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PubMed1990

[The Pharmacia CAP system as a new measure of specific IgE. Application in the diagnosis of hypersensitivity to the venom of the Vespula wasp].

Every improvement can make the measurement of specific IgE more effective. This is the case with the CAP system, a new solid phase technique which uses an Immunocap made from hydrophilic polymer, encased in a capsule, for the fixation of anti-IgE or allergen. The Immunocap fixes 3 times more protein than paper discs. Another improvement is that of calibration of the CAP System by the WHO standard for IgE, with the results expressed in KUI/I: 0.35 to 100 KUI/I for specific IgE and 2 to 2,000 KUI/I for total IgE. The study was of patients who had an anaphylactic or systemic reaction after wasp stings. The subjects were divided into two groups A and B and compared with a reference Groupe C. The study also included subjects who were sensitive to mites, grass pollens and egg. Finally, the technique itself was submitted to a reproducibility test. The results confirm that the sensitivity and specificity of the test were both equal to unity. The CAP System therefore offers the possibility of detecting the lowest amounts of IgE.

Anaphylaxis↗
PubMedDate not supplied

[Prevalence of sensitization to Blomia in Gran Canaria].

In the present study we assessed the prevalence of sensitizations to Blomia in our environment. Perennial climatological conditions in the Canarian Islands facilitates the habitat to mites. The Blomia is found in tropical and subtropical homes of Europe, USA and Asia like the Dermatophagoides. We have designed a transversal and prospective study on 100 patients who consulted our Unit for the first time. A Prick-test for the common allergens, including Blomia Kulagini, and in vitro determination of total and specific IgE to Dermatophagoides Pteronyssinus and Farinae, was performed in those patients showing the same or higher levels than class 3 CAP System RAST-Fesa against Blomia Tropicalis. Results achieved in our environment and our patients are the following: 1. Prevalence of sensitizations to BK in patients who consult for the first time is 50%. 2. The specific IgE immunologic response to BK is 56% (45 patients). 3. Scarce correlation in the papule areas and specific IgE levels between BT and DTP (p = 0.22020); BT and DFA (p = 0.09063). Table VI. Fig. I and II. 4. These results and previous studies of allergenization suggest poor crossed reactivity between Blomia and Dermatophagoides. 5. We think that Blomia is a new etilogic agent of the allergic respiratory disorders in our environment and therefore it should be included in the standard set of allergens from a diagnostic and therapeutical point of view. We keep studying its identification and count in dust samples from our homes, as well as the crossed reactivity, by RAST inhibition, with other acari.

Adolescent↗
PubMed1986

Postnatal development of dendritic reticulum cells and their immune complex trapping ability.

The postnatal development of dendritic reticulum cells in the rat popliteal lymph nodes was electron microscopically investigated in relation to the appearance of immune complex trapping capacity. The popliteal lymph nodes of neonatal rat consisted of loosely arranged fibroblastic reticulum cells. In the following stage, the peripheral cortex and paracortex became distinguishable. The former was made up of an accumulation of small lymphocytes, scattered within a framework of reticulum cells. On te 28 th day, the first primary follicle appeared in the peripheral cortex. Simultaneously the immune complex could be trapped on the cytoplasmic membrane of reticulum cells, which were located in the central portion of the primary follicles. The early image of germinal centers appeared corresponding to immune complex trapping areas. In the well-developed secondary follicles, the immune complex trapping cells were mainly localized in the cap area. Their cytoplasmic membranes formed the dendritic processes, on which the distinct ability of trapping of the immune complex was recognized. It was demonstrated that the fibroblastic reticulum cells, forming the stroma of lymph nodes, were transformed into the typical dendritic reticulum cells with labyrinth structures in the cap area. Desmosomal junctions were often found, not only between the dendritic reticulum cells themselves, but also between the dendritic reticulum cells and lymphocytes. We suggest that the desmosomal junctions play a role as the channel for a transmission of immunological information.

Animals↗
PubMed1978

Detection of mouse alloantibodies by rosetting with protein A-coated sheep red blood cells.

Staphylococcal protein A has an affinity for the Fc portion of the IgG molecule of different species and can therefore be used to detect cell-bound immunoglobulin. Using this property, protein A coupled to sheep red blood cells via chromic chloride can detect alloantibodies to mouse H-2, Thy-1, Ly-1, 2, 4, 5, 6, and 7, and Ia antigenic specificities bound to the surface of lymphocytes by the formation of rosettes. In comparison with other rosetting and cytotoxicity assays, the protein A assay shows a greater sensitivity than does cytotoxicity using spleen cells as the target, as does the sheep anti-mouse Ig rosetting assay, whereas cytotoxicity shows greater sensitivity with some antisera on thymocytes. The major advantages of the protein A assay are that constant low reproducible backgrounds are obtained, there is no need to remove surface Ig by capping prior to antiserum treatment, and that viable cells can be recovered.

Animals↗
PubMed1975

Characterization of an antibody directed against a surface component of normal and pleomorphic cells of Streptococcus sanguis.

Whole cells of Streptococcus sanguis were utilized as an immunoadsorbent to purify large quantities of an antibody (S1) directed against a cell surface component. The S-1 antibody was isolated from antisera to normal (N) and pleomorphic (O) cells by a similar adsorption-elution procedure. The S-1 antibody isolated from antisera to N cells reacted in gel diffusion in identify with the S-1 antibody to O cells, indicating that the antigen which binds S-1 antibody (Ag-1) may not be radically altered when cells become pleomorphic. The S-1 antibodies directed against both N and O cells had restricted heterogeneity, indicating that for both types of cell Ag-1 may have a simple repeating structure. However, N cells were agglutinated to a greater extent by S-1 antibody than O cells. In addition the distribution of the bound S-1 antibody became altered as the cells became pleomorphic. Utilizing the technique of indirect immunofluorescence we observed that the S-1 antibody was distributed evenly on the surface of N cells. As the cells became pleomorphic, the antibody appeared to bind preferentially at the cell poles (capping). Later, as the cells became more grossly deformed, additional bands of immunofluorescence appeared to bisect the cells. Electron microscopic analysis indicated that the bound antibody was not associated with septal notches. The results suggest that the arrangement rather than the immunological properties of Ag-1 became altered as cells became pleomorphic.

Agglutination↗
PubMed1986

Effects of anti-schistosomal chemotherapy on immune responses, protection and immunity. I. Changes in cellular and humoral responses.

The cellular and humoral immune responses of CF1 and C57BL/6 mice with schistosomiasis mansoni were evaluated before and after chemotherapeutic cure of their infections by praziquantel. Mice were infected for either 10 or 20 weeks prior to treatment and followed until 10 weeks after treatment. Peripheral blood eosinophilia, without concomitant general leukocytosis, was observed within 3 days of treatment and persisted for up to 4 weeks. By 6 and 10 weeks after treatment schistosomal-associated hepatosplenomegaly had greatly decreased. Delayed-type hypersensitivity to a soluble adult worm extract (SWAP) was modulated over 20 weeks of infection, and in C57BL/6 mice this modulation was alleviated by cure. In parallel studies of pulmonary egg granuloma formation, granuloma modulation was not effectively reversed. Antibodies against egg (SEA), cercarial (CAP) and adult worm (SWAP) extracts generally decreased by 10 weeks after chemotherapy of mice that were previously infected for 10 weeks. Mice infected for 20 weeks and then treated, generated increased levels of antibodies to SWAP and CAP by 10 weeks after treatment. Immunoglobulin isotypic analyses largely reflected the results of total antibody studies. These data demonstrate that the duration of infection prior to treatment is a determining factor in subsequent expression of immune reactivity, and provide the immunological background for experiments on resistance following chemotherapy of experimental murine schistosomiasis mansoni.

Animals↗
PubMed1993

Bond strength of resin to dentin treated with calcium phosphate desensitizer.

In a previous study we proposed a new method utilizing immediate precipitation of calcium phosphate (CaP) in situ for treatment of dentin hypersensitivity. Sequential application of 5% disodium phosphate followed by 10% calcium chloride on patent dentin surface resulted in instant occlusion of dentin tubules and immediate relief from the hypersensitivity. To guarantee long-lasting effectiveness of the CaP treatment, resin coverage of the treated dentin surface will be the next practical approach. However, the CaP covering the surface may possibly affect bonding of resins. This study determined the effect of the CaP treatment on tensile bond strength of adhesive resins to dentin. Independent of the type of resin used, a bond strength of 4-5 MPa was obtained; cohesive failure occurred within the resin-CaP hybridized composite layer rather than at the dentin surface. The CaP treatment produced a significant increase in the bond strength of one resin.

Animals↗
PubMed2005

Community-acquired pneumonia: new facets of an old disease--Hantavirus pulmonary syndrome.

It seems that with climatic and geoecologic changes, Hantaviruses have re-emerged as human pathogens related to increases in interaction between humans and rodent reservoirs. Infection with SNV in North America and the Andes virus in South America can produce infection manifest initially as a flu-like illness. In the setting of a history of possible exposure to rodents or their excreta, clinical symptoms and laboratory clues such as thrombocytopenia should raise the suspicion of HPS. Clinical deterioration can be rapid, so patients should be hospitalized and transported to tertiary care centers where mechanical ventilation is available if necessary. Presumptive treatment for other forms of sepsis should be considered before confirmation of diagnosis. Survival seems to be determined in part by viral and host factors. Canadian and South American data suggest that there may be species variations influencing clinical manifestations and course of disease. Because the pathogenesis seems to be based on immunologic injury, future treatments will likely focus on interventions other than antiviral medications. Prevention strategies should be emphasized, particularly when recognized climatic conditions favor rodent abundance. Physicians should remain alert to the possibility of such a diagnosis when evaluating a patient with CAP and should request appropriate serology while supporting the patient in a closely monitored setting. The declining mortality rates seen over the past decade may be a consequence of improved medical management or better recognition of cases, including those less severe than originally described.

Adult↗
PubMedDate not supplied

Food allergy: a challenge for the clinician.

Adverse reactions to food resulting in gastrointestinal symptoms and due to immunologic reactions (allergy) are discussed: their pathogenesis, the prevalence of food allergens and the clinical digestive expressions of food allergy in children and adults are reviewed. In IgE-mediated food allergy, the usefulness of the biological available tests is considered, mainly CAP tests, for proceeding to the diagnosis and the monitoring of the allergic disease. Finally, the best actual diagnostic tools in food allergy are considered (clinical history, skin tests, biological tests and food oral challenges), with their limitations and indications.

Adolescent↗
PubMed1995

Morphological fate and sequelae of human atherosclerosis: evaluation of immune mechanisms in atherogenesis through immunohistological and ultrastructural analysis.

Modern techniques of investigation have revealed several similarities between atherosclerosis and chronic inflammation, and that immune mechanisms seem to operate in the incipient and subsequent phases of atherosclerosis. In the present study, the fate and morphogenesis of human atherosclerosis was considered, and the immune aspects of atherogenesis were analysed, using fresh human aorta obtained from autopsy cases. One of the earliest changes in the grossly normal, lesion-prone area of the aorta from young cases (prelesional changes) was the infiltration of blood-borne T lymphocytes and monocytes/macrophages beneath the endothelium. Cell-populated lesions abounding in T lymphocytes and macrophages, often bearing signs of activation, with or without cytoplasmic lipids were found in the fatty streaks, cap and shoulder regions of more advanced atheromatous plaques. The ultrastructural observation of cell-rich areas suggested that cognate cell-to-cell interaction plays a pivotal role in atherosclerosis, as well as cytokine-mediated paracrine or autocrine mechanisms. From an immunological perspective, the areas where both cell types are especially numerous and in close proximity are considered to be the areas with an index of disease activeness or progressiveness. Also, the present authors show evidence of clonal expansion of T lymphocytes. It is most likely that the increase of intimal cells was caused by the recruitment of immunocompetent cells from the blood-stream into the intima and by the clonal expansion of T lymphocytes. In addition, dead or dying cells were identified in areas of different stages ranging from prelesional areas to atheromatous plaques. Thus, the initiation and progression of human atherosclerosis appears to be punctuated by brief episodes of immunological events related to cell infiltration, proliferation and death.

Adolescent↗
PubMed1995

Monophosphoryl lipid A behaves as a T-cell-independent type 1 carrier for hapten-specific antibody responses in mice.

It is known that the lipopolysaccharide (LPS) of gram-negative bacteria, in addition to being a potent adjuvant, is an effective carrier for covalently associated haptens. However, the toxic nature of most forms of LPS precludes their use as adjuvants or carriers for human vaccines. 4'-Monophosphoryl lipid A (MLA), a derivative of LPS with attenuated toxicity, is currently being tested in humans as an immunological adjuvant. In this study, MLA was tested for its ability to function as a carrier for a small hapten, the trinitrophenyl group (TNP). MLA was first modified by addition of 6-aminocaproic acid to the 6' position of the disaccharide backbone (Cap-MLA). TNP was then attached to Cap-MLA via the free amino group, yielding TNP-Cap-MLA. Immunization of normal mice with TNP-Cap-MLA resulted in high-titer anti-TNP responses of immunoglobulin M and all immunoglobulin G subclasses. Furthermore MLA, like other T-cell-independent type 1 (TI-1) carriers, induced responses in athymic and X-linked immunodeficient mice. In all cases, immunization with either MLA alone or TNP-Cap plus MLA failed to induce measurable anti-TNP antibodies of any isotype, indicating that covalent association of MLA and hapten was necessary for MLA's carrier activity to be manifested. These properties of MLA make it a potential candidate as a carrier for vaccine subunit components, such as small peptides, especially for situations in which T-cell help is impaired, as occurs following human immunodeficiency virus type 1 infection.

Animals↗
PubMed1996

Effect of captopril on intracellular free calcium ([Ca2+]i) of thymocytes in spontaneously hypertensive rats.

Hypertension is associated with dysfunction of the immune system. Intracellular free calcium ([Ca2+]i) was reportedly increased in immune cells, such as lymphocytes and thymocytes, both in hypertensive patients and in animals. To evaluate if captopril has an effect on the immune system, [Ca2+]i in thymocytes was monitored during the development of hypertension from 4-24 weeks, and after treatment with captopril for 4 and 12 weeks in spontaneously hypertensive rats (SHR). SHR (n = 24) were treated with captopril at a dose of 20 mg/kg/day at 12 weeks of age after establishment of hypertension. Untreated SHR (n = 17) and normotensive Wistar Kyoto (WKY n = 14) served as control. [Ca2+]i of thymocytes was measured using Fura-2. It was found that [Ca2+]i of thymocytes decreased both in SHR and WKY from 4-12 weeks of age. From 12-24 weeks, [Ca2+]i in thymocytes remained at a constant level in WKY. However, [Ca2+]i in thymocytes increased significantly in SHR after 12 weeks. Captopril (CAP) treatment significantly lowered [Ca2+]i in thymocytes derived from SHR at 24 weeks. This treatment was associated with a moderate reduction in systolic blood pressure. Established hypertension in SHR is associated with increased [Ca2+]i in thymocytes. This may be indicative of immunological dysfunction. Captopril inhibits the increase in [Ca2+]i of thymocytes in SHR.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1975

Stimulation and inhibition of anti-hapten responses in guinea pigs immunized with hybrid liposomes.

Guinea pigs were immunized with liposomal model membranes containing phosphatidylethanolamine (PE) or glycerophosphorylethanolamine (GPE) derivatives in which the amino function was substituted with either dinitrophenylaminocaproyl (Dnp-Cap) or mono(p-azobenzenearsonic acid)tyrosyl (ABA-Tyr) residues. Previous studies have demonstrated that hapten-specific antibodies are elicited by DNP-Cap-PE or ABA-Tyr-PE sensitized liposomes and that cell-mediated immunity is induced by ABA-Tyr-PE (but not Dnp-Cap-PE) sensitized liposomes. These liposomes differ from conventional immunogens in which haptens are covalently attached to immunogenic carriers. This investigation describes two new aspects of liposomal immunogenicity in animals immunized with hybrid liposomes containing both Dnp-Cap-PE and ABA-Tyr-PE. (1) Stimulation of the anti-Dnp response by incorporation of increasing amounts of ABA-Tyr-PE; (2) inhibition of anti-ABA antibody formation by incorporation of increasing amounts of DNnp-Cap-PE. The two phenomena are dependent on the presence of each determinant in the same lipid bilayer. Thus, entrapment of the water-soluble deacylated derivative of ABA-Tyr-PE (i.e., ABA-Tyr-GPE) in a aqueous compartments of Dnp-Cap-PE sensitized liposomes does not enhance anti-Dnp antibody production. Similarly, entrapment of the non-amphipathic derivative of DNP-Cap-PE (i.e., Dnp-Cap-GPE) within ABA-Tyr-PE sensitized liposomes does not suppress anti-ABA antibody formation. Furthermore, mixtures of Dnp-Cap-PE sensitized liposomes and ABA-Tyr-PE sensitized liposomes neither stimulated nor inhibited the anti-hapten responses. These results indicate that preparation of hybrid liposomes with different N-substituted PE derivatives provides an extremely convenient method for controlling hapten and/or immunologic carrier determinant density.

Aminocaproates↗
PubMed1997

[When the fluoro-immuno-enzymatic (FEIA) measurements turn out to be more sensitive than radioimmunologic (RIA) measurements. Application to the measurement of serum tryptase].

The measurement of Tryptase by the Fluoro-Immuno-Enzymatic (FEIA) method is nowadays possible on the Pharmacia CAP system (automatic UniCAP). This measurement is more comprehensive as it measures the release of serum tryptase from both the tissue mastocytes (MCTC) as well as the mucosal mastocytes (MCM). Technically the measurements are comparable with those made by the method of radio-immunology (RIA), are absolutely reproducible and surprisingly at 100%. It has also been possible to evaluate the two techniques of FEIA and RIA on negative and positive pools. This new FEIA technique for serum tryptase is applicable: to anaphylactic and/or anaphylactoid accidents at the time of induction of anesthesia, in general conditions such as haemorrhagic recto colitis (RCH), Crohn's disease, and mastocytosis. Finally these measurements can be used during nasal and bronchial provocation tests, as the measurements may be made on nasal and bronchial lavage liquids. The sensitivity and the very good reproducibility of this new technique of FEIA for tryptase is of very great interest and avoids use of radio-active isotopes.

Anaphylaxis↗
PubMed2000

Mitochondrial F(0)F(1) ATP synthase. Subunit regions on the F1 motor shielded by F(0), Functional significance, and evidence for an involvement of the unique F(0) subunit F(6).

Studies reported here were undertaken to gain greater molecular insight into the complex structure of mitochondrial ATP synthase (F(0)F(1)) and its relationship to the enzyme's function and motor-related properties. Significantly, these studies, which employed N-terminal sequence, mass spectral, proteolytic, immunological, and functional analyses, led to the following novel findings. First, at the top of F(1) within F(0)F(1), all six N-terminal regions derived from alpha + beta subunits are shielded, indicating that one or more F(0) subunits forms a "cap." Second, at the bottom of F(1) within F(0)F(1), the N-terminal region of the single delta subunit and the C-terminal regions of all three alpha subunits are shielded also by F(0). Third, and in contrast, part of the gamma subunit located at the bottom of F(1) is already shielded in F(1), indicating that there is a preferential propensity for interaction with other F(1) subunits, most likely delta and epsilon. Fourth, and consistent with the first two conclusions above that specific regions at the top and bottom of F(1) are shielded by F(0), further proteolytic shaving of alpha and beta subunits at these locations eliminates the capacity of F(1) to couple a proton gradient to ATP synthesis. Finally, evidence was obtained that the F(0) subunit called "F(6)," unique to animal ATP synthases, is involved in shielding F(1). The significance of the studies reported here, in relation to current views about ATP synthase structure and function in animal mitochondria, is discussed.

Adenosine Triphosphate↗
PubMed2000

Calcium phosphate nanoparticle adjuvant.

Vaccination to protect against human infectious diseases may be enhanced by using adjuvants that can selectively stimulate immunoregulatory responses. In a murine model, a novel nanoparticulate adjuvant composed of calcium phosphate (CAP) was compared with the commonly used aluminum (alum) adjuvants for its ability to induce immunity to herpes simplex virus type 2 (HSV-2) and Epstein-Barr virus (EBV) infections. Results indicated that CAP was more potent as an adjuvant than alum, elicited little or no inflammation at the site of administration, induced high titers of immunoglobulin G2a (IgG2a) antibody and neutralizing antibody, and facilitated a high percentage of protection against HSV-2 infection. Additional benefits of CAP include (i) an insignificant IgE response, which is an important advantage over injection of alum compounds, and (ii) the fact that CAP is a natural constituent of the human body. Thus, CAP is very well tolerated and absorbed. These studies were performed with animal models. By virtue of the potency of this CAP adjuvant and the relative absence of side effects, we believe that this new CAP formulation has great potential for use as an adjuvant in humans.

Adjuvants, Immunologic↗
PubMed1990

Isolation of a novel tumor protein that induces resistance to natural killer cell lysis.

The human metastatic tumor cell line CAP-2, produces a soluble factor that induces resistance to NK lysis of K-562 susceptible leukemia cell line, and does not inhibit the cytotoxic capacity of effector cells. The use of sequential HPLC, hydrophobic interaction chromatography, and reverse phase chromatography, coupled with cytotoxic assays, resulted in the isolation and separation to homogeneity of a novel protein responsible for this biologic activity. Size estimation studies based on TSK HPLC columns showed that this protein has a mass of 8 to 12 kDa. The amino acid composition analysis of the CAP-2 protein calculated from HPLC chromatograms shows that this protein contains around 108 amino acids. Subsequent gas phase sequence analysis, however, was hampered because the N terminus of this protein was blocked and therefore unsuitable for sequencing by Edman degradation. The functional studies showed that the NK lysis-resistance activity of the CAP-2 protein is mediated by interaction with and nonspecific binding to NK target cells. The lymphokine-activated killer and macrophage-mediated cytotoxicity and mitogen-induced proliferation is not affected. Unexpectedly, the CAP-2 protein appears to be mitogenic to its own cell line. Thus, the induction of NK lysis-resistance and the mitogenic activity showed by CAP-2 protein could contribute to the tumor growth and metastatic establishment.

Amino Acids↗