Interactions between human lymphocytes and paramyxovirus-infected cells: adsorption and cytotoxicity.

The capacity of human lymphocytes to adhere to paramyxovirus-infected monolayers and their capacity to kill paramyxovirus-infected cells was investigated. A large fraction of human lymphocytes was found to adhere firmly to the paramyxovirus-infected monolayers. Predsorption of lymphocytes on mumps virus-infected cells impaired their adsorption to a second cell monolayer of the same type.

A new rat lymphocyte surface marker: characterization and separation of cells with receptors for Helix pomatia hemagglutinin.

Thirty-two percent of neuraminidase-treated DA rat spleen lymphocytes and 48% of lymph node lymphocytes possess receptors for Helix pomatia hemagglutinin (HP). Moreover, these HP-receptor-bearing cells can be separated from B cells by affinity chromatography on HP-Sepharose columns. The virtual absence of immunoglobulin (Ig) receptors and the close correlation with reported T-cell content of these lymphoid tissues suggest that HP-receptor lymphocytes are probably T cells and that HP may provide a convenient marker, for both the identification and the purification of rat T lymphocytes.

Complement-dependent cytotoxicity against hepatoma cells mediated by IgM antibodies in serum from tumor-bearing rats.

Complement-dependent cytotoxic antibodies in syngeneic serum from rats carrying transplanted aminoazo dye-induced hepatomas (D23 and D33) and from rats immunized with irradiated tumor cell or homogenates were studied by a short-term 51Cr release assay. The tumor-bearer sera (TBS) were subjected to chromatography on unsolubilized protein A and Sepharose 4B. The cytolytic activity of D23 TBS was recovered in the IgM molecular weight region, whereas no such activity was obtained in the IgG fraction.

Immunological studies in ulcerative colitis. VIII. Antibodies to colon antigen in patients with ulcerative colitis, Crohn's disease, and other diseases.

Sera from patients with ulcerative colitis or Crohn's disease had elevated titers to colon antigen from germ-free rats significantly more often than sera from patients with gastroenteritis, irritable colon, non-gastrointestinal diseases, and healthy controls. Elevated anticolon titers in significant frequency were also found in patients with liver cirrhosis, urinary tract infections, and in polyposis coli and their relatives. Females with ulcerative colitis had, on an average, higher titers than men especially in the age group 30 years and over.

Fractionation of human T lymphocytes on wheat germ agglutinin-sepharose.

T cells from human peripheral blood was purified by fractionation on columns charged with human immunoglobulin and rabbit anti-human immuno-globulin. When assayed with 125I- or fluorescein isothiocyanate-labeled wheat-germ agglutinin (WGA), a weakly binding and a strongly binding subpopulation could be distinguished. These T-cell subpopulations were fractionated on columns charged with WGA, convalently bound to Sepharose 6MB.

Analysis by a plaque assay of IgG- or IgM- dependent cytolytic lymphocytes in human blood.

When monolayers of bovine erythrocytes (Eb) were exposed to purified human blood lymphocytes and either IgG or IgM fractions of rabbit anti-Eb serum, clear zones (plaques) appeared when Eb had been lysed by antibody-dependent effector cells (K cells). IgG-dependent plaque formation was complete by 20 h of incubation, while the IgM-dependent reaction required 40 h. The estimated minimal numbers of plaque forming cells (PFC) were 5.

Receptors for Helix pomatia A haemagglutinin on leukaemic lymphocytes from patients with chronic lymphocytic leukaemia (CLL).

Blood lymphocytes from thirteen patients with CLL were studied for surface-bound Ig (SIg), Fc receptors (EA rosettes), receptors for sheep erythrocytes (E rosettes) and receptors for Helix pomatia A haemagglutinin (HP), a carbohydrate-binding protein with specificity for N-acetyl-D-galactosamine and related sugars. Fluorescein-labelled HP binds to subpopulations of human peripheral blood lymphocytes (PBL) treated with neuraminidase. In normal peripheral blood, HP binds to the T-lymphocytes while the majority of the B cells bearing surface-bound immunoglobulin do not have receptors for HP.

Interaction of K lymphocytes with myeloma proteins of different IgG subclasses.

Human myeloma proteins of the four IgG subclasses and their Fc, F(ab)2, and Fab fragments were tested for their ability to inhibit antibody-dependent human K lymphocyte-mediated cytotoxicity to chicken erythrocytes (CRBC) sensitized with specific rabbit antibodies. In addition, the adsorption of K cells onto glass bead columns coated with myeloma proteins was investigated. Myeloma proteins and their Fc fragments of all four subclasses inhibited K cell activity.

Fractionation of human blood lymphocytes on Helix pomatia A haemagglutinin coupled to sepharose beads.

Treatment of human blood lymphocytes with neuraminidase has previously been shown to uncover receptors for the A haemagglutinin of the snail Helix pomatia (HP). Neuraminidase-treated lymphocytes were now fractionated on columns charged with large Sepharose particles to which HP had been coupled covalently. HP-receptor negative (HP-) lymphocytes passed the columns while HP-receptor positive (HP+) lymphocytes were retained.

Purification, fractionation and assay of antibody-dependent lymphocytic effector cells (K cells) in human blood.

In this article we present methods for the purification and fractionation of human blood lymphocytes, which have been used in our laboratory to characterize antibody-dependent cytotoxic effector cells (K cells). The assay system consists of highly purified lymphocytes, 51Cr-labelled chicken erythrocytes (Ec) and IgG rabbit anti-Ec in high dilutions. Various ways of comparing K-cell potentials of different lymphocyte preparations in this system are discussed.

Differential cytotoxicity of activated lymphocytes on allogeneic and xenogeneic target cells. III. Species specificity of lymphocyte target cell recognition in vitro.

Cytotoxic effector lymphocytes were produced by stimulation of human peripheral blood or mouse spleen lymphocytes with PPD, PHA or PWM in vitro. The specificity of the lymphocyte target cell interaction was studied in vitro. The specificity of the lymphocyte target cell interaction was studied by adsorption of effector cells on various target cell monolayers.

The interaction of nonmitogenic and mitogenic lectins with T lymphocytes: association of cellular receptor sites.

The relationship between the surface receptors on neuraminidase-treated human blood lymphocytes for the mitogenic lectins Phaseolus vulgaris leukoagglutinin (La), concanavalin A (Con A) and soy bean agglutinin (SBA) and the non-mitogenic lectin Helix pomatia A hemagglutinin (HP) was investigated. Two different techniques, co-capping with different fluorochrome-labeled lectins and cell binding-inhibition experiments with 125I-labeled lectins, were used. The results demonstrated that the nonmitogenic lectin HP and the mitogenic lectins SBA, La and Con A bind either to the same macromolecule (s) or to different but physically linked macromolecules on the surface of human T lymphocytes.

Effect of radiotherapy on lymphocyte cytotoxicity in vitro.

The cytotoxic functions of highly purified blood lymphocytes from patients with breast cancer were studied before and after radiotherapy. Addition of PHA or of rabbit antibodies to target cells (chicken erythrocytes) were chosen as two means of inducing lymphocyte cytotoxicity in vitro. The proportion of T and non-T-lymphocytes was determined by means of E and EAC rosette tests.

Cellular immunity to transitional cell carcinoma of the urinary bladder. III. Effects of hydrostatic pressure therapy.

The effects of hydrostatic pressure therapy on in vitro cellular cytotoxicity responses have been studied in 19 patients with transitional cell carcinoma of the bladder (TCC). Cytotoxicity was quantitated against allogeneic targets in a microplate assay or by 51chromium isotope release. Two types of reactivity were detected, the most common being a differential cytotoxicity for targets derived from TCC, in either short-term or long-term tissue culture.

Human lymphocyte cytotoxicity against mumps virus-infected target cells. Requirement for non-T cells.

Subpopulations of human lymphocytes were tested for their capacity to kill mumps virus-infected target cells in a 51-chromium release asaay. Using two different cell fractionation techniques, lymphocytes were fractionated into T cell-enriched (primarily T cells) and T cell-depleted (primarily B cells) subpopulations. Filtration of lymphocytes through columns coated with human immunoglobulin and rabbit anti-human-immunoglobulin (Ig-anti-Ig) rendered the resulting T-cell preparation inactive as effector cells against target cells carrying mumps virus antigens.

Soluble and membrane-bound enzyme-active antigens of rat-liver lysosomes.

Secondary lysosomes were isolated from rat liver and separated into a soluble and a membrane fraction. Plasma membranes and microsomes were also isolated and antisera against the various fractions were prepared in rabbits. Lysosomal content and detergent-solubilized membrane fractions were analysed in two-dimensional immunoelectrophoresis (crossed immunoelectrophoresis).

Cytolytic lymphocytic cells with complement receptor in human blood. Induction of cytolysis by IgG antibody but not by target cell-bound C3.

Human blood lymphocytes were fractionated on glass bead columns charged with sheep erythrocyte (Es) membranes-bearing human C3b (7,000-10,000 molecules/Es). In the passaged cells the proportion of C receptor lymphocytes was strongly reduced, in parallel with the capacity to lyse chicken erythrocytes (Ec) in the presence of IgG-rabbit anti-Ec antibody. In other experiments, lymphocytes forming rosettes with Es bearing activated rabbit complement [C(ra)] from C6-deficient rabbits were removed by centrifugation through human serum albumin-gelatine mixtures.

In vitro target cell lysis mediated by normal human lymphocytes. Influence of molecular properties and affinity of inducing antibody.

Lysis of DNP-coated chicken erythrocytes by human blood lymphocytes (K cells) was induced by means of rabbit anti-DNP antibodies. Antisera were prepared by injecting the animals with DNP-conjugated proteins emulsified in Freund's complete adjuvant. An ammonium sulphate precipitation technique was used for assay of antibody concentration and affinity.

Destruction of dextran-coated target cells by normal human lymphocytes and monocytes. Induction by a human anti-dextran serum with IgG antibodies restricted to the IgG2 subclass.

A human anti-dextran serum, EAK, with IgG antibodies restricted to subclass IgG2, was tested for its capacity to induce lysis of dextran-coated chicken erythrocytes by normal human lymphocytes or monocytes. Another human anti-dextran serum, RGM, with most antibodies belonging to sublass IgG1, and a hyperimmune rabbit anti-dextran serum were used for reference. In lymphocyte-mediated erythrolysis, serum EAK gave rise to 51-Cr release varying from 20% to 80% in different experiments.

A plaque technique for assay and characterization of antibody-dependent cytotoxic effector (K) cells.

Using monolayers of erythrocytes as target cells, a plaque assay was developed by which individual antibody-dependent cytolytic lymphoid cells (K cells) could be identified at the cellular level. The extent of plaque formation depended on the concentration of sensitizing antibody, time of incubation, and number of lymphocytes added. Most of the plaque-forming cells (50% to 70%) were shown to possess receptors for activated complements, a small but variable fraction (5% to 25%) had surface Ig detectable by indirect immunofluorescence, and 5% to 10% bound sheep erythrocytes at 5 degrees C.

Contamination of anti-immunoglobulin reagents with antibodies to beta 2-microglobulin and other unrelated antigens. Effects on immunofluorescence staining of human lymphocytes.

IgG fractions from three of four rabbit antisera to Bence Jones proteins of chi-type were found to contain antibodies to beta 2-microglobulin and to stain 80%-100% of human blood lymphocytes by indirect immunofluorescence. Antibody fractions from these sera, which contained anti-beta 2-microglobulin but not anti-Ig, stained all lymphocytes, whereas the isolated anti-Ig antibodies (anti-chi) stained only a minor cell population. In both instances, the specificity of the staining was confirmed by absorption experiments.