bluuebunny/crossref_metadata_2025 huggingface dataset:
Reactivity of Antihuman Thymocyte Serum with Acute Leukemic Blasts
Authors: Mills Bonnie , Sen Luisa , Borella Luis
Published: December 2022

The presence of receptors for sheep erythrocytes (E) on some leukemic lymphoblasts is suggestive but not conclusive evidence for a thymic origin of these lymphoid cells. Normal and leukemic lymphoid populations were therefore examined to determine if the formation of spontaneous E rosettes correlated with the presence of thymus-associated cell surface antigens.

Lymphoblasts from four of 12 children with acute lymphoblastic leukemia (ALL) formed spontaneous E rosettes at 4°C and 37°C (E+) and bound rabbit antihuman thymocyte serum, as determined by indirect immunofluorescence and by an indirect radiolabeled antibody assay. The blasts from the other eight children did not form E rosettes at 4 or 37°C (E-) and did not bind antithymocyte serum. Absorption of the antiserum with peripheral blood leukocytes removed all detectable reactivity with normal peripheral blood lymphocytes, normal bone marrow cells, and E- bone marrow cells, but did not remove all reactivity with thymus cells or E+ leukemic blasts. Absorption of the antiserum with thymus from one donor removed all reactivity against thymus cells from another donor. After absorption with normal peripheral blood leukocytes, the antithymocyte serum reacted with E+ leukemic blasts but not with remission lymphocytes from the same patients. Thus, there are at least two distinct thymus-associated antigens—one that is present on T lymphocytes from peripheral blood as well as thymus and another that is present only on normal thymus and on ALL blasts that are identified by their ability to form E rosettes at 37°C.
Stimulation of Human Lymphocytes by Galactose-Specific Abrus and Ricinus Lectins
Authors: Closs Otto , Saltvedt Erling , Olsnes Sjur
Published: December 2022

Human lymphocyte cultures were incubated with the nontoxic abrus agglutinin and with ricin B chain, and the incorporation of 3H thymidine was measured. Abrus agglutinin stimulated strongly the thymidine incorporation whereas ricin B chain had a much lesser effect.

When galactose or lactose was added to the cultures together with the lectins, the abrus agglutinin and ricin B chain induced thymidine incorporation was strongly reduced. There was a linear relationship between the concentration of lectin and the concentration of lactose required for inhibition of lymphocyte stimulation. N-acetyl-galactosamine had a much lesser inhibiting effect and α-methyl-mannoside did not cause any inhibition.

The abrus agglutinin induced thymidine incorporation was not demonstrable before 36 to 40 hr and reached its maximum after 2 to 5 days. If lactose was added within the first 4 hr of incubation with abrus agglutinin no stimulation was observed.
Rosette Formation of Human Lymphoid Cells with Monkey Red Blood Cells
Authors: Pellegrino M. A. , Ferrone S. , Theofilopoulos A. N.
Published: December 2022

Studies of human peripheral lymphocytes (HPL) from healthy subjects and of human lymphoblastoid cell lines (HLC) revealed spontaneous formation of rosettes with Macaca speciosa monkey red blood cells (MRBC). The percentage of HPL forming rosettes with MRBC paralleled the percentage of HPL exhibiting markers ascribed to human B cells. The MRBC-rosetting cells were positive for membrane-bound immunoglobulin (MBIg). Furthermore, by employing a mixed rosette method, lymphocytes carrying both EAC3dmo and FITC-MRBC were encountered. In contrast, no lymphocytes carrying both SRBC and FITC-MRBC were observed. Ninety-six per cent of purified B cells obtained at the interface after interaction of HPL with SRBC and Ficoll-Isopaque centrifugation formed rosettes with MRBC. In contrast, only 8% of an enriched population of T cells obtained at the interface after interaction with EAC3dmo and Ficoll-Isopaque centrifugation formed MRBC-rosettes. Incubation of HPL with MRBC and subsequent centrifugation on Ficoll-Isopaque resulted in a highly enriched T cell population at the interface. Several lymphoblastoid cell lines with B cell characteristics, but not two cell lines with T cell characteristics, formed spontaneous rosettes with MRBC. Inhibition experiments showed that the structure on the lymphoid cell surfaces mediating the adherence of MRBC was independent of the MBIg as well as the receptors for IgGFc, C3b, and C3d. It appears that rosetting with MRBC will provide a new detection marker for human cells classified as B type and a technique for obtaining an enriched T cell population.
Immunologic Properties of Mast Cells from Rats Infected with Nippostrongylus Brasiliensis
Authors: Ishizaka Teruko , König Wolfgang , Kurata Munetsugu , Mauser Linda , Ishizaka Kimishige
Published: December 2022

The concentration of IgE in the serum of Sprague-Dawley rats increased after infection with Nippostrongylus brasiliensis (NB). The IgE concentration in normal rats was less than 1 µg/ml. After re-infection with NB, the concentration increased to 100 to 300 µg/ml. Mast cells were purified from peritoneal cells of both normal and NB-infected animals. Purified mast cells from the infected animals released histamine upon exposure to NB antigen. The antibody specific for IgE released histamine from purified mast cells of both normal and infected animals. Dose-response curves of histamine release suggested that mast cells from NB-infected animals bear more IgE molecules than normal mast cells. Binding of 125I-labeled rat E myeloma protein with normal mast cells was demonstrated by autoradiography. Under the same experimental conditions, mast cells of infected animals were not labeled with 125I-IgE. Mast cells from both normal and infected animals failed to combine 125I-labeled IgG. The number of IgE molecules bound per mast cell was determined by incubating 125I-labeled IgE with purified mast cells. When mast cells were incubated in 0.6 to 2 µg/ml of IgE, the number of IgE molecules combined with the mast cells from infected animals was about 10% of that bound with normal mast cells. The results indicated that a large proportion of IgE receptors on mast cells of infected animals was occupied by their own IgE. No significant difference was observed between normal mast cells and those of infected animals with respect to histamine content and intracellular levels of cyclic nucleotides.
Synergy Between Subpopulations of Normal Mouse Spleen Cells in the in Vitro Generation of Cell-Mediated Cytotoxicity Specific for “Modified Self” Antigens
Authors: Hodes Richard J. , Hathcock Karen S. , Shearer Gene M.
Published: December 2022

Responding lymphoid cells cultured in vitro with irradiated trinotrophenyl (TNP)-modified syngeneic spleen cells develop direct cell-mediated cytotoxicity which is specific for target cells bearing both the TNP moiety and histocompatibility determinants of the modified sensitizing cell. Two subpopulations of normal mouse spleen cells have been shown to synergize in the in vitro generation of specific cell-mediated cytotoxicity to these “modified self” antigens. The synergizing populations are nylon wool column-adherent and column-nonadherent fractions of normal mouse spleen. When mixtures of these two cell populations are cultured in vitro with irradiated TNP-modified syngeneic spleen cells, greater cytotoxicity is generated than would be expected from the sum of cytotoxic activities generated in the two populations sensitized separately. The synergizing cell in the column-adherent population is resistant to lysis by rabbit anti-mouse brain serum, is distinct from the cytotoxic effector T lymphocyte, and is unresponsive to phytohemagglutinin; its synergizing function could not be replaced by peritoneal cells. These results suggest that it is a non-T cell which may be distinct from the macrophage.
Inhibition of Macrophage-Lymphocyte Interaction by Cytochalasin B During Antigen Recognition by T Lymphocytes
Authors: Rosenthal Alan S. , Blake J. Thomas , Lipsky Peter E.
Published: December 2022

The effects of cytochalasin B on functional and physical macrophage-lymphocyte interaction have been examined. Cytochalasin B, an inhibitor of a variety of membrane activities blocks antigen-dependent binding of immune lymphocytes to macrophages and antigen-triggered lymphocytes proliferation if added at the initiation of culture. Cytochalasin B becomes progressively less inhibitory if addition is delayed by increasing intervals from the onset of culture. Under these conditions neither antigen handling by macrophages nor the proliferative response of lymphocytes to PHA is inhibited by cytochalasin B. These data are interpreted to suggest that cytochalasin B inhibits antigen-specific macrophage-lymphocyte interaction either by inhibition of an initial antigen-independent phase of macrophage-lymphocyte interaction or by interfering with a lymphocyte membrane event necessary for the interaction of the antigen-specific lymphocyte receptor with the macrophage-bound antigenic signal.
Decreased Antibody Response in the Offspring of Immunized High Responder Rats
Authors: Davis Bridgett K. , Gill Thomas J.
Published: December 2022

The antibody response to poly(Glu52Lys33Tyr15) in the rat is genetically controlled. Aggregation of this antigen with methylated bovine serum albumin decreases the amount of antibody made by high responder strains and increases that made by low responder strains. Immunization of female high responder rats with aggregated antigen before mating leads to a decrease in antibody formation in their offspring. Aggregation may protect the antigen from degradation and facilitate its transplacental passage so that it can interact with the developing lymphoid tissue of the fetus and alter its immune capabilities.
Lymphocyte Populations of AKR/J Mice III. Changes in the Preleukemic State
Authors: Zatz Marion M.
Published: December 2022

These studies suggest that the leukemogenic process in AKR mice selectively affects subpopulations of PHA and Con A responsive lymphocytes in the periphery and in the thymus, but that there is no generalized major deficit of T cells before leukemia development. These differential changes in spleen and thymus cell reactivity to mitogens may reflect a differential susceptibility of PHA and Con A responsive cells to infection by murine leukemia viruses.
An Analysis of the Defective Response of CBA/N Mice to T-Dependent Antigens
Authors: Janeway Charles A. , Barthold David R.
Published: December 2022

CBA/N mice fail to respond to T-independent antigens, and give a diminished response to T-dependent antigens. The cellular basis of the diminished response to T-dependent antigens in this strain was analyzed by means of adoptive transfers of various primed cell populations along with cells from the immunologically normal, histocompatible strain CBA/CaJ. CBA/N mice behaved normally as recipients and had normal levels of spleen helper cell activity. However, CBA/N spleen cells, used as a B cell source, produced only 56% as much antibody as CBA/CaJ cells. Since CBA/N mice have a relative frequency of immunoglobulin-bearing lymphocytes roughly half that of normal mice, these results suggest that CBA/N mice have a quantitative defect in spleen B cells responsive to T-dependent antigens.
Antigen-Enhanced Glucosamine Incorporation by Peritoneal Macrophages in Cell-Mediated Hypersensitivity
Authors: Hammond M. Elizabeth , Selvaggio Salvatore S. , Dvorak Harold F.
Published: December 2022

The interaction between sensitized lymphocytes and specific antigen occurring in classic delayed hypersensitivity causes guinea pig peritoneal macrophages to incorporate increased amounts of glucosamine into TCA precipitable, membrane-associated, cell surface material. Antigen-induced stimulation of glucosamine also occurred in peritoneal exudate cells (PEC) isolated from animals primed for cutaneous basophil hypersensitivity with certain strong antigens (KLH, vaccinia virus) in incomplete Freund's adjuvant (IFA), and lymphocytes from such animals elaborated MIF when cultured with specific antigen. Thus, the use of complete Freund's adjuvant is not obligatory for the induction of sensitized lymphocytes capable of secreting MIF or stimulating macrophage glucosamine incorporation; however, the potency of the immunogen employed is a critical variable since lymphocytes from animals primed with weaker antigens (HSA, BGG) in IFA did not have these capabilities.

Significantly enhanced incorporation of radioactive glucosamine by macrophages occurred when normal PEC were cultured in lymphokine-containing supernatants, but the magnitude of incorporation was smaller than that of sensitized PEC stimulated by antigen. The final 24 hr of macrophage culture was critically important because lymphokines were equally effective in promoting glucosamine incorporation when present for only this interval. The kinetics of this response are thus very similar to those reported for macrophage “activation”. The mechanism by which sensitized lymphocytes and their products stimulate glucosamine incorporation is not established, but at least part of the increment may be attributed to enhanced transport of glucosamine across the macrophage plasma membrane.

The plant lectins Con A and PHA stimulated unsensitized plastic-adherent cells to increased glucosamine incorporation and exerted a further additive stimulation on sensitized PEC when nonadherent sensitized lymphocytes were present. It is likely that these mitogens stimulate glucosamine incorporation by two distinct mechanisms, one involving sensitized nonadherent lymphocytes and a second involving only adherent cells (macrophages and/or plastic adherent lymphocytes).
Antigenic Determinants of the 70,000 Molecular Weight Glycoprotein of Woolly Monkey Type C RNA Virus
Authors: Hino Shigeo , Stephenson John R. , Aaronson Stuart A.
Published: December 2022

The 70,000 molecular weight glycoprotein (gp70) of a type-C RNA virus originally isolated from a woolly monkey has been partially purified and immunologically characterized. Evidence that this viral protein is viral coded was derived from studies showing its antigenic properties to be unaltered by virus passage in cells of different species. A broadly reactive competition immunoassay was developed utilizing antiserum prepared against feline leukemia virus to precipitate 125I-labeled woolly monkey virus gp70. Gibbon and woolly viruses, as well as feline and several mouse type-C viruses, all reacted with equal efficiency in this assay. In contrast, an endogenous virus of the baboon failed to cross-react, suggesting that viruses of this latter group are less immunologically related to the others. In a homologous competition immunoassay for the woolly viral glycoprotein, the woolly virus was readily distinguished from otherwise closely related viruses of gibbon apes. These findings demonstrate the pronounced type-specific antigenic determinants possessed by this viral protein. The antigenic determinants of gp70 responsible for neutralization have also been investigated.
Synthesis but not Secretion of J Chain by Variant Mouse Myeloma Cells Which Lose α-Chain-Synthesizing Ability
Authors: Mosmann Timothy , Baumal Reuben
Published: December 2022

We have devised a rapid method for obtaining large amounts of J chain from IgA in the ascitic fluid of mice bearing the MOPC 315 tumor. The J chain was released by reduction from the MOPC 315 IgA adsorbed onto a DNP-lysyl-Sepharose column, and was further purified by DEAE Sephadex chromatography. The mouse J chain was characterized as to its electrophoretic mobility, amino acid composition, apparent size, presence in different immunoglobulin classes, and reactivity with an antiserum containing anti-J chain activity.

Variant cell lines have been selected from the IgA-producing mouse myeloma cell line MOPC 315. The variants did not synthesize detectable quantities of α heavy chains but continued to synthesize and secrete light chains. J chain was synthesized by both parent and variant cell lines but only secreted by the parent cells. It is postulated that J chain synthesis is not dependent on α heavy chain synthesis, but that secretion of J chain by MOPC 315 cells occurs only because of its attachment to the IgA molecule.
Antigenic Analysis of Chlamydiae by Two-Dimensional Immunoelectrophoresis
Authors: Caldwell Harlan D. , Kuo Cho-Chou , Kenny George E.
Published: December 2022

Two-dimensional immunoelectrophoresis was utilized to study precipitins in hyperimmune rabbit serum made against chlamydiae and from patients with chlamydial infections. An antigen of Triton X-100-solubilized L2/434/Bu organisms with an electrophoretic mobility of 0.65 relative to bovine serum albumin at pH 8.6 was excised from the agarose gel of electrophorograms as antigen-antibody complexes and used to immunize rabbits. A monospecific antiserum to antigen 0.65 was obtained that reacted with Trachoma-LGV strains L2/434/Bu, B/TW-5/OT, and K/UW-31/Cx, but not with the mouse pneumonitis (Nigg) strain or the psittacosis strain meningopneumonitis (Cal-10). The Trachoma-LGV specificity of antigen 0.65 was further shown by indirect immunofluorescence straining with the monospecific antiserum of chlamydial inclusions in infected HeLa cells. Precipitins with a specificity for antigen 0.65 were identified in 15 of 18 sera from patients with diagnosed Chlamydia trachomatis infections LGV, trachoma, nongonococcal urethritis, and nongonococcal cervicitis by using monospecific antiserum to antigen 0.65 in the peak suppression test. Thus, antigen 0.65 appears to be a Trachoma-LGV-specific antigen that has considerable promise for serodiagnosis.
Alloantiserum-Mediated Suppression of Ir Gene Controlled Antigen-Induced Lymphocyte Activation: Effect on Uridine and Leucine Incorporation
Authors: Greenberg Philip D. , Bluestein Harry G.
Published: December 2022

Antisera to guinea pig histocompatibility antigens specifically suppress Ir gene-controlled antigen-stimulated, DNA-synthetic responses in vitro. To define further the mechanisms of alloantiserum-mediated suppression and to utilize this suppression as a probe of the cellular events occurring during lymphocyte activation, we have examined the effects of alloantisera on 14C-leucine and 3H-uridine incorporation, earlier events after antigen stimulation. The RNA and protein synthetic responses of peritoneal exudate lymphocytes from immunized guinea pigs to DNP-GL (controlled by a strain 2-linked Ir gene) are 40 to 50% of maximum by 24 hr in culture and at or near maximum by 48 hr. DNA synthesis was only 2% of maximum at 24 hr and maximum at 72 hr. Comparisons of the degree of stimulation of the incorporation of all three precursors reveals an excellent correlation between leucine and uridine but poor correlation between leucine and thymidine.

Despite the observed differences in time course of incorporation and magnitude of response, the susceptibility to suppression by alloantisera of all three responses was virtually identical. Anti-2 sera, added at the initiation of culture, completely suppress all three responses. If addition was delayed 8 hr, 50 to 60% suppression was still demonstrated, whereas only minimal suppression was evidenced if addition was delayed 18 hr. These results suggest that by 8 to 10 hr of incubation with antigen, the responding cells have undergone the necessary biochemical and structural changes to eventuate in an immune response; and, further - more, that alloantisera do not suppress by blocking antigen access to, or release from, its lymphocyte membrane receptor, but rather by a dynamic alteration of the lymphocyte membrane surface which interferes with the stimulus being generated by the antigen-receptor complex.
Lymphoid Cell Subpopulations
Authors: Howe Michael L. , Cohen Lewis
Published: December 2022

Mixtures of isogeneic lymph node cells (LNC) and thymocytes (TC) exhibit far greater responsiveness in the murine MLI, as measured by proliferation and development of cytotoxic effector cells, than either cell type cultured alone. Pretreatment of either LNC or TC with mitomycin-C or ultraviolet irradiation completely abolished their synergistic interaction. Administration of cortisone acetate to cell donors 20 hr before sacrifice reduced the capacity of LNC and enhanced the capacity of TC to synergize. The LNC and TC populations participating in synergy were found to be thymus dependent. LNC were shown to be responsible for the bulk of proliferative and effector activity observed in synergizing cultures, whereas TC appeared to amplify the activation of LNC. These findings provide the basis for a three cell model of MLI responsiveness.
Peanut Agglutinin, A New Mitogen That Binds to Galactosyl Sites Exposed after Neuraminidase Treatment
Authors: Novogrodsky Abraham , Lotan Reuben , Ravid Amiram , Sharon Nathan
Published: December 2022

Peanut agglutinin, purified by affinity chromatography, agglutinates lymphocytes from mouse, rat, guinea pig, and man only after their treatment with neuraminidase. However, it stimulates only neuraminidase-treated rat and human cells. A similar number of cell surface receptors for peanut agglutinin was found on neuraminidase-treated rat and mouse lymphocytes although the latter cells were not stimulated by the lection. Galactose specifically inhibited the agglutination and stimulation of lymphocytes by peanut agglutinin. Sequential treatment of lymphocytes with neuraminidase and β-galactosidase markedly reduced the response of the cells to stimulation by peanut agglutinin, soybean agglutinin, and galactose oxidase. It is suggested that the same galactosyl residue may be the target for the initial step in triggering lymphocytes by the above mentioned mitogens.
Anomalous Reactions of Mouse Alloantisera with Cultured Tumor Cells
Authors: Nowinski Robert C. , Klein Paul A.
Published: December 2022

Certain alloantisera prepared in mice against H-2 region membrane antigens were found to be unexpectedly cytotoxic for murine sarcoma and leukemia cells in culture. This anomalous cytotoxicity was shown to be the result of antibody in these alloantisera directed against the p15 and gp70 envelope proteins of MuLV which were present on the surface of the tumor target cells.

Sera from aged unimmunized mice of strains used for the preparation of alloantisera also contained antibodies against MuLV proteins p15 and gp70 that were cytotoxic for sarcoma and leukemia cells, which indicates that these antibodies occurred naturally in mice. These results independently confirm earlier findings of the widespread occurrence in mouse serum of antibodies reactive with MuLV.

The presence of antibody against MuLV in mouse serum which can cause cytotoxic reactions with tumor cells points to the fact that particular caution should be used during the typing of murine sarcomas or leukemias for cell surface antigens, since mouse antisera may yield cytotoxicity (or other serologic reactions) based on anti-MuLV specificities, rather than on anticipated antigens.
T Lymphocyte-Enriched Murine Peritoneal Exudate Cells
Authors: Schwartz Ronald H. , Jackson Lula , Paul William E.
Published: December 2022

The in vitro activation of murine thymus-derived (T) lymphocytes by soluble protein and synthetic antigens has been difficult to assess because of the lack of a specific and reliable proliferation assay. The present report describes the development of an assay system which overcomes these problems by making use of a population of nylon wool column-purified T lymphocytes obtained from thioglycol-late-induced peritoneal exudates of immunized mice. PETLES (peritoneal exudate, T lymphocyte-enriched cells) were composed mainly of T lymphocytes, eosinophils and small numbers of macrophages. Contamination with bone marrow-derived (B) lymphocytes averaged only 2%. When PETLES from immunized mice were stimulated in microtiter cultures with the immunizing antigen, large degrees of proliferation ensured as measured by incorporation of 3H-methyl-thymidine 5 days after initiation. As few as 1.25 × 104 cells and as little as 50 ng/ml of antigen gave significant stimulation. Maximum responses were obtained with 2 × 105 cells and about 100 µg/ml of antigen; dinitrophenylated-ovalbumin (DNP5OVA), in a series of 10 experiments under these optimal conditions, gave a mean incorporation of 70,900 cpm while the controls cultured without antigen showed only 3,600 cpm. PETLES from nonimmunized mice or from mice immunized to other antigens did not respond to DNP5OVA although they did respond to mitogens. The antigen-induced proliferation was shown to require the presence of immune T lymphocytes by two criteria: elimination of the response by treatment with anti-Thy 1.2 serum plus complement and failure to reconstitute the response when the few remaining immune B lymphocytes left after anti-Thy 1.2 treatment were added to nonimmune T lymphocytes. In addition, the system exhibited carrier specificity. Because of the paucity of B lymphocytes in the population, their contribution to the overall magnitude of the proliferative response was negligible as demonstrated by the small response to B cell mitogens. Thus, the assay appears to be a quantitative as well as a qualitative assay for one aspect of T lymphocyte function. This technique should prove useful for the study of murine T lymphocytes in vitro.
Cell-Mediated Immunity to Friend Virus-Induced Leukemia
Authors: Ting Chou-Chik , Bushar Grace Shiu , Rodrigues Dennis , Herberman Ronald B.
Published: December 2022

Cell-mediated cytotoxic reactivity of C57BL/6 mice against syngeneic FBL-3 cells, a Friend virus-induced leukemia, was measured by the 125IUdR release assay with tumor target cells in suspension. The tests could be performed either with Linbro tissue culture plates (16-mm wells) or with Microtest II plates (6-mm wells). The former could only be harvested manually; the latter could be harvested mechanically by an automatic harvesting apparatus which permitted larger scale tests. With either plate, it was found that careful preparation of the target cells and of the attacker cells has important effects on the results obtained. When performed under optimal test conditions, the 125IUdR assay can be used as a very simple, objective, and reproducible assay for testing cell-mediated cytotoxicity.
Initiation of C3 Cleavage in the Alternative Complement Pathway
Authors: Fearon Douglas T. , Austen K. Frank
Published: December 2022

The capacity of C3 and 125I-C3 to interact with B and D̄ to generate a C3 convertase was recognized by identification of C3b and Bb on immunoelectrophoresis and 125I-C3b on disc gel electrophoresis. Since the C3 was devoid of C3b as assessed by immunoelectrophoresis, alkaline disc gel electrophoresis, and isoelectric focusing, the initial C3 cleavage was not by the C3b-dependent C3 convertase. In addition, on isoelectric focusing C3 hemolytic activity and the capacity to permit B cleavage by D̄ were isoelectric at pH 6.4, whereas the capacity of C3b to permit cleavage by D̄ was isoelectric at pH 5.65. Comparison of the dose-response effects of C3b and C3 for B inactivation by D̄ revealed linear and sigmoidal relationships, respectively, consistent with formation of an initial C3 convertase independent of C3b followed by generation of the C3b-dependent convertase in the reaction initiated with native C3. Further, preincubation of C3 with C3bINA did not diminish its subsequent capacity to permit B inactivation by D̄ as compared to the introduction of C3bINA during the assay, thus supporting the view that native C3, B, and D̄ can form a convertase capable of generating initial C3b.
In Vivo Effects of Cortisone on the B Cell Line in Chickens
Authors: Rusu V. M. , Cooper M. D.
Published: December 2022

The effects of a single dose of cortisone acetate (5 or 10 mg/100 g body weight) on B cells were examined in young chickens. A dose-dependent increase in numbers of circulating B lymphocytes and a change in their Ig-class distribution were followed by parallel increase in splenic plasma cells and serum immunoglobulins. The higher dose of cortisone produced changes in Bµ and Bγ cells, whereas the lower dose primarily affected Bµ cells. These steroid-induced changes were preceded by lymphocyte depletion in the cortical regions of bursal follicles, and priorbursectomy prevented steroid-induced increases in circulating B lymphocytes and tissue plasma cells. The results suggest that cortisone can induce bursal lymphocytes to migrate from the bursa and to settle subsequently in peripheral lymphoid tissues where they become mature plasma cells.
Reappraisal of Lymphocyte Classification by Means of Surface Morphology
Authors: Baur Paul S. , Thurman Gary B. , Goldstein Allan L.
Published: December 2022

Prior scanning electron microscope studies have indicated that T (thymus derived) lymphocytes appear “smooth” while human B (thymus independent) lymphocytes appear “hairy” with numerous microvilli covering their surfaces. A recent report (24) has raised some doubt as to the validity of classifying lymphocyte subpopulations on this purely morphologic basis.

We have examined lymphocytes taken from congenitally athymic “nude” mice (lacking T cells) and compared them with those taken from normal littermates and CBA/J mice (both having T cell populations). We have determined that there are no distinguishable differences between teh distribution of surface morphologies of lymphocytes from animals either with or without T cells. Our studies indicate that the classification of T and B cells, based on cell surface features, requires more than just finding out whether or not the cells are smooth or hairy.
The Ontogeny of Murine B Lymphocytes
Authors: Hammerling Ulrich , Chin Ann F. , Abbott Joan , Scheid Margrit P.
Published: December 2022

When bone marrow and spleen cells of 4 week-old mice are fractionated on a discontinuous BSA gradient, a small fraction of Ia- cells is obtained which can be induced in vitro to express the Ia alloantigen within 2 hr. This is in precise parallel to the prothymocyte induction system of Komuro and Boyse. Ia specificity is ascertained by the use of two reciprocal antisera, A.TH anti-A.TL (anti-Iak) and A.TL anti-A.TH (anti-Ias), which yield the expected reaction pattern on induced bone marrow cells of (B6 × A)F1 (Iak) and SJL/J (Ias) mice. Induction can be effected by a number of agents, such as catecholamines, prostaglandin PGE1, cAMP, bacterial endotoxin, lipid A, ubiquitin, and thymopoietin. The last requires a 100-fold higher concentration for Ia+ induction as compared to prothymocyte induction, thus implying a lower affinity for the B cell receptor than for the thymocyte receptor.

Ia- to Ia- conversion involves cells different from prothymocytes, as indicated by: 1) the specific cytolytic effect of our anti-Ia sera which were shown to be free of activity against thymocytes; 2) an additive cytolytic effect of anti-Ia and anti-Thy-1 sera; and 3) the fact that the Ia inducible cells are sensitive to pretreatment with anti-immunoglobulin and C. The finding that Ia- precursor cells are already Ig+ is of interest for the B cell ontogeny, as it implies that Ig expression precedes Ia expression.
Niridazole A Potent Long-Acting Suppressant of Cellular Hypersensitivity
Authors: Pelley Ronald P. , Pelley Robert J. , Stavitsky Abram B. , Mahmoud Adel A. F. , Warren Kenneth S.
Published: December 2022

Niridazole, an anthelminthic drug, has been shown to be a potent long-acting suppressant of cell-mediated immune responses both in man and in experimental animals. In the present study the effect of niridazole on the magnitude and kinetics of the primary and secondary antibody responses of mice to sheep erythrocytes, human serum albumin, and keyhold limpet hemocyanin was investigated. The therapeutic dose of niridazole for murine schistósomiasis moderately and transiently inhibited the primary antibody response to these antigens, but had no effect on ongoing antibody production against keyhold limpet hemocyanin or on secondary responses to any of the antigens. Single doses of niridazole (which suppress cell-mediated immune responses) had no effect whatsoever on the most strongly inhibited of the primary antibody responses, that to alumprecipitated polymerized human serum albumin.
Goat Antibody Directed Against a Human Leb Blood Group Hapten, Lacto-N-Difucohexaose I
Authors: Zopf David A. , Ginsburg Ann , Ginsburg Victor
Published: December 2022

A goat immunized with lacto-N-difucohexaose I (Fucα1-2Galβ1-3GlcNAcβ1-3Galβ1-4Glc) conjugated to polylysine produces antibody directed against the oligosaccharide. The antibody specifically precipitates Leb blood group substance and specifically binds lacto-N-difucohexaose I . Antibody purified by affinity chromatography is a 6.8S immunoglobulin of 158,000 ± 6,000 molecular weight.
Human Leukemia-Associated Antigens: Detection on Cells of Established Lymphoblastoid Lines
Authors: Mohanakumar T. , Pauly John L. , Sokal Joseph E. , Metzgar Richard S.
Published: December 2022

Primate and rabbit antisera to different morphologic classes of human leukemia cells, after appropriate absorptions, detected leukemia-associated antigens present on cultured lymphoblastoid cell lines derived from leukemia patients. The primate antisera distinguished antigens on cells derived from myeloid leukemia patients from those on cells derived from lymphocytic leukemia patients. Of particular interest was the fact that antigens of myeloid leukemia, but not of lymphatic leukemia, were detected on lymphoid cell lines established from blood of patients with myeloid leukemia. One of four lymphoblastoid cell lines derived from normal donors expressed antigens of lymphatic leukemia. Leukemia-associated antigens were not found on the HRIK lymphoblastoid line derived from a Burkitt's lymphoma patient on skin fibroblasts or HeLa cells. Expression of these antigens on cultured cells derived from leukemia patients could not be related to the presence of the EB virus or the EB virus genome. Rabbit antisera detected antigens common to cells from patients with myeloid and lymphocytic leukemia. Absorption experiments demonstrated that the antigens detected on cell lines derived from leukemia patients are similar to those detected by the primate and rabbit antisera on fresh peripheral blood leukemic cells. The serologic detection of leukemia-associated antigens on lymphoblastoid cell lines indicates that some of these cultures contain cells with antigenic properties similar to those of human peripheral blood leukemic cells.
Cytotoxic T Cell Activity is Strain-Specific in Outbred Mice Infected with Lymphocytic Choriomeningitis Virus
Authors: Zinkernagel Rolf M. , Dunlop Malcolm B. C. , Doherty Peter C.
Published: December 2022

The cytotoxic T cell response in outbred mice infected with lymphocytic choriomeningitis virus (LCMV) is strain specific. The same is true for adoptive transfer of fatal LCM disease. The response of individuals within an outbred strain is completely cross-reactive, as shown by using immune lymphocytes and virus-infected macrophage targets from individual mice. Reciprocal exclusion of cytotoxic T cell activity between inbred and outbred mouse strains is the rule, the exception being one strain (H) known to have some C57BL ancestry. Immune T cells from one of 7 H mice specifically lysed LCMV-infected C57BL macrophages. Experiments with inbred mice have shown that only one allele need be shared at either the H-2K or H-2D locus for cytotoxic T cell activity to be manifest. Adoptive transfer protocols may thus be considered in outbred situations, providing that T cells are effective before allograft rejection occurs. Also, the LCMV cytotoxic T cell assay may be useful for determining the degree of H-2 variability in wild mouse populations, as novel H-2 types can be detected and mice need not be congenic.
Rapid Isolation of Antigens from Cells with A Staphylococcal Protein A-Antibody Adsorbent: Parameters of the Interaction of Antibody-Antigen Complexes with Protein A
Authors: Kessler Steven W.
Published: December 2022

The Cowan I strain of the bacterium Staphylococcus aureus has been used as an adsorbent for antibodies complexed with radiolabeled antigens from cell lysates. This application is advanced as a superior alternative to other methods of immune precipitation for the isolation of antigens. It exploits the high adsorption capacity for IgG molecules by protein A molecules on the cell walls of certain strains of staphylococci, along with the advantageous sedimentation properties of the bacteria. The interaction of immune complexes with the adsorbent was defined initially using a model system of bovine serum albumin with a high excess of rabbit anti-bovine serum albumin antibodies (IgG). The uptake of immune complexes under these conditions was extremely rapid, occurring within seconds, whereas maximum binding of free IgG was much slower. In addition, once bound the complexed antigen could not be displaced from the adsorbent either by large amounts of normal IgG or by extra free antibody. Antigen could be eluted almost completely from the inert adsorbent for analytic or preparative purposes with a variety of solvent systems, such as the detergent SDS in combination with urea and high temperature, and neutral salts with strong lyotropic salting in properties.

The efficacy of the protein A-antibody adsorption technique was tested in direct comparisons with a conventional double antibody precipitation method for the isolation of mouse lymphocyte IgM. The bacterial adsorbent not only had a distinct advantage in speed of antigen isolation, but analyses by polyacrylamide gel electrophoresis in SDS also revealed consistently higher antigen recoveries, lower levels of background radioactivity, and an absence of other cell components which may nonspecifically bind to and complicate analyses using conventional immune precipitates.
Effects of Anti-Ia Serum on Mitogenic Responses
Authors: Niederhuber John E. , Frelinger Jeffrey A. , Dugan Elizabeth , Coutinho A. , Shreffler D. C.
Published: December 2022

Specific anti-Ia serum in combination with rabbit complement removed the B lymphocyte population normally responsive to LPS mitogenic stimulation. Significant inhibition of this proliferative response was also obtained by brief periods of cell pretreatment with anti-Ia antibodies, but not with specific anti-H-2K antibodies. The population of splenic B cells resistant to anti-Thy-1.2 serum and complement was blocked in a very similar manner by anti-Ia serum, but not by anti-H-2 serum.
Functional Deficiency of Splenic Adherent Cells in New Zealand Black Mice
Authors: McCombs Candace , Hom Joanne , Talal Norman , Mishell Robert I.
Published: December 2022

New Zealand Black (NZB) mice hyporespond after in vitro immunization to sheep erythrocytes but not after in vivo immunization. The difference between in vitro and in vivo immunization was found to involve a non-antigen-specific change in the spleen which occurs in vivo soon after priming. Adherent spleen cells from young NZB or control strain mice were compared in their ability to cooperate with nonadherent cells in the induction of a primary immune response to sheep erythrocytes in vitro. The results support the hypothesis that there is a functional deficiency of adherent cells in the spleens of unprimed NZB mice.
Chlamydia Trachomatis Immunotype J
Authors: Wang San-Pin , Grayston J. Thomas
Published: December 2022

A new immunotype J is proposed for a group of Chlamydia trachomatis strains which are related to trachoma type C in the microimmunofluorescence typing test. Ten immunologically identical strains have been identified from four separate areas of the world. The strains were either isolated from the genital tract or from eye infection originating from the genital tract. They grow readily in HeLa 229 cell culture but poorly in egg culture. Whereas two-way cross-reactions were found between types C and J, tests with mouse antisera cross-absorbed with heterologous antigens showed that type J is completely separable from type C. The J strain caused typical follicular conjunctivitis in monkey eyes. Both humans and monkeys infected with J strains developed type-specific micro IF antibody patterns similar to immunized mouse antisera. C antisera showed broader and higher titer heterologous reactions than J antisera.

Two immunologically identical strains, UW-61/Cx and 469/OC, related to both C and J, remain unclassified. They immunologically bridge the two types but are more closely related to type C immunologically and at the same time resemble type J epidemiologically and biologically.
Partial Characterization of Murine Migration Inhibitory Factor (MIF)
Authors: Kühner Alice L. , David John R.
Published: December 2022

These studies describe the production of murine migration inhibitory factor (MIF)3 in sufficient quantities to allow its partial characterization by physiochemical and enzymatic methods. MIF was obtained from murine spleen cell cultures (C57BL/6 strain) stimulated with concanavalin A (Con A). Characterization of murine MIF was performed using Sephadex G-100 gel chromatography, isopycnic centrifugation in a CsCl density gradient, polyacrylamide disc electrophoresis, heat stability, and enzymatic treatment. MIF-containing and control fractions were assayed on normal C57BL/6 peritoneal exudate cells by using a microcapillary tube assay.

Peak MIF activity was found in a Sephadex G-100 fraction containing molecules the size of albumin and slightly smaller, molecular weight 67,000 to 48,000. Murine MIF was stable to heating at 56°C for 30 min but lost its activity at 80°C for 30 min. Incubation of G-100 fractions containing MIF with water insoluble chymotrypsin destroyed the activity of MIF, indicating its protein nature. CsCl density gradient centrifugation revealed that murine MIF had a buoyant density greater than protein, consistent with its being a glycoprotein. Further, when subjected to disc electrophoresis on polyacylamide gels, murine MIF migrated in a region cathodal to albumin. Thus, mitogen stimulation of murine spleen cells produced MIF in quantities which allowed its partial characterization and purification, and its comparison with human and guinea pig MIF; this makes it feasible to analyze the role of murine MIF in cellular immunity and in its relationship to lymphocyte mediators which regulate humoral immune responses.
The Effect of Heat-Killed Streptococci on the Survival of Heart Grafts in Inbred Strains of Mice
Authors: Davis William C. , Pachman Lauren M.
Published: December 2022

Studies with inbred strains of mice revealed that exposure to type 12, Group A, β-hemolytic streptococci affects the host response to heart grafts implanted in the ear. Intraperitoneal injection of streptococci 10 days before grafting led to curtailed survival of syngrafts without altering the normal rejection time of allografts. Similar sensitization, combined with local injection of streptococci into the graft site at the time of grafting, was followed by rapid rejection of both syngrafts and allografts. The time interval between exposure and grafting was critical. Injection with streptococci 5 days before grafting led to a prolonged survival of allografts and no demonstrable effect on syngrafts. In contrast, injection of streptococci 15 days before grafting did not alter survival of either type of graft. The data indicate heart grafts implanted in the ear may serve as a useful model for the study of the host responses to streptococcal antigens.
The Effects of External K+ and Na+ on the Chemotaxis of Rabbit Peritoneal Neutrophils
Authors: Showell Henry J. , Becker Elmer L.
Published: December 2022

Extracellular K+ enhances the chemotactic responsiveness and spontaneous movement of rabbit peritoneal neutrophils but is not required for these functions. Other monovalent cations act the same; the rank order of their effectiveness is K+ = NH4 τ; Rb+ Cs+ τ; Li = Na. The K+ specific ionophore, valinomycin (10-7 M) inhances chemotaxis in the presence of K+ but not in its absence; another K+-specific ionophore, nigericin (10-7 M) inhibits chemotaxis in the absence of K+ but not in its presence. Ouabain (5 × 10-6 M) prevents the enhancing effects of K+ on chemotaxis. Removing the Na+ of the buffer and substituting it with K+, choline or glucose greately enhances spontaneous motility but depresses chemotactic activity. One hypothesis suggested by the above results is that as a part of their action, chemotactic factors stimulate a net influx of K+ into the neutrophil; and alternative or additional hypothesis is that chemotactic factors stimulate a net efflux of Na+ from the neutrophil.
In Vitro Immune Response to the 2,4,6-Trinitrophenyl Determinant in Aged C57BL/6J Mice: Changes in the Humoral Immune Response to, Avidity for the TNP Determinant and Responsiveness to LPS Effect with Aging
Authors: Kishimoto Susumu , Takahama Toshikatsu , Mizumachi Hachiro
Published: December 2022

An in vitro anti-TNP response of the spleen cells from aged C57BL/6J mice showed approximately 4-fold less PFC than did that from young adult mice. Anti-θ serum-treated young spleen cells gave an anti-TNP response that was definitely greater than the response of the anti-θ serum-treated aged spleen cells in the presence of the exogenous activated thymus cells as helper cells. These results suggested that the deficits in B cells may be partly responsible for the imparied anti-TNP response of the aged spleen cells.

To examine further the capacity of stem cells in the bone marrow to generate B cells responsible for anti-TNP response in the spleen, we injected i.v. 1.5 to 2.0 × 107 bone marrow cells from young or aged mice into lethally irradiated syngeneic recipients that had previously been thymectomized. Four to 6 weeks later, 107 spleen cells from the two groups of these recipient mice were immunized with TNP-SRBC in the presence of the exogenous activated thymus cells and assayed for anti-TNP PFC. The response of the aged marrow-derived B cells was approximately one-half of that of the young marrow-derived B cells.

The avidity for TNP determinant of the antibodies produced by the PFC was determined by the plaque-inhibition technique. The avidity of the antibodies produced by the aged mice was approximately 33 times lower than that by the young mice.

Anti-TNP response of the young spleen cells were markedly enhanced by the addition of LPS to the cultures, whereas no or little enhancement of the response was induced in the aged spleen cells even in the presence of high concentration of LPS. In contrast, DNA synthesis of both the young and aged spleen cells was comparably stimulated by 1 µg/ml and 10 µg/ml of LPS, however, it was rather less in the aged spleen cells at a concentration of 100 µg/ml.

Mechanisms responsible for the changes in avidity and responsiveness to LPS with aging are discussed.
K Cell Cytotoxicity against Antibody-Coated Chicken Erythrocytes in Tumor-Bearing Mice: Its Development with Progressively Growing Tumor and the Effect of Immunization against the Tumor
Authors: Ghaffar A. , Calder Elizabeth A. , Irvine W. J.
Published: December 2022

Antibody-dependent (K cell) cytotoxic activity of spleen cells from mice bearing a chemically induced fibrosarcoma has been studied by using antibody-coated chicken erythrocytes as target cells. Spleen cells from tumor-bearing animals caused a significantly greater degree of target cells destruction than did those from control animals. The elevated cytotoxic activity in tumor-bearing animals increased with time after the tumor inoculation and correlated directly with the size of the tumor. The development of increased cytotoxic activity could be circumvented by surgical removal of the tumor. Mice that received x-irradiated tumor cells or x-irradiated tumor cells followed by a live challenge did not show a tumor growth and also failed to show increased K cell cytotoxic activity. It has been concluded that the increased K cell activity results directly from the active growth of the tumor. The role of K cells in immunosurveillance has been discussed.