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Biomedical subjects

R D MacGregor

Publications and source records attributed to R D MacGregor.

11 recordsLinked to original sources
PubMed1994

Characterization of the oligodeoxynucleotide-mediated inhibition of interferon-gamma-induced major histocompatibility complex class I and intercellular adhesion molecule-1.

The major histocompatibility complex (MHC) Class I and II genes and intercellular adhesion molecule-1 (ICAM-1) are regulated by interferon-gamma in a variety of cell types. We have previously shown that the oligodeoxynucleotide 5'-GGG GTT GGT TGT GTT GGG TGT TGT GT-RNH2 (oligo I) inhibits the interferon-gamma-mediated enhancement of MHC Class I and ICAM-1 proteins in the K562 cell line. We have now investigated the mechanism of action of oligo I and report that it acts by inhibiting the binding of interferon-gamma to cells. We also show that the dose-response curves, the selectivity profile, and the kinetics of oligo I are consistent with this novel mechanism of action. The dose-response curves for oligo I, obtained using antibodies against the MHC Class I heavy chain, beta 2-microglobulin, or ICAM-1, are almost superimposable at each observation time. MHC Class I induction by 6400 units/ml interferon-alpha or interferon-beta or ICAM-1 enhancement by 800 units/ml tumor necrosis factor-alpha is not inhibited by oligo I. However, the synergistic induction of MHC Class I by mixtures of tumor necrosis factor-alpha and interferon-gamma is inhibited. Oligo I belongs to a class of active oligodeoxynucleotides that inhibits interferon-gamma-induced MHC Class I and ICAM-1 in K562 cells. The activity and potency is sequence-dependent, but remarkably different sequences can have comparable effects. The activity of oligo I in the HeLa S3 cell line inhibits the interferon-gamma-mediated enhancement of both ICAM-1 and MHC Class II DR and the interferon-gamma-mediated reduction in transferrin receptor expression. Thus, oligo I appears to specifically inhibit interferon-gamma-induced changes in protein expression, which is consistent with oligo I acting at an early step(s) in the induction process. Taken together, our results show that oligo I exerts its effects by inhibiting the association of interferon-gamma with the cell surface, which is a novel mechanism of action for oligodeoxynucleotides.

Base Sequence↗
PubMed1994

Inhibition of interferon-gamma-induced major histocompatibility complex class I expression by certain oligodeoxynucleotides.

We report that certain oligonucleotides are capable of inhibiting cell surface induction of the major histocompatibility complex class I (MHC-I) proteins by interferon-gamma in K562 cells. The inhibition by oligodeoxy-nucleotide I 5' GGG GTT GGT TGT GTT GGG TGT TGT GT-RNH2 is dose-dependent, with an EC50 24 hr after dosing of approximately 4 microM for 800 U/ml interferon-gamma. The reverse complement II 5' AC ACA ACA CCC AAC ACA ACC AAC CCC-RNH2 did not show activity. Oligodeoxynucleotide I inhibits induction of MHC-I by interferon-gamma, but does not inhibit induction by either interferon-alpha or interferon-beta. Four other oligodeoxynucleotides were also evaluated, and three showed activity against interferon-gamma at 25 microM.

Base Sequence↗
PubMed1994

Characterization of apocytochrome C binding to human erythrocytes.

The binding of 125I-labeled apocytochrome c to human erythrocytes was determined for free apocytochrome c concentrations at 10(-10)-10(-6) M. At about 2 x 10(-9) M, maximum cell association of apocytochrome c occurs at 50 mM NaCl and at 22 degrees C. Intact erythrocytes at 22 degrees C have three classes of apocytochrome c binding sites: one high-affinity noncooperative site (n1 = 728 per cell, Kd1 = 1.5 x 10(-9) M) and two positively cooperative sites (n2 = 3.7 x 10(4) per cell, Kd2 = 1.2 x 10(-7) M, alpha 2 = 2.0, and n3 = 2.5 x 10(5) per cell, Kd3 = 7.1 x 10(-7) M, alpha 3 = 12). Erythrocytes at 37 degrees C, and erythrocyte ghosts at 22 degrees C, also have three classes of apocytochrome c binding sites, and most sites are positively cooperative.

Apoproteins↗
PubMed1993

Predictions of effect for intracellular antisense oligodeoxyribonucleotides from a kinetic model.

We have analyzed the implications of a simple two-compartment mathematical model (Hargrove and Schmidt, 1989) to anticipate the limits of antisense oligodeoxyribonucleotide action within single cells. The steady-state equations are derived for four special cases representing the following mechanisms: (i) ribosome blockage, (ii) mRNA cleavage by RNase H, (iii) concurrent ribosome exclusion and RNase H action, and (iv) decreased delivery of mature mRNA to the cytoplasm due to transcriptional blockage, interference with nucleocytoplasmic transport, or splicing. Dose-response relationships have been derived for these mechanisms under ideal conditions. Our results indicate that frequently translated mRNAs producing stable proteins are the most attractive antisense targets because these protein levels are sensitive to the changes in the mRNA levels that can be effected using antisense oligodeoxyribonucleotides. The nonsteady-state solutions show that both mRNA and protein half-life can determine the kinetics of antisense oligonucleotide action. A rapid onset of effect will be observed when the mRNA is rapidly degraded and slowly translated and when the translated protein is rapidly degraded. When the protein is slowly degraded, the kinetics of effect are limited by protein half-life. When the translational rate constant is large compared to the absolute difference between the mRNA and protein degradation rate constants, the kinetics of antisense action are determined by both degradation rate constants but are limited by the slower of the two degradative processes. We also show that the steady-state and nonsteady-state solutions may be used to design experiments that discriminate among mechanisms of antisense action.

Algorithms↗
PubMed1991

Comparison and critique of two models for regional drug delivery.

A simple stochastic recirculatory formalism is used to compare models of regional drug delivery due to Hunt et al. and Boddy and Aarons. It is shown that these two models are equivalent when regional delivery is ideal. The latter model has the advantage of simplicity. However, the former model appears more useful in relating predictions to experimentally accessible quantities. Neither model is sufficiently general to cover all possible topologies of regions associated with drug response and toxicity. Knowledge of this topology is essential in determining the drug targeting index. The underlying assumptions of the models are discussed, and situations where these assumptions may break down are identified. Finally, it is noted that the analysis of regional delivery may also apply to metabolite and prodrug kinetics.

Drug Administration Schedule↗
PubMed1990

Artificial red cells. A link between the membrane skeleton and RES detectability?

Factors governing nonspecific reticuloendothelial system (RES)-detectability are largely unknown. Will a liposome that mimics the lipid composition of the outer leaflet of the erythrocyte membrane be invisible to the RES? On both experimental and theoretical grounds we believe the answer is no, in part because 1) sorption of proteins is believed to be important in determining RES uptake, 2) a membrane skeleton is apparently necessary to inhibit protein sorption into erythrocyte membranes and 3) Neohemocytes (a liposome encapsulated hemoglobin product) currently lack a membrane skeleton. Neohemocytes with erythrocyte outer leaflet lipid composition do have extended circulation half-times, but these are at least two orders of magnitude shorter than the circulation half-times of erythrocytes. How might a membrane skeleton modulate RES-detectability? Can avoidance of opsonization result in part from the properties of the membrane skeleton? If so, then how? To explore and quantify such questions we have developed a theoretical, statistical-thermodynamic model of protein binding into membranes. It predicts that the membrane area available for rapid lateral diffusion is critically important in controlling the amount of sorbed protein per unit area, and that a membrane skeleton can reduce a protein's sorption by several orders of magnitude. Based on theoretical results, we offer a speculative model for the detection of non-self lipid bilayers by the RES.

Blood Substitutes↗
PubMed1985

Synthesis and evaluation of a prototypal artificial red cell.

A new process allows microencapsulation of purified human hemoglobin and 2,3-diphosphoglycerate to form neohemocytes. The microcapsule membrane is composed of phospholipids and cholesterol. Neohemocytes are substantially smaller than erythrocytes, contain 15.1 grams per decaliter of hemoglobin, and have a P50 value (the partial pressure of oxygen at which the hemoglobin is half-saturated) of 24.0 torr. All rats given 50-percent exchange transfusions survived with only limited evidence of reversible toxicity. Normal serum glutamate-pyruvate-transaminase values at 1, 7, and 30 days after transfusion were consistent with minimal hepatotoxicity. The concentration of blood urea-nitrogen was elevated by 35 percent after 1 day but returned to normal by day 7. However, histopathology revealed normal kidneys on day 1 as well as on days 7 and 30. Neohemocytes cleared from the circulation of transfused rats with an apparent half-life of 5.8 hours.

Alanine Transaminase↗