B. M. Stadler

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Monoclonal antibody against human interleukin 2 (IL 2). I. Purification of IL 2 for the production of monoclonal antibodies.

The Journal of Immunology (1982)

B. M. Stadler Elsa H. Berenstein R P Siraganian J J Oppenheim

Human interleukin 2 (IL 2) was produced under serum-free conditions by stimulating mononuclear cells with concanavalin A (Con A) in the presence of phorbol myristate acetate (PMA) and hydroxyurea. The IL 2 was partially purified by sequential chromatography by using phenyl-Sepharose, DEAE Sephacel, and AcA 54 gel filtration. This partially purified material was used to immunize BALB/c mice. After immunization with a total of 48,000 U (spec. act. approximately 10(5) units/mg protein), the spleen cells were adoptively transferred into x-irradiated syngeneic mice and the animals were boosted with another 12,000 U of IL 2. Four days later their spleen cells were hybridized with plasmacytoma cells. Supernatants of the hybridoma cultures were screened for their capacity to inhibit the IL 2-induced proliferation of the CT6 cell line. After expansion and cloning eight different lines were selected for ascitic antibody production. The monoclonal antibodies inhibited the proliferation of the IL 2-dependent cell line in response to either human crude or purified IL 2, as well as rat and mouse IL 2. However, these anti-IL 2 antibodies did not inhibit the proliferation of human T cell lines capable of producing IL 2. Monoclonal antibodies coupled to Sepharose 4B absorbed IL 2 crude culture supernatant, confirming that they react directly with IL 2. The absorbed IL 2 could, for the most part, be eluted by using sodium dodecyl sulfate, thus providing a means for further immunoaffinity purification of IL 2.

Sepharose

10.4049/jimmunol.128.4.1620

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Citations (49)

Transcription and translation dependent induction of interleukin 2 (IL-2) and IL-2 receptors

Immunology Letters (1984)

Y. Wang Christoph Walker B. M. Stadler A.L. de Weck

Transcription

Puromycin

10.1016/0165-2478(84)90083-x

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Citations (4)

[Relationship between T-subsets and clinical aspects of HIV-associated diseases].

PubMed (1991)

M. Fuchs B. M. Stadler R. Malinverni A.L. de Weck Werner J. Pichler

HIV predominantly infects the CD4+ T cells, which during the progression of the disease are eliminated, causing an immune deficiency which renders the patients more susceptible to infections. To evaluate the relevance of the CD4+ T cell elimination and thus the clinical usefulness of CD4/CD8 subset determinations in HIV infected persons, we investigated whether analyses of 667 subset determinations of 365 patients correlated with clinical stages of HIV-infection (CDC classification). Progress of HIV related disease was accompanied by a fall in CD4+ cells and an increase in CD8+ cells, leading to a drastically reduced CD4/CD8 ratio. This change of T-cell subset values correlated well with the clinical classification (CDC). It was, however, only statistically significant if percent values were used, but not if absolute CD4 cell counts, calculated from the peripheral lymphocyte count, were considered. While patients in CDC stage IVC2 (mainly Candida stomatitis) did not differ from stages IIB, IIIB, IVA, we found statistically lower CD4 values if the patients had stage IVA plus IVC2. Stage IVC1 (mainly Pneumocystis carinii pneumonia [PcP, n = 20]) had even lower CD4 values, as PcP appeared almost exclusively in patients with CD4 counts below 20% or 200/microliter. The lowest CD4 counts were observed in patients with Kaposi sarcoma (n = 11) with CD4 cells less than 10% and significant elevated values of CD8 cells (greater than 50%). While the total lymphocyte count correlated with the absolute counts of CD4 and CD8 cells, it was impossible to estimate the T-subset distribution from the absolute lymphocyte count. Our investigations show that a decreased number of circulating CD4 cells correlates well with an increased tendency to develop infections, and thus support the relevance of CD4 cell measurements for the optimal care of asymptomatic HIV infected persons in particular. They also show that the percent values correlate better with clinical stage than the absolute CD4 cell count.

Clinical Significance

Source

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Citations (2)

Front & Back Matter

International Archives of Allergy and Immunology (2012)

Yan Xia Thomas S. Harris Alastair G. Stewart Graham A. Mackay W.L. Wang

10.1159/000346401

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Citations (0)

Cross‐reactions vs co‐sensitization evaluated by in silico motifs and in vitro IgE microarray testing

Allergy (2011)

Pascal B. Pfiffner B. M. Stadler Chiara Rasi Enrico Scala Adriano Mari

To cite this article: Pfiffner P, Stadler BM, Rasi C, Scala E, Mari A. Cross‐reactions vs co‐sensitization evaluated by in silico motifs and in vitro IgE microarray testing. Allergy 2012; 67 : 210–216. Abstract Background and objective: Using an in silico allergen clustering method, we have recently shown that allergen extracts are highly cross‐reactive. Here we used serological data from a multi‐array IgE test based on recombinant or highly purified natural allergens to evaluate whether co‐reactions are true cross‐reactions or co‐sensitizations by allergens with the same motifs. Methods: The serum database consisted of 3142 samples, each tested against 103 highly purified natural or recombinant allergens. Cross‐reactivity was predicted by an iterative motif‐finding algorithm through sequence motifs identified in 2708 known allergens. Results: Allergen proteins containing the same motifs cross‐reacted as predicted. However, proteins with identical motifs revealed a hierarchy in the degree of cross‐reaction: The more frequent an allergen was positive in the allergic population, the less frequently it was cross‐reacting and vice versa. Co‐sensitization was analyzed by splitting the dataset into patient groups that were most likely sensitized through geographical occurrence of allergens. Interestingly, most co‐reactions are cross‐reactions but not co‐sensitizations. Conclusions: The observed hierarchy of cross‐reactivity may play an important role for the future management of allergic diseases.

Cross-reactivity

10.1111/j.1398-9995.2011.02743.x

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Citations (18)

Increased Level of Antibodies Cross-Reacting with Ves v 5 and CRISP-2 in MAR-Positive Patients

International Archives of Allergy and Immunology (2012)

S. Brunner-Agten Rodoljub Pavlovic Lorenz K. Müller Michael P. Horn Andreas Huber

Anti-sperm antibodies (ASA) have been described to be involved in immunological infertility. A possible antigen for ASA is the human cysteine-rich secretory protein 2 (CRISP-2), a sperm surface protein important in sperm-oocyte interaction. Furthermore, anti-CRISP-2 antibodies were shown to decrease fertility rates in vitro. Recently, we have reported cross-reacting antibodies recognizing CRISP-2 and antigen 5 from yellow jacket venom (Ves v 5) in human serum.Here, we investigated anti-Ves v 5 and CRISP-2 antibodies in sera from two groups of donors: MAR+ and MAR- patients.A higher incidence of allergy against hymenoptera venom was found in MAR+ patients. Interestingly, affinity-purified ASA from MAR+ patients' sera reacted against both Ves v 5 and CRISP-2, leading to sperm immobilization. Immunofluorescence analysis showed that ASA bound to the sperm surface, including the head part where CRISP-2 is localized.Taken together, these results showed a higher incidence of antibodies cross-reacting with Ves v 5 and CRISP-2 in MAR+ patients. This leads to the hypothesis that MAR+ patients may have a higher risk to develop wasp allergy.

Immunofluorescence

10.1159/000338946

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Citations (8)

Effects of drinking water monochloramine on lipid and thyroid metabolism in healthy men.

Environmental Health Perspectives (1993)

Robert G. Wones Colleen Deck B. M. Stadler Suzanne Roark Elizabeth Hogg

The purpose of this study was to determine whether a 4-week consumption of 1.5L per day of drinking water containing monochloramine at a concentration of 2 ppm (ppm = mg/L) or 15 ppm under controlled conditions would alter parameters of lipid or thyroid metabolism in healthy men. Forty-eight men completed an 8-week protocol during which diet (600 mg cholesterol per day, 40% calories as fat) and other factors known to affect lipid metabolism were controlled. During the first 4 weeks of the protocol, all subjects consumed distilled water. During the second 4 weeks, one-third of the subjects were assigned randomly to drink 1.5 L per day of water containing 2 ppm of monochloramine, to drink 1.5 L per day of water containing 15 ppm monochloramine, or to continue drinking distilled water. Four blood samples were collected from each subject at the end of each 4-week study period. Subjects drinking monochloramine at a concentration of 2 ppm showed no significant changes in total cholesterol, triglycerides, HDL cholesterol, LDL cholesterol, apolipoproteins A1, A2, or B when compared to the distilled water group. Parameters of thyroid function also were unchanged by exposure to monochloramine at this concentration. However, subjects drinking monochloramine at a concentration of 15 ppm experienced an increase in the level of apolipoprotein B. Other parameters of lipid and thyroid metabolism did not change. We conclude that consumption of drinking water containing 2 ppm of monochloramine does not alter parameters of lipid and thyroid metabolism in healthy men. Consumption of water containing 15 ppm monochloramine may be associated with increased levels of plasma apolipoprotein B.

Distilled water

10.1289/ehp.9399369

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Citations (5)

Contents Vol. 145, 2008

International Archives of Allergy and Immunology (2007)

Rudolf Valenta Barbara Bohle Kurt Blaser Davos Chapman Va Hayglass

10.1159/000121448

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Citations (0)

Contents, Vol. 103, 1994

International Archives of Allergy and Immunology (1994)

Georg Wick Beatrix Grubeck‐Loebenstein Boris Albini Roy Sir Cambridge Caine

10.1159/000236647

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Citations (0)

Natural anti‐IgE auto‐antibodies interfere with diagnostic IgE determination

Clinical & Experimental Allergy (1990)

C. C. Vassella Anja Weck B. M. Stadler

Summary A simple method is described which enables measurement of anti‐IgE antibodies in free form as well as in immune complexes of IgE and anti‐IgE. Anti‐IgE antibodies were purified from serum of one selected blood donor with highly elevated levels of such auto‐antibodies. These purified anti‐IgE auto‐antibodies inhibited the measurement of myeloma IgE. Purification also revealed that 98% of the subject's serum IgE was masked by anti‐IgE auto‐antibodies. Our data suggest that IgE determinations in sera containing anti‐IgE antibodies might be underestimated.

10.1111/j.1365-2222.1990.tb02687.x

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Citations (33)