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M D Boyle

Publications and source records attributed to M D Boyle.

At least 73 records · Page 4Linked to original sources
PubMed1991

Isolation and partial characterization of a type IV bacterial immunoglobulin binding protein.

A series of bovine G streptococcal isolates were screened for expression of type IV Fc binding proteins. An isolate expressing high levels of type IV binding proteins was selected and expanded by use of a colony selection technique. A variety of different extraction procedures were compared and the optimal solubilization procedure was found to be hot acid extraction of the bacteria. The extracted protein could be affinity purified on a column of immobilized rabbit IgG. The type IV Fc binding protein was found to be antigenically unrelated to the type I, II or III bacterial Fc binding proteins and displayed the lowest affinity and most limited range of species and subclass reactivity of any bacterial Fc binding protein thus far characterized.

Bacterial Proteins↗
PubMed1991

A simple preparative procedure to extract and purify protein G from group G streptococci.

A rapid method for the solubilization of the bacterial type III Fc binding protein, protein G, from a group G streptococcus is described. Treatment of intact bacteria with cyanogen bromide results in the solubilization of a homogeneous Mr approximately 50,000 protein which retains IgG and human serum albumin binding properties. The solubilized protein could be purified to homogeneity by molecular sieving chromatography and retained all of the functional properties of the native protein.

Amino Acid Sequence↗
PubMed1991

Group B streptococcal C protein-associated antigens: association with neonatal sepsis.

The c protein (Ibc) of group B streptococci (GBS) is associated with at least four antigens (alpha, beta, gamma, delta). To assess the virulence potential of these antigens, 255 GBS isolates recovered from septic neonates, healthy neonates, and pregnant women were serotyped and surveyed for reactivity with sera to c protein and the four associated antigens. A radioimmunoassay using intact bacteria was used to detect the GBS antigens. In contrast to earlier reports, most (66%) of the type III strains expressed the c protein. Except for the gamma antigen, none of the other c protein-associated antigens showed an increased association with pathogenic strains independent of the polysaccharide antigens. The gamma antigen was expressed by 15 of 41 c protein-positive early-onset strains and by 4 of 38 c protein-positive late-onset strains (P = .007). This association was independent of the type-specific antigen, suggesting a potential role for the gamma antigen as a virulence factor in GBS strains causing early-onset sepsis.

Antigens, Bacterial↗
PubMed1991

Comparison of albumin receptors expressed on bovine and human group G streptococci.

The albumin receptor expressed by bovine group G streptococci was extracted and affinity purified. The protein was characterized for species reactivity, and monospecific antibodies were prepared to the purified receptor. The bovine group G albumin receptor was compared functionally, antigenically, and for DNA homology with the albumin-binding protein expressed by human group G streptococci. In agreement with previous reports, the albumin-binding activity of human strains was mediated by a unique domain of the type III immunoglobulin G-Fc-binding molecule, protein G. The albumin receptor expressed by bovine group G strains was found to lack any immunoglobulin G-binding potential but displayed a wider profile of species albumin reactivity than protein G. Both albumin receptors could inhibit the binding of the other to immobilized human serum albumin, and each displayed similar binding properties. Antigenic comparison of the two albumin receptors demonstrated a low level of cross-reactivity; however comparison at the DNA level, using an oligonucleotide probe specific for the albumin-binding region of protein G, demonstrated that the two albumin receptors expressed by human and bovine group G streptococcal strains do not display significant homology.

Albumins↗
PubMed1991

Protein B: a versatile bacterial Fc-binding protein selective for human IgA.

Protein B, a selective bacterial IgA Fc-binding protein isolated from group B streptococci, has been used to quantify fluid phase and immobilized human IgA. Protein B detects both human IgA1 and IgA2 subclasses and is also reactive with secretory IgA. Protein B can be used immobilized to microtiter plates to capture IgA or following biotinylation as a tracer for fluid phase or immobilized human IgA. The studies presented here suggest protein B will prove to be a valuable reagent for quantitative immunochemical procedures involving human IgA antibodies and facilitate a variety of studies of IgA responses in man.

Antigens, Bacterial↗
PubMed1990

Isolated DNA repeat region from fcrA76, the Fc-binding protein gene from an M-type 76 strain of group A streptococci, encodes a protein with Fc-binding activity.

The DNA repeat region of fcrA76, the gene encoding a group A streptococcal Fc-binding protein, was subcloned in-frame into an Escherichia coli plasmid expression vector. The expressed protein product displayed the same Fc-binding properties as the full-length Fc-binding protein expressed from fcrA76. The affinity-purified, full-length Fc-binding protein was found to compete with staphylococcal protein A or streptococcal protein G for binding to beads coated with human IgG. These results are consistent with earlier studies suggesting that the binding sites on human IgG for protein A, protein G and the type II Fc-binding protein from group A streptococci are located at the interface of the CH2 and CH3 domains of the Fc region.

Amino Acid Sequence↗
PubMed1990

Colloidal gold immunolabeling of immunoglobulin-binding sites and beta antigen in group B streptococci.

We have characterized the immunoglobulin A (IgA)-Fc-binding properties and beta-antigen expression of several strains of group B streptococci by using ultrastructural immunocytochemistry. Colloidal gold-labeled tracers were used with intact and sectioned bacteria in order to gain information regarding the location and distribution of cell surface and cytoplasmic IgA-Fc-binding molecules and beta antigen. Colloidal gold (5- or 15-nm particles) was conjugated to IgA to characterize IgA-binding properties and to IgG to test for IgG binding. Rabbit anti-beta antiserum was reacted with the bacteria and then with protein G labeled with 15-nm colloidal gold particles. A double-labeling technique was used for simultaneous localization of IgA-Fc- and anti-beta-antibody-binding properties on sectioned bacteria. The data corroborated previous results which indicated that (i) IgA-Fc-binding and IgA-Fc-nonbinding forms of beta antigen can be secreted by strains which do not express beta antigen on the cell surfaces (HG806, VC75); (ii) differences in levels of expression of beta antigen and/or IgA-Fc-binding proteins can be detected among various group B isolates; (iii) group B streptococci do not express human IgG-Fc-binding proteins; and (iv) not all forms of beta antigen are capable of binding human IgA.

Animals↗
PubMed1989

Immunological applications of type III Fc binding proteins. Comparison of different sources of protein G.

Protein G, a type III bacterial IgG Fc receptor isolated from certain group C or G streptococci, shows a wider range of species and subclass immunoglobulin reactivity than staphylococcal protein A and has been shown to be more useful than protein A for many immunochemical applications. Recently, two forms of wild type protein G and three forms of recombinant protein G have become commercially available. Each form of protein G was tested for reactivity with a variety of species of immunoglobulin and albumin. Additionally, one form of wild type protein G and two forms of the recombinant protein G were examined for their ability to stimulate in vitro proliferation of human peripheral blood leukocytes (PBL). Similar IgG species reactivity was observed for all forms of unlabeled protein G. By contrast, considerable variability in the relative IgG binding potentials of different protein G preparations was observed following radioiodination. Binding to human serum albumin was observed with one of the wild type protein G samples, however, the IgG binding activity of this protein was not affected by the presence of excess human serum albumin. In the human PBL proliferation assays, wild type protein G was weakly mitogenic and one form of recombinant protein G was shown to be a potent mitogen, while another form of recombinant protein G displayed no mitogenic potential. Differences in both functional and biological reactivities were observed among the various sources of protein G. These differences may lead to confusion if investigators consider all sources of protein G as a single entity with common properties.

Antigen-Antibody Complex↗
PubMed1989

Group A streptococci bind human plasmin but not other structurally related proteins.

Certain group A streptococci are known to possess a receptor for the human enzyme plasmin. Plasmin is a member of a super gene family that includes other serine proteases and kringle containing proteins. In this study we have examined the interaction of a group A streptococcus with structurally related proteins, including plasmin, glu-plasminogen, tissue plasminogen activator, kallikrein, factor XII, prothrombin, thrombin, trypsin, and urokinase. Our studies indicate that only the key fibrinolytic enzyme, plasmin, demonstrates significant binding activity to the group A streptococcus.

Animals↗
PubMed1989

Identification of non-immunoglobulin A-Fc-binding forms and low-molecular-weight secreted forms of the group B streptococcal beta antigen.

The beta antigen expressed on the surfaces of certain strains of group B streptococci has been reported to bind to the Fc region of human immunoglobulin A (IgA). In this study, we screened 100 isolates of group B streptococci for expression of both beta antigen and IgA-Fc-binding activity. We identified two isolates which expressed the beta antigen but could not bind human IgA Fc fragments and also observed variability in IgA-Fc-binding activity among other beta-antigen-expressing strains. Novel low-molecular-weight forms of beta antigen were secreted by four beta-antigen surface-negative isolates and included IgA-Fc-binding (Mrs, 55,000 and 53,000) and non-IgA-Fc-binding (Mr, 38,000) molecules. These results suggest that the IgA-Fc-binding site represents a unique domain of the beta antigen. The 55,000- and 53,000-Mr forms of secreted beta antigen were functionally and antigenically representative of the size-heterogeneous (Mr, up to 145,000) beta-antigen molecules expressed by surface-positive strains. The cell surface-localized IgA-Fc-binding molecules could bind only human serum IgA efficiently; however, once solubilized, these molecules could bind both human serum and secretory IgAs.

Antigens, Bacterial↗
PubMed1989

Mapping of the human plasmin domain recognized by the unique plasmin receptor of group A streptococci.

A high-affinity surface receptor for human plasmin has been reported on certain group A streptococci. To map the region of the plasmin molecule that binds to the bacterial receptor, isolated domains of plasmin were tested for their ability to inhibit the binding of intact radiolabeled plasmin to receptor-positive bacteria. Complete inhibition of binding of labeled plasmin to bacteria by isolated heavy chains was achieved, but this inhibition was not as efficient on a molar basis when compared with that of unlabeled plasmin. By contrast, a conformationally altered form of native plasminogen was found to bind to bacteria and was as efficient a competitive inhibitor as intact plasmin was. The results of this study indicate that the selective binding of human plasmin to a group A streptococcus is dependent on structures present in the conformationally altered form of native plasminogen or plasmin that are not found on the native zymogen, the plasminogen with NH2-terminal glutamic acid.

Amino Acid Sequence↗
PubMed1988

Subunit structure of high molecular weight mouse nerve growth factor.

Studies from several laboratories have shown that mouse submandibular glands and mouse saliva contain nerve growth factor (NGF) as part of a high molecular weight oligomeric macromolecule composed of three different subunits, termed alpha, beta, and gamma. The beta-subunit is the nerve growth-promoting protein. The gamma-subunit is a serine protease class enzyme of highly restricted substrate specificity. The alpha-subunit has no known function. This high molecular weight form of nerve growth factor is also a Zn(II)-containing metalloprotein. In the present study, measurements of multiple physicochemical parameters have been used to deduce the subunit structure of high molecular weight NGF. Results demonstrate that it contains two alpha-, one beta- and one gamma-subunit together with one tightly bound Zn(II) ion per molecule.

Animals↗
PubMed1988

Purging tumor cells from bone marrow by use of antibody and complement: a critical appraisal.

This review highlighted several problems associated with the use of antibody and complement in the elimination of tumor cells from bone marrow that was to be used for transplantation, and it discussed some of the difficulties encountered in developing this approach in model systems. These problems should be seriously considered by any clinician contemplating this method for bone marrow purging.

Antibodies↗
PubMed1988

Enzyme-labeled type III bacterial Fc receptors. A versatile tracer for immunoassay.

The type III bacterial Fc receptor isolated form a group C streptococcus has been conjugated to alkaline phosphatase and used as a tracer in a variety of direct and indirect immunoassays. These immunoassays have utilized specific antibodies prepared in species whose immunoglobulins are poorly reactive with the type I Fc receptor, staphylococcal protein A. The value of the type III Fc receptor as a tracer for immunoassays utilizing antibodies produced in sheep and goats is documented.

Alkaline Phosphatase↗
PubMed1988

Characterization of the interaction of human plasmin with its specific receptor on a group A streptococcus.

Certain Group A beta-hemolytic streptococci express a receptor that is capable of specifically binding the human plasma protease plasmin. Once bound, plasmin remains enzymatically active and is unregulated by its naturally occurring inhibitor alpha-2-antiplasmin (Lottenberg, R., C. C. Broder and M. D. P. Boyle, 1987. Infect. Immun. 55: 1914-1918). In this study certain characteristics of the interaction between plasmin and the receptor expressed on a group A beta-hemolytic streptococcus, strain 64/14, were examined. Binding occurred optimally at physiologic pH and ionic strength. The KD was 5 x 10(-11) M and there were approximately 800 receptors per bacterium. Mouse passage of strain 64 had no significant effect on the KD of the receptor. Binding of plasmin to the bacteria was inhibited by lysine and epsilon-aminocaproic acid in a concentration dependent manner. Similarly these amino acids would displace pre-bound plasmin from the bacteria. These findings suggest a role for plasmin's high affinity lysine binding site in the interaction of plasmin with the bacteria.

Aminocaproic Acid↗
PubMed1988

Two novel antigens associated with group B streptococci identified by a rapid two-stage radioimmunoassay.

A two-stage radioimmunoassay (RIA) has been developed that identifies type-specific antigens on the surfaces of group B streptococci. In addition to detecting the type-specific carbohydrate antigens Ia, Ib, II, and III, the RIA identified four unique antigens reactive with the Centers for Disease Control's c-protein typing antiserum. Analysis of hot-acid extracts by immunoelectrophoresis confirmed that two of these reactivities corresponded to the reported alpha and beta antigens of the c-protein marker. In addition, the identification of two heretofore unidentified antigens, gamma and delta, by the two-stage RIA is detailed in this report. The assay uses intact bacteria and does not require hot-acid extraction, a procedure thus enabling detection of acid-labile antigens in their native unmodified form. The semiquantitative RIA requires less typing reagent than does precipitin testing and is more objective, reproducible, and rapid. The assay described here could be applied to the detection of any cell surface antigen for which a monospecific antiserum is available.

Antigens, Bacterial↗
PubMed1988

Selective colony blotting to expand bacterial surface receptors: applications to receptors for rat immunoglobulins.

Many bacterial surface receptors demonstrate a heterogeneous expression pattern among individual colonies. Methods have been developed to select bacteria expressing high levels of a stable surface receptor. This process is illustrated using a Streptococcus zooepidemicus isolate demonstrating a high level of Fc receptors for rat immunoglobulins. This strain was selected and expanded to obtain a bacterial isolate demonstrating approximately 100 fold greater reactivity with rat immunoglobulins than protein A positive Staphylococcus aureus or 30-40 fold higher reactivity for rat IgG than type III Fc receptor positive streptococcal group G strains. The optimal pH for rat IgG binding and the reactivity with rat IgG subclasses and certain rat monoclonal antibodies is described. The potential application and limitations of the selected rat Fc receptor positive bacterial strain to immunoassays based on the specificity of rat monoclonal and polyclonal antibodies is discussed.

Adsorption↗