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Biomedical subjects

P Perlmann

Publications and source records attributed to P Perlmann.

At least 253 records · Page 14Linked to original sources
PubMed1976

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. However, there were wide variations within a given subclass and IgG2 and IgG4 proteins usually inhibited less than IgG1 and IgG3 proteins. Aggregation of the weakly inhibitory proteins with bis-diazotized benzidine increased their inhibitory effect. An IgG1 half-molecule with a deletion in the Cgamma3 domain was weakly inhibitory. Passage of lymphocytes through glass bead columns coated with IgG1 and IgG3 proteins removed K cell activity. In contrast, columns coated with IgG2 and IgG4 proteins, even when aggregated with BDB, failed to absorb K cells but removed significant numbers of SIg positive B lymphocytes. An enhancement of the antibody-dependent cytotoxicity was observed in 34% of the inhibition experiments in the presence of low concentrations of the weakly inhibitory proteins, usually IgG2 and IgG4. This enhancement occurred more frequently (53% of the experiments) with Fc fragments independent of the subclass. Moreover, addition of IgG2 and IgG4 but not IgG1 and IgC3 fragments induced a dose-dependent cytotoxicity to CRBC in the absence of anti-CRBC antibodies. These data indicate that IgG2 and IgG4 proteins have a lower affinity to K cells than IgG1 and IgG3 proteins and are compatible with an earlier hypothesis that proposes that more than one site on the Fc fragment can react with Fc receptors. The present results suggest in addition that there may be functionally different sites, one having a triggering function in K lymphocyte lysis that may be localized on the second constant domain and one being responsible for high affinity binding of IgG to cell receptors that is probably localized on the third constant domain.

Animals↗
PubMed1976

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. In untreated CLL, HP binds to 90-100 percent of the neuraminidase-treated PBL. Almost all of the SI-G-POSITIVE CELLS IN CLL patients also have receptors for HP. Two groups of patients were found: in one the total fraction of SIg+ cells was less than or equal to 50 percent and about 30 percent of these lost their Ig during incubation at 37 degrees C. No such loss of SIg was revealed in the remaining patients where total SIg+ fraction was approximately 70 percent. These patients usually had higher blood lymphocyte counts, probably reflecting a more advanced disease. CLL patients in remission with low numbers of leukaemic cells also had low numbers of blood lymphocytes carrying both SIg and HP-receptors. It is concluded that leukaemic cells carry both HP receptors and SIg. Testing of this combination therefore provides a valuable new tool for monitoring patients with CLL.

Aged↗
PubMed1976

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. The latter cells were eluted by addition of the competitive hapten N-acetyl-D-galactosamine (D-GalNAc). The total yield of cells recovered after fractional was 60-80% Surface marker studies indicated that there was no selective loss of any of the major lymphocyte subpopulations. The fraction that passed the columns (fraction I) consisted of approximately 10% of all lymphocytes. It contained approximately 1% HP+cells and approximately 3% of all lymphocytes forming rosettes which sheep erythrocytes (E+ cells) present before fractionation. 50-55% of the lymphocytes in this fraction had surface-bound immunoglobulin (SIg+ cells) and complement receptors (EAC+ cells). Of the SIg+ cells, approximately 60% were true B cells while the remaining 40% had IgG adsorbed to their surface. The majority of the B cells were recovered in this fraction. The lymphocytes of this fraction responded poorly to T-cell mitogen but had an enhanced K-cell activity to chicken erythrocytes. Elution of the cells retained on the column with 0.1 mg/ml D-GalNAc gave a fraction II, consisting of approximately 15% of all lymphocytes. This fraction had a mixed composition. The majority of the cells (approximately 45%) were recovered by subsequent elution with 1.0 mg/ml D-GalNAc. This fraction III was strongly enriched with HP+ and E+ cells (T cells). About 10% of the HP+ cells in this fraction were SIg+. However, on the majority of these cells this surface-bound immunoglobulin was probably externally absorbed IgG. These HP+-SIg" cells were also EAC+ and had Fc receptors, as shown by rosette formation with IgG-coated bovine erythrocytes. The lymphocytes of fraction III responded most strongly to T-cell mitogen while their K-cell activity was weak.

Cell Membrane↗
PubMed1976

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. In contrast, only part of beta2-microglobulin (beta2-m) or beta2-m-bearing complexes, appear to be physically linked to the lectin receptor complex(es). On the lectin-binding substance(s) at least two saccharide structures were recognized, one of which binds both HP and SBA and another which binds SBA and La (and probably also Con A) but not HP.

Binding Sites↗
PubMed1975

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. This reaction is operationally termed "tumor-specific". Less frequently, a general cytotoxicity for targets of diverse histogenic origins was observed. For the present, this is termed "non-specific". Nine patients were tested before pressure therapy and of these only two gave a specific reaction and one a non-specific reaction, while six were non-reactive. Eighteen patients were tested at varying intervals after treatment and of these 11 gave a specific reaction and one a non-specific effect, while six were non-reactive. Three individuals who were non-reactive prior to therapy had a specific reaction post therapy. Two who reacted specifically before therapy became non-reactive post therapy. The results of serial in vitro testing for cytotoxicity are presented with individual case histories, tumor staging and grading and the clinical outcome of hydrostatic pressure therapy.

Adult↗
PubMed1975

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). The immunoprecipitates were stained by histochemical procedures for different enzyme activities such as phosphatases, non-specific esterase, arylsulphatase, glycosidases and L-leucyl-beta-naphthylamidase. When lysosomal content was tested against its corresponding antiserum, 17 different precipitates could be seen. Most of the enzyme activities tested were shown to reside separately in one or a few precipitates each. In contrast, when the membrane extracts were investigated, a more polymorphic pattern of enzyme-active precipitates appeared. Thus, when lysosomal membrane extracts were reacted with homologous antiserum 11 precipitates with acid phosphatase activity were obtained. Several of the antigens were electrophoretically different and immunologically non-identical. As expected from the biology of secondary lysosomes, many of their antigens were also found in microsomes and/or plasma membranes, but several antigens unique for lysosomes were detected concomitantly. Closer analysis of these results indicated that several seemingly identical enzyme-active proteins occurred both in soluble and membrane-associated forms. However, while many of the membrane antigens expressed 2-4 different enzyme activities, only one activity was detected in individual precipitates of the lysosomal content. Thus, acid phosphatase activity was found together with esterase activity in three membrane-associated antigens. The precipitates formed by two of these also stained for arylsulphatase and nucleoside tri-, di- and monophosphatase activities. L-Leucyl-beta-naphthylamidase activity was found in one additional acid-phosphatase-active precipitate.

Animals↗
PubMed1975

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. This procedure also depleted the lymphocyte preparations of antibody-dependent cytolytic effector cells. The results suggest that rations of antibody-dependent cytolytic effector cells. The result suggest that such effector cells have receptors for human C as well as for C(ra). Lymphocytes were not able to lyse erythrocytes bearing either human C3b (similar to 30,000 molecules/Ec) or activated C(ra) in the absence if IgG antierythrocyte antibodies. Under the same experimental conditions these target cells were efficiently lysed in the presence of small amounts of IgG antitarget cell antibodies. This suggests that the interaction between the cellular Fcreceptors and the Fc part of the inducing antibodies is of special significance for the triggering of the cell-mediated lytic reaction. However, although target cell-bound C did not trigger cytolysis, it seemed to potentiate antibody-dependent cytolysis, probably by enhancing effector cell-target cell contacts.

Animals↗
PubMed1975

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. The hyperimmune rabbit serum was 100 to 1000 times more active, whereas serum RGM was consistently negative. These results correlated well with the concentration of anti-dextran antibodies in these sera. In monocyte-mediated erythrolysis serum EAK had a somewhat higher titer than in lymphocyte-mediated lysis, and serum RGM had a weak but significant activity at low dilutions. Serum EAK also induced erythrophagocytosis by monocytes. Ultracentrifugation did not significantly decrease the inductive capacity of this serum. The results show that antibodies of human sublass IgG2 are efficient inducers of effector functions in both lymphocytic and monocytic cells. Myeloma proteins of the four IgG subclasses were tested for inhibitory capacity in lymphocyte- or monocyte-mediated erythrolysis. Either serum EAK or the rabbit reference serum was used for induction of erythrolysis. Individual myeloma proteins within and between the subclasses varied considerably in inhibitory power. However, whereas IgG1, IgG2, and IgG3 proteins inhibited lymphocyte-mediated erythrolysis induced by either type of antiserum, the two IgG4 proteins tested were essentially negative. These results suggest a lack of specificity of the Fc receptor for subclasses IgG1, IgG2, and IgG3 in both heterologous and homologous inhibition. In monocyte-mediated erythrolysis, IgG1 and IgG3 were strong inhibitors, whereas inhibition by IgG2 and IgG4 was weak and inconsistent. This pattern was seen regardless of whether and inducing antiserum was of rabbit or human origin. Similar results were obtained in monocyte-induced erythrophagocytosis induced by serum EAK. These and previous results suggest that effector cells of the lymphocytic (K cell) variety have Fc receptors different from those of monocytic cells. However, the basis for the differences observed in the inhibition tests remains to be elucidated.

Animals↗
PubMed1975

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. One antiserum to the constant half of lambda-type Bence Jones protein also contained antibiodies to beta 2-microglobulin and stained all lymphocytes. Four other anti-lambda reagents contained no antibodies to beta 2-microglobulin and stained at most about half of the lymphocytes. The antigen responsible for this staining is unknown. The isolated anti-immunoglobulin antibodies (anti-lambda) stained only 5%-10% of the lymphocytes. Antisera to serum IgG or its fragments were free of antibodies to beta 2-microglobulin and stained only 10%-25% of the lymphocytes. This staining was in all instances due to antibodies to human immunoglobulin. Five of eight undiluted sera from normal rabbits with no detectable antibodies to human immunoglobulin or beta 2-microglobulin stained 25%-60% of the lymphocytes. This staining rapidly disappeared on dilution.

Antibodies↗
PubMed1975

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. Sephadex G-200 chromatography indicated that 90% of the DNP antibodies were 7S in the bleedings on days 10-16, whereas 99.8% were 7S in later bleedings. 7S antibodies induced K-cell lysis at high dilutions, whereas 19S antibodies were essentially negative. Antibody fractions obtained by DEAE- or CM-cellulose chromatography were used to establish possible heterogeneities in the capacity of 7S antibodies to induce either K-cell- or complement-mediated target cell lysis. No such heterogeneities were founnd. Fifteen IgG preparations containing antibodies of different affinities were compared with regard to their capacity to induce K-cell-mediated lysis. A statistically significant correlation was found between antibody affinity and efficiency in K-cell-mediated lysis. In a similar study of complement-mediated lysis the correlation was not significant at the 5% level but was significant at the 10% level.

Animals↗
PubMed1975

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. The plaque-forming cells appeared as villous, small to medium-sized lymphocytes when studied by transmission or scanning electron microscopy. Under standardized experimental conditions the minimal number of active, plaque-forming K cells in purified lymphocyte preparations could be estimated.

Antibody Formation↗
PubMed1975

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. The antibody-induced cytotoxocity of lymphocytes decreased following radiotherapy while that mediated by PHA remained unchanged. There was some reduction in the percentage of EAC rosette-forming cells. These results, as well as our earlier observations, suggest that the decrease in the peripheral blood of the proportion of lymphocytes with receptors for activated complement is responsible for changes in the antibody-mediated lymphocyte cytotoxicity.

Adult↗
PubMed1975

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. In the second technique, lymphocytes were fractionated by centrifugation into two fractions according to their ability to form spontaneous rosettes with sheep erythrocytes (E). The E-rosette-forming population (primarily T cells) was shown to lack cytotoxic activity against mumps virus-infected target cells. This activity was present in the nonrosetting population. The results suggest that the effector cells involved in this cytotoxic system are of a non-T variety.

Animals↗
PubMed1974

Lymphoid cells mediating tumor-specific cytotoxicity to carcinoma of the urinary bladder. Separation of the effector population using a surface marker.

Peripheral lymphocytes from patients with urinary bladder carcinoma and controls have been separated on the basis of rosette formation with sheep erythrocytes. The fractions were tested for tumor-specific cytotoxicity. The E rosette-forming cells of purity 90% respond well in PHA-induced cytotoxicity but are totally inactive in the tumor assay. The non-E rosette-forming cells (purity 91%) give enhanced activity in the tumor-specific cytotoxicity as well as in antibody-mediated target cell lysis in a model system. These data support the notion that the effector cells in cell-mediated immunity to carcinoma of the urinary bladder are members of the nonthymus-derived population of peripheral lymphocytes.

B-Lymphocytes↗