bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of monoclonal antibodies to detect antigens of Toxoplasma gondii in serum and other body fluids
Authors: Araujo F G , Handman E , Remington J S
Published: January 2020

Monoclonal antibodies to Toxoplasma gondii were used in an enzyme-linked immunosorbent assay to detect antigens of the parasite in toxoplasma lysate, in peritoneal fluid of mice, and in sera from humans acutely infected with T. gondii. Four of the six monoclonal antibodies were able to detect antigens of toxoplasma in these specimens. Control sera from individuals not infected with T. gondii and from individuals chronically infected with the parasite were negative in the enzyme-linked immunosorbent assay. Sera from individuals not infected with T. gondii but with positive titers from rheumatoid factor were also negative; 2 or 10 sera from individuals not infected with T. gondii but with positive titers for antinuclear antibodies reacted with the monoclonal antibodies. When the results of the enzyme-linked immunosorbent assay with monoclonal antibodies and with the F(ab)2 fraction of an immunoglobulin G from a rabbit infected with T. gondii were compared, it was noted that the F(ab)2 was more active in detecting parasite antigens than were the monoclonal antibodies. Thus, although monoclonal antibodies can be used to detect antigens of T. gondii in sera and other body fluids, polyvalent antibody (such as the F(ab)2 fraction) appears to be more satisfactory for this purpose.
T-cell-mediated cytotoxic response to mumps virus in humans
Authors: Tsutsumi H , Chiba Y , Abo W , Chiba S , Nakao T
Published: January 2020

Mumps-specific T-cell-mediated cytotoxic activity against virus-treated autologous lymphocytes was studied after peripheral blood lymphocytes of sensitized subjects had been incubated with ultraviolet light-inactivated virus antigen. Generation of the cytotoxic activity in vitro was associated with an antecedent lymphoproliferative response to mumps virus. The virus specificity of the effector cells was demonstrated by a lack of lysis of type 1 parainfluenza virus (HVJ)-treated as well as of type A influenza virus-treated autologous target cells. This activity was largely associated with E-rosette-forming T lymphocytes as revealed by negative selection of a population from cultured whole lymphocytes. In addition, sequential investigations for subjects with a natural mumps virus infection clearly demonstrated individual characteristics of the cytotoxic response. Therefore, the assay described could be used to reflect mumps virus-specific T-cell-mediated immunity in humans.
Formation of intraperitoneal abscesses by Staphylococcus aureus
Authors: Kapral F A , Godwin J R , Dye E S
Published: January 2020

Certain Staphylococcus aureus strains, when inoculated into the peritoneal cavity of mice, were clumped and surrounded by a thick layer of leukocytes. After being enclosed with a connective tissue capsule, the structures histologically resembled staphylococcal abscesses. Of four strains examined, all were destroyed within abscesses, although at different rates. Abscess homogenates possessed bactericidal activity toward staphylococci, and this activity was associated with the sedimentable fraction of the homogenates. Leukocytes did not appear to be responsible for the bactericidal activity. Appreciable quantities of alpha toxin accumulated in these abscesses even without multiplication of the organisms. This model infection offers opportunities for studying some aspects of staphylococcal host-parasite interactions occurring in localized lesions.
Specificity of immunoglobulin M antibodies in normal human serum that participate in opsonophagocytosis and intracellular killing of Bacteroides fragilis and Bacteroides thetaiotaomicron by by human polymorphonuclear leukocytes
Authors: Bjornson A B , Bjornson H S , Kitko B P
Published: January 2020

Studies were performed to determine the specificity of immunoglobulin M (IgM) antibodies in normal human serum that participate in opsonophagocytosis and intracellular killing of Bacteroides fragilis 1365 and Bacteroides thetaiotaomicron 1343 by human polymorphonuclear leukocytes. Purified normal human IgM was adsorbed with washed heat-killed cells of the homologous strains and heterologous strains of B. fragilis, B. thetaiotaomicron, Bacteroides vulgatus, Bacteroides distasonis, and Bacteroides asaccharolyticus and with erythrocytes coated with outer membrane complex prepared from the homologous strains. Hypogammaglobulinemic serum was supplemented with the adsorbed IgM preparations, and the ability of the supplemented sera to support opsonophagocytosis and killing of B. fragilis 1365 and B. thetaiotaomicron 1343 by human polymorphonuclear leukocytes was measured in vitro under anaerobic conditions. Normal IgM adsorbed with heat-killed cells of B. fragilis 1365 and B. thetaiotaomicron 1343 or with erythrocytes coated with outer membrane complex prepared from these strains failed to restore the ability of hypogammaglobulinemic serum to support opsonophagocytosis and intracellular killing of the homologous strain. In contrast, adsorption of normal IgM with heat-killed cells of the heterologous strains did not alter its opsonophagocytosis-promoting activity for either test strain. These results indicated that the IgM antibodies in normal human serum that participate in opsonophagocytosis and intracellular killing of B. fragilis 1365 and B. thetaiotaomicron 1343 are directed against strain-specific antigenic determinants contained in the outer membrane complex.
Interactions of human neutrophils with leukotoxic streptococci
Authors: Sullivan G W , Mandell G L
Published: January 2020

Most strains of Streptococcus pyogenes contain a toxin which can kill neutrophils. Previous workers failed to show any correlation between leukotoxin content and virulence of animals or humans. We examined the in vitro interactions of a leukotoxic streptococcus and a nonleukotoxic variant with human neutrophils. At ratios of 200 streptococcal colony-forming units per neutrophil, the toxic strain killed 92.8 +/- 2.0% of neutrophils, and the nontoxic strain killed only 9.0 +/- 1.2%. Despite this, ingestion of the two strains was equal. Postphagocytic oxidative metabolism was equivalent with low numbers of either toxic or nontoxic streptococci but depressed with high numbers of leukotoxic streptococci. At 20 min, neutrophils were able to kill leukotoxic (99.6 +/- 0.3% killed) and nonleukotoxic streptococci (99.5 +/- 0.2% killed) equally efficiently (P = 0.42). Thus, leukotoxicity does not interfere with the ability of neutrophils to destroy streptococci. This may explain why leukotoxicity does not appear to be an important factor in streptococcal virulence.
Mycoplasma-dependent activation of normal mouse lymphocytes: requirement for functional T lymphocytes in the cytotoxicity reaction mediated by Mycoplasma arthritidis
Authors: Cole B C , Aldridge K E , Sullivan G J , Ward J R
Published: January 2020

Syngeneic and allogeneic target cells were killed in the presence of CBA mouse lymphocytes and viable Mycoplasma arthritidis. Medium supplementation had no effect on the response. Nonviable M. arthritidis was also capable of stimulating lymphocytotoxicity, although to a much lesser extent. Cytotoxicity was shown to be largely dependent upon the lymphocytes, since lymphocytes preincubated with mycoplasmas and treated to remove remaining organisms were highly toxic to target cells, whereas supernatants prepared from lymphocyte/mycoplasma mixtures exhibited minimal effects. A 6-h exposure of lymphocytes to mycoplasmas at a ratio of 100:1 was sufficient for commitment to target cell killing. Functional lymphocytes were required for the reaction, since gamma-irradiated lymphocytes did not develop cytotoxic potential despite the fact that the mycoplasmas replicated equally well in the presence of these and untreated lymphocytes. Furthermore, lymphocytes already activated with mycoplasmas lost cytotoxic potential after disruption. The kinetics and degree of lymphocytotoxicity induced by M. arthritidis and phytohemagglutinin toward 51Cr-labeled syngeneic fibroblasts were similar. Removal of most B cells and other adherent cells by column separation did not abrogate the cytotoxic effect. Lymphocyte suspensions treated with anti-Thy 1 antiserum and complement exhibited a marked decrease in their cytotoxic potential when added to labeled target cells in the presence of M. arthritidis. We conclude that the cytotoxic reaction is dependent upon the T-lymphocyte subpopulation.
Adjuvant activities of quinonyl-N-acetyl muramyl dipeptides in mice and guinea pigs
Authors: Saiki I , Tanio Y , Yamawaki M , Uemiya M , Kobayashi S , Fukuda T , Yukimasa H , Yamamura Y , Azuma I
Published: January 2020

The adjuvant and tumor-suppressive activities of the quinonyl [2,3-dimethoxy-5-methyl-6-(9'-carboxynonyl)-1,4-benzoquinone (QS-10)] derivatives of N-acetyl muramyl dipeptides were examined. N-Acetyl muramyl-L-valyl-D-isoglutamine (MurNac-L-Val-D-isoGln), QS-10-MurNAc-L-Val-D-isoGln, and their methyl esters were shown to have potent adjuvant activity on the induction of delayed-type hypersensitivity to monoazobenzenarsonate-N-acetyl-L-tyrosine in guinea pigs and on the primary immune response against sheep erythrocytes in vitro; however, only QS-10-MurNAc-L-Val-D-isoGln methyl ester, i.e., QS-10-MurNAc-L-Val-D-Glu(OCH3)NH2 (quinonyl-MDP-66), was shown to be an active adjuvant for the induction of allogeneic killer T cells in mice and the suppression of tumor growth in syngeneic mice when it was administered as a suspension in phosphate-buffered saline. The effectiveness of the chain length of the quinonyl moiety in quinonyl-MDP-66 and the replacement of the L-valine residue with L-serine or L-threonine were also examined in comparison with the adjuvant and tumor-suppressive activities of quinonyl-MDP-66.
Protective Effect of Immunization of Rats with Holotoxin or B Subunit of
Escherichia coli
Heat-Labile Enterotoxin
Authors: Klipstein Frederick A. , Engert Richard F.
Published: January 2020

The relative immunogenicities of three forms of the
Escherichia coli
heatlabile enterotoxin (LT), the holotoxin, its B subunit, and the polymyxin-release form (PM LT) were compared by immunizing rats with various dosages of each given exclusively by the parenteral (IP/IP) or peroral (PO/PO) routes or by a combination of the two (IP/PO). The degree of protection was evaluated by challenge in ligated ileal loops, and the serum antitoxin response was determined by an enzyme-linked immunosorbent assay with homologous antigens. When given by the PO/PO route, each LT antigen provided only weak protection against the toxin and virtually none against viable LT-producing strains; serum antitoxin titers were not significantly increased. When the toxins were given after a parental primary immunization by either the IP/IP or the IP/PO routes, each LT antigen provided a dose-related increase in serum antitoxin titers and in the degree of protection against the toxin as well as against viable strains which produce LT alone (LT
+
/ST
−
) or in combination with the heat-stable toxin (LT
+
/ST
+
). The degree of protection against viable bacteria, particularly the LT
+
/ST
+
strain, was stronger in animals which received booster immunizations by the PO route. When expressed on the basis of molar equivalents, holotoxin provided significant protection (a protection index of >5 against toxin challenge and >50% reduced secretion with bacterial challenge) with 4 to 15 times fewer moles than PM LT and up to 50 times fewer moles than the B subunit. These observations indicate that, on the basis of molar equivalents, the holotoxin (which contains one A plus five or six B subunits) is a more potent immunogen than either PM LT (which contains one A and probably one B subunit) or the B subunit.
Immunochemical characterization of Brucella lipopolysaccharides and polysaccharides
Authors: Moreno E , Speth S L , Jones L M , Berman D T
Published: January 2020

Purified lipopolysaccharide (LPS) extracted with phenol-water from smooth Brucella abortus was hydrolyzed with 1% acetic acid at 100 degrees C. The degraded polysaccharide (AH) released gave reactions of identity with the native polysaccharide hapten (NH) in phenol-water- or trichloroacetic acid-extracted endotoxin preparations of B. abortus and with the polysaccharide (poly B) extracted by trichloroacetic acid from rough B. melitensis strain B115. Poly B was present in the soluble cytoplasmic fraction but not in the membrane fraction, of disrupted B115 cells. It could not be extracted from three rough mutants of B. abortus or from B canis or B. ovis cells. Both AH and NH shared determinants present on smooth LPS and missing from poly B. Sugars found in purified LPS, NH, and AH included mannose, glucose, quinovosamine, glucosamine, and 2-keto-3-deoxyoctonate. Poly B contained only a trace amount of quinovosamine and no 2-keto-3-deoxyoctonate detectable by the thiobarbiturate assay. Sera from some rabbits immunized with pure smooth LPS and some, but not all, cows infected with field strains of B. abortus recognized the determinants missing from poly B. A subclass-specific enzyme-linked immunoassay showed that most of the antibody in sera from infected cows which binds to smooth LPS and to NH is of the immunoglobulin G1 subclass.
Characterization of murine antibody response to Salmonella typhimurium by a class-specific solid-phase radioimmunoassay
Authors: Metcalf E S , O'Brien A D
Published: January 2020

A heavy-chain class-specific, solid-phase radioimmunoassay was developed to characterize the murine antibody response to Salmonella typhimurium. The specificity of the assay was verified by quantitation of the extent of binding of anti-S. typhimurium antibodies to other bacterial genera and species and by cross-adsorption studies. The sensitivity of the procedure was also examined, and it was determined to be substantially more sensitive than either the passive hemagglutination or the whole-cell agglutination technique. The method was subsequently used to analyze th murine antibody response to S. typhimurium. Groups of mice were prebled and then immunized with live S. typhimurium via different routes. The animals were bled weekly for 12 weeks, and then sera were assayed for antibodies directed against whole bacteria or purified lipopolysaccharide. Anti-Salmonella antibodies of the immunoglobulin M class appeared in the serum approximately 2 to 3 weeks after immunization, and then immunoglobulin G anti-Salmonella antibodies appeared which constituted the major part of the long-term response. Immunoglobulin A was not a major component of the serum antibody response. The antibodies were primarily directed against the lipopolysaccharide determinants, but a small percentage of the response was directed against other cell surface components. Qualitatively and quantitatively similar anti-Salmonella antibody responses were observed in sera of outbred and inbred strains of mice.
K88-mediated binding of Escherichia coli outer membrane fragments to porcine intestinal epithelial cell brush borders
Authors: Middeldorp J M , Witholt B
Published: January 2020

We have examined the interactions between various radiolabeled membrane fractions obtained from an enterotoxigenic Escherichia coli strain and brush borders isolated from porcine intestinal epithelial cells. Outer membrane fragments containing the K88 attachment factor bound tightly to brush borders, whereas cytoplasmic membrane vesicles did not. Three different types of outer membrane preparations were tested: (i) cellular outer membranes isolated from lysozyme spheroplasts, (ii) medium vesicles or outer membrane fragments released into the medium during growth, and (iii) periplasmic vesicles, or outer membrane fragments which were released from the cells during spheroplast formation and were therefore isolated in the periplasmic fraction. Of these fractions, which were heterogeneous, it was always the outer membrane subfraction which bound tightly to brush borders. This binding, which was K88 dependent, may have some physiological significance in view of the association between outer membrane fragments and enterotoxin. Thus, released outer membrane fragments equipped with attachment factors may function as enterotoxin carriers which increase the efficiency with which enterotoxin can be delivered to intestinal epithelial cells.
Inhibition of phagolysosome fusion is localized to Chlamydia psittaci-laden vacuoles
Authors: Eissenberg L G , Wyrick P B
Published: January 2020

Intracellular survival of Chlamydia psittaci is in part dependent on the ability of the organism to thwart phagolysosome formation. Circumvention of phagolysosome fusion could be either localized to chlamydia-laden vacuoles or generalized to all phagosomes in the host cell. To determine which of these modes is in operation the ability of chlamydia elementary and reticulate bodies to protect Saccharomyces cerevisiae from degradation in macrophage phagolysosomes was examined via acridine orange and Giemsa staining. No statistically significant difference was evident between the amount of fusion observed in coinfected macrophages and those infected with yeast cells alone. This was ot dependent on some unique interaction between the chlamydia and the yeast cells since viable count studies to determine the protection of a second organism, Escherichia coli, also failed to show significantly different amounts of inactivation of the bacteria by macrophages in the presence of C. psittaci. Therefore, the inhibition of phagolysosome fusion is localized to chlamydia-laden phagosomes.
Trapping and Killing of
Candida albicans
by
Corynebacterium parvum
-Activated Livers
Authors: Sawyer R. T. , Moon R. J. , Beneke E. S.
Published: January 2020

Corynebacterium parvum
vaccination significantly increased the number of leukocytes adherent to hepatic vessels. Perfused
C. parvum
-treated livers killed significantly more
Candida albicans
than did livers not treated with
C. parvum
, an effect reversed by the macrophage inhibitors silica, phenylbutazone, and iodoacetate.
Murine cytomegalovirus infection in the spleen and its relationship to immunosuppression
Authors: Loh L , Hudson J B
Published: January 2020

An attempt was made to correlate murine cytomegalovirus-induced immuno-suppression with the presence of virus-infected cells in the spleen. For this purpose, SWR/J mice were infected with murine cytomegalovirus, and spleens were removed periodically and assayed for the presence of infectious centers in different cell populations and for their capacity to respond to mitogens. The maximal degree of immunosuppression correlated with the peak in adherent cell infectious centers. This was followed by a rapid rise in the number of cells per spleen (splenomegaly), a more gradual return to normal responsiveness to mitogens, and a rapid disappearance of infectious centers. These results add more support to the hypothesis that murine cytomegalovirus-induced immunosuppression is caused indirectly by infection of adherent spleen cells.
Induction of immunoenhancing factors for murine splenocyte cultures by Salmonella typhimurium ribosome and ribonucleic acid extracts
Authors: Butler R C , Friedman H , Specter S C , Eisenstein T K
Published: January 2020

Ribosomal and ribonucleic acid (RNA)-rich preparations derived from Salmonella typhimurium were examined for their ability to enhance the primary in vitro antibody response of normal mouse spleen cell cultures to sheep erythrocytes. Both of these fractions were consistently more active in elevating the antibody response of normal mouse splenocytes from lipopolysaccharide (LPS) responder mice than was LPS. Furthermore, injection of mice with either the ribosomal or RNA-rich fraction induced antibody response helper factor activity in 2-h post-treatment serum similar to that induced by LPS. Endotoxin low-responding C3H/HeJ mice were stimulated to release helper factors by ribosomes and the RNA extracts but not by LPS. Treatment of the ribosomes and RNA fractions with ribonuclease destroyed their ability to stimulate the production of the helper factor in serum of treated mice. Therefore, it appears likely that ribosomes and RNA-rich fractions stimulated an intermediate helper factor due to the presence of RNA and not LPS.
Oligonucleotide Fingerprinting of Ribonucleic Acids of Infectious Bronchitis Virus Strains
Authors: Clewley J. P. , Morser J. , Avery R. J. , Lomniczi B.
Published: January 2020

A total of 11 distinct oligonucleotide fingerprints were obtained in studies of the ribonucleic acids of 13 isolates of infectious bronchitis virus. Different serotypes had distinct fingerprints, but so did varieties within a serotype, allowing a greater degree of strain differentiation than was previously possible. Some conclusions can be drawn from the fingerprints concerning theories of origin and spread of infectious bronchitis virus.
Phagocytosis and Killing of
Streptococcus pyogenes
by Human Alveolar Macrophages
Authors: Gemmell C. G. , Peterson P. K. , Regelman W. , Schmeling D. , Hoidal J. R. , Quie P. G.
Published: January 2020

In contrast to human polymorphonuclear leukocytes and monocytes, alveolar macrophages were able to readily phagocytose and kill an M protein-positive
Streptococcus pyogenes
strain after opsonization in normal human serum.
Adjuvant Activity of 6-Amino-6-Deoxy-Muramyldipeptides and Their Acylamino Derivatives on the Induction of Delayed Hypersensitivity to Azobenzenearsonate-
N
-Acetyl-
l
-Tyrosine in Guinea Pigs
Authors: Azuma Ichiro , Okumura Hiroyuki , Saiki Ikuo , Tanio Yoshiro , Kiso Makoto , Hasegawa Akira , Yamamura Yuichi
Published: January 2020

The 6-amino-6-deoxy-
N
-acetylmuramyldipeptides and their 6-acylamino derivatives were shown to be active as adjuvants on the induction of delayed-type hypersensitivity to azobenzenearsonate-
N
-acetyl-
l
-tyrosine in guinea pigs. However, 6-acylamino-6-deoxy-
N
-(acyl)muramyldipeptides were inactive as adjuvants.
Inhibition of human herpesviruses by combination of acyclovir and human leukocyte interferon
Authors: Levin M J , Leary P L
Published: January 2020

Human leukocytes interferon in low concentrations (1 to 5 U/ml) enhanced the antiviral effect of acyclovir against herpes simplex virus, varicella-zoster virus, and cytomegalovirus grown in human fibroblasts. This occurred without additive inhibition of the division of human fibroblasts or proliferation of peripheral blood mononuclear cells. The combined antiviral effect was additive against clinical isolates of cytomegalovirus and was synergistic against clinical isolates of the other two viruses. The magnitude of the effect with cytomegalovirus was the same when laboratory and wild-type virus were compared. The persistence of varicella-zoster virus in the presence of acyclovir in infected human cells was also reduced by the addition of interferon.
Proliferative responses of central and peripheral rat lymphocytes elicited by cord factor (trehalose 6,6'-dimycolate)
Authors: Kierszenbaum F , Walz D R
Published: January 2020

Cord factor--a mixture of 6,6'-diesters of alpha, alpha-D-trehalose with natural mycolic acids--which is purified from mycobacteria and other microorganisms, is known to have adjuvant activity as well as to enhance nonspecific resistance to infections and tumor development. In this work, trehalose 6,6'-dimycolate (TDM) was found to induce proliferative responses in rat thymus and lymph node cells. With the thymus cells, TDM responses were greater after removal of the adherent cell subpopulation. Consistent with this observation was the finding that addition of phagocytic cells purified from peritoneal or lymph node cell suspensions to nonadherent thymocytes abrogated the response of thymocytes to TDM. With the lymph node cells, the presence or removal of adherent cells had no major consequence on the TDM-induced proliferative response, since similar increases in deoxyribonucleic acid synthesis were observed with unfractionated and nonadherent cells. The difference between the sensitivities of thymus cells and lymph node cells to regulation by adherent cells indicated the existence of more than one type of TDM responder cell in rats. TDM also displayed marked stimulatory activity on thymus and lymph node cells from germ-free rats, ruling out the possibility that TDM might have triggered a specific, secondary, in vitro immune response. Expansion of a selected cell population(s) triggered by TDM may be involved in the manifestation of adjuvant activity and possibly other immunological properties of cord factor.
Comparative neurovirulence of selected vesicular stomatitis virus temperature-sensitive mutants of complementation groups II and III
Authors: Rabinowitz S G , Huprikar J , Dal Canto M C
Published: January 2020

Weanling mice were inoculated intracerebrally with selected vesicular stomatitis virus (VSV) complementation group II and III temperature-sensitive (ts) mutants. Of the VSV ts mutants studied, only ts G32, a group III complementation mutant, appeared neurovirulent. Interestingly, neither the capacity to replicate in central nervous system tissue nor the ability to replicate in certain neurally derived continuous cell lines at semipermissive or nonpermissive temperatures appeared different among the VSV ts mutants employed. Finally, the pathological alterations in central nervous system tissue produced by VSV ts G32 were entirely different than those produced by G31 VSV ts in the group III mutant. These studies support the hypothesis that both the virological and neuropathological features produced by different VSV ts mutants are dependent upon the unique characteristics of each mutant, rather than upon a common biochemical defect shared by all members of a complementation group.
Complement-fixing antibody to the AG-4 antigen in herpes simplex virus type 2-infected patients
Authors: Arsenakis M , May J T
Published: January 2020

Sera collected from confirmed herpes simplex virus type 2 (HSV-2) patients were found to be devoid of complement-fixing antibody to the AG-4 antigen at the time of the herpes lesion outbreak in 10 out of 13 cases. However, 1 to 4 weeks after HSV-2 lesion appearance, 28 out of 30 patients acquired complement-fixing antibody to the AG-4 antigen. The sera of these patients contained immunoglobulin M antibody activity and the ability to immunoprecipitate a 160,000-molecular weight early HSV-2 antigen (the AG-4 antigen). Also, these sera were used to show that a variety of anti-herpes virus compounds had a negligible effect on AG-4 production in HSV-2-infected HEp-2 cells. The majority of the compounds tested (including acycloguanosine and phosphonoformate) are known to inhibit late antigen production, suggesting that the AG-4 antigen is an early antigen. It is probably an immediate early antigen (alpha) as it is formed in the presence of cycloheximide and actinomycin D, a treatment which is used to accumulate alpha proteins.
Pathogenicity of Spiroplasma sp. strain SMCA in rabbits: clinical, microbiological, and histological aspects
Authors: Kirchhoff H , Heitmann J , Trautwein G
Published: January 2020

Newborn rabbits inoculated intracerebrally with early-passaged broth cultures of Spiroplasma strain SMCA (suckling mouse cataract agent) either died or developed eye disease. Death occurred 4 to 12 days after infection. Rabbits which died showed hemorrhages throughout the brain, especially in areas leading to the optic nerve, and hemorrhages in the liver. SMCA could be reisolated from brain, liver, and eyes. Rabbits with eye disease did not open their eyes during an observation time of 3 to 4 months. Pathological examination revealed marked microophthalmia and severe cataract formation characterized by complete degeneration of lens fibers and liquefaction and mineralization of the lens. Other ocular changes were chronic panophthalmitis with complete destruction of the retina. Neither eye disease nor death could be induced by inoculating SMCA subcutaneously into newborn rabbits. In adult rabbits, no disease occurred after intravenous or subcutaneous injection or after inoculation into the conjunctival sac.
Basis of the specificity of rodent trypanosomes for their natural hosts
Authors: Albright J W , Albright J F
Published: January 2020

The strong natural insusceptibility of mice to infection with the rat-specific Trypanosoma lewisi was investigated in a variety of mouse strains. Parasite elimination by all strains was similar involving a lag phase of about 7 h of constant parasitemia followed by rapid clearance (half-times of 55 and 130 min in different strains). In vitro cultures were utilized to determine whether mouse cells and serum were inherently deficient in supporting T. lewisi or, alternatively, toxic for the parasite; neither was found to be the case. In attempts to prolong T. lewisi survival and growth, mice were treated with various preparations, including T. lewisi sonicates, rabbit antiserum against mouse macrophages, silica dust, normal rat and rabbit sera and crude immunoglobulin fractions, and combinations of silica dust and serum or immunoglobulin fractions. The most striking effects were obtained with combinations of silica dust and serum, resulting in extensive T. lewisi growth and death of a portion of the infected mice. These results, together with microscopic observations, suggested that the principal mechanism responsible for murine resistance to heterologous trypanosomes is a type of antibody-dependent, cell-mediated immunity involving granulocytes and (probably) platelets.
Comparative virulence of Absidia corymbifera strains in mice
Authors: Kitz D J , Embree R W , Cazin J
Published: January 2020

The comparative virulence of six different strains of Absidia corymbifera for cortisone-treated and untreated Swiss mice was determined. Spores of the six strains were inoculated into mice by the intravenous, intraperitoneal, and intranasal routes. All six strains were found to be virulent in cortisone-treated and untreated mice by the intravenous route. Up to a 16-fold difference in strain virulence was observed for cortisone-treated mice and up to a 10-fold difference for untreated mice. When spores were administered by the intraperitoneal route, 50% lethal dose values could be calculated only for the cortisone-treated mice, although a few deaths were seen in untreated mice challenged with 10(7) spores. Each of the six isolates of A. corymbifera, when administered in an intranasal dosage of 10(6) spores, produced death in some cortisone-treated mice. Studies made to determine the viability of spores produced by each strain revealed that germination was 90% or greater on Littman and YpSs agars at an incubation temperature of 40 degrees C in less than 12 h.
Plasmid-chromosomal transition of genes important in staphylococcal enterotoxin B expression
Authors: Dyer D W , Iandolo J J
Published: January 2020

Experiments were performed to further elucidate the genetic mechanisms underlying the synthesis of staphylococcal enterotoxin B (SEB). Our laboratory has previously shown that, in strains of Staphylococcus aureus which harbor a 1.15-megadalton plasmid (pentB or pSN2), the plasmid appears to be required for SEB synthesis; in other S. aureus strains, designated chromosomal SEB producers, this 1.15-megadalton plasmid is conspicuously absent. We report here than in both Bacillus subtilis minicells and a coupled translational assay, pSN2 codes for a polypeptide of 18,000 daltons. This product is not immunologically reactive with purified anti-SEB globulin. Nevertheless, pSN2 is necessary but not sufficient for SEB synthesis in strains which harbor the plasmid. Further, the data provide a reasonable link between plasmid-bearing and chromosomal SEB producers: transformational analysis indicates that both require functions specified (in plasmid-bearing strains) by pSN2 for SEB synthesis. The combined genetic and biochemical data suggest that pSN2 is not the reservoir for the SEB structural gene, but that the pSN2-specific functions required for SEB synthesis are regulatory in nature.
Primary acute histoplasmosis in guinea pigs exposed to aerosolized Histoplasma capsulatum
Authors: Schlitzer R L , Chandler F W , Larsh H W
Published: January 2020

Guinea pigs were examined as a possible in vivo model for human histoplasmosis. Guinea pigs were exposed to an aerosol of viable microconidia and mycelial fragments of Histoplasma capsulatum generated in a Henderson apparatus. Colonization and infection of the lungs occurred, with subsequent involvement of the regional lymph nodes and reticuloendothelial organs. Cultural recovery of the fungus from the nasopharynx and bronchoalveoli was initially high, but decreased with time. The mean number of colonies recovered from the lungs gradually increased, reaching a peak at 2 weeks, with involvement of the regional lymph nodes. Extrathoracic dissemination to the liver and spleen occurred in only a few animals. After 4 weeks, all tissues except the cervical and tracheobronchial lymph nodes were culturally negative; all specimens were culturally negative after 8 weeks. Disappearance of H. capsulatum from tissues appeared to correlate inversely with the development of hypersensitivity as measured by skin test reactivity. Histopathological studies supported cultural results and were similar to those described for primary human and canine histoplasmosis.
Virulence of Three Clinical Isolates of
Vibrio cholerae
Non-O- 1 Serogroup in Experimental Enteric Infections in Rabbits
Authors: Madden Joseph M. , Nematollahi Wales P. , Hill Walter E. , [ill] Barbara A. , Twedt Robert M.
Published: January 2020

Three
Vibrio cholerae
, [ill] isolates, pathogenic in both adult rabbit ligated ileal loop and infant rabbit assays, produced no heat-labile or heat-stable toxins. Their pathogenicity did not correlate with the presence of plasmids.
Defective bactericidal reaction by the alternative pathway of complement in atopic patients
Authors: Siccardi A G , Fortunato A , Marconi M , Dos Santos C , Grassi F , Sacchi F , Fietta A
Published: January 2020

Atopic sera were shown to be defective in serum bactericidal activity by a reaction dependent upon the integrity of the alternative pathway of complement activation. In normal human sera, the efficiency of such a reaction could be modulated by the addition of thermostable serum fractions with inhibitory and stimulatory activity, as demonstrated on two different assay systems. Atopic sera contained the inhibitory activities, but no stimulatory activity. The bactericidal defect of atopic sera was shown to be familial, but its hereditary mechanism could not be unambiguously interpreted.
Polymorphonuclear neutrophil chemotaxis induced and inhibited by Bacteroides spp
Authors: Adamu S A , Sperry J F
Published: January 2020

A possible virulence factor for Bacteroides subcutaneous abscesses has been found. The effect of Bacteroides culture filtrate and outer membrane on chemotaxis of rabbit peritoneal polymorphonuclear (PMN) neutrophils was assayed in Boyden chambers and via exocytosis of N-acetyl-beta-glucosaminidase. Both Bacteroides culture filtrate and outer membrane elicited some chemotaxis, as measured in the Boyden chamber; however, they had little effect upon exocytosis in cytochalasin B-treated PMN neutrophils. In the presence of serum complement, they completely abolished PMN neutrophil movement in the Boyden chamber assay, yet they gave a definite positive response for the exocytosis assay in the presence of serum complement or activated complement fragment.
Chronological Development of Cellular Immunity in Human Toxoplasmosis
Authors: Johnson Warren D.
Published: January 2020

A dichotomy between the resolution of clinical illness in acute toxoplasmosis and the responsiveness of B and T lymphocytes to toxoplasma antigens is described.
Effect of pH on the production of the Kanagawa hemolysin by Vibrio parahaemolyticus
Authors: Cherwonogrodzky J W , Clark A G
Published: January 2020

Production of the Kanagawa hemolysin by patient strains of Vibrio parahaemolyticus was found to respond to the pH rather than to the type of carbohydrate present in the growth medium. Regardless of the carbohydrate present, hemolysin production in peptone broth cultures occurred only when the pH was between 6.5 and 5.5. Mannitol, the sugar used in the Wagatsuma agar, lowered the pH to within this range, thus providing optimal conditions for hemolysin production. Glucose and mannose, although readily metabolized, lowered the pH below this range, inhibiting growth and hemolysin production. Alkaline cultures either without carbohydrates or containing non-metabolizable sugars showed little hemolytic activity because the pH always remained alkaline. In pH-stat cultures maintained at pH 6.2, higher hemolysin yields were produced irrespective of the presence or absence of mannitol. We conclude that the production of the Kanagawa hemolysin is under pH control. Marine strains of V. parahaemolyticus, which are Kanagawa negative, did not express detectable amounts of hemolysin under those conditions shown to stimulate hemolysin production by Kanagawa-positive strains.
Idiotypes and biological activity of murine monoclonal antibodies against the hemagglutinin of measles virus
Authors: Gheuens J , McFarlin D E , Rammohan K W , Bellini W J
Published: January 2020

Three hybridomas, designated C2, V17, and B2, were produced from BALB/c mice after immunization with measles virus. All three were directed against the virus hemagglutinin (HA). The HA is a structural peptide of the virus and constitutes a major target for the host immune response during measles infection. The monoclonal anti-HA antibodies have biological functions such as (i) measles virus neutralization in vitro, (ii) binding to acutely and persistently infected cells, and (iii) inhibition of HA-mediated Rhesus monkey erythrocyte agglutination. Different idiotypes, designated HAMM-1, HAMM-2, and HAMM-3, were defined on C2, V17, and B2, respectively, by syngeneic anti-idiotype sera against those three monoclonal antibodies. A limited cross-reactivity with the HAMM-1 idiotype was detected in sera from some BALB/c mice immunized with measles virus. The anti-idiotype sera could significantly inhibit the biological functions of the HAMM-1 and HAMM-3 idiotypes bearing monoclonal anti-HA-antibodies. This suggests a possible role for auto-anti-idiotypes in the immune response after infection with measles virus.
Nonspecific suppression of primary antibody responses and presence of plastic-adherent suppressor cells in Toxoplasma gondii-infected mice
Authors: Suzuki Y , Watanabe N , Kobayashi A
Published: January 2020

The effect of Toxoplasma infection on primary antibody responses to both T-dependent and T-independent antigens was examined in mice. Drastic suppression of primary responses to sheep erythrocytes (SRBC) occurred when mice were immunized 7 days after infection. The suppression was observed in both 2-mercaptoethanol-sensitive and -resistant hemagglutinin responses. Anti-dinitrophenol (DNP) immunoglobulin E and G1 responses to DNP-conjugated keyhole limpet hemocyanin were also suppressed by infection. It was suggested that the suppressive effect is nonspecific for the antigens and immunoglobulin classes produced. Anti-DNP responses to DNP-Ficoll, a T-independent antigen, were suppressed by infection, but the suppressive effect was weaker than that on the responses to SRBC. This suggests that both T and B cells are suppressed by infection. In vitro responses of infected mouse spleen cells to SRBC and DNP-Ficoll confirmed the results of in vivo experiments. In addition, the presence of plastic-adherent suppressor cells was demonstrated in the spleen cells of infected mice, which suppressed the ability of normal mouse spleen cells to mount an SRBC-specific plaque-forming cell response. These plastic-adherent suppressor cells appeared to be a major cause of nonspecific suppression of primary antibody responses in Toxoplasma-infected mice.
Fractionation of Schistosoma japonicum soluble egg antigen glycoproteins by hydrophobic interaction chromatography
Authors: Long G W , Lewert R M , Pelley R P
Published: January 2020

We are currently studying the soluble egg antigens of Schistosoma japonicum in an attempt to determine which antigens are potent immunogens. Previously, we demonstrated by Ouchterlony immunodiffusion and inhibition of the circumoval precipitin test that the glycoprotein fraction of soluble egg antigens contains the antigens which are most immunogenic in natural infections. The soluble egg antigen glycoproteins have now been further fractionated via hydrophobic interaction chromatography on phenyl Sepharose. We found that there were at least two antigens involved in the circumoval precipitin reaction. Both the hydrophilic antigen which we call japonicum antigen glycoprotein II (JAG II) and a mixture of hydrophobic antigens (JAG III and the JAG IV complex) were capable of causing a 50% inhibition of the COP reaction around S. japonicum eggs. JAG II was not a major serological antigen of S. japonicum since it gave only a weak precipitin line upon Ouchterlony immunodiffusion analysis with pooled sera from Filipino patients with chronic S. japonicum infections. Hydrophobic interaction chromatography yielded preparations which were sufficiently pure for use in radioimmunoassays. By radioimmunoassay, the best antigens among the glycoproteins were moderately hydrophobic JAG III and the JAG IV complex. They had large amounts of antibody directed toward them in patients with schistosomiasis japonica and exhibited little reactivity with S. mansoni. The hydrophilic glycoproteins JAG I and II were poor immunogens and extensively cross-reacted with S. mansoni. This cross-reactivity means that diagnostic tests with crude soluble egg antigens would run the risk of potential false-negative results in patients with other trematode infections.
Interferon induction in primates by stabilized polyriboinosinic acid-polyribocytidylic acid: effect of component size
Authors: Levy H B , Riley F L , Lvovsky E , Stephen E E
Published: January 2020

Two series of interferon-inducing complexes containing polyriboinosinic and polyribocytidylic acids, poly-L-lysine, and carboxymethyl cellulose were prepared. One series contained carboxymethyl cellulose, 27,000-molecular-weight poly-L-lysine, and either 4S, 6S, or 9S polyriboinosinic and polyribocytidylic acids. The other series contained carboxymethyl cellulose, 9S polyriboinosinic and polyribocytidylic acids, and poly-L-lysine, whose molecular weights ranged from 2,000 to 27,000. The homogeneity of these double-stranded polynucleotide complexes was confirmed by single-step thermal denaturation profiles and by single peaks in sucrose gradient velocity sedimentation. The complexes have a greater resistance to hydrolysis by ribonuclease than does polyriboinosinic-polyribocytidylic acid. The resistance to ribonuclease increased with the increasing size of polynucleotide homopolymers and poly-L-lysine. In monkeys and, to a lesser extent, in mice, serum interferon levels induced by the different complexes were related to the degree of resistance of the complexes to hydrolysis by ribonuclease. In mice, 4S, 6S, and 9S complexes of polyriboinosinic-polyribocytidylic acid, poly-L-lysine, and carboxymethyl cellulose had a higher level of toxicity than did polyriboinosinic-polyribocytidylic acid as measured by 50% lethal dose. The toxicity was parallel to the ribonuclease resistance of the complexes. It was concluded that an increase in the size of the polynucleotides and the polyamino acids in these complexes leads to higher resistance to hydrolysis by ribonuclease and to greater interferon responses in mice and rhesus monkeys.
Characterization of Micropolyspora faeni antigens
Authors: Kurup V P , Ting E Y , Fink J N , Calvanico N J
Published: January 2020

Culture filtrate antigens of Micropolyspora faeni grown in a synthetic medium in a stirred fermentor were characterized. The culture filtrate antigens were fractionated by preparative isoelectric focusing with a pH gradient of 3.5 to 5.5. The fractions were pooled according to their reaction with rabbit anti-M. faeni sera. A pool containing two major antigens which were resolved by analytical isoelectric focusing and polyacrylamide disc gel electrophoresis was obtained. One antigen was stainable with Coomassie blue and periodic acid-Schiff stain and was determined to have a mass of 51,000 daltons. The other antigen was stainable only with Coomassie blue and was determined to have a mass of 29,000 daltons. When used at 1 mg/ml, this pool reacted with the sera from all patients with farmer's lung disease by immunodiffusion but failed to react with control sera.
Lipopolysaccharides from Bacteroides fragilis are mitogenic for spleen cells from endotoxin responder and nonresponder mice
Authors: Joiner K A , McAdam K P , Kasper D L
Published: January 2020

The lipopolysaccharides (LPS) from Bacteroides fragilis are structurally atypical and give weak responses in most tests of endotoxic activity, but the mitogenic activity of LPS from B. fragilis has not been tested. We prepared LPS from B. fragilis 23745 by three methods and compared their mitogenic activity for murine spleen cells with that of LPS from Escherichia coli K235 prepared by similar techniques. LPS extracted from B. fragilis with hot phenol-water, with butanol-water, or by detergent separation from the outer membrane were mitogenic for spleen cells from C57BL/10ScN, C57BL/10ScCR, and C3H/HeJ mice. The outer membrane, the outer membrane protein-polysaccharide complex, and the capsular polysaccharide from B. fragilis were also mitogenic for spleen cells from the same murine strains. LPS extracted from E. coli K235 with hot phenol-water, butanol-water, or sodium deoxycholate were mitogenic for C57BL/10ScN spleen cells, but only the LPS extracted with butanol and deoxycholate were stimulatory for spleen cells from C57BL/10ScCR and C3H/HeJ mice. Two types of LPS varying in the 2-keto-3-deoxyoctonate-to-carbohydrate ratio were isolated from E. coli K235 with sodium deoxycholate; both endotoxins contained protein which was typical of lipid A or endotoxin protein. These results indicate that the LPS from B. fragilis is a potent mitogen for spleen cells from endotoxin responder and endotoxin nonresponder mice.
Role of macrophage oxidative metabolism in resistance to vesicular stomatitis virus infection
Authors: Rager-Zisman B , Kunkel M , Tanaka Y , Bloom B R
Published: January 2020

The role of oxygen metabolites in mediating virucidal activity was studied in two cloned macrophage-like cell lines. The parental cell line, J774.16, upon appropriate stimulation with either phorbol myristate acetate (PMA) or aggregated immunoglobulin, is induced to oxidize glucose via the hexose monophosphate shunt and produce O2- and H2O2. A variant derived from it, clone C3C, is defective in oxidative metabolism and cannot be stimulated to produce O2- or H2O2. Significant differences in yields of vesicular stomatitis virus (VSV) between stimulated clone 16 cells and unstimulated cells could be obtained only when low multiplicities were used for infection. Under the same conditions, PMA stimulation of the variant clone C3C produced no reduction in yields. The effect of PMA on virus yields in clone 16 was short-lived and dose dependent. PMA stimulation of either cell line had no effect on the number of infectious centers, suggesting that the antiviral effect was likely to be an extracellular, rather than an intracellular, one. Using glucose oxidase plus aglucose to generate H2O2 in solution, we observed that H2O2 alone is capable of killing limited amounts of VSV. The inactivation of VSV, both by H2O2 in solution and by activated clone 16 cells, could be inhibited by catalase. We conclude that intracellular resistance to VSV is primarily mediated through nonoxidative mechanisms, since activated macrophages can kill only a limited number of infectious virus particles extracellularly by means of secreted H2O2.