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Biomedical subjects

J E Butler

Publications and source records attributed to J E Butler.

At least 55 records · Page 3Linked to original sources

Dimeric mouse IgA is transported into rat bile five times more rapidly than into mouse bile.

The kinetics of the biliary dimeric IgA transport in rats is well documented from studies on animals with cannulated bile ducts, while in mice such information rests on the analysis of samples periodically removed from the gall bladder and on serum disappearance rates. We have quantitatively compared the kinetics of IgA transport in these two rodents by using affinity-purified radiolabelled monomeric and dimeric M315 IgA which specifically binds dinitrophenyl (DNP). Bile collection procedures, sample analysis, and administration of the radiolabelled M315 were the same for both species, although in mice the cystic duct was ligated to prevent reflux of any bile or IgA from the gall bladder into the common bile duct. A pronounced selection for transport of dimeric M315 was seen in both species. When dimeric M315 was administered, it was recovered in the dimeric form from the bile of both species, and 60-80% could specifically bind to DNP-gelatin. When monomeric M315 was administered, very little was transported, and the IgA recovered in bile had a much lower capacity to bind DNP-gelatin and appeared to be of low molecular weight. Approximately 60% of the intravenously administered dimeric IgA was transported into bile of both species but biliary transport of the heterologous dimeric IgA into rat bile was five times more rapid than the transport of the dimeric M315 into its homologous species, the mouse. Hence, IgA transport in these species is similar in specificity, integrity of the transported IgA and in the percentage cleared, while differing significantly in the rate of transport. This cannot be ascribed to the species source of the IgA.

Animals

Preliminary evidence for a diagnostic immunoglobulin G1 antibody response among culture-positive cows vaccinated with Brucella abortus strain 19 and challenge exposed with strain 2308.

The sera of cows inoculated with Brucella abortus have a characteristically high titer of immunoglobulin (Ig) G1 antibodies to a soluble brucella antigen compared with sera of noninoculated vaccinated cattle. Concentrations of antigen-specific IgG1 were greater than 10-fold higher than those for IgG2, even though total IgG2 concentrations were higher than total IgG1 concentrations. Increases in IgG1 antibodies to Brucella abortus soluble antigen were detected shortly after vaccination in those cows from which strain 19 was isolated and by 28 weeks in cows from which strain 2308 was isolated. Increases in specific antibodies were not paralleled by increases in either total IgG1 or total IgG2 concentrations. Rather, there was a 15-fold to greater than 200-fold increase in specific activity, with up to 16% of the IgG1 specific for the brucella antigen used in the assay. Thus, measurement of changes in total IgG1 concentrations is not a reliable method to identify brucellosis-associated anti-Brucella abortus soluble antigen activity. Only one cow in a panel of 10 selected for detailed study showed a false-positive IgG1 titer, whereas some serologic assays showed as many as 4 or 5 false-positives. Results of the complement-fixation test, among the battery of serologic tests used for detection of brucellosis, best agreed with the occurrence of increased IgG1 antibody levels.

Animals

Identification and characterization of a hapten-modifiable TEPC 15 cross-reactive idiotype in swine.

Rabbits and swine immunized with TEPC 15 IgA, goats immunized with T15-positive IgM and swine immunized with affinity-pure swine anti-phosphorylcholine (PC) all produce antibodies which recognize a hapten-inhibitable idiotypic determinant on swine anti-PC. The similarity in reactivity and order of inhibitability with various PC analogs of the heterologous (swine anti-TEPC 15) and isologous (swine anti-swine anti-PC) reagents indicates that they recognize a related idiotype and suggest it may be the predominant idiotype expressed on swine anti-PC antibodies. The heterologous and isologous anti-idiotypic reagents generated in this study recognize swine and mouse anti-PC but not normal swine IgM, IgG or MOPC 460. Only reactions with swine anti-PC and mouse T15-positive anti-PC proteins are hapten-inhibitable. The greater inhibitory capacity of trimethylammonium and acetylcholine than PC suggests that the idiotope(s) recognized on swine anti-PC by the anti-idiotypic reagents is integral rather than peripheral to the anti-PC binding site. The nearly exclusive IgM anti-PC response of swine to Streptococcus pneumoniae R36A and PC-Brucella have so far hindered attempts to study the isotypic distribution of the idiotype.

Animals

Basophil chemotaxis under agarose: variability among guinea pig donors and response to sera of different species.

Guinea pig basophils were tested against chemotactic factor(s) generated from homologous and heterologous sera. An "under agarose" method of quantitating leukocyte chemotaxis was adapted to measure serum-derived basophil chemotactic factor(s) (BCF). The method was found to be simple, reproducible, and sparing of time and reagents and it provided simultaneous controls. Because of the economy of reagents compared with the Boyden chamber technique, basophils from individual guinea pigs could be studied and were found to be variable in responsiveness. A definite difference in the response of guinea pig basophils to zymosan-activated sera of various other species was demonstrated. Guinea pig basophils responded to zymosan-activated serum from guinea pig and rat but failed to respond to zymosan-activated serum from human, hamster, rabbit, or cow. Optimal conditions were determined for cell numbers, serum activation and dilution, and migration times for studies of guinea pig BCF. The under agarose technique provides a practical alternative to chamber methods for the study of basophil chemotaxis. Although guinea pig basophils do not respond to human serum-derived BCF, the economy of reagents with this technique gives promise toward adaptation of the method to studies of human basophil chemotaxis.

Animals

Immunologic aspects of breast feeding, antiinfectious activity of breast milk.

The most apparent immunologic role of the mammary gland is supply to antibodies to the neonate. In cattle the gland must be able to secrete large quantities of IgG antibodies over a short time period to supply the offspring with protection against systemic pathogens. This is accomplished by selective transfer of IgG from serum to the gland followed by eventual absorption by the neonate gut. In all mammals, the mammary glands provide IgA antibodies specific for pathogens or antigens which enter or invade the neonatal gut. An entero-mammary cell circulation provides the mechanism for conveying such specificity to the lacteal IgA antibodies. Some IgA antibodies may also be derived from the circulation so that the quantitative significance of serum derived versus locally produced IgA in different species requires clarifications. IgG and IgG lacteal antibodies ingested by the neonate, provide short-term systemic and long-term enteric humoral immunity to the neonate. In addition to providing passive immunity, at least swine IgG appears to have a regulatory role in the development of the systemic humoral immune system of the neonate. Such a phenomenon may be general for IgG antibodies transferred in colostrum or in utero. While passive antibodies and immunoglobulins may be most important for the neonate, the many other potentially anti-infectious elements transferred in colostrum and milk may also play important roles. 'Bifidus' factor particularly, but also lysozyme and lactoferrin are probably all important although more convincing experimental data will be needed to support this assumption. Studies of cells of the lymphoid and reticuloendothelial systems in milk are more recent and their role in the neonate remains to be convincingly demonstrated. In summary, the immunologic and anti-infectious roles of the mammary glands are (1) Supply of IgA antibodies against enteric antigens to the neonate on a 'long-term' basis throughout lactation; (2) Short-term supply of IgG (and IgA) in Group II and III mammals for eventual absorption into neonatal serum; (3) The supply of numerous nonspecific factors such as 'bifidus factor,' lactoferrin, and lysozyme throughout lactation; (4) Regulation of the development of humoral immunity by an apparent feedback mechanism involving maternal IgG; (5) Self-protection of the gland by sensitized T-lymphocytes acting directly or using lymphokines on macrophages; and (6) Self-protection of the gland by secreted antibodies that may act in complement-independent cytolysis, as opsoins for polymorphonuclear-leukocytes or directly as agents preventing colonization.

Animals

The differential enzyme susceptibility of bovine immunoglobulin G1 and immunoglobulin G2 to pepsin and papain.

Purified bovine immunoglobulins IgG1 and IgG2 were subjected to enzymatic degradation with pepsin and papain. Results were monitored using density gradient ultracentrifugation, acrylamide electrophosesis and immunodiffusion employing subclass- and light chain-specific antisera. The results indicated a marked enzymatic susceptibility of IgG1 to digestion with pepsin. This differential susceptibility can also be demonstrated in unfractionated bovine gamma-globulin. No striking differences between the two subclasses were observed during treatment with papain in the presence of cysteine and after 24 h, most IgG1 and IgG2 was degraded to Fc and Fab fragments. The pepsin Fc fragment generated from IgG2 was larger than that generated from IgG1 although the F(ab')2 fragments were simialr in size. These results are consistent with the hypothesis that the Fc region of IgG1 contains multiple cleavage sites for pepsin whereas IgG2 has few. Rabbits immunized with the first elution peak from a 30 h pepsin digest of bovine gamma-globulin fractionated on Sephadex G-150, responded primarily to common gamma-chain and IgG2-specific determinants. Thus, the differential susceptibility of bovine subclasses to pepsin provides a method for stimulating IgG2-specific antibodies in rabbits.

Animals

Amplification of the enzyme-linked immunosorbent assay (ELISA) in the detection of class-specific antibodies.

A modification of the standard enzyme-linked immunosorbent assay (ELISA) is described which circumvents the requirement for specifically purified antibodies from which antibody-enzyme complexes are made. The assay utilizes the principle of a soluble anti-alkaline phosphatase immune complex (AP-A-AP) and has been called the amplified ELISA. Methods for preparing and evidence for the specificity of rabbit anti-rat gamma-FC, IgM (mu) and IgA (alpha) are presented. These reagents are used to measure anti-DNP antibodies belonging to classes IgG, IgM and IgA in rat serum. Using antiglobulin and anti-enzyme reagents prepared in guinea pigs, anti-ovalbumin antibodies are measured in rabbit serum. Titration curves are similar when the amplified ELISA is compared to the standard ELISA. A change in slope suggesting an effect of saturation of antigen sites, occurs at the same input antibody concentration for both assays. Determination of the anti-DNP concentration of unknown sera by extrapopulation from titration graphs of a known serum suggests that the value is overestimated, i.e., amplified when the amplified ELISA is used. In addition, the amplified ELISA has an improved ability to detect low levels of antibody. Evidence is presented which illustrates how the use of optimally conjugated DNP-proteins, age of conjugates, and optimal dilutions of secondary antiglobulins and the AP-A-AP reduce non-specific binding in the amplified ELISA. The amplified ELISA is capable of detecting 2.4 ng of antibody to ovalbumin in a one: one million dilution of rabbit serum with high reproducibility and low background.

Animals

Incomplete inhibition of the immunoglobulin G-binding behavior of nitrated protein A.

Nitration of protein A with tetranitromethane incompletely blocks its Fc-binding ability for immunoglobulin G when measured by more sensitive methods of immunoprecipitation and when tested in the enzyme-linked immunosorbent assay. In addition, binding of protein A and blockage by nitration is heterogeneous and depends on the species origin of the immunoglobulin G.

Animals

Evaluation of enzyme-linked immunosorbent assay for quantitation by subclass for bovine antibodies.

The enzyme-linked immunosorbent assay (ELISA) was evaluated for use in the quantitative measurement of bovine immunoglobulin IgG1 and IgG2 antibodies. A method for standardization was devised in which IgG1 or IgG2 was directly adsorbed to polystyrene tubes and the actual degree of binding was calculated by using different input amounts of 125I-labeled IgG1 or IgG2. Values for quantity of IgG1 antibodies to human serum albumin were only slightly higher when measured by the ELISA than when measured by quantitative precipitation although the value measured by the ELISA for IgG2 antibodies was twice that determined by quantitative precipitation. This discrepancy could result from conjugate cross reactivity, differences in affinity between antibodies of the 2 subclasses, or the occurrence of IgG2 nonprecipitating antibodies. The danger of overlooking subclass anti-globulin cross reactivity because of the failure to detect it by immunoprecipitation, also is illustrated. In addition, only enzyme-antibody conjugates prepared with specifically purified antibodies were effective, and reproducibility of individual data points required that 4 replicate determinations be performed. Advantages, pitfalls, and limitations of the ELISA are discussed.

Animals

Tickborne oculoglandular tularemia: case report and review of seasonal and vectorial associations in 106 cases.

A patient acquired tickborne oculoglandular tularemia in early summer in rural Virginia. Tick exposure may be a clue to the diagnosis of tularemia in the eastern as well as the western United States, especially in summer months. A review of the experience with tularemia in Virginia for the last 13 years shows a bimodal seasonal incidence of tularemia with an associated vector exposure in 77.4% of 106 cases. The majority of cases occurring during winter months have been associated with rabbit exposure, while those in summer months are often associated with tick exposure.

Aged

Two immunochemically and physicochemically distinct secretory components from rat exocrine secretion.

Two proteins were isolated from rat milk by using ion-exchange and gel filtration chromatography and shown to be secretory components by the criteria of: a) occurrence as free proteins in exocrine secretions, as well as bound to exocrine IgA while absent from serum and serum IgA, b) immunohistochemical localization in the epithelial cells of the gut, c) dissociability from and in vitro binding to 11 S SIgA, d) cross-reactivity with antisera to mouse and bovine secretory components, and e) significant peptide homology with bovine secretory component. The secretory components were immunochemically and physicochemically distinct as shown by: a) the lack of cross-reactivity between their specific antisera and differential reactivity with anti-bovine FSC and anti-mouse SC, b) different electrophoretic mobilities, c) DEAE elution profiles, d) carbohydrate contents, e) molecular weights, and f) apparent polypeptide chain constitution. Another interesting difference in the two is that only one of the components dissociates spontaneously from SIgA. Possible functions of two secretory components are discussed.

Animals

The immunochemistry of sandwich ELISAs--V. The capture antibody performance of polyclonal antibody-enriched fractions prepared by various methods.

Studies compare the performance of antibody-enriched serum fractions prepared by various methods, when adsorbed on polystyrene microtiter wells as capture antibodies (CAbs) and tested against multivalent antigens. The criteria of performance in the RIA used included antigen capture capacity (AgCC) and the nmol of functional capture sites per microtiter well (CAbt). Affinity purified polyclonal (pAb) and monoclonal antibodies (mAb) were employed as reference CAbs. AgCC was highest for enriched fractions prepared using caprylic acid and a high-pressure SpG affinity column. The performance of capture antibodies is expressed by an equation which was empirically derived and experimentally tested; CAbt x AgCC/ng adsorbed IgG. In terms of this parameter, CAb-enriched fractions prepared with caprylic acid performed best. The data reported also provide insight into solid-phase ligand immunochemistry. Adsorbed polyclonal CAb performed with remarkable homogeneity in percent bound and in Scatchard plots. Values obtained for CAbt from Steward-Petty plots were directly correlated with the length of the LBR of log-log percent bound plots but indicated that less than 10% of the potential capture sites of polyclonal CAbs remained functional after adsorption; mAb showed a more serious loss of activity. The loss of CAbt was a general phenomenon for all preparations tested although relative to their antibody content, certain antibody-enriched fractions retained a higher proportion of CAbt than their affinity-purified counterparts. Comparative studies in which the activity of adsorbed mAb and pAb was compared to the same antibodies immobilized by a non-adsorptive procedure, indicated that adsorbed CAbs also express lower affinity. The studies we report offer a single parameter criterium for comparatively evaluating CAb performance while simultaneously revealing the need to develop immobilization procedures that can preserve CAbt and antibody affinity so that immunoassays with wide dynamic ranges and high AgCC can be developed without waste of antibody.

Adsorption