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Immune reactivity during ageing. III. Removal of peanut-agglutinin binding cells from ageing mouse spleen cells leads to increased reactivity to mitogens.

Spleen cells from ageing (24--30-month-old) mice, manifesting low response to concanavalin A (Con A) and phytohemagglutinin (PHA), were separated by peanut agglutinin (PNA) into agglutinable (PNA+) and unagglutinable (PNA-) fractions. The PNA+ cells were found to suppress the response of young (2--3-month-old) mouse spleen cells to the mitogens Con A and PHA. On the other hand, PNA- cells of the ageing mice expressed a response to these mitogens, at levels higher than those of the unseparated cells. Re-mixing of the PNA- and PNA+ cells of the ageing mouse spleens at various proportions indicated that the PNA- cells are indeed suppressible by the PNA+ cells. Anti Thy-1.2 treatment to the PNA+ fraction suggested that this suppression was not exerted by T cells. It thus appears that at least part of the reduced lymphoid cell function in ageing is related to an effect of suppressor cells, interfering with the expression of available reactive cells.

Aging↗

Agglutination of Staphylococcus saprophyticus: a structural and cytochemical study.

Staphylococcus saprophyticus was shown to be agglutinated by wheat germ agglutinin, wheat germ agglutinin-biotin and bovine serum albumin-p-aminophenyl-N-acetyl-beta-D-glucosaminide (GlcNAc-BSA), and sheep red blood cells. In these agglutinations, filamentous or amorphous structures radiating from the surface of S. saprophyticus were demonstrated by electron microscope observation. Cytochemical analyses of the agglutination revealed the binding sites of wheat germ agglutinin in S. saprophyticus and the binding sites of GlcNAc in the sheep red blood cells and S. saprophyticus. Since GlcNAc-BSA contains N-acetylglucosamine to which wheat germ agglutinin can bind, it is most likely that an interaction between a wheat germ agglutinin-bindable substance in S. saprophyticus and an N-acetylglucosamine-bindable substance in sheep red blood cells is involved in the agglutination.

Acetylglucosamine↗

Characterization of a subpopulation in neonatal thymus which suppress the graft-vs.-host reaction.

Thymus cells from neonatal and infant mice were found to have a high capacity to prevent mortality from acute graft-vs.-host disease as compared with spleen cells from stable radiation chimeras. This suppressive capacity of thymocytes decreases with age after birth as was demonstrated by semi-quantitative cell titrations. This suppressor activity is restricted to syngeneity of the graft-vs.-host disease-including cells. The thymic suppressor cells are Thy-1+ and Lyt-1+ and IgG- and IgM-. They do not agglutinate with peanut agglutinin and have a high electrophoretic mobility. In vitro irradiation experiments showed that the suppressor cells are radiation sensitive. These results are compared with the available information on cells suppressing delayed-type hypersensitivity reactions and those suppressing B cell responses.

Animals↗

1,25-Dihydroxyvitamin D3 inhibits selectively the mitogenic stimulation of mouse medullary thymocytes.

Mouse thymocytes were separated into cortical and medullary subpopulations by differential agglutination with peanut agglutinin. A high-affinity receptor for 1,25-dihydroxyvitamin D3 is present in medullary immunocompetent mouse thymocytes and is absent from cortical immature cells. 1,25-dihydroxyvitamin D3, at physiological concentrations, inhibits the mitogenic response of the medullary cells to phytohemagglutinin and interleukin-2, but has no effect on the cortical subpopulation. Other less active metabolites of vitamin D had little or no effect on medullary cell stimulation.

Animals↗

Studies on the possible chemical, immunochemical and morphological differences at the cell surfaces of immunosensitive and immunoresistant, Moloney virus-induced, lymphoma cell-lines.

Comparison was made of several cell-surface parameters in the immunosensitive, Moloney virus-induced, mouse lymphoma, YAC, and its immunoresistant variant, YACIR. The characteristics of the two cell lines appeared to be similar by most of the criteria employed. The poly(acrylamide)-gel electrophoresis (with sodium dodecyl sulfate) patterns, after staining with Coomassie Brilliant Blue, of detergent-solubilized materials, appeared to be identical. After elution from a gel filtration column, no major differences were observed in the protein profiles of material cleaved from viable cells by proteolysis. Scanning and transmission electron microscopy revealed no major differences between the YAC and YACIR cells. The concentration of the lectins, Ricinus communis agglutinin, concanavalin A, wheat-germ agglutinin, and Solanum tuberosum (potato) agglutinin, required to agglutinate viable cells of the two lines were not significantly different. Neither cell was agglutinated by the lectins from Dolichos biflorus or Vicia graminea. Significant differences were, however, observed in the concentrations of lectin from Arachis hypogaea (peanut) needed to agglutinate the two cells. Although similar amounts (184-188 micrograms/10(9) cells) of sialic acid were released from viable cells by neuraminidase (V. cholerae), striking differences were observed in the composition of this material: 48% of N-glycolylneuraminic acid for YAC and 15% for YACIR. The remainder was N-acetylneuraminic acid for each cell line.

Agglutination↗

Cyclophosphamide-induced suppressor cells in mice: suppression of the antibody response in vitro and characterization of the effector cells.

It was found earlier that nonspecific suppressor cells obtained from the spleen of mice injected with cyclophosphamide (Cy) at a dose of 200 mg/kg body wt are nonadherent, surface Ig negative, and Thy-1 negative. It is now reported that Cy-induced suppressor (Cy-S) cells suppress the in vitro primary and secondary humoral responses in a dose-dependent manner. Suppressor activity is a property of a low density cell that is found in the spleen from 5 to 11 days after Cy administration. Cy-S cells are negative for a number of T-cell markers and are nonadherent to plastic. They are not agglutinated by peanut agglutinin and do not acquire the Thy-1 antigen after treatment with thymosin, suggesting that they are not T-cell precursors. While NK activity was detected in the Cy-S cell preparations, this activity was removed by treatment with antibodies specific for the Qa-5 antigen and complement without affecting the suppressor activity. Cy-S cells are, therefore, distinct from B cells, T cells, NK cells, and macrophages.

Animals↗

Selective activation of murine V beta 8.2 bearing T cells by Pseudomonas exotoxin A.

We have determined that Pseudomonas aeruginosa exotoxin A (PE) can selectively stimulate the proliferation of V beta bearing T lymphocytes. Murine thymocytes were fractionated by selective agglutination with peanut agglutinin (PNA) and the PNA- thymocytes, which represent mature thymocytes, were shown to be responsive to PE stimulation. In addition, mature peripheral T lymphocytes (nylon wool nonadherent splenocytes) were also observed to respond to PE stimulation. Both CD4+ and CD8+ splenic T lymphocyte populations proliferated in response to PE. Flow microfluorimetry analysis of PNA- thymocytes stimulated with PE indicated that V beta 8.2 bearing T cells were preferentially expanded. Thus, our data indicate that PE represents a microbial super antigen which stimulates murine thymocytes which bear the V beta 8.2 element of the T cell receptor.

ADP Ribose Transferases↗

Expression of blood group-related antigens and Helix pomatia agglutinin in malignant pleural mesothelioma and pulmonary adenocarcinoma.

In an attempt to differentiate malignant pleural mesothelioma from pulmonary adenocarcinoma by histochemical and immunohistochemical means, the glycoconjugate profiles of five reactive mesothelial lesions, 29 mesotheliomas (20 epithelial, three biphasic, and six fibrous types), and 38 well-differentiated pulmonary adenocarcinomas (34 papillary, two tubular, and two bronchioloalveolar types) were tested with ABH blood group-related antigens (BGR-Ag) antibody and Helix pomatia agglutinin (HPA) which agglutinates human type A erythrocytes. Formalin-fixed, paraffin-embedded sections were stained by the avidin-biotin-peroxidase complex method. Reactive mesothelial lesions and malignant mesothelioma of the pleura were not stainable with BGR-Ag antibody or HPA, irrespective of the blood group type. In pulmonary adenocarcinoma, however, the test with BGR-Ag antibody showed a high positive rate with the compatible blood group type, especially in type O cases (83%). Using HPA, reactions of adenocarcinoma with types A and AB also demonstrated high positive results (94% and 100%, respectively), but even with types B and O positive reactions occurred in 80% and 33% of cases, respectively. The findings suggest that positive reactions with either BGR-Ag antibody or HPA can be indicative of pulmonary adenocarcinoma.

ABO Blood-Group System↗

Heterogeneity of splenic natural suppressor cells induced in mice by treatment with cyclophosphamide.

Administration of high dose cyclophosphamide (CY, 200 mg/kg body weight) to adult mice induces transient, nonspecific suppressor activity in the spleen of treated animals. Characterization of the CY-induced natural suppressor (NS) cells which inhibit mixed lymphocyte reactions revealed a heterogeneous population of lymphocytes expressing the CD8 T cell marker and the B220 B cell marker, as well as cells bearing the granulocyte-monocyte marker CD11b. On a cell per cell basis the most potent of these suppressors were found to be positive for CD11b. Inhibitory activity was also detected in the CD8-, CD11b-, B220- compartment of CY-spleen, suggesting the presence of null NS cells. The fact that several phenotypically distinct cell populations contribute to the overall inhibitory effect of CY-spleen cells indicates that natural suppression defines an activity rather than a specific cell type. Interestingly, NS activity was observed to reside solely within the fraction of CY-spleen that is agglutinable with soybean agglutinin or wheat germ agglutinin, suggesting that expression of receptors for these plant lectins is a universal characteristic of CY-induced NS cells, regardless of their lineage. CY-spleen cell-mediated suppression of lymphoproliferative responses was found to be partially dependent on DNA synthesis and totally dependent on protein synthesis, but did not require cell-cell contact, indicating the production of soluble suppressor factor(s).

Animals↗

Biology of the group E streptococci: a review.

Group E streptococci are identified by a group antigen consisting of a cell wall rhamnose-glucose polymer. Other specific cell wall polysaccharides separate the group into at least six serotypes (II, IV, V, VI, VII and VIII). No species name has been accepted for the group E Streptococcus, although the organism is often designated as Streptococcus infrequens or S. lentus. Recently, the name S. porcinus has been proposed for a species that would include streptococci of groups E, P, U and V. Group E streptococci are fairly widely distributed in animals, especially swine, and are occasionally isolated from bovine milk. They cause streptococcal lymphadenitis of swine, an economically important disease, particularly in the United States. They do not appear to be the primary cause of any other disease or condition. The organisms enter the swine host through the mucosa of the pharyngeal or tonsillar surfaces, and are carried to the lymph nodes, primarily of the head and neck region, where abscesses are formed. Cell wall antigens induce the formation of serum agglutinins; a microtitration agglutination test, based upon reactions involving the type IV antigen, is considered a reliable test for detection of infection in animals. Antigenic components include group and type antigens, extra-cellular enzymes, and an antiphagocytic factor which may be associated with virulence of the organism and with protective immunity to streptococcal lymphadenitis of swine. This factor, a surface protein, develops on cells cultured in media fortified with serum, rendering the cells resistant to phagocytosis by porcine leukocytes. The classification, morphology, physiology, biochemistry and nutrition of the group E streptococci, and methods for their isolation, cultivation, and identification are reviewed.

Animals↗

Cold agglutinin related acrocyanosis and paroxysmal haemolysis.

The clinical features of severe cold agglutinin related paroxysmal agglutination and haemolysis could be confused with a number of conditions. A 61-year old lady presented with features of acute peripheral ischaemia who had recently been started on treatment for Raynaud's disease. Haematological investigations revealed the presence of potent cold haemagglutinins to be the cause of her symptoms but no definite cause for the raised titres were found. She was managed conservatively by keeping her hands and feet warm and regular chlorambucil for a suspected underlying lymphoproliferative disorder. It is important to differentiate the features of Raynaud's disease from those of severe haemagglutination so that identification of any underlying disorder can be made and appropriate treatment instituted.

Anemia, Hemolytic, Autoimmune↗

Immunoreconstitution in severe combined immunodeficiency after transplantation of HLA-haploidentical, T-cell-depleted bone marrow.

Immunological reconstitution by transplantation of HLA-haploidentical, paternal bone marrow was attempted in four infants with severe combined immunodeficiency who lacked HLA-identical donors. To prevent graft-versus-host disease, T lymphocytes were removed from the grafts by agglutination with soybean agglutinin and rosette formation with sheep red blood cells. None of the patients received conditioning treatment before transplantation. Normal, donor-derived cellular immune functions developed in all four patients within several months of transplantation. Threatening complications of graft-versus-host reactions were not seen. All four patients remain in excellent health 12-15 months after discharge home, with persisting normal T-cell functions.

Bone Marrow↗

[Analysis of results in studies on anti-erythrocyte auto-sensitization in 2400 patients].

The authors report their experience since 1970 in the laboratory investigation of anti-erythrocyte auto-immunization. 2400 tests were performed, and the positive results were divided into 2 groups: --341 patients with a positive direct antiglobulin test; --502 patients without positive direct antiglobulin test, but with various abnormalities: positive elution, increased titer of cold agglutinin, auto-agglutinability of protease treated red cells... The immunohaematological and etiological data are described for each group. It appears: --that there is no significant difference in etiological data between the two groups; --that the direct antiglobulin test is not sufficient in investigation of anti-erythrocyte auto-immunization, but some other immunohaematologic tests are indispensable.

Anemia, Hemolytic, Autoimmune↗

In vivo and in vitro anti-tumor effect of 14F7 monoclonal antibody.

The 14F7 monoclonal antibody (MAb) is an IgG(1) antibody that reacts specifically with GM3 (NeuGc) and with tissue sections of human tumors. We demonstrated here that this MAb is agglutinin that specifically agglutinated horse erythrocytes. Additionally, the capacity of 14F7 MAb to mediate cytotoxicity against GM3 (NeuGc)-positive murine myeloma cells, in vitro and in vivo, was evaluated. High concentrations of 14F7 MAb were needed to induce complement-dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) against the murine myeloma cells. The most relevant finding was the ability of this MAb to directly kill the target cells without participation of complement. This cytotoxicity was dependent on the temperature and MAb concentration and the number of the target cells. In vivo, the passive treatment with 14F7 MAb produced a strong anti-tumor activity, similar to the anti-tumoral response obtained with standard chemotherapy treatment.

Adjuvants, Immunologic↗

Sugar composition of O-antigenic lipopolysaccharides isolated from Vibrio parahaemolyticus.

Vibrio parahaemolyticus, a causative bacterium of food poisoning unique for its particular primary association with sea products, is now divided serologically into 11 or 12 O-forms based on agglutination and agglutinin-absorption tests. We determined the sugar composition of the somatic O-antigens, i.e., lipopolysaccharides (LPS), of representative strains of each O-form. Of particular interest is the absence of evidence for the presence of 2-keto-3-deoxy-octonic acid (KDO), a regular sugar component of gram-negative bacterial LPS, in any LPS examined, with the exception of 06. Furthermore, 07 and 012 LPS contained a KDO-like compound that is, however, not identical with KDO. Glucose, glucosamine, and L-glycero-D-mannoheptose were found as common sugar constituents. Three unidentified amino sugars, designated here as P1, P2, and P3, were found. Various combinations of each of these unidentified amino sugars, and of galactose, fucose, arabinose, D-glyucero-D-mannoheptose, galactosamine, KDO, and the KDO-like substance were detected in accordance with the O-form of LPS. On the basis of the sugar composition, LPS of the 12 O-forms of V. parahaemolyticus can be classified into nine chemotypes, because 03, 05, and 011 LPS belong to the same chemotype and 07 and 012 to another chemotype.

Antigens, Bacterial↗

Characterisation of 20-kDa lectin-spermagglutinin from Arum maculatum that prevents Chlamydia pneumoniae infection of L-929 fibroblast cells.

A novel lectin from the root of Arum maculatum was isolated by saline extraction and purified by cold ethanol precipitation and subsequent fractionation on Superose 6 column. The lectin named A. maculatum agglutinin is a non-glycosylated protein with 20-kDa molecular mass agglutinating human ejaculated spermatozoa, but not human erythrocytes. The agglutination was blocked in the presence of N-acetylneuraminic acid indicating that the lectin is sialoglycoprotein specific. Chlamydia pneumoniae strain AR-39 showed considerable potential to grow in murine L-929 fibroblast cells. Pretreatment of the cell monolayers with purified lectin reduced the entry and intracellular replication of C. pneumoniae. These results suggest that the isolated lectin prevents attachment by binding to a C. pneumoniae specific sialoglycoprotein receptor expressed on the surface of L-929 fibroblast cells.

Agglutination Tests↗

Gamma-A cold agglutinin: importance of disulfide bonds in activity and structure.

Cleavage of disulfide bonds caused by mild reduction with mercaptoethanol produces a reversible loss of cold agglutinin activity and antigen-binding ability in a gamma-A polymer cold agglutinin. Return of agglutination and antigen-binding ability occur only in those molecules that have not been depolyimerized by reduction. Loss of activity becomes irreversible if reduction is followed immediately by alkylation.

Amides↗

A new pathogenic Leptospira, not readily cultivated.

Alexander, Aaron D. (Walter Reed Army Institute of Research, Washington, D.C.), Herbert G. Stoenner, Garnett E. Wood, and Robert J. Byrne. A new pathogenic Leptospira, not readily cultivated. J. Bacteriol. 83:754-760. 1962.-A pathogenic Leptospira was isolated from water of the Grand River, (S.D.) that differed significantly from other known leptospirae in that it could not be cultivated in conventional leptospiral media. Growth was promoted in Fletcher's medium modified to contain 20% rabbit serum. The isolate, after several serial passages, was lethal for hamsters. It could not be adapted to grow in the chick embryo. Guinea pigs and calves inoculated with the isolate developed febrile and antibody responses but showed no other overt signs of disease. The strain was identified on the basis of cross-agglutination and agglutinin-adsorption tests as a new subserotype of Leptospira naam and was therefore designated as L. naam, subserotype dakotii.

Animals↗