bluuebunny/crossref_metadata_2025 huggingface dataset:
HLA and Lymphocyte Histamine Sensitivity in Mastocytosis
Authors: Olafsson Jon H. , Roupe Gösta , Rydberg Lennart
Published: August 2009

HLA-ABC and DR typing was performed on 50 patients with mastocytosis. Lymphocyte stimulation with concanavalin A has earlier been performed in 22 of these patients. No phenotypic aberrations were detected in the patient group. 8 patients had the HLA-B12 phenotype. Lymphocytes from the HLA-B12 phenotype stimulated with a lymphocyte mitogen have earlier been shown to react with decreased inhibitory effect to histamine. 2 patients in this study had the HLA-B12 phenotype and systemic mastocytosis with high histamine turnover, but these patients reacted with a low mitogen response of the lymphocytes.
In vitro Effects of Enzymes of Polymorphonuclear Leukocytes on the Antigenicity of Stratum corneum
Authors: Dabski Krzysztof , Glinski Wieslaw , Beutner Ernst H. , Jablonska Stefania
Published: August 2009

Anti-stratum corneum (SC) antibodies of human serum bind in vitro to the SC of frozen sections of normal skin of psoriatic patients and of healthy controls. Pretreatment of skin sections with selected dilutions of acid extracts from polymorphonuclear leukocytes (PMN) prior to the incubation with serum containing anti-SC autoantibodies enhances this binding in comparison with binding to untreated sections. Pretreatment of at least some specimens with higher concentrations of the same PMN extracts brings about a reduction in their reactivity with anti-SC autoantibodies. These findings indicate that enzymes of PMN are capable of altering SC antigens by first increasing and then decreasing their reactivity with SC autoantibodies.
A Reappraisal of IgE Levels in Various Human Secretions by Particle Counting Immunoassay Combined with Pepsin Digestion
Authors: Magnusson C.G.M. , Masson P.L.
Published: August 2009

We have assayed IgE in various human secretions by particle-counting immunoassay (PACIA). The introduction of a pepsin digestion step of the sample prior to the assay not only eliminates possible interfering factors but also releases from IgE the protease-resistant FcXXXε fragment. Since this fragment could represent the residual form of IgE in certain secretions rich in proteases it should be a suitable calibrator for the assay of IgE in secretions. The concentrations of IgE that we found in most fluids (urine, saliva, tears, milk, aqueous humour, cerebrospinal fluid, bronchoalveolar washings, intestinal fluids and faecal extracts) were up to 100 times lower than those reported by others who have used the radioimmunosorbent test (RIST). We suspect that the high values found with RIST are related to the susceptibility of this test to various interfering factors and not to a local production of IgE in most secretions of normal individuals.
Effects of Serum Amyloid P Component on Human Lymphocytes
Authors: Levo Y. , Wollner S.
Published: August 2009

The P component of amyloid is a normal serum protein designated SAP. SAP has substantial homology with C-reactive protein (CRP). Recent studies have established the structure, tissue distribution and binding reactivities of SAP; however, as yet, very minimal insight into its function has been achieved. Recent studies, though somewhat controversial, have suggested a regulatory role for CRP on the immune response. In view of these studies, we wanted to evaluate the in vitro effects of SAP on several immunological properties of human lymphocytes. We found that SAP had a marked inhibitory effect on the proliferative response of lymphocytes to a variety of T-dependent mitogens. In addition, SAP markedly enhanced the formation of active E rosettes, a marker of activated T cells.
A Monoclonal Antibody-Based Radioimmunoassay for the in vitro Production of IgE by Lymphocyte Cultures
Authors: Zaunders John J. , Ziegler John B. , Penny Ronald , Cooper David A.
Published: August 2009

Very sensitive radioimmunoassay systems have been described for the measurement of IgE produced in cultures of human peripheral blood mononuclear cells. However, differing results have been reported when cultures from non-atopic donors are stimulated with pokeweed mitogen, which may be due to cross-reactivity of anti-IgE antibodies with IgG. A monoclonal antibody specific for the Fc region of human IgE, and two polyclonal affinity-purified antibodies to IgE were tested for binding to 125I-labelled IgE myeloma proteins and polyclonal IgG in a sensitive double antibody precipitation assay. The monoclonal antibody and one of the polyclonal antibodies bound only IgE, whereas the other polyclonal antibody bound a significant proportion of labelled IgG. A solid phase radioimmunoassay was developed which combined the specificity of the monoclonal antibody with the sensitivity of the first polyclonal antibody as radioactive tracer. A second assay system was also tested using the cross-reacting antibody as tracer. Supernatants of pokeweed mitogen-stimulated peripheral blood mononuclear cell cultures from non-atopic donors were examined for IgE synthesis using both assays. The assay based on the monoclonal antibody did not detect IgE synthesis, whilst the second assay, based on the cross-reacting antibody indicated that spurious IgE had been produced in the same cultures. This study shows that protein-binding assays provide a simple means for checking the specificity of antibodies in solid phase radioimmunoassays, and confirms that pokeweed mitogen does not stimulate IgE production by cells from non-atopic donors when measured by a specific radioimmunoassay.
Immunoglobulin Production in Epstein-Barr Virus-Transformed Lymphoblastoid Cell Lines in Patients with Ataxia Telangiectasia
Authors: Ariga Tadashi , Sakiyama Yukio , Matsumoto Shuzo , Okano Motohiko , Osato Toyoro
Published: August 2009

Peripheral blood lymphocytes from 7 patients with ataxia telangiectasia were transformed by Epstein-Barr virus, and lymphoblastoid cell lines (LCL) were established. Immunoglobulin levels in the supernatants of the LCL were measured in order to analyze immunological abnormalities, especially the low levels of serum IgA in these patients. We found that the geometric means of IgA production in the supernatants of the LCL from the 7 patients were significantly lower than those from 9 healthy controls (5.0 vs. 834 ng/ml, p < 0.001). These results indicate that the low levels of serum IgA in ataxia telangiectasia are attributable at least in part to an intrinsic abnormality of B cells.
Characterization of Allergens and Patient Sera by a Nitrocellulose Immunoprint Technique
Authors: Bengtsson A. , Rolfsen W. , Einarsson R.
Published: August 2009

A rapid and convenient protein-gel blot technique for qualitative detection of antigens/allergens in pollen allergen extracts and IgE/IgG antibodies in patient sera has been developed. The antigens were separated by isoelectric focusing in agarose gel and transferred to nitrocellulose by capillary migration. After incubation of the nitrocellulose strips with serum from allergic patients, the binding of the patient’s specific IgE or IgG antibodies was analyzed by using isotope-labelled or enzyme-labelled anti-IgE or anti-IgG. The time needed for detection with isotope-labelled antibody was approximately 20 h and with enzyme-labelled antibody 2 h. The immunoprint technique is easy to use, which renders it suitable for routine use in allergy research and quality control.
Effects of Pneumococcal Vaccination on Tonsillo-Pharyngitis and Upper Respiratory Tract Flora
Authors: Christensen Poul , Hovelius Birgitta , Prellner Karin , Rosén Christer , Christensen Karen Kvist , Kurl Daya N. , Larsson Lennart , Stjernquist-Desatnik Anna , Schalén Claës
Published: August 2009

The effect of a 14-valent pneumococcal vaccine on upper respiratory tract infections and carriage of beta-haemolytic streptococci was studied in a double-blind prospective study of 405 children 0.5–5 years of age. In the children under 2 years of age at vaccination, the cases of acute tonsillitis were more frequent among the vaccinees than among the controls (p < 0.01). In contrast, among the children over 2 years of age at vaccination, fewer episodes of acute tonsillitis were reported among the vaccinees than among the controls (p < 0.01). In the older age group, the total rate of upper respiratory tract infections was also significantly reduced. Similarly, an increase in asymptomatic carriage of group A streptococci was registered in children vaccinated at 2–5 years of age (p < 0.01). Inversely, the carriership of group C streptococci diminished among the vaccinated children (p < 0.05). Some possible mechanisms underlying these unexpected findings, including non-specific mitogenic features and cross-protection due to antigenic similarities, were explored.
Detection of a Thermostable Brain Antigen in the Circulation of Patients after Cerebrovascular Accident
Authors: Chen H. , Soria E. , Milgrom F.
Published: August 2009

Enzyme immunoassay was developed with a thermostable antigen of human brain and its corresponding antiserum of rabbit origin. Inhibition of this assay proved to be a sensitive technique for detection of this antigen in the circulation. Using the inhibition test, we tested 170 serum samples originating from 42 patients with cerebrovascular accident (CVA), 166 sera from 166 patients with other neurological disorders, and 56 sera from 56 normal subjects. Fourteen patients with CVA were found to be ‘positive’ and 28 ‘negative’. Positive results in inhibition tests were first noted within 1–2 days after CVA and they appeared to decline to negative values within 3–5 weeks after CVA. None of the sera from subjects without CVA were positive.
In vitro Natural Killer and Killer Cell Functions in Uremia
Authors: Langhoff E. , Ladefoged L.
Published: August 2009

In uremia, mitogen responses of lymphocytes are decreased as is their resistance to the immunosuppressive effect of steroids. We examined the functions of natural killer (NK) and killer (K) cells from uremic patients and their sensitivity to in vitro immunosuppression by methylprednisolone. NK- and K-cell activity was as high as that of control subjects, and remained so at different titrations of effector cells. The functions of NK and K cells from uremic patients tended to be more resistant to methylprednisolone than those of control NK and K cells. Thus, while uremia impairs T-cell functions and T-cell sensitivity to steroids in vitro, the cytotoxic effector cell functions which are not dependent on cell proliferation remain unaffected.
Binding and Activation of Human Precursor C1 by Soluble Aggregates of Human and Rabbit IgG
Authors: Doekes G. , van Seggelen-van Zijp A.C.M. , van Es L.A. , Cats A. , Daha M.R.
Published: August 2009

The capacities of soluble human and rabbit IgG aggregates to bind and to activate human C1 were compared. Aggregates prepared by incubation of purified IgG at 63 °C were fractionated by gel filtration and hemolytic assays were used to measure the binding and activation of isolated human precursor C1. The C1 binding and activation capacities of both human and rabbit IgG aggregates were highly dependent on their size. Human IgG aggregates had a slightly higher binding avidity for human C1 than rabbit IgG aggregates of comparable size, but no clear differences were found between their capacities to activate C1. Experiments with nonaggregated IgG also indicated that although human IgG binds human C1 somewhat more avidly, human and rabbit IgG do not differ in their capacities to initiate fluid-phase activation of the human classical complement pathway.
Glycoprotein Allergens in Pollen of Timothy
Authors: Haavik Svein , Paulsen Berit Smestad , Wold Jens K.
Published: August 2009

A basic glycoprotein allergen has been isolated from pollen of timothy by a combination of chromatography on columns of CM-Sepharose CL-6B, Bio-Gel P-30, DEAE-Sepharose CL-6B and Ultrogel AcA 44. The allergen which is immunologically identical with the allergen formerly denoted as antigen 30 had an isoelectric point of 9.45. The MW of the glycoprotein was found to be 54,000 and 38,000 by SDS-PAGE and gel filtration, respectively. The allergen contains arabinose (3.1%), fucose (0.7%), xylose (0.7%), mannose (2.1%), galactose (2.8%) and glucose (0.2%). In rocket radioimmunoelectrophoresis the allergen bound IgE from 57 (78%) of 73 patient sera tested. The allergen did not bind Yariv artificial galactosyl- and glucosyl-carbohydrate-binding antigens or a monoclonal antibody against ryegrass pollen glycoprotein allergens.
Further Characterization Including Preliminary Chemical Analysis of Antigen MLW1 from Mycobacterium leprae
Authors: Reitan Liv Jorun , Closs Otto , Jantzen Erik
Published: August 2009

MLW1, an antigen preparation from Mycobacterium leprae previously shown to have a high content of M. leprae antigen No. 7 (ML7), was found to contain the typical cell wall constituents arabinose, galactose and mannose. The fatty acid composition of MLW1 was largely comparable to that of undisrupted cells. The capacity of MLW1 to stimulate lymphocytes was further studied. Good correlation was obtained between the in vitro lymphocyte responses to MLW1 and human-derived M. leprae, indicating similar specificity of the two antigen preparations in this test. The stimulatory activity of MLW1 was not significantly influenced by batch to batch variations, was well-preserved during storage and most of it was heat-stable. Attempts to remove the ML7 antigen indicate that this component plays a dominant role in inducing in vitro lymphocyte stimulation.
Grass Pollen Allergens: Antigenic Relationships Detected Using Monoclonal Antibodies and Dot Blotting Immunoassay
Authors: Singh M.B. , Knox R.B.
Published: August 2009

Monoclonal antibodies against major rye-grass pollen allergens have been used to detect cross-reactive determinants in other grass pollen extracts. Antibody binding was detected by the dot blotting immunoassay in which alkaline phosphatase-conjugated anti-mouse IgG was used as secondary antibody. Taxonomically ordered variations were found between pollen of 22 grass species representing all major natural groups. One of the antibodies, that bound to the 28 to 30Kd allergen, showed high species specificity; immunoblotting showed binding to similar polypeptides in Festuca elatior but to none of the other grasses tested. This simple assay has applications in standardizing grass pollen extracts.
Regulation of Natural Killer Cell Activity and Interferon Production in the Rat Lung Following Aerosol Challenge
Authors: Farrell H.E. , Holt P.G. , Shellam G.R.
Published: August 2009

High levels of natural cytotoxicity were detected in vitro in cells from the lung interstitium of the rat following collagenase digestion of lung tissue. In contrast, alveolar cells exhibited negligible levels of natural cytotoxicity and furthermore suppressed the natural cytotoxicity of interstitial lung leukocytes. This suppression was alleviated following in vivo administration of indomethacin. The natural cytotoxicity of cells from both the lung and spleen was transiently suppressed following exposure of normal rats to aerosols of Escherichia coli lipopolysaccharide or of ovalbumin-sensitized rats to an ovalbumin aerosol. Parenchymal lung leukocytes, like those from the spleen and peripheral blood, showed enhanced cytotoxicity when treated with interferon in vitro. In addition interstitial leukocytes were capable of producing interferon when cocultured with P815 cells and, as was the case with the spleen, low density cells produced the most interferon. However, alveolar cells did not produce interferon in this system. These studies suggest that the lung is capable of self-regulation of the high levels of natural cytotoxicity present in interstitial tissue; alveolar cells or their products may suppress interstitial natural killer cells whilst interstitial leukocytes have the capacity to stimulate natural killer cells by producing interferon.
Enhancement of Hydrophobic Interaction, Negative Charge and Phagocytosis by Dinitrophenyl Ligand Coupling to Salmonella typhimurium 395 MS
Authors: Edebo Lars , Richardson Neil
Published: August 2009

Dinitrophenylation of Salmonella typhimurium bacteria increased their hydrophobicity, negative charge, and interaction with PMNL in vitro, effects which are similar to the consequences of S-R mutation in S. typhimurium. The effects are ascribed to the hydrophobicity of the dinitrophenyl (DNP) ligand. Coupling of DNP-groups to the bacteria also enhanced their clearance by the reticuloendothelial system after intravenous injection into mice. Extensive DNP-coupling to the bacteria led to rapid deposition of injected bacteria outside the reticuloendothelial system. The kinetics of DNP-coupling showed saturation at 4–5 × 107 DNP groups per bacterium, similar for S and R bacteria, boiled and nonboiled. After 2 h of reaction with fluoro-2,4-dinitrobenzene (FDNB) more than 80% of the total DNP coupling observed after 4h had occurred. The capacity of MOPC-315 IgA myeloma protein, which possesses anti-DNP activity, to bind to dinitrophenylated bacteria paralleled the number of DNP groups per bacterium, but smaller numbers of IgA molecules were bound. Approximately one IgA molecule bound per thousand DNP ligands. Only when bacteria were boiled prior to DNP coupling was agglutination observed after the addition of anti-DNP-IgA.
Fragments of IgE Antibodies in Human Feces
Authors: Kolmannskog Svein , Marhaug Gudmund , Haneberg Bjørn
Published: August 2009

The gel filtration profile of immunoglobulin E (IgE) in extracts of feces from 2 children was compared with IgE myeloma protein which had been exposed to proteolytic digestion by chymotrypsin. The peak of the chymotrypsin-digested IgE myeloma protein was found to be similar to that of fecal IgE after an elution volume between those of albumin and myoglobin, corresponding to a molecular weight of approximately 40,000 daltons. In the fractions where the peak of fecal IgE was found, no evidence for the presence of specific IgE antibodies (measured by RAST) could be detected. Fecal IgE could be purified by using an immunosorbent column to which rabbit antihuman IgE was coupled. Sufficient amounts of fecal IgE could thus be obtained and used in autoradiographic experiments. The IgE-containing fractions could also be detected with 125I-labelled second antibodies to visualize the IgE precipitates.
Demonstration of Antibodies to Mosquito Antigens in Man by Immunodiffusion and ELISA
Authors: Ailus Kirsi , Palosuo Timo , Brummer-Korvenkontio Markus , Rantanen Tapio , Reunala Timo
Published: August 2009

Cutaneous reactions to mosquito bites are believed to be immunologically mediated, but knowledge of the underlying mechanisms is very limited. In the present work, comprising 598 subjects, we show that precipitating and ELISA-reactive IgG-class antibodies occur frequently in the adult population. The operating antigen(s) were found in whole-body extracts of Aedes mosquitoes and also in a continuous mosquito cell line culture. The appearance of antimosquito antibodies seems to be correlated with the exposure to mosquito bites; however, massive exposure at an early age may result in a suppressed response.
Immunochemical Characterization of Antigens of Parietaria judaica Pollen
Authors: Geraci Domenico , Billesbølle Kirsten Bache , Cocchiara Roberta , Løwenstein Henning , Ipsen Henrik
Published: August 2009

A crude extract of Parietaria judaica pollen was obtained by means of extraction, centrifugation and dialysis, and studied by means of quantitative immunoelectrophoresis. Crossed immunoelectrophoresis, using a high-titer purified rabbit antibody fraction, showed that the pollen extract contained at least 26 antigens of which 18 moved towards the anode, 6 moved towards the cathode and 1 moved both towards the anode and the cathode. The allergens in the extract were identified by means of crossed radio immunoelectrophoresis. Nine of the 26 antigens were able to bind specific human IgE to their corresponding immunoprecipitates, and 4 of these antigens can be classified as major allergens. The apparent molecular weights of 17 antigens were determined by a combination of size exclusion chromatography and immunochemical analysis. Ten antigens had Kav values corresponding to molecular weights in the range of 10–40 kilodaltons, 1 antigen possessed an apparent molecular weight of less than 10 kilodaltons, and six antigens had molecular weights above 40 kilodaltons. Most of the antigens, as analysed by column isoelectric focusing, had pi values between 4 and 7. Crossed line immunoelectrophoresis using an extract of Parietaria officinalis shows that the antigens of this extract exhibit a high degree of identity with those of P. judaica.
Augmentation of Natural Killer Cytotoxicity by Alpha or Gamma Natural and Recombinant Interferons and Interferon Inducers
Authors: Platsoucas Chris D. , Fernandes Gabriel , Good Robert A. , Gupta Sudhir
Published: August 2009

We investigated the effect of human peripheral blood monocytes on the augmentation of natural killer cytotoxicity by α or γ natural and recombinant interf erons (IFN) and certain interferon inducers. We observed that: (1) in the majority of the donors examined (75%) human peripheral blood monocytes do not affect natural killer cytotoxicity, determined by a 4-hour chromium-51 release assay, against target cells from hemopoietic human tumor cell lines. (2) Monocytes are not required and do not affect the augmentation of natural killer cytotoxicity by Escherichia coli-derived IFN-γ, natural human IFN-γ, E. Coli-derived IFN-α2 or natural human IFN-α. E. Coli-derived IFN-/and natural human IFN-γhave been reported to activate monocyte cytotoxicity determined in 72-hour assay. (3) Monocytes are not required for the augmentation of natural killer cytotoxicity against target cells from hemopoietic tumor cell lines by polyinosinic acid-polycytidylic acid or staphylococcal enterotoxin A.
Kinetics of Cells in the Intestinal Mucosa of Mice following Oral Infection with Ancylostoma ceylanicum
Authors: Carroll S.M. , Grove D.I. , Heenan P.J.
Published: August 2009

A variety of cell types in the intestinal mucosa of mice has been observed following oral infection with Ancylostoma ceylanicum and the changes in their numbers quantified. The number of lamina propria mast cells increased following infection and reached a peak at 4 days; by 21 days the numbers had returned to normal levels. Globule leucocytes displayed a similar marked response but the numbers returned to within normal limits 7 days after infection. Granular intraepithelial lymphocytes did not alter significantly during the period of observation whereas non-granular intraepithelial lymphocytes declined in numbers following infection and were significantly decreased by 3 weeks. Goblets cells showed a similar response to infection as lamina propria mast cells and reached a peak level at 4 days. Following initial fluctuations in eosinophil numbers there was a substantial increase in the numbers of these cells with peak levels being reached at 21 days; eosinophil numbers then remained elevated for the duration of observation. Partial villous atrophy was seen 4 days after the arrival of larvae in the small intestine with maximum reduction in the length of villi at 7 days. Thereafter, there was a return to normal length followed by a significant elongation in villus length 28 days after infection. The influences of these various cell types on the expulsion of intestinal parasites are discussed.
Phthalic Anhydride-Induced Occupational Asthma
Authors: Wernfors Margareta , Nielsen Jörn , Schütz Andrejs , Skerfving Staffan
Published: August 2009

Out of 118 workers exposed to phthalic anhydride (PA) dust for 2 months or more in four plants producing alkyd and/or polyunsaturated polyester resins, 28 (24%) suffered from work-related rhinitis, 13 (11 %) from chronic productive bronchitis, and 21 (28%) from work-associated asthma. Asthma was generally preceded by rhinitis and was mostly of late type. 3 out of 11 asthmatics had a PA-positive skin test. In 2 subjects the presence of antibodies was demonstrated by the Prausnitz-Küstner test. 4 out of 25 heavily exposed subjects without asthma had a nonspecific bronchial hyperreactivity. The results did not indicate any significant ‘healthy worker selection’ as regarding rhinits and asthma. Chronic productive bronchitis was common and was more prevalent among former workers than among present employees, indicating a selection of nonreacting subjects in the plant. The time-weighted average breathing zone PA levels in two plants were between 3 and 13 mg/m3 during different direct PA handling operations and less than 0.3 mg/m3 at other kinds of work. In 2 subjects, positive bronchial provocation was obtained by short-term exposure to 0.5 and 6 mg PA/m3, respectively.
Autoantibodies of the IgM Class against a Human Myeloma Protein IgE(DES). I. Occurrence
Authors: Magnusson C.G.M. , Vaerman J.P.
Published: August 2009

We report on the natural occurrence of human serum antibodies with specificity for a human monoclonal myeloma IgE(DES). These antibodies were of the IgM class, based on their susceptibility to reduction, sedimentation in sucrose gradients, gel filtration and inhibition of agglutination by anti-IgM antiserum. Auto-antibody levels were studied in several groups of patients by particle-counting immunoassay using latex particles to which purified monoclonal IgE(DES) was coupled. Only sera of patients suffering from parasitosis had significantly higher levels (p < 0.0005) than those of healthy blood donors. Cord sera had very low levels, followed by an age-dependent increase during early infancy. There was no relation (p > 0.10) between serum IgE and IgM antibody level. On the other hand, significant relations between IgM anti-IgE(DES) levels and serum IgM (p < 0.0005), serum IgA (p < 0.001) and serum IgG (p < 0.05) were observed suggesting that high levels were caused by or related to polyclonal activation of the immune system.
Tolerogenic Properties of Human IgG Subclasses in Mice
Authors: Huchet R.
Published: August 2009

The tolerogenic properties of the purified myelomatous human IgG subclasses IgG1, IgG2, IgG3 and IgG4 were investigated in mice, when given at a dose of 1 or 5 mg either, chemically nonmodified or trinitrophenylated. A complete tolerance at the helper T cell level was obtained with IgG1, IgG2 and IgG3, and a partial tolerance with IgG4 at the dose of 5 mg. At a dose of 1 mg, tolerance was observed only with IgG3. The ability of these human IgG subclasses to induce tolerance at the helper T cell level was modified after trinitrophenylation. Each subclass induced a complete tolerance when given at a dose of 1 mg. At higher doses (5 mg), a complete tolerance was obtained at the B cell level, using trinitophenyl (TNP) IgG1, TNP IgG2, TNP IgG3 or TNP IgG4 as carriers.
Paradoxical Augmentation of Tuberculin-Like Hypersensitivity, but Not Jones-Mote or Contact Hypersensitivity, in Cyclosporin A Treated Guinea Pigs
Authors: Aldridge R.D. , Thomson A.W.
Published: August 2009

Administration of cyclosporin A (CsA; 25 mg/kg) orally to guinea pigs from the time of immunization with ovalbumin (OVA) in complete Freund’s adjuvant, followed by drug withdrawal 4 days later, resulted in marked potentiation of classical, tuberculin-like delayed-hypersensitivity skin responses to OVA. However, no such augmentation of delayed-type hypersensitivity (DTH) to purified protein derivative (PPD) was demonstrated. The enhancing effect of CsA was also dependent on the dose of OVA used for both immunization and skin testing and on the interval between drug withdrawal and the elicitation of DTH. A single intraperitoneal injection of CsA (200 mg/kg) given 2 days before immunization also had an augmentary effect on 14-day responses to OVA. Similar treatment protocols, however, did not enhance Jones-Mote (cutaneous basophil) hypersensitivity to OVA or contact sensitivity reactions to dinitrofluorobenzene. Longer courses of CsA (25 mg/kg per os) between sensitization and skin testing severely depressed all three categories of type IV hypersensitivity reactions. Our observations may have important cautionary implications for the prospective management of immunologically mediated diseases of intermittent activity, including certain autoimmune disorders, where short courses of CsA might be contemplated.
Augmentation of Human Polymorphonuclear Leukocyte Adherence by Interferon
Authors: Seow Kim , Thong Y.H.
Published: August 2009

Augmentation of human polymorphonuclear leucocyte adherence by alpha and gamma interferons occurred as early as 2 min after incubation. Enhancement of adherence occurred at optimal concentrations of 100–1,000 IU/ml. There was synergism between alpha and gamma interferons, but not between two subtypes of alpha interferon, on augmentation of adherence, indicating that alpha and gamma interferons act on different receptors on the polymorphonuclear leucocyte. Heat treatment at 65°C for 30 min abolished the effect of interferon on adherence.
Dispersion and Characterisation of Mast Cells from Human Skin
Authors: Benyon Christopher , Church Martin K. , Clegg Stephen , Holgate Stephen T.
Published: August 2009

A technique is described for the enzymatic dispersion of mast cells in high yield from human infant foreskin. Dispersed mast cells exhibit high viability as assessed by light microscopy, low spontaneous histamine release, and survival in culture. Dispersed mast cells release histamine in response to immunological stimulation and synthetic secretagogues including ionophore A23187, compound 48/80 and poly-L-lysine. Reactivity to these stimuli indicates that cutaneous mast cells differ in their properties from human pulmonary mast cells.
Activation of Effector Cells of Experimental Allergic Encephalomyelitis in Lewis Rats: Comparison of T-Cell Lines with Primary Cultured Lymph Node Cells
Authors: Namikawa Tadashi , Yamamura Takashi , Sakai Koichiro , Kunishita Tatsuhide , Tabira Takeshi
Published: August 2009

We studied the mechanism of activation of effector cells in experimental allergic encephalomyelitis (EAE) by using T-cell lines (EAE/TL) reactive against guinea pig myelin basic protein generated from in vivo primed lymph node cells (LNC) of Lewis rats. EAE-effector cells are activated in the presence of a specific antigen and antigen-presenting cells (APC) with a compatible la antigen. The antigen presentation occurs during the first 18 h. EAE-effector cells cannot be activated by allogeneic stimulator cells, but a nonspecific T-cell mitogen, concanavalin A, can activate the effector cells in the presence of syngeneic as well as allogeneic APC.
Induction of c-myc Expression Early in the Course of B-Cell Activation: Studies in Normal Humans and Patients with Systemic Lupus erythematosus
Authors: Suzuki Hiroshi , Nakanishi Kenji , Steinberg Alfred , Green Ira
Published: August 2009

The proliferative response of B lymphocytes to stimulation with anti-IgM antibodies and B-cell growth factors was studied in 27 patients with systemic lupus erythematosus (SLE) and 17 normal donors. In addition, the expression of messenger RNA of the proto-oncogene c-myc was also studied in B cells from SLE patients and normal donors. The proliferative response of lupus B cells to anti-IgM and B-cell growth factors as compared to normal B cells demonstrated a wide range of responses, 10 were lower than normal and 8 were either normal or supernormal. As compared to normals, expression of B-cell c-myc RNA from SLE patients was either normal or depressed. In general in patients with SLE there was a positive correlation between levels of c-myc expression and the degree of proliferation in B-cells after stimulation with anti-IgM and B-cell growth factors.
Induction of Autoimmune Diseases with Adjuvants: Separation of Delayed Hypersensitivity and Antibody Formation from Diseases in Experimental Thyroiditis and Aspermatogenesis with Legionella Adjuvant
Authors: McMaster P.R.B. , Wong K.H. , Gardner J.J. , Feeley John C. , Rudofsky U.R.
Published: August 2009

The induction of autoimmune diseases in animals was studied with Legionella and mycobacteria as adjuvants, emulsified in oil with antigen extracts of thyroid, testis, spinal cord, and peripheral nerve. Both adjuvants were equally effective in inducing delayed hypersensitivity and humoral antibody to the tissue antigens. The Legionella adjuvant, however, induced little or no thyroiditis and aspermatogenesis, whereas the mycobacterial adjuvant induced thyroiditis and aspermatogenesis. Both adjuvants caused allergic encephalomyelitis and peripheral neuritis. The results indicated that delayed hypersensitivity by itself may not be sufficient to cause thyroiditis and aspermatogenesis. Legionella adjuvant apparently lacked the ability to induce certain immune factor(s) which caused the disease in experimental thyroiditis and aspermatogenesis. The differential properties of Legionella adjuvant and mycobacterial adjuvant in inducing immunity to autoantigens could provide a useful means to study the pathogenic and immunoregulatory mechanisms of some experimental autoimmune diseases.
Comparative Studies on Tree Pollen Allergens
Authors: Florvaag E. , Elsayed S. , Hammer A.S.E.
Published: August 2009

Horizontal isoelectric focusing (IEF) and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2-D SDS-PAGE) combined with electrophoretic transfer (ET) and immunoautoradiography (I AR) were used to characterize the crude alder (Alnus incana) pollen extract (AI crude) and fraction AI 34. IEF resolved AI crude and AI 34 into 34 and 39 distinct protein bands, respectively, when stained with Coomassie brilliant blue. The band patterns were similar and the majority of bands were located in the pH region 4.0–7.0. After ET and IAR, the distribution of antigenic reactivities was similar to that of the protein stain. Allergenic reactivity was detected in the pH region 4.6–5.2 only, where two bands at pH 4.9 demonstrated dominating IgE-binding properties. The 2-D reference maps of AI crude and AI 34 consisted of 40 and 45 protein spots, respectively. The autoradiogram of AI crude demonstrated 35 spots capable of binding rabbit IgG. The main IgE-binding zone was located at pH 4.6–5.2 and 20 kilodaltons with an area of weaker autoradiographic signals at pH 4.6–5.2 and 40 kilodaltons. ET from polyacrylamide gels to nitrocellulose (NC) membranes was quantitative and IAR gave significant radiostaining of spots containing 0.01 and 0.1 μg of AI crude in the IgG and IgE systems, respectively. For the characterization of the alder pollen extract, horizontal 2-D SDS-PAGE combined with ET and IAR provided a powerful supplement to conventional crossed immuno- and radioimmunoelectrophoretic techniques (CIE/CRIE).
Serum Factors Which Suppress Natural Cytotoxicity in Cancer Patients
Authors: Holmes E. , Sibbitt, Jr. W.L. , Bankhurst A.D.
Published: August 2009

Spontaneous cytotoxicity mediated by natural killer (NK) cells is potentially an important mechanism of immunosurveillance against tumor or virus-infected cells. NK activity is impaired in many cancer patients. This study investigated the possibility that humoral factors are responsible for depressed NK activity in cancer patients and examined whether these factors were anti-lymphocyte antibodies (ALA) or immune complexes. The degree of NK suppression induced by 20 cancer patients’ sera was determined by preincubating normal peripheral blood mononuclear cells with cancer sera. Eight of the 20 serum samples from cancer patients had NK-suppressive humoral factors. Elimination of immune aggregates by ultracentrifugation did not remove the inhibitory factors. The degree of NK suppression induced by cancer sera correlated with the extent of NK impairment in the serum donors (p < 0.05). The cancer sera was examined for the presence of ALAs using flow cytometry. All cancer sera tested contained only low ALA reactivity to NK cell-enriched suspensions (less than 15%). This is in marked contrast to previous reports regarding NK-inhibitory systemic lupus erythematosus sera which contained large amounts of large granular lymphocyte (LGL) reactive ALAs. This study demonstrates that certain cancer sera suppress NK cell function. These inhibitory serum factors do not appear to be LGL-reactive ALAs or immune aggregates.
T Cell Responses to a Dermatophagoides farinae Allergen Preparation in Allergies and Healthy Controls
Authors: Halvorsen Ragnhild , Bosnes Vidar , Thorsby Erik
Published: August 2009

We have studied possible differences between mite-allergic individuals and healthy controls in their T cell proliferation and T cell surface markers after in vitro stimulation with a Dermatophagoides farinae allergen preparation. No significant differences were found between the two groups in 3H-thymidine incorporation in T cells after 7 days in culture with allergen- and antigen-presenting cells. The frequencies of allergen-reactive T cells determined by limiting dilution were also similar in the two groups. Finally, no differences were found in surface markers on allergen-activated T cells from mite-allergic and healthy blood donors using a variety of monoclonal antibodies reactive with T cell subsets and activation antigens. Our conclusion is that with the D. farinae preparation employed in this investigation and using T cells prepared from peripheral blood no significant differences were seen between mite allergies and controls.
Immune Response Potential of Mast Cell-Deficient W/Wv Mice
Authors: Ha Tai-You , Reed Norman D. , Crowle Patricia K.
Published: August 2009

Immune responses of mast cell-deficient WBB6F1-W/Wv mice and their mast cell-sufficient littermates (LM: WBB6F1-W/+, WV/+ and +/+) were compared. After a single intravenous injection of sheep erythrocytes (SE), polyvinylpyrrolidone or bacterial lipopolysaccharide, the antigen-specific IgM plaque-forming cell (PFC) response of W/Wv mice was similar to or greater than the response of LM mice. When both primary and secondary injections of SE or chicken γ-globulin were given to mice and antigen-specific IgG PFC responses quantified, the response of W/Wv again was similar to or greater than that of LM mice. Serum titers of antigen-specific IgE were higher in W/Wv than in LM mice after injections of ovalbumin in alum or infections of Nippostrongylus brasiliensis. Ovalbumin-sensitized W/Wv and LM mice developed active systemic anaphylaxis after ovalbumin challenge. The ability of W/Wv mice to be sensitized for and elicit contact sensitivity (CS) reactions was studied using picryl chloride or dinitrofluorobenzene as sensitizing and challenge agents and quantifying 24-hour reactions by change in ear thickness. SE or methylated bovine serum albumin was used to sensitize and challenge mice for delayed-type hypersensitivity (DTH) reactions which were quantified at 24 h by change in foot pad or ear thickness. CS and DTH reactions of W/Wv and LM mice were similar. No evidence of immune deficiency of W/Wv mice was found.
Isolation of a Kentucky Blue Grass Pollen Allergen Using a Murine Monoclonal Antibody Immunosorbent
Authors: Ekramoddoullah A.K.M. , Kisil F.T. , Sehon A.H.
Published: August 2009

We have previously reported the production of murine monoclonal antibodies to the retentate fraction of Kentucky Blue Grass pollen (KBG-R). In the present study, one of these monoclonal antibodies (Mab 27) was used in a combination of methods employing Western and immunoblot analysis to establish its reactivity to various antigenic components present in KBG-R. Thus, Mab 27 reacted predominantly with an antigen having a MW of 30 kD and to a lesser extent with other antigenic components with MW of 35 and 17 kD. Fractions of KBG-R obtained by preparative isoelectrofocusing, revealed on analysis by ELISA, that Mab 27 reacted with several components differing in charge. From these observations it was concluded that KBG-R contained a group of related antigens detectable with Mab 27 and differing in size and charge. A reverse immunosorbent, prepared by coupling Mab 27 to CNBr-activated Sepharose 4B, was used to absorb KBG-R. Bound material was recovered by acid elution and designated as Ag 27. SDS-PAGE analysis revealed that Ag 27 consisted of at least two components with molecular weights of 30 and 17 kD. Analysis by RAST using sera from humans allergic to KBG indicated that Ag 27 was highly allergenic.
Sensitization Induced in Guinea Pigs with Beta-Propriolactone-Treated Serum Albumin: Experimental Evidence for the Cause of Allergic Reactions in Humans Receiving Human Diploid Cell Rabies Vaccines
Authors: Levenbook Inessa S. , Merritt Babette A. , Fitzgerald Edward A. , Elisberg Bennett L.
Published: August 2009

β-Propiolactone-treated (BPL-T) homologous serum albumin caused anaphylaxis in guinea pigs with a frequency and severity equal to that of guinea pigs inoculated with human albumin. Untreated guinea pig serum albumin did not cause any reactions in these animals. Some recipients of current rabies vaccine produced in human diploid cells available in the USA develop systemic allergic reactions, usually following booster immunization. The BPL-T human albumin component of the vaccines was believed to be the cause of the complications. Our studies support this conclusion.
Monocyte-Mediated Suppression of T Lymphocyte Blastogenesis and Its Reversal by Deoxyguanosine
Authors: Yamane Kazuhide , Kono Ichiro , Kabashima Teizo , Sakurai Tetsushi , Kashiwagi Heihachiro
Published: August 2009

We investigated monocyte-mediated suppression of T lymphocyte blastogenesis in normals and patients with systemic lupus erythematosus (SLE). When monocytes from normals were cocultured with auto-logous T lymphocytes with a ratio of 1:1 and stimulated with phytohemagglutinin (PHA), 3H-thymidine incorporation by T lymphocytes was suppressed. This monocyte-mediated suppression was reversed by purine nucleoside phosphorylase substrate, deoxyguanosine. In SLE patients, both monocyte-mediated suppression and its reversal by deoxyguanosine were defective. The defective function was observed both in patients with active and inactive diseases. The defective function was studied sequentially before and after change in the clinical status of patients. The defects remained unaffected regardless of the disease activity. The defects in monocyte-mediated suppression and its reversal by deoxyguanosine in SLE patients as demonstrated in our study suggest the presence of intrinsic monocyte dysfunction in SLE.
Labile Disulfide Bonds and Free Thiol Groups in Human IgG I.
Authors: Schauenstein E.S. , Dachs F. , Reiter M. , Gombotz H. , List W.
Published: August 2009

The content of free SH groups (about 0.24) and labile S-S bonds (about 0.64) per mole human IgG can be differentially determined by the reaction with 5,5′-dithio(2,2′-dinitro)benzoate (DTNB) for 30 min and 24 h, respectively. Highly significant linear correlations were found between the number of labile S-S and the percentage of IgG1, and the number of free SH and the percentage of IgG2 of the total IgG fraction. It is concluded that the IgG1 molecule contains one S-S bond which is opened during the 24 h interaction with DTNB by a disulfide exchange reaction. This was also confirmed by the investigation of pure monomeric IgG1. The splitting of this bond does not alter molecular weight, antigenic properties or antigen binding activity, but reduces significantly the complement binding activity of IgG. On the other hand, one free SH group could be found in the IgG2 molecule. Changes have been observed in SH and S-S levels of total IgG in patients with various malignant diseases that are to be explained by corresponding changes of the percentages of IgG1 and IgG2.
Development of Rheumatoid Factors and Anti-F(ab′)2 Antibodies in Guinea Pigs Immunized with Type II Bovine Collagen
Authors: Wolf Benjamin , Bashey Reza I. , Newton Charles D. , Jimenez Sergio A.
Published: August 2009

The purpose of this report is to present results demonstrating for the first time the development of rheumatoid factors to rabbit IgG-Fc as well as antirabbit F(ab′)2 antibodies in guinea pigs after chronic sensitization with purified native type II bovine collagen. The sensitized animals also developed antibodies to guinea pig Ig. Antibodies to rabbit Ig arose as early as 3 weeks after bovine type II collagen injection and persisted for as long as 80 weeks when the experiment was terminated. The anti-Ig antibodies did not cross-react with the type II bovine collagen. Despite development and persistence of higher titers of RF and anti-F(ab′)2 antibodies in the immunized animals, the animals failed to show clinical evidence of inflammatory polyarthritis. These results indicate that rheumatoid factors as well as antibodies to F(ab′)2 arise independently of the clinical expression of disease.
Studies on Vascular Permeability Increasing Factors Involved in 48-Hour Homologous PCA in the Mouse Ear
Authors: Inagaki Naoki , Goto Shoichi , Yamasaki Mie , Nagai Hiroichi , Koda Akihide
Published: August 2009

Several attempts were made to elucidate the possible role of histamine, serotonin, leukotrienes C4 (LTC4) and D4 (LTD4), and prostaglandin E1 (PGE1) as vascular permeability increasing factors involved in 48-hour homologous passive cutaneous anaphylaxis (PCA) in the mouse ear. Increased vascular permeability in the mouse ear caused by the mediator injection or PCA was assessed quantitatively by measuring the amount of extravasated dye. In skin reactions, all of the mediators used in the present study significantly increased vascular permeability. The most potent mediator was serotonin, which increased the vascular permeability from a concentration of 10––8 g/ml, and the activity was about 100 times higher than that of histamine on a weight basis. Vascular permeability increasing activity of LTC4 was about 10 times higher than that of histamine, and LTD4 and PGE1 were also more potent than histamine. Increases of vascular permeability caused by histamine, serotonin, LTC4 and LTD4 were significantly potentiated by injecting 10––6 g/ml of PGE1 simultaneously. Histamine-, serotonin- and LTC4-induced skin reactions in the mouse ear were suppressed significantly by the administrations of chlorpheniramine, methysergide and FPL 55712, respectively. In contrast, though chlorpheniramine and methysergide suppressed also mouse ear PCA (about 50 and 40%, respectively), neither FPL 55712, indomethacin nor BW 755C suppressed it. These results strongly suggest that the most important mediator involved in mouse ear PCA is histamine and that serotonin also plays an important role in the increase of vascular permeability caused by PCA. Despite their potent vascular permeability increasing activity LTC4, LTD4 and PGE) do not seem to play an important role in mouse ear PCA.