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Biomedical subjects

W J Halliday

Publications and source records attributed to W J Halliday.

At least 37 records · Page 2Linked to original sources
PubMed1985

Monoclonal antiidiotypic antibodies related to a murine oncofetal bladder tumor antigen induce specific cell-mediated tumor immunity.

Rat monoclonal antibody 6.10 recognizes a 175-kDa protein expressed in all BALB/c mouse transitional cell bladder carcinomas tested, in epithelial cells of the mouse embryo, and in a few epithelial cells of adult mice. The antibody was used as an immunogen to generate two mouse monoclonal antibodies, 21D9 and 43A10, which bind to idiotopes on antibody 6.10 associated with the binding site for the 175-kDa antigen. The antiidiotypic antibodies induced bladder tumor-specific, cell-mediated immunity when injected into syngeneic mice, as shown by delayed-type hypersensitivity reactions in vivo and leukocyte adherence inhibition reactions in vitro. Tumor specificity was demonstrated by employing as controls a chemically induced BALB/c fibrosarcoma, MCA-1511 (MCA, 3-methylcholanthrene), and its corresponding antiidiotypic antibody, 5.96. Lymphocytes from mice sensitized with antibody 21D9 or 5.96 specifically recognized antigens in extracts of BALB/c bladder carcinoma BTCC-1660 (BTCC, bladder transitional cell carcinoma) and sarcoma MCA-1511, respectively, as shown by leukocyte adherence inhibition reactivity. This reactivity was selectively abrogated by prior treatment of the sensitized cells with the appropriate antiidiotypic antibodies and complement. An antigen recognized in vitro by antibody 21D9-sensitized lymphocytes could be separated from BTCC-1660 extract by immunoabsorption with antibody 6.10 and elution with acidic buffer. Our findings indicate that the oncofetal antigen defined by antibody 6.10 is recognized by the immune system of syngeneic mice and suggest that antiidiotypic antibodies related to certain oncofetal antigens can be used to immunize against syngeneic tumors.

Animals↗
PubMed1985

Apoptosis as the mode of cell death in antibody-dependent lymphocytotoxicity.

A light and electron microscopic study of antibody-dependent lymphocytotoxicity was carried out with the object of elucidating the mechanisms responsible for the cell killing, the basis for the research being the relationship that has recently been shown to exist between the morphology of cell death and its pathogenesis. Chang liver cells coated with a rabbit anti-human antibody were used as targets and normal human peripheral-blood lymphocytes as effector cells. Cytotoxicity assays using release of 51Cr demonstrated extensive K-cell killing, thus validating the suitability of the model for morphological studies. Cell death displaying the features of apoptosis correlated with K-cell activity. A small amount of cell death by classical necrosis was observed, but its extent appeared to be unrelated to the presence of lymphocytes, to pre-treatment of the target cells with antibody, or to the magnitude of 51Cr release. The results support evidence indicating that lymphocytotoxicity depends on activation of a self-destruct program within the target cell. They do not favour a mechanism involving the production of plasma membrane lesions analogous to those responsible for complement-mediated immune cytolysis.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1985

Leukocyte adherence inhibition to myelin basic protein by cancer patients' T-lymphocytes in association with class II major histocompatibility antigens on monocytes.

Patients' leukocytes were shown to react consistently in tube leukocyte adherence inhibition (LAI) assays with myelin basic protein (MBP) at optimal concentration, whereas control leukocytes were nonreactive. Mononuclear cells from patients with cancer gave positive LAI reactions with MBP, but separated T-lymphocytes, monocytes, and neutrophils did not. The mononuclear cell LAI responses were blocked by monoclonal antibody (MAb) to monomorphic determinant of class II major histocompatibility complex (MHC) antigens and to T4+ (Leu-3a+) and T3+ (Leu-4+) T-cell differentiation antigens but not by antibody to class I MHC antigens or T8+ (Leu-2a+) antigens. MBP was thus recognized by helper T-cells, requiring presentation in association with class II MHC determinants on monocytes. MAb to class I and class II MHC antigens and to T8+ (Leu-2a+), T4+ (Leu-3a+), and T3+ (Leu-4+) differentiation antigens did not negate LAI mediated by peripheral blood lymphocytes to organ-specific cancer neoantigens (OSN) of crude extracts of allogeneic cancer, which had previously been shown to react with cytophilic antibody on allogeneic monocytes. When membrane OSN and leukocytes were autologous, T8+ (Leu-2a+) phenotypic T-cells also mediated LAI that was blocked by anti-T8 (Leu-2a) and anti-T3 (Leu-4). LAI induced by MBP was also negated by drugs that antagonize thromboxane-leukotriene biosynthesis, indicating that, in common with other LAI reactions, the terminal mediators of nonadherence are oxidative metabolites of arachidonic acid. In addition to clarifying the role of MBP in the cellular in vitro immunoreactivity of cancer patients, the present observations have important implications for theories of LAI. Sensitized leukocytes have different mechanisms for the recognition of antigens in different forms, and the antigen-stimulated leukocytes produce mediators that in a final common pathway induce nonadherence of surrounding cells through leukotriene-like metabolites.

Antibodies, Monoclonal↗
PubMed1984

A two-chain tumour-related suppressor factor specific for a sequence antigen.

Suppressor factors in the serum of CBA mice bearing transplanted methylcholanthrene-induced tumours suppressed the leukocyte adherence inhibition reaction between tumour-sensitized peritoneal cells and a sequenced antigen, myelin basic protein (MBP). Chromatography on Sephacryl S-200 columns demonstrated suppressive activity in low molecular weight fractions (40-50 kDa) derived from whole serum. The factors were absorbed from serum by MBP, but not by lysozyme, coupled to Sepharose gel. Cleavage with sodium dodecyl sulphate (SDS) and absorption with either MBP or anti-I-J antibody on Sepharose gels yielded inactive supernatants that regained suppressive activity when combined. Elution of these gels with acidic buffer yielded eluates that were inactive alone but suppressive when combined. These results indicate that the tumour-related suppressor factors consist of two chains separable by detergent action; one chain is antigen (MBP)-binding and the other contains an I-J determinant.

Animals↗
PubMed1984

Suppression by serum of immunoreactivity to a common tumour antigen in cancer patients.

Leucocytes from 4 cancer patients showed cellular reactivity in the leucocyte adherence inhibition (LAI) assay in the presence of the synthetic encephalitogenic peptide of human myelin basic protein. All patients exhibited reactivity at a peptide concentration of 500 ng/ml. Leucocytes from 4 non-cancer patients failed to react. Suppression of LAI was detected in all 4 cancer patients by adding their serum to reactive mixtures containing peptide and autologous leucocytes. Each serum was subjected to column chromatography on Sephacryl S-200 to determine the molecular weight distribution of suppressive (blocking) factors. The greatest suppression was found in all cases within the range 90-155 kdalton.

Aged↗
PubMed1983

Cellular origin of blocking factors from cultured spleen cells of tumor-bearing mice.

Spleen cells from mice bearing methylcholanthrene-induced tumors were cultured for 2 days without further stimulation. Blocking factors were consistently detected in culture supernatants by their ability to suppress leukocyte adherence inhibition reactions between soluble tumor antigens and peritoneal cells of tumor-bearing mice. The blocking factors were specific for individual tumors. The cellular origin of these factors was investigated by depleting the spleen cell population of various cell types before culturing. The cells involved were removed by treatment with antibodies to certain membrane markers (Thy-1, Ly-2, Ia, I-J) but not by anti-Ly-1 antibodies. Removal of adherent cells also prevented production of blocking factors, which was restored by reconstitution with syngeneic but not allogeneic cells from normal mice. The normal reconstituting cells were shown to bear Ia, but not I-J or IgM. This indicates that blocking factors (previously shown to have I-J determinants in their molecules) originate from suppressor T lymphocytes (Thy-1+, Ly-1-2+, I-J+), with macrophages (I-J-, Ia+) in the role of accessory cells.

Animals↗
PubMed1983

Antigenic specificity in the E-rosette augmentation reaction.

The antigen reactivity of murine T-lymphocytes can be detected by their formation of a lymphokine that augments the formation of E-rosettes by normal human lymphocytes and sheep erythrocytes. Aspects of the specificity of E-rosette augmentation (ERA) were investigated with spleen cells of mice immunized with proteins or contact-sensitizers. The antigen dose-response curves in ERA were bell-shaped, with peak reactivity at optimal antigen concentrations and complete inhibition with moderate antigen excesses of 5-10 times the optimum. Optimal ERA was antigen-specific, spleen cells reacting only with molecules containing the immunizing determinant. Inhibition with moderate excess was also antigen-specific but great excess of antigen (100 times the optimum) was non-specifically suppressive. The mechanism of specific suppression by moderate excess of antigen appears to be related to the mode of its presentation to lymphocytes by accessory cells. The property of T-lymphocytes, reacting with a single defined determinant, to be inhibited specifically by moderate excess of any antigen containing this determinant, suggests a method of analysis of complex antigens.

Animals↗
PubMed1983

Detection of specific murine anti-tumour immunity and serum factors by the indirect E-rosette augmentation assay.

The spontaneous interaction of sheep erythrocytes and human lymphocytes results in the formation of aggregates known as E-rosettes. The antigen reactivity of murine T-lymphocytes can be detected indirectly by the production of a lymphokine that augments the E-rosette-forming ability of human lymphocytes. An indirect E-rosette augmentation (ERA) assay was used to study cellular reactivity and tumour-related serum reactions in mice bearing methylcholanthrene (MCA)-induced tumours. The assay was used to demonstrate the antigenic specificity of two tumours and to detect blocking factors in tumour-bearer serum. Blocking factors were shown to be tumour-specific and to act on lymphocytes at an early stage before exposure to antigen.

Animals↗
PubMed1983

Immunological stability of an elapid venom, Tropidechis carinatus, and its relevance to the clinical detection of snake venom.

Using immunological techniques, cases of human snake-bite can be proven and individual snake species identified. A series of experiments is described, to test the antigenic stability and physical properties of an Elapid venom, venom properties which will have implications in field use, transport, or the laboratory procedures relating to the immunological characterization of Australian snake venoms. Using venom from the Elapid, Tropidechis carinatus, we have demonstrated significant immunological stability at a temperature of 37 degrees for 48 h at least, and for 24 h at least when incubated continuously with skin, muscle, and fat homogenates; the venom is stable at 56 degrees for 2 h at least. No loss of venom occurs over the pH range 7-10; at pH ranges less than 4 and at ranges greater than 10 only 30% of venom is detected after 30 min incubation. Sonication (10 min at 100 watts) does not destroy immunological properties of the venom; 10 freeze-thaw cycles result in some 9% loss of immunologically detectable activity. Venom is adsorbed significantly on to dacron swabs, only 13% of the applied does being detected after 48 h incubation at 28 degrees. Sixty-seven percent of applied venom can be recovered from the skin of human volunteers 6 h after a simulated bite. The venom of this Elapid is antigenically robust. With the currently available sensitive assays, venom from human skin should still be detected in material kept without special preparation under field conditions for up to 2 days at least.

Animals↗
PubMed1983

Restricted activity of serum blocking factors related to a common tumour antigen in mice.

Mice of four different inbred strains (CBA, Balb/c, C57Bl/6 and DBA/2), bearing different transplanted tumours (methylcholanthrene-induced sarcomas, B16 melanoma and P-815 mastocytoma), were tested for cellular immune reactivity to the synthetic encephalitogenic peptide of human myelin basic protein by the leucocyte adherence inhibition (LAI) assay. All exhibited reactivity at about the same optimal concentration of peptide. Normal mice of all strains and pregnant CBA mice were non-reactive. Blocking of LAI was detected with serum obtained 10 or more days after tumour transplantation. Sera from mice bearing different transplanted tumours abrogated the adherence-inhibitory effect of peptide on sensitized syngeneic peritoneal leucocytes. The blocking factors were cross-reactive between different tumours only within the same mouse strain, indicating a requirement for genetic compatibility between donors of the reactive cells and the serum blocking factors.

Animals↗
PubMed1983

Serum blocking factors in bovine ocular squamous cell carcinoma demonstrated by inhibition of erythrocyte rosette augmentation.

Blood leukocytes from cattle with bovine ocular squamous cell carcinoma (BOSCC) react with extracts of BOSCC, and blood leukocytes from cattle with cutaneous papillomas react with extracts of papillomas in the erythrocyte rosette (E-rosette) augmentation test. Sera from cattle with BOSCC, with cutaneous papillomas, and with ocular papillomas and sera from lesion-free cattle were tested for their ability to block antigen-induced E-rosette augmentation and E-rosette augmentation induced by phytohemagglutinin. Autologous serum from 16 BOSCC-affected cattle blocked E-rosette augmentation induced by BOSCC extract, and autologous sera from three cattle with cutaneous papillomas blocked E-rosette augmentation induced by papilloma extract. Sera from cattle with BOSCC did not block E-rosette augmentation induced by papilloma extract, nor did sera from cattle with cutaneous papillomas block BOSCC extract-induced E-rosette augmentation. Sera from cattle with BOSCC blocked BOSCC extract-induced E-rosette augmentation in some but not all allogeneic combinations. E-rosette augmentation was observed when blood leukocytes from all 12 BOSCC-affected and six control cattle were exposed to phytohemagglutinin. This reaction was blocked by autologous sera from 11 of the 12 BOSCC-affected cattle, from one of the three cattle with cutaneous papillomas, but from none of the three other cattle. Sera from BOSCC-affected cattle also blocked phytohemagglutinin-induced E-rosette augmentation in allogeneic combination, regardless of the tumor status of the donor of the leukocytes.

Animals↗
PubMed1982

Appearance of three lymphokines in culture supernatants harvested at different times.

Peritoneal cells from mice with methylcholanthrene-induced tumours were reacted with the corresponding tumour extract (antigen) and supernatants were prepared after incubation times of 1, 3.5 and 24 h. All the supernatants were then tested simultaneously on the same pools of normal indicator cells for their effects on leucocyte adherence, E-rosette formation and macrophage migration. The leucocyte adherence inhibition factor (LAIF) was consistently detected only in the 1-h supernatant, E-rosette augmentation factor (E-RAF) only in the 3.5-h supernatant and migration inhibition factor (MIF) only in the 24-h supernatant. It is concluded that LAIF, E-RAF and MIF are probably distinct factors, having different stabilities to degradation or inhibition under the conditions of cell culture. A less likely interpretation is that a single factor with multiple activities is affected by a series of distinct inhibitors. In practical terms, the 3 assays correlate only if the appropriate incubation times are used.

Animals↗
PubMed1982

Cellular immunity to encephalitogenic peptide in tumour-bearing mice.

Mice bearing a methylcholanthrene-induced tumour were tested for their cell mediated reactivity to the experimental allergic encephalomyelitis (EAE) peptide of human myelin basic protein (MBP) in the leucocyte adherence inhibition (LAI) test. Tested over a range of peptide concentrations, peritoneal cells (PC) from tumour-bearing mice exhibited optimal adherence inhibition at 640 ng/ml; PC from normal and parasite-infected mice were unreactive. The EAE peptide also stimulated PC from tumour-bearing mice in the E-rosette augmentation (ERA) test and in the macrophage migration inhibition (MMI) test. MMI appeared to be the most sensitive assay, in that significant reaction at peptide concentrations well below those giving significant LAI and ERA. LAI reactivity to the peptide was detected 5 days after tumour transplantation, and continued to be detectable even with very large tumours. In vitro assays were confirmed by demonstration of EAE peptide recognition in vivo, in tumour-bearing and tumour-excised mice, using the delayed-type hypersensitivity reaction. The present experiments demonstrate an antigenic determinant in murine tumours, similar to the well-characterized EAE peptide of human MBP, and establish an animal model for study and characterization of common tumour-associated antigens.

Animals↗
PubMed1982

Regulation of cell-mediated immunologic reactivity to Moloney Murine sarcoma virus-induced tumors. III. Further characterization of tumor-specific serum blocking factors.

Blocking factors in the sera of inbred CBA mice bearing either Moloney murine sarcoma virus-induced tumors or transplanted 3-methylcholanthrene-induced tumors specifically blocked in the leukocyte adherence inhibition reaction between tumor-sensitized peritoneal cells and tumor antigens. These factors were absorbed from serum by treatment with anti-Ia antibodies and could be eluted in a partially purified form. Chromatography on Sephacryl S-200 columns demonstrated blocking activity in fractions of high and low molecular weight (greater than 100,000 and 40,000--50,000, respectively) derived from whole sera or eluates. Active fractions from serum appeared to retain the tumor specificity of the original material.

Animals↗
PubMed1981

Inhibition and stimulation of leukocyte adherence by soluble factors from murine lymphocytes.

Activation of murine spleen cells with either specific antigen or mitogen resulted in the production of soluble factors capable of either inhibiting or stimulating the adherence of normal peritoneal cells. Leukocyte adherence inhibition factor (LAIF) and leukocyte adherence stimulation factor (LASF) were detected using the original leukocyte adherence inhibition (LAI) assay as performed with haemocytometers. An important factor in determining both the production and detection of adherence-modulating factors was the serum content in both the preparation and testing of supernatants favoured LAIF, whereas a lower serum content resulted in LASF predominating. With mitogen (concanavalin A) -activated cells, variable production of adherence inhibitory and stimulatory activities was found, but LASF appeared to be favoured at low concentrations of concanavalin A and serum. Adherence inhibitory and stimulatory factors in concanavalin A-generated spleen cell supernatants were separable on Sephacryl S-200 columns. LAIF had a molecular weight of less than 55,000 daltons, whereas most LASF activity was greater than 70,000 daltons. The occurrence of adherence-modulating factors with opposing effects may be a potential source of variability in LAI assays.

Animals↗
PubMed1981

Regulation of cell-mediated immunologic reactivity to Moloney murine sarcoma virus-induced tumors. I. Cell and serum activity detected by leukocyte adherence inhibition.

Cell-mediated immunity and serum regulatory factors were studied in an in vitro system involving a spontaneously regressing, virus-induced tumor. Inbred BALB/c and CBA mice were inoculated with Moloney murine sarcoma virus and their peritoneal cells were tested for reactivity in leukocyte-adherence inhibition tests with extracts of syngeneic tumors. Sera from inoculated mice were tested for their effect on this reactivity. At the optimal dilution, tumor extracts induced significant reactions with cells from tumor-bearing mice (progressors) and from mice with regressed tumors (regressors); cells from normal mice and from mice with transplanted, chemically induced tumors were unaffected. Sera from progressor mice specifically blocked the reactivity of syngeneic cells. At the time of maximal tumor development, this blocking activity disappeared and the serum became unblocking: i.e., the regressor serum neutralized the blocking factor in progressor serum. The blocking or unblocking factors were tumor-specific; no cross-reactivity occurred with similar factors related to the chemically induced tumor. Normal cells were not significantly affected by exposure to blocking and unblocking sera. The development of cellular immune reactivity and serum factors detected in vitro corresponded to the cycle of tumor progression and regression observed in vivo.

Animals↗