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Biomedical subjects

A J Crowle

Publications and source records attributed to A J Crowle.

At least 55 records · Page 3Linked to original sources
PubMed1982

Identification of transcalciferin as a major component of human parotid saliva by crossed immunoelectrophoretic mapping.

Human parotid saliva collected from Stenson's duct during sour candy-stimulated salivation was studied by crossed immunoelectrophoresis (X-IEP). Eleven antigens were identified in a pool of salivas from 10 adult, caucasoid males and females. Four were related to serum antigens, three being previously known: IgG, IgA and albumin. The fourth was identified as Gc globulin, also known by function as transcalciferin. Salivary Gc is electrophoretically different from serum Gc, migrating as an alpha 2-beta component rather than as an alpha 1 globulin. The quantities of 8 of the 11 antigens detected by X-IEP were compared for salivas from 10 subjects. These quantities and their ratios to each other were highly variable, indicating idiosyncratic secretory patterns. Only quantities of amylase were moderately consistent from donor to donor. Quantitatively, Gc is a major antigen in parotid saliva. Large proportions of a salivary antigen, alpha 1c, were found in two of the 10 subjects--adult males who never had developed caries. Among the other 8 subjects, all with caries, proportions of alpha 1c were much lower, and 2 subjects lacked it. None of the other antigens measured showed any correlation with caries resistance. The nature and function of alpha 1c are unknown.

Adult↗
PubMed1982

Crossed immunoelectrophoresis: qualitative and quantitative considerations.

Invented 20 years age, crossed immunoelectrophoresis (X-IEP) today is a technique of unusual power and myriad application. It combines very high resolution with exquisite specificity by alloying 2-dimensional electrophoresis with immunoprecipitation for symbiotic new potentialities. The consequent matchless quantitative/qualitative capabilities of X-IEP for analyzing antigens in complex mixtures, particularly by their idiomatic internal comparison, are still not widely recognized. Because of this and the supposed complications of its use and interpretation, X-IEP is more rarely used than it should be. This essay discusses contemporary X-IEP with the particular aims of demonstrating that it is not difficult to use and of explaining with selected examples why it is peculiarly powerful for analyzing antigen mixtures like the body fluids, tissue and cell extracts, and microbial homogenates.

Analysis of Variance↗
PubMed1982

Regression and differentiation of neuroblastoma tumors in mice treated with differentiating agents--prostaglandin E1 and a phosphodiesterase inhibitor, RO 20-1724.

Prostaglandin E1 (PGE) and d, 1-4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (RO 20-1724, an experimental cyclic nucleotide phosphodiesterase inhibitor) cause cell death and cyclic AMP-mediated differentiation of murine neuroblastoma (NB) (cell line C-1300) cells in vitro. We report, for the first time, that these agents can be used in vivo to cause regression of NBs in A/Jax mice. These drugs were also able to slow the growth of primary tumors, prevent metastases, and cause morphological differentiation of tumors. The similarity of the response of human and mouse NB cells to differentiating drugs indicate that these agents have the potential to replace or support conventional chemotherapy in treatment individualized to NB. The mouse model of NB provides an excellent means to study the effectiveness of alternative drug therapy in vivo.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗
PubMed1982

Antibodies against the Ag2 fimbriae of Actinomyces viscosus T14V inhibit lactose-sensitive bacterial adherence.

Monospecific antisera against the Ag1 and Ag2 fimbrial components of Actinomyces viscosus T14V were produced by immunizing rabbits with immune precipitates of each antigen harvested from crossed-immunoelectrophoresis plates. The Fab fragments prepared from these sera were used as specific reagents in immunoelectron microscopy to identify each fimbrial component on the bacterial surface and also were assayed for their abilities to prevent the coaggregation of A. viscosus T14V with Streptococcus sanguis 34, an interaction that is lactose sensitive. Each Fab preparation appeared to react with different fibrillar structures present on the actinomycete, and only the Ag2-specific Fabs blocked coaggregation. These results provide strong support for the presence of distinct types of fimbriae on A. viscosus T14V and indicate the exclusive involvement of Ag2 fimbriae in lactose-sensitive adherence.

Actinomyces↗
PubMed1982

Lymphokine-induced mycobacteriostatic activity in mouse pleural macrophages.

We have developed an in vitro assay to study the intramacrophage replication of Mycobacterium tuberculosis or Mycobacterium bovis BCG. In this assay, extracellular mycobacterial replication was avoided by using a small infecting inoculum and a new method for efficiently removing nonphagocytocytized bacilli. The ability of different mycobacterial strains to replicate in mouse pleural macrophages correlated with their virulence. The effect of lymphokines prepared by incubating spleen cells from BCG-immunized mice with mycobacterial antigens was tested with this assay. In the presence of lymphokines, mouse pleural macrophages were able to inhibit the intramacrophage replication of both BCG and M. tuberculosis. This assay is a useful in vitro model for studying antibacterial cellular immunity expressed by macrophages.

Animals↗
PubMed1982

Suppression of tuberculoimmunity in mice by Evans blue dye.

Evans blue dye, which acts as an adjuvant in mice to promote induction of cell-mediated immunity, was found earlier to apparently act contrarily and decrease tuberculoimmunity. This phenomenon was investigated here in a long-term experiment examining the persistence of the suppressed immunity. Systemic suppression after injection of only 1 mg of dye was confirmed and was found to last for more than 2 weeks but less than 11 weeks. Localized evidence for dye-caused depression of tuberculoimmunity also was seen by the development in dye-injected mice immunized with emulsified whole bacilli of caseating tubercles at sites of pulmonary infection, a very rare type of pathology for tuberculosis in mice. The experiment, in addition, detected an aging-related decline in mice of resistance to tuberculosis evident by shortened survival, enlarged pulmonary tubercles, and increased susceptibility to systemic tuberculin reaction.

Aging↗
PubMed1982

Anaphylactic and precipitating antibody responses of aging A/J female mice.

Aging A/Jax female mice were compared with young controls for abilities throughout life to make IgG1 and IgE anaphylactic antibodies and precipitins to chicken conalbumin (CA) and methylated human serum albumin (MeHSA) injected in five different regimens of immunization. These regimens consisted of various combinations of injections of antigen in water-in-oil emulsion or in saline. CA induce those antibodies well, MeHSA poorly. The mice were able to respond nearly equally at all ages, a finding differing from earlier ones for at least two reasons: (1) neither antigen used is related to any in mouse food, and (2) the regimens of immunization caused sustained antigenic stimulation together with induction of T cell responses, which could have compensated for otherwise expected age-weakening of necessary TH cell functionings. We conclude that A/J female mice can make good anaphylactic and precipitating antibody responses to new antigens throughout life.

Aging↗
PubMed1982

Culture conditions required for primary isolation and study of mouse blood monocytes.

Blood monocytes, the precursors to macrophages found in inflammatory exudates, have several important immunologic functions, and may also contribute to populations of mature macrophages found in tissues and serous cavities. Mouse blood monocytes have not been studied much because they have been difficult to culture. If plated alone, ie, in the absence of blood lymphocytes, they die in media usually used to culture mouse macrophages. Here we describe conditions for plating them alone and maintaining them for several weeks in a standard culture medium (Neuman-Tytell) supplemented only with a normal serum. Plating them at a high concentration (8 x 10(7) cells/ml) and using horse serum instead of fetal calf serum were important, and their survival was improved by adding reduced glutathione or sodium thioglycollate but not 2-mercaptoethanol. Using high concentrations of horse serum (20%) and high plating concentrations of cells both encouraged monocyte replication. We studied the morphologic and functional characteristics of these cells at succeeding intervals during several-day cultures. At the beginning, during, and shortly after replication they had the characteristics of monocytes by several criteria. Later, and when not replicating, they developed into macrophages and varying numbers of epithelioid cells. By moderate manipulation of culturing conditions we have been able, then, to observe various aspects of mouse monocyte-macrophage differentiation and replication in relatively simple medium and using blood-adherent monocytes alone.

Animals↗
PubMed1981

Crossed immunoelectrophoresis of mouse serum.

Because crossed immunoelectrophoresis (X-IEP) is quantitative and offers more information than immunoelectrophoresis (IEP), we selected X-IEP to study various physiologic and pathologic effects on the antigen composition of mouse serum. We therefore needed and X-IEP map of normal mouse serum. This paper presents one that we have developed, along with brief descriptions of how the mouse serum antigens were identified. Cross-reactivity between several mouse and human serum antigens was especially helpful. Some data from using ratios of the areas of precipitation of detected antigens to that of an internal standard antigen, alpha-macroglobulin, are presented. They show that X-IEP readily detects quantitative abnormalities among any of the several serum antigens detected by this technique, and that 'normal' values for these antigens in one population of mice can be established in the same way of using ratios with just a few analyses.

Animals↗
PubMed1981

Evans blue dye adjuvant enhances delayed hypersensitivity while blocking immunity to Mycobacterium tuberculosis in mice.

Evans blue dye functions as an adjuvant with protein antigens in saline to induce cell-mediated immunological responses in mice. But when used to help induce cell-mediated tuberculoimmunity, it decreased mouse resistance to tuberculosis instead of helping induce immunity. This paradox was investigated. As could be expected from previous work with other antigens, the dye did promote induction of delayed hypersensitivity in mice to tuberculoprotein when injected in saline with killed tubercle bacilli. Peritoneal macrophages from mice injected with the dye responded normally to migration inhibition factor. Morphologically, these cells were moderately "activated" compared with similar cells taken from untreated mice. However, such cells incubated with tuberculosis growth inhibition lymphokine in an in vitro test for tuberculoimmunity did not express tuberculoimmunity, whereas macrophages from untreated mice did. Therefore, Evans blue dye did promote induction of cell-mediated immunological responses and tuberculoimmunity in lymphocytes, but under the conditions used in these experiments, it also blocked expression of tuberculoimmunity by macrophages.

Adjuvants, Immunologic↗
PubMed1981

Preliminary demonstration of human tuberculoimmunity in vitro.

In this paper a method for studying human tuberculoimmunity in vitro is described. Results from its use support an explanation for human tuberculoimmunity that is like that for mouse tuberculoimmunity: that immune lymphocytes are stimulated by being cultured with immunizing antigen to make a lymphokine which enables syngeneic macrophages to kill intracellular tubercle bacilli. This method uses antigen-responding lymphocytes and effector monocytes taken in the same original 20-ml sample of venous blood. These cells are cultured separately, the lymphocytes for 72 h with antigen to make immune lymphokine and the monocytes for 7 days to become macrophages. The macrophages are then infected with attenuated or virulent tubercle bacilli and cultured for 7 days more in medium with or without the immune lymphokine. Without it they are unable to control intracellular replication of the bacilli, whereas with it they do. This lymphokine was produced only by lymphocytes of immune subjects, of whom there were three kinds: tuberculin positive naturally immunized, Mycobacterium bovis BCG immunized, and trypsin-extracted bacillary antigen immunized. This method for detecting human tuberculoimmunity in vitro should be useful for comparing experimental vaccines and for studying cellular and molecular mechanisms of human tuberculoimmunity under better controlled conditions than hitherto have been possible.

BCG Vaccine↗
PubMed1979

Identification of alpha1-lipoproteins in crossed immunoelectrophoresis.

Evans Blue dye binds selectively, but with different avidities, to five major antigens in human serum. The anodic mobility of the antigen-dye complexes is greater than that of the antigens alone in crossed immunoelectrophoresis, which is of practical value for identification. We used this characteristic to show that in some human sera there is a population of alpha1-lipoprotein molecules that migrates electrophoretically in the beta-lipoprotein region, where in conventional zone electrophoresis it could be mistaken for beta-lipoprotein. We also demonstrate that horses, unlike rabbits, rarely make precipitins to human serum alpha1-lipoprotein. Equine antiserum to human serum therefore generally is useless for detecting this important serum antigen in immunoelectrophoresis.

Evans Blue↗