endonucleases and shown to be a discrete entity. The plasmid, designated QpH1, is approximately 36 kilobases in size and has a molecular mass of 2.4 x 10(7) daltons. A partial restriction map of QpH1 has been constructed by using the restriction endonucleases SalI, KpnI, PstI, and XbaI. QpH1 DNA radioactively labeled by nick translation was used to show that sequences similar to the plasmid are also present in the phase II antigenic variant of C. burnetii.
Recombinant mouse interferon-gamma regulation of antibody production
Authors: Johnson H M , Torres B A
Published: January 2020

Interferon-gamma produced in monkey cells by transfection with mouse interferon-gamma cDNA suppressed the mouse in vitro antibody response in a manner similar to that of natural mouse interferon-gamma. Significant suppression was obtained with as little as 1 U of interferon. Recombinant human interferon-gamma produced by cloning in a similar fashion was not suppressive. Both the suppressive and the antiviral activities of recombinant interferon-gamma were neutralized by antibodies to mouse natural interferon-gamma. Thus, interferon-gamma was responsible for the immunosuppression. At the cellular level, the recombinant interferon-gamma was capable of activating macrophages to suppress antibody production. The data provide clear-cut evidence that interferon-gamma plays an important role in regulation of immunological processes.
Immunological evaluation of meningococcal group C polysaccharide-tetanus toxoid conjugate in mice
Authors: Beuvery E C , van Delft R W , Miedema F , Kanhai V , Nagel J
Published: January 2020

Neisseria meningitidis group C polysaccharide-tetanus toxoid conjugate was prepared to obtain the polysaccharide component in a thymus-dependent form and to preserve the immunogenic properties of the tetanus toxoid component. Biochemical and immunochemical analyses of this conjugate revealed that (i) it was composed of equal amounts of polysaccharide and protein; (ii) the antigenic activity of the polysaccharide component was greatly reduced; (iii) it contained about 10% free polysaccharide; (iv) the composition was not homogeneous; and (v) only 5% of the tetanus toxoid component was present at the surface of the conjugate molecules. In this study, the influence of these characteristics on the antibody response to both components in mice was investigated. The dose-response relationship, the persistence of antibodies, a possible antigenic competition, and the specificities of the antibodies induced were also studied. Our data suggest that the conjugate behaves as a pronounced thymus-dependent antigen, that the tetanus toxoid component is more immunogenic at lower dosages (0.8 and 20 ng) than equivalent doses of tetanus vaccine, that the presence of free polysaccharide does not influence the induction of antibodies to polysaccharide by the conjugate, and that no antibodies to new structures in the conjugate are induced. These characteristics favor the application of this conjugate as a vaccine for human use.
In vitro and in vivo properties of chemically induced temperature-sensitive mutants of Chlamydia psittaci var. ovis: screening in a murine model
Authors: Rodolakis A
Published: January 2020

Two temperature-sensitive (ts) strains, 1B and 1H, were obtained after nitrosoguanidine treatment of the wild-type virulent abortive ovine strain Chlamydia psittaci AB7. Optimum growth temperature on McCoy cells was 38 degrees C for the three strains, but at the restrictive temperature, 39.5 degrees C, ts strains differed from the parental strain in their total infective yield of chlamydiae, their efficiency of plating, and the morphology of the plaques and cytoplasmic inclusions. Their survival at 51 degrees C was also reduced compared with that of the wild field strains. The virulence of the two ts strains was attenuated for pregnant mice, but these two ts strains were able to multiply in mice and to induce a strong immunity to virulent challenge with either the parental AB7 strain or the caprine or bovine abortive field strains.
Classical complement pathway activation by antipneumococcal antibodies leads to covalent binding of C3b to antibody molecules
Authors: Brown E J , Berger M , Joiner K A , Frank M M
Published: January 2020

We have examined whether or not a physical relationship exists between antipneumococcal antibodies (Ab) and C3b when Ab activate the classical complement pathway on the surface of pneumococci (Pn). After sensitization with 125I-labeled Ab, Pn were sequentially incubated with purified C1, C4, C2, and biotinylated C3. Ab molecules were then eluted from Pn, and C3b-associated molecules were purified on avidin-Sepharose. Both 125I-labeled immunoglobulin G (IgG) and [125I]IgM bound to C3b; the association was stable to incubation in 1% sodium dodecyl sulfate at 37 degrees C. The association was only partially reversed by incubation in 1 M hydroxylamine-0.5% sodium dodecyl sulfate (pH 10.5), implying that Ab and C3b were linked by amide as well as ester bonds. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of dithiothreitol and NH2OH eluates from the avidin-Sepharose showed that C3b bound to both heavy and light chains of the Ab. Moreover, the ability to bind to erythrocyte C3b receptors could be transferred to unsensitized Pn by eluates from Pn on which Ab had activated the classical pathway. These covalent complexes of Ab and C3b may be especially important in the opsonization of Pn by Ab and complement.
Protection of calves against fatal enteric colibacillosis by orally administered Escherichia coli K99-specific monoclonal antibody
Authors: Sherman D M , Acres S D , Sadowski P L , Springer J A , Bray B , Raybould T J , Muscoplat C C
Published: January 2020

A monoclonal antibody (MCA) to enterotoxigenic Escherichia coli K99 antigen agglutinated K99+ enterotoxigenic E. coli strains B44 (O9:K30;K99;F41:H-) and B41 (O101:K99;F41:H-) grown at 37 degrees C but not at 18 degrees C. The MCA, which was characterized as immunoglobulin G1, reacted specifically with K99 antigen in an enzyme-linked immunosorbent assay and precipitated radiolabeled K99 antigen. A total of 45 colostrum-fed and colostrum-deprived calves were used in three separate trials to determine whether the orally administered K99-specific MCA would prevent diarrhea caused by strain B44. Twenty-eight calves were fed 1 ml of mouse ascitic fluid containing K99-specific MCA at 10 h of age and were orally challenged with strain B44 at 12 to 14 h of age. Control calves either received no placebo or were fed 1 ml of mouse ascitic fluid containing fibronectin-specific MCA at 10 h of age. There was no difference in the incidence of diarrhea between the two groups after challenge. However, the severity of diarrhea, as evaluated by the proportion of calves in each group that developed severe dehydration, the degree of clinical dehydration, the degree of clinical depression, the degree of weight loss, and the duration of diarrhea after challenge was significantly reduced in calves that received the K99-specific MCA. The mortality rate was also significantly lower (P less than 0.001) in the treated (29%) than in the control (82%) group. These results suggest that orally administered K99-specific MCA can prevent severe fatal enteric colibacillosis.
Distribution of immunoglobulin-containing cells in normal and neonatally thymectomized rats
Authors: Ebersole J L , Taubman M A , Smith D J
Published: January 2020

The distribution of immunoglobulin-containing cells (ICC) of the immunoglobulin A (IgA), IgG, and IgM isotypes was examined in various lymphoid and secretory tissues of rats. The effect of neonatal thymectomy of rats on T cells, B cells, and ICC in these tissues was determined by immunofluorescence. The results showed that although T cells were severely depleted in both lymphoid and secretory tissues of the thymectomized (Tx) rats, Tx and normal rats showed comparable numbers of B cells staining for IgA, IgG, and IgM. After neonatal thymectomy, IgA ICC in both lymphoid and secretory tissues were significantly decreased. However, the Tx rats exhibited a compensatory increase in IgM ICC in the identical tissues. Local injection of normal and Tx rats with Streptococcus mutans 6715 resulted in an increase in all isotypes of ICC in the secretory tissues. Although the primary increase in normal rats was due to IgA ICC, Tx rats exhibited the greatest change in the number of IgM ICC.
Malaria-induced lymphokines: stimulation of macrophages for enhanced phagocytosis
Authors: Ockenhouse C F , Shear H L
Published: January 2020

Culture supernatants from antigen-pulsed spleen cells of mice infected previously with either BCG or Plasmodium chabaudi were used to study macrophage activation as judged by phagocytosis of immunoglobulin G-sensitized erythrocytes and Plasmodium berghei- and P. chabaudi-infected erythrocytes. Resident peritoneal macrophages were incubated in vitro with spleen cell factor and then assayed for ingestion of immunoglobulin G-sensitized or parasitized erythrocytes. Macrophages activated with BCG-induced lymphokine bound and ingested two- to threefold more P. berghei parasitized erythrocytes than macrophages incubated with control spleen cell factor. Similarly, Plasmodium-stimulated spleen cells from mice infected with malaria produced a lymphokine(s) capable of activating macrophages for enhanced Fc receptor-mediated phagocytosis. The stimulation of phagocytosis by the lymphokine is nonspecific in nature, since phagocytosis of parasitized erythrocytes from one species of murine malaria is enhanced by the lymphokine prepared from a heterologous species. Nylon wool-nonadherent, malaria-sensitized spleen cells elaborated a lymphokine which stimulates macrophages for enhanced phagocytosis, whereas anti-0-treated spleen cells failed to produce the phagocytosis-promoting lymphokine. Consequently, this lymphokine appears to be elaborated by sensitized T lymphocytes. Interestingly, enhanced phagocytosis of opsonized trophozoites and schizonts, but not ring stage parasites of P. chabaudi, was displayed by macrophages activated with the lymphokine(s) prepared from P. chabaudi-recovered mice. Preincubation of the malaria parasitized erythrocytes with hyperimmune serum raised against the parasites greatly facilitated both binding and ingestion by the stimulated macrophages.
Effect of Toxoplasma infection on the sensitivity of mouse thymocytes to natural killer cells
Authors: Kamiyama T , Tatsumi M
Published: January 2020

Thymocytes from mice 2 weeks after infection with Toxoplasma gondii resisted natural killer (NK) cell-mediated cytolysis in contrast to the high sensitivity of normal mouse thymocytes. The infected mouse thymocytes also failed to form conjugates with effector cells and to compete for cytolysis of NK sensitive targets. These effects were mediated, at least in part, by interferon-gamma because normal thymocytes became NK insensitive after incubation in the infected mouse serum which contained significant amount of interferon-gamma, and pH 2 treatment of the serum abolished the effect. An alternate possibility for the reduced NK sensitivity of the infected mouse thymocytes was the elimination of NK-sensitive cells from the thymus, since histopathological studies showed marked atrophy and clearance of NK-sensitive thymocytes in the cortex of thymuses of infected mice. Although T. gondii induced augmentation followed by suppression of the host splenic NK activity, it seems unlikely that this altered NK activity was responsible for the lowered NK sensitivity of the thymocytes.
Intracellular location of Mycobacterium leprae in macrophages of normal and immune-deficient mice and effect of rifampin
Authors: Mor N
Published: January 2020

Soon after more than 10(6) Mycobacterium leprae, freshly harvested from armadillo liver or harvested and 60CO irradiated, were inoculated into the hind footpads of either normal or thymectomized and irradiated (T900R) mice, the organisms were found to reside within phagosomes of polymorphonuclear and mononuclear cells. On the other hand, 7 and 8 months after 10(4) freshly harvested M. leprae were inoculated into the footpads of normal or T900R mice and the organisms had multiplied to their maximum in the normal mice, many organisms, largely intact by electron-microscopic criteria, were found to reside free in the cytoplasm of the footpad macrophages, whereas damaged organisms were contained within phagosomes. After 11 months, many intact organisms were found to lie free in the cytoplasm of the macrophages of T900R mice, whereas only damaged intraphagosomal M. leprae cells were observed in the macrophages of normal mice. Finally, a remarkably large proportion of damaged extraphagosomal M. leprae was found in T900R mice administered rifampin for 2 days in a bactericidal dosage. It appears that M. leprae multiplies free in the cytoplasm of the footpad macrophages of infected mice, whereas the M. leprae cells resident within the phagosomes of the macrophages are dead. As the result of treatment with rifampin, the organisms appeared to have been killed in their extraphagosomal location, only afterwards being incorporated into phagosomes. However, the intracellular site in which M. leprae is killed in the course of an effective immune response remains unclear.
Immunoglobulin A-mediated hepatobiliary transport constitutes a natural pathway for disposing of bacterial antigens
Authors: Russell M W , Brown T A , Claflin J L , Schroer K , Mestecky J
Published: January 2020

The possibility that hepatobiliary transport of immunoglobulin A (IgA) immune complexes might eliminate bacterial antigens was investigated in mice with pneumococcal type III capsular polysaccharide and C carbohydrate and corresponding monoclonal antibodies. Although all isotypes of antibody caused uptake by the liver, only IgA, but not IgG or IgM, antibodies transported these substances into bile where they were detected in the form of immune complexes and as free antigens. Small doses (10 micrograms or less) of passively administered IgA antibody were sufficient to induce measurable transport of capsular polysaccharide into bile. Transport of C carbohydrate was significantly correlated with the level of naturally occurring IgA antibodies specific for the phosphocholine determinant, but not with IgM or IgG antibodies. These results suggest that the continual process of hepatic uptake of circulating polymeric IgA may function to eliminate bacterial macromolecular products that are not readily susceptible to other mechanisms of disposal.
Morphological changes and pathology of mouse glomeruli infected with a streptococcal L-form or exposed to lipoteichoic acid
Authors: Tomlinson K , Leon O , Panos C
Published: January 2020

The morphology and pathology of cultured mouse glomeruli were examined at the cellular and subcellular levels after infection with a physiological isotonic L-form of Streptococcus pyogenes type 12 or exposure to streptococcal lipoteichoic acid. These changes, as viewed by light microscopy, were identical regardless of the method used to induce glomerular cytotoxicity. They were characterized by an initial reduction in the outgrowth of cells, some cellular granulation, and later, destruction of the confluent monolayer. Once initiated, cytotoxicity could not be reversed by refeedings, and complete glomerular destruction resulted after 2 weeks. Electron microscope studies revealed that the basement membrane of intact glomeruli exposed to streptococcal lipoteichoic acid had become greatly thickened (two- to fourfold) and electron dense. Our recent biochemical findings have shown that streptococcal lipoteichoic acid increases the amount of collagen formed and retained by mouse fibroblasts in tissue culture as well as causing a reduction in the hydroxylation of proline in both intracellular and secreted collagenous material (Leon and Panos, Infect. Immun. 40:785-794, 1983). These results, together with the present findings, suggest that the thickening of the glomerular basement membrane may be due to defective collagen biosynthesis as a result of streptococcal lipoteichoic acid. The use of cultured glomeruli as a model system for studying the earliest basement membrane alterations in the absence of an immune response as a result of streptococcal lipoteichoic acid is suggested.