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Biomedical subjects

C D Myers

Publications and source records attributed to C D Myers.

32 records · Page 2Linked to original sources
PubMed1986

Isolation of antigen-binding virgin and memory B cells.

One major obstacle in studying the activation of antigen-specific B cells is the small number of B cells reactive with a particular antigenic epitope. In this report, we describe a method by which large numbers of highly purified antigen-binding cells can be obtained. We have shown that by varying the haptenation level of the erythrocytes used for rosetting, we can purify antigen-binding B cells which have different affinities for the antigenic epitope. Thus, memory cells (which have receptors of higher affinity) can be prepared and these cells are essentially free of contaminating virgin cells. The effects of varying the haptenation levels on the red cells used for purifying the B cells can, in turn, be related to the precursor frequency of secreting cells following their activation with T cells and antigen.

Animals↗
PubMed1985

Serological, biochemical, and functional identity of B cell-stimulatory factor 1 and B cell differentiation factor for IgG1.

By three criteria, we have demonstrated that B cell stimulatory factor (BSF-1) and B cell differentiation factor (BCDF-gamma) are the same lymphokine. Highly purified preparations of high performance liquid chromatography-purified or affinity-purified BSF-1 had BCDF-gamma activity but not BCDF-mu activity. A monoclonal anti-BSF-1 antibody coupled to Sepharose depleted both BSF-1 and BCDF-gamma activity but not BCDF-mu activity from two different T cell supernatants. Soluble monoclonal anti-BSF-1 blocked the BSF-1 and BCDF-gamma but not the BCDF-mu responses. These results suggest that BSF-1 acts on both resting and activated B cells to induce different effects.

Animals↗
PubMed1985

The human leucocyte-common (LC) molecule: dissection of leukaemias using monoclonal antibodies directed against framework and restricted antigenic determinants.

Monoclonal antibodies have previously been raised against two separate antigenic determinants on the human LC molecule. One, F10.89.4, recognizes a 'framework' epitope on all LC molecules; these are found on the majority of leucocytes. The other, F8.11.13, recognizes only a 'restricted' epitope present on a subset of these molecules; this subset is found on B lymphocytes and a subpopulation of T lymphocytes. LC molecules on myeloid cells do not carry the 'restricted' antigenic determinant. We have investigated the differential expression of these LC epitopes on human leukaemias, using immunofluorescence on fresh leukaemic blasts and established cell lines. Our study shows that, as on normal haemopoietic cells, LC molecules on B leukaemias bear both 'framework' and 'restricted' epitopes, while the majority of T leukaemias bear only the 'framework' determinant. The small proportion of T cells that are F8.11.13+ ('restricted' epitope) are relatively mature, being of either OKT4+ or OKT8+ phenotype, and may be in an activated state (HLA-DR+). However, in contrast to normal haemopoietic cells, some myeloid leukaemias carry both 'framework' and 'restricted' epitopes (30% AML and AMML samples are F10.89.4+, F8.11.13+), and it is within this group that all TdT+ AML and AMML cases lie. Thus, these monoclonal antibodies should be useful for studying haemopoiesis in man and for analyzing human haemopoietic malignancies.

Adolescent↗
PubMed1985

Selective expression of class-II MHC antigens during hemopoietic differentiation.

Human multipotential or "mixed" bone marrow colony-forming cells and lineage-restricted colony-forming progenitors have been analyzed by cell sorting (FACS) using a series of monoclonal antibodies. The latter all react with monomorphic class-II MHC glycoprotein determinants but differ in their reactivity or cross-reactivity with products of different loci, i.e., SB (DP), DC (DQ), DR. The results confirm that very few progenitor cells detectably express DC. Anti-DR monoclonals varied in their reactivity with progenitor cells. The majority of progenitors in all lineage categories express determinants that are shared or cross-reactive between DR and SB while fewer progenitors can be shown to bind antibodies specific for DR or SB.

Antibodies, Monoclonal↗
PubMed1984

Blockade of the galactose-binding sites of ricin by its linkage to antibody. Specific cytotoxic effects of the conjugates.

A method is described for preparing specific cytotoxic agents by linking intact ricin to antibodies in a manner that produces obstruction of the galactose-binding sites on the B chain of the toxin and so diminishes the capacity of the conjugate to bind non-specifically to cells. The conjugates were synthesised by reacting iodoacetylated ricin with thiolated immunoglobulin and the components of conjugate with reduced galactose-binding capacity were separated by affinity chromatography on Sepharose (a beta-galactosyl matrix) and asialofetuin-Sepharose. Fluorescence-activated cell sorter (FACS) analyses revealed that the fraction of a monoclonal anti-Thy1.1-ricin conjugate that passed through a Sepharose column had markedly diminished capacity to bind non-specifically to Thy1.2-expressing CBA thymocytes and EL4 lymphoma cells. The fraction of conjugate that passed through an asialofetuin-Sepharose column displayed no detectable non-specific binding. Both fractions of conjugate were potent cytotoxic agents for Thy1.1-expressing AKR-A lymphoma cells in tissue culture. They reduced the [3H]leucine incorporation of the cells by 50% at a concentration of 2-5 pM. Comparable inhibition of EL4 cells was only achieved with 3000-7500-fold greater concentrations of conjugate. By contrast, the fraction of anti-Thy1.1-ricin that retained Sepharose-binding capacity showed marked non-specific binding and toxicity to EL4 cells. A conjugate with diminished galactose-binding capacity was also prepared from the W3/25 monoclonal antibody which recognises an antigen upon helper T-lymphocytes in the rat. It elicited powerful and specific toxic effects upon W3/25 antigen-expressing rat T-leukaemia cells. This finding is of particular importance because isolated ricin A-chain disulphide-linked to W3/25 antibody is not cytotoxic. The property of the B-chain in intact ricin conjugates that facilitates delivery of the A-chain to the cytosol thus appears to be independent of galactose recognition. It is concluded that the 'blocked' ricin conjugates combine the advantages of high potency, which is often lacking in antibody-A-chain conjugates, with high specificity, which previously was lacking in intact ricin conjugates.

Animals↗
PubMed1984

An immunotoxin with therapeutic potential in T cell leukemia: WT1-ricin A.

A conjugate of the monoclonal antibody WT1 and ricin A-chain was studied for its suitability for purging marrow of leukemic T cells for autologous transplantation in T cell acute lymphocytic leukemia (T-ALL). The conjugate was powerfully cytotoxic to the human T-ALL cell line, GH1, which expresses the WT1 antigen at a high density. Treatment of the cells with the conjugate at 10(-11) M reduced their rate of protein synthesis by 50%, and the inclusion of 6 mM ammonium chloride in the cultures enhanced the potency of cytotoxic effect by 10-100-fold. Clonogenic assays indicated that less than 0.1% of GH1 cells survived 3-hr exposure to the conjugate in ammonium chloride. WT1 alone did not react with multipotent (CFU-GEMM) hematopoietic progenitors in normal human bone marrow, as measured by fluorescence-activated cell sorting. Under conditions giving maximal killing of GH1 cells, there was no toxicity to multipotential progenitors in normal human marrow.

Antibodies, Monoclonal↗
PubMed1984

A cell line secreting stimulating factors for CFU-GEMM culture.

The multipotent hemopoietic stem cell has fastidious growth requirements in vitro. Traditionally, phytohemagglutinin-stimulated leukocyte conditioned medium has been used to supply the undefined growth factors required for culture of the human multipotent hemopoietic progenitor. We describe the use of medium conditioned by the bladder carcinoma cell line, 5637, to replace PHA-LCM in CFU-GEMM cultures and show that the properties of this conditioned medium closely mimic those of PHA-LCM in two separate CFU-GEMM culture systems.

Cell Line↗
PubMedDate not supplied

Sex, gender, and blood pressure: contributions to experimental pain report.

OBJECTIVE: The current study investigated whether the relationship between sex and experimental pain report was explained by systolic blood pressure (SBP) at rest or during pain task, by gender-role socialization as assessed by the Bem Sex Role Inventory, or both. The influence of gender-role socialization on pain report is often inferred but rarely studied. METHODS: Fifty female and 54 male healthy, young adults completed the Bem Sex Role Inventory and then underwent a cold pressor task. Blood pressure was assessed before and during pain testing. RESULTS: Univariate analyses indicated significant sex-related differences in pain threshold and pain tolerance. Baseline SBP was positively related to pain tolerance but did not explain sex differences, in accord with previous research. The Bem Sex Role Inventory demonstrated a relationship with pain, but did not explain sex differences. CONCLUSIONS: We suggest that context-specific measures of gender are needed to assess gender-related pain behaviors in specific situations. Results from the current study support our contention that gender is part of sex as commonly measured. Also, blood pressure does not appear to fully account for sex-related differences in pain.

Adolescent↗
PubMedDate not supplied

High performance for expert systems: I. Escaping from the demonstrator class.

An investigation was performed to elucidate the problems of converting a demonstrator expert system for leukaemia diagnosis into a large high-performance system. Analysis of the logical completeness of the demonstrator system revealed only minor errors and a limited representation of the domain. Most of its test cases were quite simple and correction of the minor errors raised the performance to 94%. However, the remaining cases were very complex and it is doubtful whether the system could handle them without fundamental changes to its representation of the domain. Demonstrator systems can function with an imperfect view of the domain since the small sacrifice of accuracy is of little consequence; but in a high-performance system the imperfections must be corrected before extra detail is added. The use of default mechanisms for making specific conclusions was harmful because they concealed errors, and a numerical method for handling uncertainty contributed nothing of value to the system.

Diagnosis, Computer-Assisted↗