49 results on '"Winkelhake JL"'
Search Results
2. Contrast-Enhanced MRI of Tumors
- Author
-
David L. White, Jean W. Dupon, Kopplin J, Michael E. Moseley, Mats G. Wikstrom, R C Brasch, and Winkelhake Jl
- Subjects
Gadolinium DTPA ,Biodistribution ,Pathology ,medicine.medical_specialty ,CONTRAST ENHANCED MRI ,Macromolecular Substances ,Fibrosarcoma ,Gadolinium ,Vascular permeability ,Mice ,Meglumine ,Albumins ,Mole ,Extracellular fluid ,Organometallic Compounds ,medicine ,Animals ,Radiology, Nuclear Medicine and imaging ,Mice, Inbred BALB C ,medicine.diagnostic_test ,Chemistry ,Albumin ,Magnetic resonance imaging ,General Medicine ,Pentetic Acid ,Image Enhancement ,Magnetic Resonance Imaging ,Drug Combinations ,Female ,Macromolecule - Abstract
The study aim was to define potential differences and advantages in magnetic resonance (MR) patterns of tumoral contrast enhancement using either a small molecular, extracellular fluid contrast enhancer [Gd-DTPA] or a macromolecular agent [albumin-(Gd-DTPA)20], designed for primary intravascular biodistribution. MR images of 25 mice with implanted fibrosarcomas were obtained before and repeatedly for up to 120 minutes after injection of either Gd-DTPA [0.2 mmol/kg, n = 11] or albumin-(Gd-DTPA) [0.0029 mmol/kg, n = 14]. Histologically, this hypovascular tumor contained zones of viable tissue and non-viable, necrotic tissue. Using either type of contrast media, the viable portions enhanced strongly, up to 152% and the necrotic portions enhanced poorly, less than 31%. However, the time-course of enhancement differed between contrast agents. Gd-DTPA tended to provide maximal enhancement soon after administration with no significant changes over two hours. Enhancement from albumin-(Gd-DTPA) was weak initially, corresponding to tumor hypovascularity, but over two hours the signal of the viable tumor zones progressively increased in intensity. This gradual tumoral accumulation of the macromolecular agent within the tumor was considered to reflect abnormal capillary permeability, associated with neovascularity. Thus, the increasing intensity within the neoplastic tissues over time, reflecting abnormal capillary permeability for macromolecules, may serve as a useful, albeit indirect, marker of neoplasia.
- Published
- 1989
3. Studies on the mechanism of covalent incorporation of a lysergyl derivative to immunoglobulin peptides in vitro.
- Author
-
Winkelhake, JL, primary and Voss, EW, additional
- Published
- 1975
- Full Text
- View/download PDF
4. Antigenic peptide binding to MHC class II molecules at increased peptide concentrations.
- Author
-
Nag B, Mukku PV, Arimilli S, Phan D, Deshpande SV, and Winkelhake JL
- Subjects
- Amino Acid Sequence, Antibodies, Monoclonal immunology, Binding, Competitive immunology, Cell Line, Transformed, Chromatography, High Pressure Liquid methods, Electrophoresis, Gel, Two-Dimensional, Humans, Hybridomas, Molecular Sequence Data, Protein Binding immunology, HLA-DR2 Antigen immunology, Myelin Basic Protein immunology
- Abstract
Affinity-purified major histocompatibility complex (MHC) class II molecules are known to bind antigenic peptides in vitro. The percentage of MHC class II molecules occupied with such peptides is usually very low and varies significantly depending upon the sequence and size of a given antigenic peptide. The present study describes a method by which complete saturation of affinity-purified MHC class II with antigenic peptide can be achieved by simply incubating purified MHC class II molecules at neutral pH in the presence of several 100-fold molar excess of antigenic peptide. Complexes of human HLA-DR2 and a peptide analog from human myelin basic protein MBP (83-102)Y83 were selected for this study. The on-rate kinetic results showed saturation of MHC class II occupancy at 300-500-fold molar excess peptide concentrations. The specificity of the MBP (83-102)Y83 peptide binding to HLA-DR2 at higher peptide concentration was demonstrated by incubating an equivalent amount of another epitope from myelin basic protein [MBP (1-14) peptide] as well as by competitive binding assays. The quantitation of bound peptide was carried out using biotinylated-MBP (83-102)Y83 peptide which showed 100-125% occupancy of HLA-DR2 with a recovery of 100%. The presence of a single peptide entity in purified complexes was confirmed by reverse-phase narrowbore HPLC analysis of the acid extracted supernatant and by mass spectrometry analysis. Two-dimensional gel electrophoresis (IEF/SDS) of purified HLA-DR2 and DR2.MBP (83-102)Y83 complexes showed the absence of various endogenous polypeptides in 100% loaded complexes. These results demonstrate that higher peptide concentrations can be useful in generating MHC class II-peptide complexes of defined composition. Such complexes of MHC class II occupied with a single peptide may have significant clinical relevance for antigen-specific therapy of various autoimmune diseases and may provide better understanding of MHC-peptide-TCR interactions.
- Published
- 1994
- Full Text
- View/download PDF
5. Role of tumor necrosis factor species in the sequence-dependent effects of interleukin-2--tumor necrosis factor immunotherapy in mice: implications for preclinical cytokine testing.
- Author
-
Owen-Schaub LB, Santee SM, Winkelhake JL, and Zimmerman RJ
- Subjects
- Animals, Antibody Specificity, Drug Administration Schedule, Drug Synergism, Female, Immunotherapy, In Vitro Techniques, Interleukin-2 metabolism, Leukemia L1210 therapy, Lymphocyte Activation, Mice, Mice, Inbred C57BL, Mice, Inbred DBA, Receptors, Tumor Necrosis Factor metabolism, Species Specificity, Tumor Cells, Cultured, Tumor Necrosis Factor-alpha metabolism, Interleukin-2 therapeutic use, Tumor Necrosis Factor-alpha therapeutic use
- Abstract
We previously demonstrated that recombinant human tumor necrosis factor (hTNF) is synergistic with human interleukin-2 (hIL-2) for in vivo regression of murine tumors. In mice, the timing of cytokine administration is critical in achieving synergy. Because hTNF exhibits negligible binding to the type II murine TNF receptor (TNF-R), we questioned whether murine TNF (mTNF) would have therapeutic benefits, scheduling requirements, and toxic effects similar to those of the hIL-2-hTNF combination. To evaluate the biological effects of TNF-R types I and II interaction, we directly compared the effects of mTNF and hTNF in combination with hIL-2 on in vivo tumor regression and in vitro activation of murine splenocytes. Our results demonstrate for the first time that (a) the cytokine combination hTNF-hIL-2 is consistently more efficacious than mTNF-hIL-2 in in vivo murine immunotherapy models; (b) the in vivo antitumor effects of hTNF-hIL-2 and not mTNF-hIL-2 are critically dependent upon cytokine scheduling; and (c) in vitro culture of splenocytes with mTNF-hIL-2 enhances cellular proliferation, Lyt 2, and TNF-RI expression compared with hTNF-hIL-2. Collectively, these studies extend the previous findings of species-specific mTNF-R responses and reveal that optimal scheduling and efficacy of TNF-hIL-2 combination therapy in murine tumor models is critically dependent upon the TNF species employed.
- Published
- 1994
- Full Text
- View/download PDF
6. Human monoclonal antibodies to glycolipid A that exhibit complement species-specific effector functions.
- Author
-
Winkelhake JL, Gauny SS, Senyk G, Piazza D, and Stevens P
- Subjects
- Animals, Antibody Specificity, Enzyme-Linked Immunosorbent Assay, Humans, Lipopolysaccharides immunology, Opsonin Proteins immunology, Species Specificity, Antibodies, Monoclonal immunology, Complement Activation, Complement System Proteins immunology, Glycolipids immunology, Gram-Negative Bacteria immunology, Lipid A immunology
- Abstract
Two human IgM monoclonal anti-glycolipid A antibodies (MAbs) were evaluated for their abilities to bind to various endotoxins and pathogenic gram-negative bacteria and to activate complement pathways, thereby accomplishing bactericidal and opsonic effector functions. Both MAbs cross-reacted with glycolipid A mutant lipopolysaccharides from rough colony-forming gram-negative bacteria and with selected endotoxins from smooth colony-forming bacteria. However, MAb 10235 bound to all clinical isolates of Escherichia coli and Klebsiella pneumoniae tested but only very weakly to Pseudomonas aeruginosa, whereas MAb 10058 bound to all three genera. Several strains of serum-insensitive organisms were selected for evaluation of antigen-specific, complement-mediated effector functions for the two MAbs. Assessment of bactericidal and opsonic activities showed that neither MAb was able to activate complement from nonprimate species (mouse, rat, rabbit, guinea pig, or sheep). However, both MAbs were highly effective in using primate sources of serum complement to mediate these effector functions.
- Published
- 1992
- Full Text
- View/download PDF
7. Will complex carbohydrate ligands of vascular selectins be the next generation of non-steroidal anti-inflammatory drugs?
- Author
-
Winkelhake JL
- Subjects
- Animals, Cell Adhesion Molecules chemistry, Humans, Inflammation metabolism, Anti-Inflammatory Agents, Non-Steroidal, Carbohydrate Metabolism, Cell Adhesion Molecules physiology, Lectins metabolism
- Published
- 1991
- Full Text
- View/download PDF
8. Effect of production method on the systemic clearance rate of a human monoclonal antibody in the rat.
- Author
-
Gauny SS, Andya J, Thomson J, Young JD, and Winkelhake JL
- Subjects
- Animals, Antibodies, Monoclonal metabolism, Half-Life, Humans, Male, Metabolic Clearance Rate, Rats, Antibodies, Monoclonal biosynthesis, Immunoglobulin M metabolism
- Abstract
Pharmacologic studies of human immunoglobulins (IgM) in non-primate animal models, whether directed toward efficacy or toxicity, rely on pharmacokinetic parameters to achieve optimal doses and schedules. In rodents, human IgMs have an effective circulatory half-life of 11 h and a plasma clearance rate of 0.044 ml/min/kg. Studies of a new group of human monoclonal antibodies (hMAb) specific for Gram-negative bacteria and endotoxin revealed an IgM molecule, hMAb-10058, which, when purified from tissue culture medium, exhibited a suprisingly short circulatory lifetime in rodents. Investigations into possible explanations for this short circulatory half-life resulted in the development of a simple and efficient method for producing hMAbs in the immunodeficient NIH-3 mouse (bg x nu x XID). This method of production of hMAb-10058 had dramatic effects on its half-life. Whereas hMAb-10058 produced in serum-free, defined medium had a clearance rate of 14.4 ml/min/kg and an effective half-life of 0.12 h, the same hMAb-10058 raised in mouse ascites had a decreased clearance rate of 0.092 ml/min/kg and an increased effective half-life of 12 h. This 100-fold enhancement of the hMAb's half-life was not affected by the purification process. Some potential molecular structures involved in the circulatory half-life of this hMAb are discussed.
- Published
- 1991
9. Contrast-enhanced magnetic resonance imaging of tumor-bearing mice treated with human recombinant tumor necrosis factor alpha.
- Author
-
Aicher KP, Dupon JW, White DL, Aukerman SL, Moseley ME, Juster R, Rosenau W, Winkelhake JL, and Brasch RC
- Subjects
- Animals, Gadolinium DTPA, Mice, Mice, Inbred BALB C, Organometallic Compounds, Pentetic Acid, Radiography, Regression Analysis, Magnetic Resonance Imaging methods, Sarcoma, Experimental diagnostic imaging, Sarcoma, Experimental therapy, Tumor Necrosis Factor-alpha therapeutic use
- Abstract
Pharmacological effects of recombinant human tumor necrosis factor alpha (TNF) were studied in a mouse fibrosarcoma model using magnetic resonance imaging enhanced with a macromolecular contrast agent, albumin(gadolinium-diethylenetriamine pentaacetic acid)35. TNF was administered i.v. in a dose of 150 micrograms/kg, 60 to 80 min prior to imaging. Contrast-enhanced and nonenhanced magnetic resonance images of TNF-treated (n = 10) and untreated (n = 8) Meth A fibrosarcomas were obtained at 2.0 Tesla using T1-weighted spin-echo pulse sequences. Serial images spanning an interval of 60 to 120 min after TNF administration showed that the TNF-treated tumors enhanced significantly more overall than did untreated tumors (43% versus 31%). The most marked differential tumor enhancement was observed in the tumor rim (59% versus 40%). Nontumorous tissue, including muscle and brain, revealed no significant enhancement differences between TNF-treated animals and controls. The observed tumor enhancement corresponded strongly with Evans blue staining; the TNF-treated tumors stained deep blue, while untreated tumors and normal tissues observed did not stain. The different enhancement and Evans blue staining patterns between TNF-treated tumors and untreated tumors are attributed to TNF-induced changes in tumor capillary integrity. The data indicate that TNF effects on tumors include an increased capillary permeability for macromolecules at early times after administration. The ability to detect changes in capillary permeability in vivo using contrast-enhanced magnetic resonance imaging may prove to be clinically useful to monitor tumor response to TNF.
- Published
- 1990
10. Human recombinant interleukin-2 as an experimental therapeutic.
- Author
-
Winkelhake JL and Gauny SS
- Subjects
- Animals, Humans, Interleukin-2 pharmacology, Interleukin-2 toxicity, Recombinant Proteins pharmacology, Recombinant Proteins therapeutic use, Recombinant Proteins toxicity, Interleukin-2 therapeutic use, Neoplasms therapy
- Published
- 1990
11. Effects of hyperthermia on plasma glycoprotein catabolism by the isolated perfused rat liver.
- Author
-
Skibba JL, McKean LP, and Winkelhake JL
- Subjects
- Animals, Glycoproteins blood, Iodine Radioisotopes, Male, Rats, Rats, Inbred Strains, Time Factors, Glycoproteins metabolism, Hot Temperature, Liver metabolism
- Abstract
Clearance and degradation of the glycoprotein, asialofetuin (AF), by the isolated perfused rat liver at supranormal temperatures were investigated. The half-life for disappearance of AF was similar at 37, 41, and 42 degrees C, P greater than 0.05. There was a significant difference between the amount of hydrolysis of AF at 37, 41, and 42 degrees C, P less than 0.05. This indicates that there was significant retardation of lysosomal proteolysis or receptor endocytosis by the hepatocyte at elevated temperatures.
- Published
- 1983
- Full Text
- View/download PDF
12. Model immune complexes formed between monoclonal antibodies and a novel bivalent affinity label.
- Author
-
Winkelhake JL
- Subjects
- Animals, Cell Line, Immunoglobulin G, Lymphocytes immunology, Mice, Mice, Inbred BALB C, Plasmacytoma, Affinity Labels, Antibodies, Neoplasm, Antigen-Antibody Complex, Dinitrofluorobenzene analogs & derivatives, Hybrid Cells immunology, Nitrobenzenes, Pimelic Acids
- Published
- 1980
- Full Text
- View/download PDF
13. Complement C1q binding affects spin-labeled heterosaccharides of rabbit antibodies in immune but not artificial immunoglobulin G aggregates.
- Author
-
Winkelhake JL, Kusumi A, McKean L, and Mandy WJ
- Subjects
- Animals, Antigen-Antibody Complex, Complement C1q, Electron Spin Resonance Spectroscopy, Haptens, Rabbits, Spin Labels, Antibodies, Complement Activating Enzymes immunology, Epitopes analysis, Immunoglobulin G immunology, Oligosaccharides
- Abstract
IgG anti-hapten antibodies were purified from the sera of rabbits homozygous for allotypic determinants d11 and d12 in the constant region of the heavy chain. Correlative with this determinant is the absence (d11) or presence (d12) of an oligosaccharide chain just below the hinge region of the IgG molecule. Both d11 and d12 molecules contain a complex heterosaccharide chain located near the carboxyl terminus of the second constant region domain. The two populations of IgG antibodies were thus selectively labeled with the spin probe Tempamine in their second constant region domains by reductive amination primarily of terminal N-acetylneuraminic acid residues. Chemical and enzymatic cleavages showed about 80% of the attached spin labels were N-acetylneuraminic acid-associated. Analysis of probe adducts by ESR spectrometry showed the presence of slower and faster moving subcomponents. Formation of immune complexes by antigen induces slight but significant restrictions of spin label mobility for both d11 and d12 IgG molecules. This restriction is qualitatively different from that seen in glutaraldehyde-, carbodiimide-, or ethanol-induced aggregates of the same IgG antibodies. The addition of purified complement C1 subcomponent C1q to immune aggregates resulted in marked immobilization of spin labels, the rotational correlation time of which was 30-40 mu s for both d11 and d12 molecules (evaluated by saturation transfer spectroscopy). A similar spin probe immobilizing effect is not seen when C1q binds to chemically aggregated IgG antibodies (which also do not activate C1). A novel model is proposed in which C1q is hypothesized to juxtapose Fc moieties in a discrete fashion required for subsequent C1 activation processes mediated by immune complexes.
- Published
- 1984
14. Protracted circulating lifetimes of mannose-terminated glycoproteins and aggregated albumin in mice infected with LDH-elevating virus.
- Author
-
Winkelhake JL, Elcombe BM, and Chang RJ
- Subjects
- Animals, Antibodies, Immunoglobulin G immunology, Kidney metabolism, Liver metabolism, Lung metabolism, Mice, Ovalbumin metabolism, Spleen metabolism, alpha-Fetoproteins metabolism, Glycoproteins metabolism, Lactate dehydrogenase-elevating virus, Mannose, Serum Albumin, Bovine metabolism, Virus Diseases metabolism
- Abstract
Several macromolecular homeostasis-regulating mechanisms were tested for functional integrities in mice during acute and early chronic phases of infection with lactic dehydrogenase-elevating virus (LDV). Fractional catabolic rates of carbodiimide-aggregated albumin and immunoglobulin G were studied to evaluate glomerular filtration and hepatic Kupffer cell phagocytic activities. Several glycoproteins (fetuin, IgG antibodies, and ovalbumin) were also compared with their deglycosylated counterparts for fractional catabolic rates and organ distributions as a basis for evaluating virus-induced modifications of saccharide-binding "receptor functions" in vivo. Findings were that normal hepatic clearance of aggregated albumin and of ovalbumin is slowed from the onset of viremia. Fractional catabolic rates of amannosyl-ovalbumin and amannosyl-IgG are similar in uninfected animals to those seen with native ovalbumin or with mannose-terminated IgG in LDV-infected animals. Ovalbumin and aggregated albumin were also found to be mutually competitive for hepatic uptake in uninfected animals. It is proposed that LDV, which replicates in cells of the mononuclear phagocyte system (reticuloendothelial system), alters the clearance functional state of fixed tissue macrophage, thereby explaining in part the protracted circulatory longevity of several enzymes, aggregated albumin and mannose-terminated ovalbumin, and IgG in LDV-infected mice.
- Published
- 1978
15. Contrast-enhanced MRI of tumors. Comparison of Gd-DTPA and a macromolecular agent.
- Author
-
Wikström MG, Moseley ME, White DL, Dupon JW, Winkelhake JL, Kopplin J, and Brasch RC
- Subjects
- Albumins, Animals, Drug Combinations, Female, Fibrosarcoma pathology, Gadolinium, Gadolinium DTPA, Macromolecular Substances, Meglumine, Mice, Mice, Inbred BALB C, Organometallic Compounds, Pentetic Acid, Fibrosarcoma diagnosis, Image Enhancement methods, Magnetic Resonance Imaging methods
- Abstract
The study aim was to define potential differences and advantages in magnetic resonance (MR) patterns of tumoral contrast enhancement using either a small molecular, extracellular fluid contrast enhancer [Gd-DTPA] or a macromolecular agent [albumin-(Gd-DTPA)20], designed for primary intravascular biodistribution. MR images of 25 mice with implanted fibrosarcomas were obtained before and repeatedly for up to 120 minutes after injection of either Gd-DTPA [0.2 mmol/kg, n = 11] or albumin-(Gd-DTPA) [0.0029 mmol/kg, n = 14]. Histologically, this hypovascular tumor contained zones of viable tissue and non-viable, necrotic tissue. Using either type of contrast media, the viable portions enhanced strongly, up to 152% and the necrotic portions enhanced poorly, less than 31%. However, the time-course of enhancement differed between contrast agents. Gd-DTPA tended to provide maximal enhancement soon after administration with no significant changes over two hours. Enhancement from albumin-(Gd-DTPA) was weak initially, corresponding to tumor hypovascularity, but over two hours the signal of the viable tumor zones progressively increased in intensity. This gradual tumoral accumulation of the macromolecular agent within the tumor was considered to reflect abnormal capillary permeability, associated with neovascularity. Thus, the increasing intensity within the neoplastic tissues over time, reflecting abnormal capillary permeability for macromolecules, may serve as a useful, albeit indirect, marker of neoplasia.
- Published
- 1989
- Full Text
- View/download PDF
16. Effects of chemical modification of antibodies on their clearance from the circulation. Addition of simple aliphatic compounds by reductive alkylation and carbodiimide-promoted amide formation.
- Author
-
Winkelhake JL
- Subjects
- Alkylation, Animals, Antigen-Antibody Reactions, Borohydrides, Carbodiimides, Glycine metabolism, Kidney metabolism, Kinetics, Oxidation-Reduction, Protein Binding, Rabbits immunology, Swine immunology, Amides, Immunoglobulin G metabolism
- Abstract
Anti-hapten antibodies from the ascitic fluid of inbred mice were purified by immunoadsorption and characterized immunochemically for in vivo studies of their plasma clearance rates and organ distributions after chemical modification. Following sodium borohydride-promoted reductive methylation and carbodiimide-promoted amide linkage of glycine and several other simple aliphatic compounds, the antibody populations were recharacterized, radiolabeled, and introduced intravenously into syngeneic animals. Using double radioiodine labels, it was possible to show that stoichiometric combinations of additive and chemical reactant did not alter antibody survival time in the circulation or antigen-binding capabilities. However, modifications involving excess reagent (carbodiimide or sodium borohydride) resulted in significant decreases in both circulatory longevity and ligand binding capacities. Excess carbodiimide treatments resulted in immunoglobulin cross-linkage which could be detected by molecular sieve chromatography. Increased kidney localization found with carbodiimide-treated antibodies was apparently due to trapping of the cross-linked aggregates. Elevated clearance of highly methylated antibodies could not be attributed to a single organ. However, sodium borohydride-catalyzed reductive methylation resulted in antibody populations which were localized primarily in the liver and spleen. Results are evaluated in terms of the concept, developed in this paper, of essential groups for the circulatory longevity of glycoproteins.
- Published
- 1977
17. The role of oxidant injury in tumor cell sensitivity to recombinant human tumor necrosis factor in vivo. Implications for mechanisms of action.
- Author
-
Zimmerman RJ, Marafino BJ Jr, Chan A, Landre P, and Winkelhake JL
- Subjects
- Acetylcysteine administration & dosage, Animals, Female, Free Radicals, Glutathione metabolism, Humans, Lipid Peroxides toxicity, Maleates administration & dosage, Mice, Mice, Inbred BALB C, Mice, Nude, Neoplasm Transplantation, Neoplasms, Experimental pathology, Neoplasms, Experimental therapy, Rats, Rats, Inbred Strains, Neoplasms, Experimental metabolism, Oxygen toxicity, Recombinant Proteins administration & dosage, Tumor Necrosis Factor-alpha administration & dosage
- Abstract
The intracellular glutathione levels of two human tumor lines and seven murine tumor lines were determined in order to investigate the role of oxidant injury in tumor cell sensitivity to human rTNF (rhTNF). Correlations were found between high intracellular glutathione levels and in vivo tumor resistance to rhTNF, and on the other hand, low glutathione levels and rhTNF sensitivity. The transplantable murine fibrosarcoma, Meth A, a TNF-sensitive line in vivo, was less sensitive to rhTNF and host toxicity was reduced when the hosts were pretreated with uric acid, a major reactive oxygen scavenger in humans and certain other primates. Conversely, pretreatment of the tumor-bearing hosts with DL-buthionine-(S,R)-sulfoximine, an inhibitor of GSH biosynthesis, resulted in an increased sensitivity of Meth A to rhTNF. This effect was not limited to tumor-bearing mice, as rats pretreated with diethyl maleate, a compound which irreversibly binds glutathione, were more sensitive to rhTNF toxicity than control rats. On the other hand, pretreatment with N-acetyl cysteine, an oxidant scavenger, reduced the toxicity of rhTNF treatment in rats. The data are consistent with the hypothesis that tumor cell sensitivity to rhTNF in vivo is dependent on its capacity to buffer oxidative attack. In addition, host toxicity is also related to the production of reactive oxygen species. Activated effector cells such as granulocytes and macrophages are hypothesized to produce most of this damage by their respiratory burst and oxidant release, although the direct action of rhTNF may also contribute to oxidative injury in vivo.
- Published
- 1989
18. Acute phase (C-reactive) protein-like macromolecules from rainbow trout (Salmo gairdneri).
- Author
-
Winkelhake JL and Chang RJ
- Subjects
- Agglutination Tests, Animals, C-Reactive Protein biosynthesis, Chromatography, Affinity, Electrophoresis, Polyacrylamide Gel, Female, Macromolecular Substances, Male, Rabbits, Stress, Physiological chemically induced, Stress, Physiological etiology, Temperature, Turpentine, C-Reactive Protein isolation & purification, Salmonidae immunology, Trout immunology
- Abstract
A protein which reacts with the Cx-polysaccharide of Streptococcus pneumoniae and is inhibited by phosphorylcholine was isolated from the serum of rainbow trout by affinity chromatography. The protein, which exists in monomeric and oligomeric forms in non-immune trout serum, is very similar with regard to specificity and size to the Cx-reactive protein from rabbits. A semi-quantitative analytical method for evaluating bacterial agglutination with an electronic particle counter and size distribution analyzer was developed to compare natural and acute serum levels of trout and rabbit Cx-reactive proteins. Results indicate that the poikilotherm has much higher concentrations in normal serum. The trout serum protein can also be rapidly induced to yet higher levels by both chemical and physical stress. The implications for such a protein in the teleost's natural defense system and overall homeostasis are discussed.
- Published
- 1982
- Full Text
- View/download PDF
19. Schedule dependency of the antitumor activity and toxicity of polyethylene glycol-modified interleukin 2 in murine tumor models.
- Author
-
Zimmerman RJ, Aukerman SL, Katre NV, Winkelhake JL, and Young JD
- Subjects
- Animals, Drug Administration Schedule, Immunotherapy, Interleukin-2 administration & dosage, Interleukin-2 pharmacokinetics, Mice, Mice, Inbred Strains, Polyethylene Glycols, Recombinant Proteins, Interleukin-2 analogs & derivatives, Neoplasms, Experimental therapy
- Abstract
Modification of recombinant human interleukin 2 (rhIL-2) with monomethoxy polyethylene glycol has been shown to alter its pharmacokinetic properties. Therefore, we investigated the pharmacological parameters of schedule and dose in order to assess the impact on the in vivo antitumor activity of this modification. The antitumor efficacy, as well as the toxicity, of polyethylene glycol-interleukin 2 (PEG-IL-2) was compared to that of rhIL-2 in three transplantable syngeneic murine tumor models, Meth A fibrosarcoma, B16 melanoma, and Pan-02 pancreatic carcinoma. At equitoxic dose levels, the antitumor activity of PEG-IL-2 was far superior to that of rhIL-2 in all three tumor models. This efficacy of PEG-IL-2 was dose dependent and was greatest on a Q7D x 2 schedule in Meth A and B16. When the same total doses were further divided and delivered on any of several alternative schedules, either the efficacy was reduced or the toxicity of the treatments was increased. In Pan-02, a rhIL-2-resistant tumor, PEG-IL-2 treatment on either the Q7D x 2, Q4D x 3, or Q3D x 4 schedule resulted in approximately a 200% increase in lifespan; however, the toxicity of the treatment increased as the interval between doses was shortened. Simulations of the pharmacokinetic profiles of these various regimens suggested that the toxicity of PEG-IL-2 and rhIL-2 was related to the minimum plasma concentration that was obtained and the time interval between peak levels. The efficacy of the treatment was associated with the interleukin 2 plasma peak height, since a dose response was observed; however, peak plasma concentration did not appear to be the only parameter which determined efficacy. We hypothesize that this observed schedule dependence is also affected by the kinetics of the host's biological response to rhIL-2.
- Published
- 1989
20. Alcian blue as a protein stain for sodium alkyl sulfate--polyacrylamide gels: application to analysis of polypeptide components of the human platelet.
- Author
-
Kunicki TJ, Christie DJ, Winkelhake JL, and Aster RH
- Subjects
- Detergents, Electrophoresis, Polyacrylamide Gel, Humans, Peptides blood, Sodium Dodecyl Sulfate, Staining and Labeling, Alcian Blue, Blood Platelets analysis, Blood Proteins analysis, Indoles
- Published
- 1981
- Full Text
- View/download PDF
21. Mechanism of lysergic acid diethylamide interference with rabbit antibody biosynthesis.
- Author
-
Voss EW Jr and Winkelhake JL
- Subjects
- Animals, Antibodies analysis, Haptens, Immunoglobulin G biosynthesis, Iodine Radioisotopes, Isoelectric Focusing, Leucine metabolism, Rabbits, Swine, Tritium, Tryptophan metabolism, Antibody Formation drug effects, Antibody-Producing Cells drug effects, Lysergic Acid Diethylamide pharmacology
- Abstract
Lymphoid cells from hyperimmune rabbits producing antibodies to a hapten, incubated in the presence of d-lysergic acid diethylamide, continued to synthesize protein at a normal rate. Isoelectric focusing analysis of the low-molecular-weight protein secreted by the cells incubated with lysergic acid diethylamide indicated two components, with pI's of 4.9 and 5.2. Immune cells not exposed to lysergic acid diethylamide secreted only 7S IgG molecules with an average pI of approximately 7.0.
- Published
- 1974
- Full Text
- View/download PDF
22. Organ specificity of blood-borne tumour metastasis determined by cell adhesion?
- Author
-
Nicolson GL and Winkelhake JL
- Subjects
- Animals, Cell Aggregation, Cell Line, Cell Separation, Centrifugation, Density Gradient, Erythrocytes immunology, Immunity, Cellular, Kidney Neoplasms immunology, Liver Neoplasms immunology, Lung Neoplasms immunology, Mice, Mice, Inbred C57BL, Neoplasms, Experimental immunology, Splenic Neoplasms immunology, Cell Adhesion, Melanoma immunology, Neoplasm Metastasis immunology, Organ Specificity
- Published
- 1975
- Full Text
- View/download PDF
23. Induction in rainbow trout of an acute phase (C-reactive) protein by chemicals of environmental concern.
- Author
-
Winkelhake JL, Vodicnik MJ, and Taylor JL
- Subjects
- Animals, Cytochrome P-450 Enzyme System biosynthesis, Female, Male, Methylcholanthrene pharmacology, Microsomes, Liver drug effects, Microsomes, Liver metabolism, C-Reactive Protein blood, Insecticides pharmacology, Salmonidae blood, Trout blood, Water Pollutants, Water Pollutants, Chemical, Water Pollution, Chemical
- Abstract
1. Rainbow trout, Salmo gairdneri, produce elevated amounts of a serum acute phase (C-reactive) protein (CRP) when administered a variety of chemicals of environmental importance. 2. Compounds administered in doses which induce the cytochrome(s) P450 catalytic enzymes in trout hepatic microsomes also induce serum CRP. 3. However, an interferon-inducing virus does not induce CRP. Interferon induction by the virus is not significantly inhibited by chemicals which induce trout cytochrome(s) P450. 4. Simultaneous administration of chemicals and virus or virus alone results in depression of P450 protein production and only minor induction of CRP. 5. Thus, as with mammals, a reciprocating relationship appears to exist between the hemeprotein monooxygenase and immune systems of this freshwater teleost, and C-reactive protein appears to fit the reciprocating scheme closer to the cytochromes P450 response.
- Published
- 1983
- Full Text
- View/download PDF
24. Effects of pH treatments and deglycosylation of rabbit immunoglobulin G on the binding of C1q.
- Author
-
Winkelhake JL, Kunicki TJ, Elcombe BM, and Aster RH
- Subjects
- Animals, Antigen-Antibody Complex, Complement C1q, Erythrocytes immunology, Glycoside Hydrolases, Hydrogen-Ion Concentration, Immunoglobulin Fab Fragments, Immunoglobulin Fc Fragments, Kinetics, Protein Binding, Rabbits immunology, Sheep, Complement Activating Enzymes, Immunoglobulin G
- Published
- 1980
25. A dynamic continuum model for molecular regulation of the humoral immune response.
- Author
-
Winkelhake JL
- Subjects
- Antibody Formation, Antibody Specificity, Antibody-Producing Cells, Antigens, Binding Sites, Antibody, Cell Membrane immunology, Epitopes, Immunologic Memory, Immunity, Models, Biological
- Published
- 1976
- Full Text
- View/download PDF
26. Evolution of antibody structure and effector functions: comparative hemolytic activities of monomeric and tetrameric IgM from rainbow trout, Salmo gairdnerii.
- Author
-
Elcombe BM, Chang RJ, Taves CJ, and Winkelhake JL
- Subjects
- Animals, Antibody Formation, Antigen-Antibody Complex analysis, Chromatography, Affinity, Haptens, Immunodiffusion, Immunoglobulin M genetics, Immunoglobulin M isolation & purification, Macromolecular Substances, Peptide Fragments analysis, Biological Evolution, Epitopes analysis, Hemolysis, Immunoglobulin M physiology, Salmonidae immunology, Trout immunology
- Abstract
Monomeric and tetrameric IgM anti-haptin antibodies isolated from the sera of rainbow trout (S. gairdnerii) by immunoaffinity chromatography were compared both immunochemically and with regard to their functional abilities to lyse haptenated trout erythrocytes in the presence of trout complement. The two populations had similar binding affinities for hapten and apparently identical L chains, but differed in H chain peptide maps and immunoreactivity with rabbit anti-trout H chain serum. These differences could not be attributed to J-chain. The abilities of the two antibody subpopulations to activate C to lyse haptenated trout erythrocytes also differed dramatically. Such functional differences are not simply explained by the greater avidity of the tetrameric form since preliminary studies show that the monomeric form of trout IgM activates C via an alternative pathway mechanism while the tetrameric form activates both classical and alternative pathway mechanisms. Results suggest divergent evolution of antibody structures involved in the familiar effector functions (C activation, transport, etc.).
- Published
- 1985
- Full Text
- View/download PDF
27. Purification of calciferol-binding proteins from kidney: physicochemical and immunological properties.
- Author
-
Ghazarian JG, Hsu PY, Girotti AW, and Winkelhake JL
- Subjects
- Amino Acids analysis, Animals, Binding, Competitive, Carrier Proteins immunology, Dihydroxycholecalciferols metabolism, Immunoelectrophoresis, Molecular Weight, Rats, Ultracentrifugation, Carrier Proteins isolation & purification, Hydroxycholecalciferols metabolism, Kidney metabolism
- Abstract
The calciferol-binding system of rat kidney cytosol has been purified and is shown to consist of two proteins, each capable of binding either 25-hydroxy-vitamin D3 (25-OH-D3) or 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). The two proteins, designated A and B, have similar sedimentation coefficients (S20w) of 5.2 S. Component A binds 25-OH-D3 with a dissociation constant (Kd) of 10(-7) M while component B binds 1,25-(OH)2D3 with a Kd of 1.6 x 10(-8) M. The estimated molecular weights (Mr) of the two proteins are 105,000 for component A and 250,000 for component B. Amino acid analyses revealed that glutamic acid is the most abundant residue in both proteins, comprising 12% of the total number of amino acid residues. Immunodiffusion test using commercial anti-human serum group-specific protein antiserum gave a precipitin reaction when purified rat serum calciferol-binding protein was used as an antigen, but no reactions could be detected with proteins A and B. This result significantly eliminated the possibility of the presence of the rat serum binding protein in either of the purified kidney proteins. In contrast, anti-rat serum calciferol-binding protein antiserum prepared in rabbits interacted with the rat serum and kidney proteins. This result suggests that the antigenic determinants recognized by the antiserum against the rat serum calciferol-binding protein appear to be similar to those recognized in the kidney proteins A and B. Immunoelectrophoresis of the three rat proteins demonstrated dissimilar electrophoretic mobilities with the serum protein showing the least mobility, a property consistent with its higher lysine content relative to proteins A and B.
- Published
- 1978
28. Aglycosylantibody. Effects of exoglycosidase treatments on autochthonous antibody survival time in the circulation.
- Author
-
Winkelhake JL and Nicolson GL
- Subjects
- Acetylglucosaminidase, Animals, Blood Circulation Time, Galactosidases, Immunoelectrophoresis, Kinetics, Lactoperoxidase metabolism, Neuraminidase, Protein Binding, Rabbits immunology, Solubility, Streptococcus pneumoniae enzymology, Swine, Time Factors, Antibodies metabolism, Glycoside Hydrolases, gamma-Globulins
- Abstract
Rabbit anti-hapten antibodies were purified by affinity chromatography and characterized immunochemical for in vivo studies of their blood clearance rate and organ distribution after treatment with various glycosidases. Following sequential removal of sialic acid, galactose, and N-acetylglucosamine with the appropriate cellulose-immobilized exoglycosidases, the antibody populations were recharacterized, radiolabeled, and introduced intravenously into the original animals. Using double radioiodine lables it was possible to demonstrate alterations in purified antibody survival times in the circulation and altered organ distribution after glycolytic cleavage. Removal of terminal sialic acid resulted in rapid blood clearance and enhanced localization of asialoantibody in the liver. Subsequent removal of penultimate galactose residues returned both antibody survival time in the circulation and organ distribution to near normal. Removal of subpenultimate N-acetylglucosamine moieties resulted in aglycosylantibody survival values which were intermediate between asialo- and asialoagalactoantibodies. Removal of the three saccharide also increased kidney localization. The results are evaluated based on current concepts of the biological roles of protein-linked carbohydrate and plasma glycoprotein survival time in the circulation.
- Published
- 1976
29. Light chain peptide secreted by rabbit antibody producing and murine MOPC 315 myeloma cells in the presence of D-lysergic acid diethylamide (LSD).
- Author
-
Voss EW Jr, Winkelhake JL, and Cobb NE
- Subjects
- Animals, Antibody Specificity, Cell Line, Chromatography, Affinity, Mice, Plasmacytoma immunology, Polyribosomes drug effects, Polyribosomes immunology, Polyribosomes metabolism, Rabbits immunology, Antibody-Producing Cells, Immunoglobulin Light Chains biosynthesis, Lysergic Acid Diethylamide pharmacology, Plasmacytoma metabolism
- Published
- 1975
- Full Text
- View/download PDF
30. Synergistic effects of combination therapy with human recombinant interleukin-2 and tumor necrosis factor in murine tumor models.
- Author
-
Winkelhake JL, Stampfl S, and Zimmerman RJ
- Subjects
- Animals, Drug Synergism, Female, Glycoproteins administration & dosage, Humans, Immunotherapy, Interleukin-2 administration & dosage, Lung Neoplasms secondary, Lung Neoplasms therapy, Melanoma, Experimental secondary, Melanoma, Experimental therapy, Mice, Mice, Inbred C57BL, Mice, Inbred DBA, Neoplasm Transplantation, Recombinant Proteins administration & dosage, Recombinant Proteins therapeutic use, Tumor Necrosis Factor-alpha, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Glycoproteins therapeutic use, Interleukin-2 therapeutic use, Neoplasms, Experimental therapy
- Abstract
Human recombinant interleukin 2 (IL-2) and tumor necrosis factor (TNF) were evaluated individually and in combination for their antitumor efficacy in vivo, using five s.c. murine tumors: L1210 leukemia, P815 mastocytoma, B16 melanoma, EL-4 lymphoma, and the methylcholanthrene-induced sarcoma, Meth A. While only the s.c. methylcholanthrene-induced tumor exhibited regression and/or cures in response to immunomodulatory therapy with either agent alone, the simultaneous administration of a maximally tolerated dose of TNF and IL-2 given daily from within 1 day (B16 melanoma), 3 days (L1210 leukemia and P815 mastocytoma) or 5 and 10 days (EL-4 lymphoma and methylcholanthrene-induced sarcoma) after tumor cell implant resulted in no tumor takes (growth). The TNF dose was apparently rate limiting in that reduction of the amount of TNF in the combination by 50% resulted in the loss of curative effects, while IL-2 doses could be reduced by 90% (depending upon tumor type) and still result in an efficacious combination. The synergy seen in combination IL-2 and TNF therapy appeared to be dependent upon tumor burden, but somewhat independent of tumor location. For example, no tumors were seen in the artificial pulmonary metastasis model of the B16 melanoma, and the percentage of extension of median lifetime (test versus control) greater than 150% was seen in the i.p. B16 melanoma, as well as several other i.p. models of the five tumor types. On the other hand, no significant extension of lifetime (greater than 150%) was seen with either lymphokine alone when administered i.p. at maximally tolerated dose for any of the five tumors tested here. Results are discussed in relation to potential immune modulatory events which may be occurring during combination treatments.
- Published
- 1987
31. Immunoglobulin structure and effector functions.
- Author
-
Winkelhake JL
- Subjects
- Animals, Antigen-Antibody Complex, Antigen-Antibody Reactions, Carbohydrates immunology, Complement C1 immunology, Complement Pathway, Classical, Disulfides, Female, Homeostasis, Humans, Immunoglobulin Constant Regions, Immunoglobulin E, Immunoglobulin Variable Region, Macrophages immunology, Maternal-Fetal Exchange, Pregnancy, Protein Conformation, Secretory Component metabolism, Immunoglobulins biosynthesis
- Published
- 1978
- Full Text
- View/download PDF
32. Preparation of protease-free neuraminidase by affinity adsorption on fetuin-derivatized cellulose.
- Author
-
Winkelhake JL and Nicolson GL
- Subjects
- Chromatography, Affinity, Sepharose, Cellulose, Fetal Proteins, Neuraminidase isolation & purification, Peptide Hydrolases, alpha-Fetoproteins
- Published
- 1976
- Full Text
- View/download PDF
33. Radioimmune assay and characteristics of antibodies to macromomycin (NSC 170105).
- Author
-
Winkelhake JL and Buckmire FL
- Subjects
- Antibiotics, Antineoplastic metabolism, Antibiotics, Antineoplastic pharmacology, Binding Sites, Cell Membrane metabolism, Cells, Cultured, DNA, Neoplasm biosynthesis, Leucine metabolism, Neoplasms, Experimental drug therapy, Neoplasms, Experimental immunology, Neoplasms, Experimental metabolism, Radioimmunoassay, Thymidine metabolism, Antibiotics, Antineoplastic immunology, Antibodies analysis
- Abstract
A radioimmune assay for the antitumor agent, macromomycin, using purified, radioiodine-labeled macromomycin and antisera raised in rabbits against a carbodiimide-catalyzed macromycin-Limulus polyphemus hemocyanin complex has been developed. Radiolabeled macromomycin was prepared by direct iodination of the polypeptide antibiotic with the use of iodine monochloride or solid-state lactoperoxidase. Antibody-bound drug was isolated from free macromomycin with dextran-coated, activated charcoal. The standard curve of the sequential saturation assay was linear on a logit-log plot and indicated a lower limit of sensitivity of approximately 100 pg macromomycin. The radioimmune assay was suitable for measuring macromomycin in the presence of other antitumor drugs, and detection of macromomycin was quantitative when it was added to normal human serum or urine. Drug binding to melanoma and mammary carcinoma cell surfaces could be inhibited by preincubating macromomycin with affinity-purified antimacromomycin antibodies. However, once the drug was bound to cell surfaces, addition of antimacromomycin antibodies did not result in removal of the drug from cell surfaces or in reversal of macromomycin-induced inhibition of thymidine incorporation into cellular DNA. Antimacromovide useful tools for developing pharmacokinetic and toxicity studies of macromomycin, as well as for analyzing the mechanism(s) of action of the drug.
- Published
- 1977
34. Determination of adhesive properties of variant metastatic melanoma cells to BALB/3T3 cells and their virus-transformed derivatives by a monolayer attachment assay.
- Author
-
Winkelhake JL and Nicolson GL
- Subjects
- Animals, Cell Aggregation, Cell Line, Mice, Mice, Inbred BALB C, Temperature, Cell Adhesion, Cell Transformation, Neoplastic, Melanoma pathology, Neoplasm Metastasis pathology
- Abstract
The hypothesis that abnormalities in intercellular adhesion are a property of metastatic tumors was examined in vitro with B16 melanoma variants that were selected in vivo for increased metastatic behavior. The adhesive characteristics of low (B16-F1), intermediate (B16-F5), and high (B16-F10) metastatic lines were determined by quantitative adhesion assays that measured the rate and degree of attachment of single cells to confluent monolayers of melanoma, BALB/3T3, or virus-transformed 3T3 cells. Intercellular adhesions were monitored by loss of single cells from suspension and adherence of intraperitoneally grown 125I-5-iodo-2'-deoxyuridine-labeled cells to the monolayers, and were affected by time, temperature, and serum concentration. Although there was little difference in adhesive properties between the untransformed and transformed 3T3 cell lines, the more metastatic melanoma variants exhibited higher relative rates and extents of homotypic and heterotypic monolayer attachment compared with lower metastatic lines (B16-F10 greater than B16-F5 greater than B16-F1). The correlation between in vivo and in vitro tumor cell adhesive properties and metastasis was discussed.
- Published
- 1976
- Full Text
- View/download PDF
35. Fc:Fc interactions revealed by spin-labeled IgG heterosaccharides in model immune complexes.
- Author
-
Kusumi A and Winkelhake JL
- Subjects
- Animals, Antibodies, Monoclonal, Dinitrobenzenes immunology, Electron Spin Resonance Spectroscopy, Glycoproteins, Macromolecular Substances, Mice, Oligosaccharides, Spin Labels, Antigen-Antibody Complex, Immunoglobulin Fc Fragments, Immunoglobulin Fragments, Immunoglobulin G
- Abstract
Dynamic properties of spin-labelled heterosaccharides in the Fc-region of murine monoclonal antihapten immunoglobulin G were studied in model immune complexes (IC) as a function of the IC size. Model IC dimers, trimers and oligomers were formed using bivalent photoaffinity antigens. The ESR spectrum exhibits two components. The rotational correlation time of the less-immobilized species is shorter than 10(-10) sec, and that of the more-immobilized component is in the order to 10(-9) approximately 10(-8) sec depending on the IC size. Fraction of the more-immobilized spin labels increases, and the mobility of this component decreases with increase in IC size (i.e., mobility: monomers approximately equal to dimers greater than trimers much greater than immune-complex precipitates). These data strongly suggest the existence of Fc:Fc interactions in IC, and provide the basis for a model in which such interactions underlie the initial mechanism by which the information of antigen binding to Fab region is transferred into organized Fc:Fc association structure for IgG effector activities.
- Published
- 1986
- Full Text
- View/download PDF
36. Ferredoxin of 25-hydroxyvitamin D3-1alpha-hydroxylase. Anatomical distribution in the chick as determined by double-antibody radioimmunoassay.
- Author
-
Kulkoski JA, Peterson BL, Elcombe B, Winkelhake JL, and Ghazarian JG
- Subjects
- Animals, Kidney enzymology, Liver enzymology, Mitochondria enzymology, Radioimmunoassay methods, Tissue Distribution, Vitamin D Deficiency enzymology, 25-Hydroxyvitamin D3 1-alpha-Hydroxylase metabolism, Ferredoxins metabolism, Steroid Hydroxylases metabolism
- Published
- 1979
- Full Text
- View/download PDF
37. Cell surface changes and Rous sarcoma virus gene expression in synchronized cells.
- Author
-
Hale AH, Winkelhake JL, and Weber MJ
- Subjects
- Animals, Biological Transport, Active, Cell Division, Cell Membrane metabolism, Cell Membrane ultrastructure, Cells, Cultured, Chick Embryo, Deoxyglucose metabolism, Fibroblasts, Genes, Mannitol metabolism, Microscopy, Electron, Scanning, Surface Properties, Thymidine metabolism, Tritium, Uridine metabolism, Avian Sarcoma Viruses growth & development, Cell Transformation, Neoplastic
- Abstract
We have investigated whether cell surface changes associated with growth control and malignant transformation are linked to the cell cycle. Chicken embryo cells synchronized by double thymidine block were examined for cell-cycle-dependent alterations in membrane function (measured by transport of 2-deoxyglucose, uridine, thymidine, and mannitol), in cell surface morphology (examined by scanning electron microscopy), and in the ability of tumor virus gene expression to induce a transformation-specific change in membrane function. We reach the following conclusions: (a) The high rate of 2-deoxyglucose transport seen in transformed cells and the low rates of 2-deoxyglucose and uridine transport characteristic of density-inhibited cells do not occur in normal growing cells as they traverse the cell cycle. (b) Although there are cell cycle-dependent changes in surface morphology, they are not reflected in corresponding changes in membrane function. (c) Tumor virus gene expression can alter cell membrane function at any stage in the cell cycle and without progression through the cell cycle.
- Published
- 1975
- Full Text
- View/download PDF
38. Bactericidal activity of macromomycin, a antitumor antibiotic.
- Author
-
Buckmire FL and Winkelhake JL
- Subjects
- Bacillus drug effects, Microbial Sensitivity Tests, Pseudomonas aeruginosa drug effects, Staphylococcus aureus drug effects, Antibiotics, Antineoplastic pharmacology, Bacteria drug effects
- Abstract
Susceptibility testing by the broth dilution method showed that all the gram-positive but only some of the gram-negative bacteria tested were susceptible to the antitumor antibiotic, macromomycin (MCR; NSC 170105). The minimal inhibitory concentration for the susceptible organisms was less than 3 mug/ml. The action of MCR was bactericidal; however, at very high concentrations (50 mug/ml and above) some organisms occasionally escaped death. None of the escaped organisms was resistant to MCR. In combination with other commonly used antibiotics, MCR displayed partial synergy for Pseudomonas aeruginosa (from a minimal inhibitory concentration of >100 to 12.5 mug/ml with 100 mug of chloramphenicol per ml) and for Bacillus pumilus and Staphylococcus aureus (from 1.6 to 0.4 mug/ml and below) with polymyxin B. As with mammalian cells, (125)I-labeled MCR was irreversibly bound to both gram-positive and -negative bacteria. Treatment with trypsin of the (125)I-labeled MCR-exposed cells did not release the bound MCR or reverse its lethal effect. When in solution in a protective buffer at 4 degrees C, MCR was stable for up to 45 days; at 37 degrees C, however, 25% of its bactericidal activity was lost in 72 h. Loss of activity was enhanced 16-fold in the presence of both heated and unheated pooled human sera. Urine had no effect on the activity of MCR.
- Published
- 1977
- Full Text
- View/download PDF
39. Comparative inhibitory action of D- and L-tryptophan on the effect of d-lysergic acid diethylamide in vitro.
- Author
-
Winkelhake JL, Voss EW Jr, and Lopatin DE
- Subjects
- Ammonium Sulfate, Animals, Antibodies analysis, Antibody Formation drug effects, Antibody-Producing Cells drug effects, Binding, Competitive, Carbon Radioisotopes, Cattle, Cells, Cultured, Chromatography, Gel, Fluoresceins, Hydrolysis, Immunization, Indoleacetic Acids, Iodine Radioisotopes, Leucine metabolism, Lysergic Acid Diethylamide metabolism, Pronase metabolism, Proteins metabolism, Rabbits immunology, Serum Albumin, Bovine, Stereoisomerism, Swine, Time Factors, Tritium, gamma-Globulins, Lysergic Acid Diethylamide antagonists & inhibitors, Tryptophan pharmacology
- Published
- 1974
40. Human aglycosyl-IgG exhibits increased hydrophobicity. Binding/fluorescence studies with 8-anilinonaphthalene-1-sulfonic acid (ANS).
- Author
-
Taves CJ, Kusumi A, and Winkelhake JL
- Subjects
- Anilino Naphthalenesulfonates, Cell Line, Chromatography, Gel, Fluorescent Dyes, Humans, Hydrogen-Ion Concentration, Immunoglobulin G isolation & purification, Molecular Weight, Protein Binding, Tunicamycin pharmacology, Antibodies, Monoclonal, Immunoglobulin G genetics
- Abstract
Human monoclonal, aglycosyl-IgG produced in vitro in the presence of tunicamycin, was compared with its native and acid pH-altered counterparts for their respective abilities to bind the fluorescent hydrophobicity probe, 8-anilinonaphthalene sulfonate. A novel technique based on continuous-flow dynamic dialysis (Sparrow et al., 1982, Anal. Biochem. 123:255-264) allowed binding studies under non-equilibrium conditions. While the native IgG conformation exhibits two, weak ANS binding sites (ca. 10(3) l/mol), aglycosyl-IgG has one weak and one moderate affinity (least squares average Ka = 2 X 10(4) l/mol) site, and the acid conformer binds yet another two ANS molecules with moderate affinity (4 X 10(4) l/mol). Increases in affinity and in the number of sites correlate roughly with increased relative percent fluorescence by conventional fluorimetry. The fluorescence lifetime of ANS bound to altered IgGs is about 10% longer (T2 = 15 nsec by time-resolved fluorimetry) than that for native IgG. All populations also exhibit a rapid decay component (T1 = 3 nsec) analogous to that seen for ANS in 50% aqueous dioxane. Results are discussed in relation to structural role(s) for IgG-linked heterosaccharides.
- Published
- 1984
- Full Text
- View/download PDF
41. Antitumor activity of an immunotoxin in a nude mouse model of human ovarian cancer.
- Author
-
FitzGerald DJ, Bjorn MJ, Ferris RJ, Winkelhake JL, Frankel AE, Hamilton TC, Ozols RF, Willingham MC, and Pastan I
- Subjects
- Animals, Cell Line, Dose-Response Relationship, Drug, Female, Humans, Mice, Mice, Nude, Neoplasm Invasiveness, Neoplasm Transplantation, Ovarian Neoplasms pathology, Transplantation, Heterologous, Immunotoxins therapeutic use, Ovarian Neoplasms therapy, Receptors, Transferrin immunology, Ricin therapeutic use
- Abstract
An immunotoxin composed of an antibody to the human transferrin receptor (454A12) and ricin A chain (RTA) was shown to inhibit the growth of NIH:OVCAR-3 tumors in a nude mouse model of human ovarian cancer. Inhibition of tumor growth by 454A12-RTA was related to the dose administered. The antitumor activity of the immunotoxin was blocked by coinjection of excess antibody with immunotoxin. An immunotoxin made using 454A12 and recombinant ricin A chain (rRTA) had an activity similar to that made with native RTA. The administration of 10 micrograms or greater of the immunotoxin 454A12-RTA/rRTA had significant antitumor activity. The injection of 30 micrograms of an irrelevant immunotoxin, MOPC21-RTA, or 30 to 500 micrograms of the 454A12 antibody had no antitumor activity.
- Published
- 1987
42. Immunological block to synthetic alpha-melanocyte-stimulating hormone: melanocyte interaction by antibodies isolated from cell-column immunoadsorbents.
- Author
-
Winkelhake JL, Elcombe BM, and Hodach A
- Subjects
- Animals, Antigens, Neoplasm, Cell Membrane immunology, Chromatography, Affinity, Melanocyte-Stimulating Hormones metabolism, Melanoma metabolism, Mice, Mice, Inbred C57BL, Neoplasms, Experimental immunology, Antibodies, Neoplasm isolation & purification, Immunity, Melanocyte-Stimulating Hormones immunology, Melanocytes immunology, Melanoma immunology
- Abstract
Antibodies to formalin-fixed, syngeneic melanoma cells were prepared in mice, purified by immunoaffinity chromatography, and tested for binding activity to viable melanoma cells. The radiolabeled antibodies detected congruent to 9 X 10(6) melanoma antigenic sites/cell. The calculated average association constant (Ka) for the antibody population was 7 to 10 X 10(7) M-1. The antibody was shown to block the binding of melanocyte-stimulating hormone in competitive cell surface binding studies. Results are discussed conceptually in terms of the potentially important role that the humoral immune response may play in the phenomenon of tumor progression.
- Published
- 1979
43. Sequence dependence of administration of human recombinant tumor necrosis factor and interleukin-2 in murine tumor therapy.
- Author
-
Zimmerman RJ, Gauny S, Chan A, Landre P, and Winkelhake JL
- Subjects
- Animals, Antineoplastic Combined Chemotherapy Protocols, Drug Administration Schedule, Drug Synergism, Immunologic Deficiency Syndromes immunology, Mice, Recombinant Proteins therapeutic use, Time Factors, Interleukin-2 administration & dosage, Neoplasms, Experimental therapy, Tumor Necrosis Factor-alpha administration & dosage
- Abstract
Simultaneous administration of recombinant human tumor necrosis factor (rhTNF) and interleukin-2 (rhIL-2) has been shown to block tumor take in murine models. We investigated the effects of sequence and schedule of administration as a function of tumor burden with two tumor models (B16 and Meth A). rhTNF followed by rhIL-2 had extraordinary antitumor efficacy, but rhIL-2 followed by rhTNF was much less effective. Sequential rhTNF/rhIL-2 therapy resulted in complete tumor regression, whereas simultaneous therapy resulted in complete tumor regression, whereas simultaneous therapy resulted in only reduced growth rate. Experiments with genetically immunodeficient mice suggested that T cell factors may be required for synergistic antitumor activity.
- Published
- 1989
- Full Text
- View/download PDF
44. Immunochemical similarity between polysaccharide antigens of Escherichia coli 07: K1(L):NM and group B Neisseria meningitidis.
- Author
-
Kasper DL, Winkelhake JL, Zollinger WD, Brandt BL, and Artenstein MS
- Subjects
- Animals, Antibody Formation, Autoanalysis, Bacterial Proteins analysis, Carbon Isotopes, Chromatography, Gel, Clostridium perfringens enzymology, Hemagglutination Inhibition Tests, Hexosamines analysis, Hexoses analysis, Humans, Immunodiffusion, Neuraminidase metabolism, Rabbits, Antigens, Bacterial analysis, Escherichia coli immunology, Neisseria meningitidis immunology, Polysaccharides, Bacterial analysis
- Published
- 1973
45. Incorporation of a lysergic acid diethylamide intermediate into antibody protein in vitro.
- Author
-
Voss EW Jr, Metzel P, and Winkelhake JL
- Subjects
- Animals, Antibody-Producing Cells metabolism, Carbon Isotopes, Centrifugation, Density Gradient, Dialysis, Leucine metabolism, Ligands, Lymphoid Tissue immunology, Puromycin pharmacology, Rabbits, Serum Albumin, Bovine, Serum Albumin, Radio-Iodinated, Spleen immunology, Sucrose, Tritium, Tryptophan metabolism, Antibody Formation, Lysergic Acid Diethylamide metabolism
- Published
- 1973
46. Affinity chromatography of anti-meningococcal antiserum.
- Author
-
Winkelhake JL and Kasper DL
- Subjects
- Acrylamides, Adsorption, Animals, Antigens, Bacterial, Bacterial Vaccines, Blood Bactericidal Activity, Carbon Isotopes, Complement System Proteins, Dextrans, Electrophoresis, Polyacrylamide Gel, Hemagglutination Inhibition Tests, Hemagglutination Tests, Immunoelectrophoresis, Rabbits immunology, Serum Albumin, Bovine, Antibodies, Bacterial isolation & purification, Chromatography, Immune Sera, Neisseria meningitidis immunology
- Published
- 1972
47. Antigenic specificity of bactericidal antibodies in antisera to Neisseria meningitidis.
- Author
-
Kasper DL, Winkelhake JL, Brandt BL, and Artenstein MS
- Subjects
- Animals, Antigen-Antibody Reactions, Bacterial Proteins, Blood Bactericidal Activity, Carbon Isotopes, Complement System Proteins, Cross Reactions, Hemagglutination Inhibition Tests, Hemagglutination Tests, Immune Sera, Male, Polysaccharides, Bacterial, Rabbits immunology, Serotyping, Antibodies, Bacterial analysis, Antibody Specificity, Antigens, Bacterial, Epitopes, Meningitis, Meningococcal blood, Neisseria meningitidis immunology
- Published
- 1973
- Full Text
- View/download PDF
48. In vitro effect of d-lysergic acid diethylamide on immunoglobulin synthesis.
- Author
-
Voss EW Jr, Babb JE, Metzel P, and Winkelhake JL
- Subjects
- Amino Acids metabolism, Animals, Antibody-Producing Cells drug effects, Antibody-Producing Cells metabolism, Carbon Isotopes, Centrifugation, Density Gradient, Dialysis, Fluorescent Antibody Technique, Hydrogen-Ion Concentration, In Vitro Techniques, Iodine Isotopes, Isoelectric Focusing, Isomerism, Molecular Weight, Rabbits, Sucrose, Swine, Tritium, Tryptophan antagonists & inhibitors, Immunoglobulins biosynthesis, Lysergic Acid Diethylamide pharmacology
- Published
- 1973
- Full Text
- View/download PDF
49. Recognition of carrier residues adjacent to hapten by anti-trinitrophenyl antibodies.
- Author
-
Winkelhake JL and Voss EW Jr
- Subjects
- Absorption, Animals, Antibody Specificity, Antigen-Antibody Reactions, Binding Sites, DNA, Dialysis, Fluorescence, Immune Sera, Lysine, Nucleosides, Rabbits, Spectrophotometry, Tritium, Antibodies isolation & purification, Haptens, Nitrophenols
- Published
- 1970
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.