Glycoprotein Preparations Research Articles

One-step purification of chicken growth hormone from a crude pituitary extract by use of a monoclonal immunoadsorbent.

The immunization of mice with an affinity-purified glycoprotein preparation from chicken pituitary tissue yielded several monoclonal antibodies towards the recently described glycosylated variant of chicken GH. As all these antibodies recognize the classical (non-glycosylated) GH molecule equally well, they provide a suitable tool for the development of both a specific immunoadsorbent and an assay method. This paper deals with the surprising purification power of the immunoadsorbent that was produced with one of the monoclonal antibodies. The resulting preparation was more than 99% pure as assessed by reversed phase high-performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis, so that no further purification steps were needed before the determination of the amino acid sequence of the material. The efficiency of the purification protocol as determined by a homologous, monoclonal antibody-based radioimmunoassay was virtually absolute. Moreover, the affinity-purified GH preparation was a mixture representing the multiple molecular forms of pituitary chicken GH, including both oligomeres and glycosylated GH. The purified preparations were finally used to demonstrate the hepatic 5'-monodeiodinase-stimulating activity of GH in the chicken embryo (results not shown), in order to prove that the biological activity of the molecule had not been damaged by elution from the immunoadsorbent.

Interleukin-2 production in vitro: a new approach to the study of rabies vaccine immunogenicity as appraised by testing different glycoprotein presentations

When injected as an immunosome presentation (molecules anchored to preformed liposome), rabies glycoprotein (GP) is capable of protecting animals against rabies either before or after viral infection. The presentation of the GP molecules in the correct form seems to be essential for the induction of antirabies protection. This condition must be taken in account in the making-up of a rabies subunit vaccine. In order to study the relationship between the immune responses induced by the rabies GP and its protective activity, different presentations of the GP were prepared. Purified glycoprotein molecules were associated under different physical forms: liposome-anchored, self-aggregated (rosettes) and associated with the viral lipids (virosomes). These presentations appeared different on electron microscopy. They also exhibited differences in the expression of an immunodominant epitope and in their protective activity. The non-specific immune response, as appraised by interferon production and natural cytotoxicity, was induced at a high level only by the purified viral particles. Specific immune responses (namely virus neutralizing antibody and interleukin-2 production) was induced at high levels only by the viral particle and by the liposome-anchored glycoprotein. A parallelism has previously been established between protection by glycoprotein preparations and interleukin-2 production in primed mice splenocytes. This suggests that the measure of interleukin-2 production in vitro could be used to evaluate the capability of a rabies antigen to induce a T-cell response and to confer protection.

Intranasal immunization of hamsters with envelope glycoproteins of human parainfluenza virus type 3.

Envelope glycoproteins of human parainfluenza virus type 3 (PIV-3) were selectively solubilized with n-octyl beta-D-glucopyranoside and reconstituted into lipid vesicles by dialysis of the detergent. The efficacy of the glycoprotein preparation as a subunit vaccine when administered to hamsters intranasally or subcutaneously was compared. Animals receiving four intranasal immunizations with 5 micrograms of the glycoprotein preparation were completely resistant to challenge infection. Only partial protection, however, was observed in animals immunized subcutaneously with the same dose of antigen. The local glycoprotein-specific IgA response was significantly higher in intranasally immunized animals and was implicated in resistance to challenge infection.

Human antibody responses to Schistosoma mansoni: the influence of epitopes shared between different life‐cycle stages on the response to the schistosomulum

Sera from 120 Kenyan schoolchildren who were infected with S. mansoni were individually examined, using an enzyme-linked immunoabsorbent assay (ELISA), for the presence of IgG and IgM antibodies reactive with antigens derived from adult worms, the outer membrane of the schistosomulum or from the parasite egg. In addition, antibodies against more purified egg antigens, an egg stage-specific glycoprotein preparation and a polysaccharide egg antigen known to share epitopes with the schistosomular surface were measured in ELISA, as were antibodies reactive with trichloroacetic acid-soluble and periodate-insensitive antigens derived from the outer membrane of schistosomulum and antigens shed when schistosomula were cultured in vitro. IgG subclass responses to the unfractionated egg antigen were also measured. The results from each of these assays were compared with the results of each other assay and with the number of parasite eggs excreted by each child, using Spearmans rank correlations. These comparisons revealed a number of statistically significant positive correlations. IgG4 anti-egg antibodies correlated better with intensity of infection than did other IgG subclasses. Total IgG responses against polysaccharide antigens did not correlate with intensity of infection as well as IgG responses against other antigens; epitopes shared between the schistosomulum surface and the adult worm were different to those shared with the parasite egg; and, there was antigen-directed restriction of IgG subclass responses to some egg and adult worm antigens which carried these shared epitopes. It is argued that this might have a qualitative effect on the nature of the antibodies directed against the schistosomulum by infected individuals and therefore have important consequences for the outcome of a subsequent exposure to infection with the same parasite.

Pilus-mediated interactions of the Escherichia coli strain RDEC-1 with mucosal glycoproteins in the small intestine of rabbits

Escherichia coli strain RDEC-1 (serotype 015:NM) is an effacing adherent enteropathogen that binds to the intestine of rabbits in a manner morphologically identical to the binding of human enteropathogenic E. coli strains to human intestine. The rabbit enteropathogen adheres to mucosal enterocytes in vivo and to microvillus membranes in vitro. Binding of RDEC-1 to ileal brush borders and to M cells overlying Peyer's patches is mediated by pili (fimbriae) expressed on the cell surface of bacteria. To examine whether similar binding occurs to glycoproteins present in the intestinal lumen, RDEC-1 was fed to rabbits and the intestinal luminal contents were examined for in vivo colonization by RDEC-1. In addition, preparations of rabbit luminal glycoproteins were tested for their ability to agglutinate RDEC-1 in vitro. After the infection of rabbits, RDEC-1 organisms were found in the intestinal lumen associated with glycoproteins, as defined by positive histochemical staining of luminal material by periodic acid-Schiff and Alcian blue. In vitro aggregation of RDEC-1 by luminal glycoproteins of rabbit intestine, a luminal glycoprotein fraction purified on column chromatography and rabbit ileal microvillus membranes, was determined using a microtiter plate assay and an aggregometer. Nonpiliated RDEC-1 did not interact with either of the intestinal glycoprotein preparations or microvillus membranes. RDEC-1 expressing mannose-resistant AF/R1 pili or mannose-sensitive type 1 pili coaggregated with both rabbit luminal glycoprotein preparations and rabbit ileal microvillus membranes at equivalent protein concentrations. These studies demonstrate that RDEC-1 are associated with luminal glycoproteins during in vivo infection of rabbits, and that piliated RDEC-1, but not nonpiliated, coaggregate with rabbit glycoprotein preparations in vitro. Luminal glycoproteins contained within the mucus layer may serve as a site for replication and colonization of bacteria before bacterial enteroadherence.

Identification of terminal N-acetylglucosamine residues of highly branched asparagine-linked oligosaccharides as immunoreactive domains of a chicken heterophile antigenic determinant

Chicken heterophile antigenic determinant (CHAD-1) has been previously found in medullary lymphocytes of the bursa and thymus as well as in some non-lymphoid cells by the immunoperoxidase method, using rabbit antiserum to a complete Freund's adjuvant (CFA) as the first antibody. In this work we demonstrated that absorption of anti-CFA serum with highly purified preparations of hen egg white glycoproteins (ovomucoid, ovoinhibitor, ovalbumin) or chicken orosomucoid completely blocked immunoperoxidase staining for CHAD-1. Treatment of these glycoproteins with β- N-acetylglucosaminidase suppressed their capacity to inhibit this staining. Absorption of anti-CFA serum with asparagine-linked glycopeptides which have the mannose α1,3 arm disubstituted by GlcNAc residues and which have another GlcNAc residue linked β 1,4 to the β-linked mannose of the core also inhibited staining for CHAD-1. These data indicated that highly branched asparagine-linked oligosaccharides with terminal GlcNAc residues β-linked to mannose represent immunoreactive domains of CHAD-1.

Immunization with envelope glycoprotein of an avian RNA tumor virus protects against sarcoma virus tumor induction: role of subgroup

Avian RNA tumor virus envelope glycoprotein protects against sarcoma development by an avian sarcoma virus of the same subgroup. Avian RNA tumor viruses, members of the retrovirus family, induce various malignancies in fowl (Weiss et al. (eds.), 1982, RNA Tumor Viruses, Cold Spring Harbor, N.Y.). These viruses consist of a genomic RNA core surrounded by an envelope with embedded glycoproteins, of 85 and 37 kDa. The 85 kDa glycoprotein is antigenically specific for each subgroup as determined by neutralization. The envelope glycoprotein can be removed from the virion with retention of its antigenicity (Duesberg et al., 1970, Virology 41, 631–646). Two fractions of 4-6S and 8S, separated by sedimentation, were shown to retain antigenicity by interference of neutralization of virus by antibody. Thus, the 4-6S and 8S preparations could possibly serve as immunogens. The objective of this study was to determine if such envelope glycoprotein preparations could function as potential vaccines, and if so, whether the protection afforded would be subgroup specific.

Native and recombinant herpes simplex virus type 1 envelope proteins induce human immune T-lymphocyte responses.

The abilities of whole herpes simplex virus type 1 (HSV-1) antigen (HSV-ag) and purified HSV-1 native and recombinant envelope proteins to stimulate in vitro T-lymphocyte responses were compared in patients with recurrent herpes labialis. Immunochemically purified preparations of native glycoproteins B, C, and D (ngB, ngC, ngD) from cultured HSV-1 as well as expressed recombinant plasmid preparations of gD (rgD-1t, rgD-45K) elicited lymphocyte proliferation (LT) and production of gamma interferon (IFN-gamma) and interleukin-2 (IL-2) only in seropositive individuals. The IFN-gamma induced by rgD-1t correlated with the time to the next herpetic lesion in 19 volunteers followed to recurrence (r = 0.69, P less than 0.008), although the magnitude and frequency of LT and IFN-gamma responses were lower with either recombinant or native purified antigens than with the whole-virus antigen. Combinations of ngB plus ngD or ngB plus ngC plus ngD stimulated more IFN-gamma, equivalent to whole-virus-antigen responses. Recombinant-derived human IL-2 also specifically increased LT and IFN-gamma responses in antigen-driven cultures. ngD stimulated IL-2 and LT responses similar to those of whole-virus antigen and higher than those of ngC. HSV-ag and ngB induced significantly higher titers of total IFN than could be accounted for by IFN-gamma; this was not seen for the other antigens, which induced only IFN-gamma. HSV-ag-driven Leu 2a-, plastic-nonadherent blood cells, unlike whole peripheral blood mononuclear cells, showed evidence of an increase and then a decline in the frequency of HSV-responsive cells after a lesion recurrence. These studies suggest that HSV-1 envelope proteins are capable of stimulating an immune T-helper-cell response which is associated with the prevention of human herpes simplex lesion recurrence. Although the whole virus probably contains additional important antigens, increasing concentrations or combinations of certain purified glycoproteins or the addition of nonspecific enhancers of T-lymphocyte function can drive in vitro immune responses to the same level as the complete set of viral antigens.

Antigenic pattern of human brain glycoproteins as described by monoclonal antibodies.

A battery of monoclonal antibodies was raised against a preparation of lentil lectin-binding membrane glycoproteins from human brain. Out of 26 established hybridomas, nine produced antibodies against the human Thy-1 antigen. For the remaining 17 lines, reactivity with at least six other antigens could be identified after immunoprecipitation and immunoblotting. Several of the antigens were di- or trimeric, mainly in the molecular weight range of 60-120 kDa. Two of the antibodies were reactive with high-molecular-weight aggregates and four targets for the antibody reactivity were not identifiable by immunoprecipitation of iodinated antigens. Three of the identified antigens were shown by quantitative enzyme-linked immunosorbent assay tests on various human tissues to be specifically expressed in the brain.

Alpha 1 acid glycoprotein in streptozotocin diabetic rats. Some aspects of sugar moiety structure and metabolism.

The pure alpha 1 acid glycoprotein (AGP) preparation from streptozotocin diabetic rat sera showed diminished sialic acid content and lower ratios of galactose to mannose and galactose to fucose. In vivo incorporation of 14C-leucine and 3H-galactose into AGP of control and diabetic animals was studied 30, 60 and 120 min after administration of the radioisotopic precursors. The changed 14C/3H ratio in AGP of diabetic rats suggested disturbed glycosylation of AGP in this disease. Similar activity of UDP-galactose 4-epimerase in control and diabetic rat liver was found suggesting that this enzyme has no influence on the decreased level of serum protein bound galactose.

Purification of individual varicella-zoster virus (VZV) glycoproteins gpI, gpII, and gpIII and their use in ELISA for detection of VZV glycoprotein-specific antibodies

We have utilized monoclonal antibodies in immune affinity chromatography to purify each of the 3 major glycoproteins of varicella-zoster virus (VZV), gpI, gpII, and gpIII, in immunologically active form. Upon injection into guinea pigs, each preparation elicited the production of specific antibodies capable of immunoprecipitating the homologous glycoprotein and of neutralizing VZV infectivity in vitro. Also, total glycoproteins from VZV-infected cells have been purified by lectin affinity chromatography. Each of the individual purified glycoproteins, as well as total VZV glycoproteins and appropriate uninfected cell protein controls, have been employed as solid-phase reagents in enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies directed against specific VZV glycoproteins. The specificity of the purified glycoproteins as ELISA reagents was verified by the ability of individual monoclonal antibodies to bind specifically to individual glycoprotein preparations. We have demonstrated the utility of the glycoprotein-specific ELISA by detecting antibodies in sera from post-zoster and post-varicella patients. The assay detects antibodies directed against each of the 3 major glycoproteins and is sensitive enough to detect antibodies in a 1:320000 dilution of some sera. This assay, as well as the purified individual glycoproteins per se, should prove to be very useful reagents in understanding the role of each of gpI, gpII, and gpIII in immunity to VZV.

Sexual Agglutination in the Unicellular Green Alga Chlamydomonas eugametos

Gametes of the unicellular green alga Chlamydomonas eugametos agglutinate via their flagella. The mating type plus agglutination factor was solubilized by relatively mild treatments such as a short pH shock or an osmotic shock indicating that it is an extrinsic membrane component. It was also extracted in the nonionic detergent Triton X-100. A simple two-step procedure consisting of gel filtration over Sepharose 4B-cross-linked followed by anion exchange chromatography of the void volume yielded an electrophoretically pure preparation of a single high molecular weight glycoprotein. The agglutination factor sedimented as a 9.3 S particle (assuming a density of 1.50) in sucrose gradients. This low value, compared with the high apparent molecular weight seen during gel filtration and electrophoresis, suggests that the agglutination factor is a rod-like molecule. This was confirmed by viewing rotary-shadowed preparations in the electron microscope. A population of long slender molecules was revealed (328 +/- 20 nanometers), many of which had a knob at one end and a flexible region about one fourth of the length from the other end.

A membrane glycoprotein involved in teratocarcinoma cell adhesion to substratum

An antiserum was raised against glycoproteins isolated from teratocarcinoma OTT6050 by affinity chromatography on Ricinus communis agglutinin-1 (RCA-1). The antiserum inhibited attachment and spreading of the teratocarcinoma cells on plastic tissue culture plates. Embryonal carcinoma cells (F9 and N4-1) and PYS-2 parietal endodermal cells were also affected. The antiserum was also effective in preventing adhesion of trophoblasts to substratum. Furthermore, the antiserum caused rounding and detachment of the cells previously spread on the substrates, and the process could be reversed by removing the antiserum. Fab fragments isolated from the antiserum also inhibited cell-substratum adhesion, but not cell-cell adhesion, indicating that the factor involved is specific for cell-substratum adhesion. The antiserum inhibited adhesion of F9 and PYS-2 cells not only to plastic dishes but also to those coated with plasma fibronectin, laminin, and plant lectins such as concanavalin A (conA). Therefore, it is likely that the antiserum inhibited cell-substratum adhesion by blocking the function of an intrinsic component of cell-substratum adhesion rather than interfering with specific receptors for fibronectin or laminin. The glycoprotein with the antibody-blocking activity was partially purified from particulate fraction of teratocarcinoma OTT6050 by extraction with Triton X-100 and affinity chromatography on RCA-1-agarose and conA-agarose Upon Western blot analysis of the glycoprotein preparation, the antibody reacted only with a glycoprotein of the apparent molecular weight (MW), 125 000 (GP-125). An antibody preparation was isolated by affinity chromatography on Sepharose 4B coupled with the glycoprotein fraction and was shown to have all of the activities described above. Molecules resembling GP-125 could be also isolated from cultured F9 and PYS-2 cells by solubilization with Triton X-100 and indirect immunoprecipitation.

Immune responses of cattle and mice to the G glycoprotein of vesicular stomatitis virus.

A subunit vaccine for vesicular stomatitis was developed from a purified vesicular stomatitis virus preparation by selectively removing the immunogenic G glycoprotein of the virus with the dialyzable, nonionic detergent, beta-D-octylglucoside. Cattle immunized intramuscularly with a single dose of 112 micrograms of G glycoprotein preparation in complete Freund's adjuvant did not develop vesicular disease following challenge by intralingual inoculation of 400 times the infectious dose of the virus. Similarly, mice vaccinated subcutaneously with a single dose of 10 micrograms of G glycoprotein preparation, with or without complete Freund's adjuvant, were protected from lethal encephalitis caused by vesicular stomatitis virus. A subunit vaccine for vesicular stomatitis of cattle, horses, and swine avoids the hazards associated with attenuated and inactivated vaccines, such as vaccine breaks, reversion to virulence, or introduction of virus into potential wild reservoirs or arthropod hosts. Further, it is possible to distinguish serologically animals vaccinated with the subunit preparation from those that have had the clinical disease or that have been vaccinated with whole virus. This is an essential consideration both for epidemiological studies and for disease control or establishment of quarantine programs.

Acidic Glycoproteins from Bovine Compact Bone

Several acidic glycoproteins with apparent molecular weights of 85,000 (bone sialoprotein, BSP), 80,000, 67,000, 60,000, 40,000 (osteonectin) and 30,000 were isolated from adult bovine compact bone. About 20-25% of the 67 K preparation was serum albumin. This contaminant was removed by passing the protein through Blue Sepharose CL-6B; the material that emerged in the void volume of this affinity column contained the bone 67 K glycoprotein free of serum albumin. All six of the proteins had high amounts of aspartic and glutamic acid. The amino acid compositions of 80 K, 67 K, 40 K and 30 K glycoproteins were very similar. The 60 K glycoprotein had an amino acid content that differed from the others with notably higher proline, alanine and valine values. All six protein samples contained mannose, galactose, glucosamine, galactosamine and sialic acid, but no fucose. The carbohydrate contents varied from about 10% for osteonectin to 38% for BSP. Antibodies were raised against electrophoretically purified osteonectin and used in enzyme-linked immunosorbent assays (ELISA). In addition to reactivity with osteonectin, the antibodies displayed strong reactivity to 80 K and 30 K glycoprotein preparations, but did not react with 67 K or 60 K bone glycoproteins or bovine serum albumin. None of the acidic glycoproteins cross-reacted with antibodies to bovine serum albumin, bovine serum proteins or human alpha 2-HS-glycoprotein.

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The specificity of complement-fixing, cytotoxic antibodies against the YAC lymphoma in sera of normal young adult (A X C57BL)F1 mice was studied. In vivo-maintained, immunoselected sublines of the YAC lymphoma expressed low amounts of the natural antibody (NAb) target structure. These cell lines were also resistant to natural killer (NK) cell-mediated lysis. After 2-3 weeks of in vitro culture the immunoselected cell lines became NK sensitive, but they remained resistant to NAb. When several independently derived variants selected for low NK sensitivity were tested for their ability to absorb NAb, the degree of absorption varied considerably among the variants. NAb could be inhibited by purified C-type virus particles and also by bacterial sonicates and various glycoprotein preparations. Treatment of target cells with tunicamycin, an inhibitor of asparagine-linked glycosylation, decreased the sensitivity to NAb lysis but had no impact on NK sensitivity. Thus the results indicated that a) NAb and NK cells recognized separate target structures and b) the target structure(s) for NAb but not for NK cells were saccharides of the N-glycosidic type.

Structural studies on a sulphated glycoprotein preparation isolated from human saliva

The identity of the sulphated component in a high-molecular-weight glycoprotein preparation (GP) from human whole saliva was investigated. A t 0.5 of 96 min for inorganic sulphate release in 0.25 M HCl indicated the presence of O-substituted sulphate, probably associated with hydroxyl groups of sugar residues. This possible association of ester was also inferred from infra-red data of the intact GP, where major absorption bands were detected at 1230–1250 and 828cm −1. Further a sulphatedmonosaccharide derivative was released during acid hydrolysis in 0.1 M HCl at 100 ° C for 75 min and isolated by gel-filtration on Sephadex G-15. This derivative was separated from a range of non-sulphated sugars by thin-layer chromatography in various solvent systems and possessed the Chromatographic characteristics of N-acetylgalactosamine-6- O -sulphate.

Recognition by a mouse monoclonal antibody of a glycoprotein antigen of rat brain which is expressed intracellularly by neurons

We describe a previously uncharacterised glycoprotein antigen of rat brain. The antigen was localised by immunofluorescence on 10 μm cryostat tissue sections, and was found to be present intracellularly in neurons. No other cell types or structures within the brain were stained. The antigen is recognised by a mouse monoclonal antibody called NGP41. The antibody was produced after immunising a mouse with glycoproteins purified by lentil lectin affinity chromatography of solubilised rat brain membranes. Spleen cells from the immunised mouse were fused with the myeloma P3X63Ag8. The antigen is expressed by neurons in all brain regions, and also in the dorsal root ganglion neurons of the peripheral nervous system. In all brain regions, the large projection neurons are the most intensely stained by immunofluorescence, but some small neurons also express the antigen. Although dendrites were not stained, sections of sciatic nerve were stained by NGP41, suggesting that the antigen is expressed by axonal processes. The cell bodies of neurons in the inferior olive were stained by NGP41, but their terminals on Purkinje cell dendrites in the cerebellar cortex were not stained, suggesting that the antigen is absent or expressed below the limit of detection in terminals. Both crude brain membranes and a lentil lectin affinity purified brain glycoprotein fraction absorbed the antibody, suggesting that the antigen is a membrane bound glycoprotein. In immunoblotting experiments, the antigen was detected in homogenates of brain and spinal cord membranes, where it appeared as a triplet of bands with molecular weights of 41K, 38K and 36K. Antigen was not detected by immunoblotting in homogenates of six different tissues of non-nervous origin. The antigen was enriched in glycoprotein fractions from adult and juvenile cerebellum as assessed by immunoblotting. Adult brain glycoprotein preparations had a triplet structure similar to that in the homogenates, although most of the antigenic activity of the juvenile preparation was found in a position corresponding to the upper two bands of the triplet.

Common conditions for high-performance liquid chromatographic microdetermination of aldoses, hexosamines, and sialic acids in glycoproteins

All aldoses known to be present in animal and plant glycoproteins were separated in partition mode on a column of a highly crosslinked cation-exchange resin (Shodex DC-613, H + form) and sensitively monitored by photometric detection after postcolumn labeling with 2-cyanoacetamide. Linearity for the sample amount ranging from 0.1 to 80 nmol was observed with high reproducibility. Hexosamines could be determined as their N-acetyl derivatives under the identical conditions with approximately one order of magnitude higher range of linearity. Sialic acid could be estimated as N-acylhexosamines produced by the action of N-acetylneuraminate pyruvate-lyase. The usefulness of these common conditions was demonstrated by analyzing component monosaccharides of some glycoprotein preparations.

Analysis of human nasal mucous glycoproteins

Human nasal turbinates were cultured in the presence of 3H-glucosamine, which is incorporated into nasal mucous glycoproteins. Nasal mucous glycoprotein was then characterized biochemically, and the effects of various neurohormones and immunologic stimulation on mucous glycoprotein release were analyzed. Fractionation of nasal mucous glycoprotein by gel filtration chromatography revealed a molecular size range of 2 to 200 X 10(5) (as judged by protein markers) but displayed a single, acidic charge, as reflected both in a narrow elution pattern from DEAE-cellulose and a sharp isoelectric focusing point of 2.6. Highly enriched nasal mucous glycoprotein preparations consisted of 80 per cent carbohydrate and 20 per cent protein (by weight) and included enzymatically cleavable carbohydrate side chains with molecular weights of 1,600 to 1,800. Thus, nasal mucous glycoproteins are a family of molecules that express uniform acidic charge characteristics and a wide range of molecular sizes. Cholinergic stimulation of atropine-inhibitable muscarinic receptors increased nasal mucous glycoprotein release in a dose-related manner, as did alpha-adrenergic stimulation. However, beta-adrenergic stimulation did not affect mucous glycoprotein release. Immunologic stimulation of nasal mast cells by either reversed anaphylaxis or antigen challenge after passive sensitization caused both histamine release and increased mucous glycoprotein release. Thus, nasal turbinates provide an accessible source of tissue for the analysis of nasal mucus secretion and mast cell degranulation and may provide a model for the study of pharmacologic approaches to the universally experienced discomfort of rhinorrhea.