Enhancement of sheep red blood cell human lymphocyte rosette formation by the sulfhydryl compound 2-amino ethylisothiouronium bromide

Pellegrino, M. A.; Ferrone, Soldano; Dierich, Manfred P.; Reisfeld, Ralph A.

doi:10.1016/0090-1229(75)90019-7

Cited by 386 publications

(80 citation statements)

References 16 publications

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“…In the remaining eight subjects, low-dose irradiation resulted in decreased specific antibody production. These experiments were initially designed to determine if the antigen-specific suppression of in vitro antibody production seen at high (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) ug/ml) in vitro concentrations of antigen was due to the generation or activation of suppressor T cells. In contrast to the previously reported findings of others working in human antigen-specific systems (10, 30), we have been unable to demonstrate a T cell contribution to this suppressor phenomenon.…”

Section: Discussionmentioning

confidence: 99%

Antigen-induced human T cell help. Precursor frequency, radiation sensitivity, and allogeneic effects.

Lane¹,

Whalen²,

Fauci³

1983

J. Clin. Invest.

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A B S T R A C T We have recently noted marked differences between the in vitro responses of human B lymphocytes to stimulation with soluble antigens vs. stimulation with mitogens. In the present study, these differences were analyzed in terms of the precursor frequencies for the T cells and B cells involved and in terms of the radiation sensitivity of the T cells providing help in the two systems. Marked differences were found between antigen-induced and mitogen-induced systems with regard to T cell precursor frequencies and radiation sensitivity. In contrast, the precursor frequencies for the B cells involved in the two systems were approximately the same. In addition, having developed a system for the study of human antigen-specific B cell responses, we were interested in delineating the nature of the allogeneic effects that might be operative in this system. Marked allogeneic effects, both positive and negative, were noted in this system and will need to be taken into account in any studies that try to address the question of the genetic restriction, if any, that exists in human antigen-specific T cell-B cell collaboration.Appreciation of the marked differences between the antigen-specific and mitogen-induced activation and immunoregulation of human B cell responses will be of importance in understanding the relationship between specificity and nonspecificity of antibody production in normal and disease states. INTRODUCTIONT cell regulation of B cell reactivity is an area of major interest and importance in both animal and human systems. With the recent availability of several systems Address all correspondence to Dr. H. Clifford Lane. Received for publication 20 January 1983 and in revised form 30 March 1983. to study human B cell function in vitro, considerable interest has centered around the precise nature of the T cell regulation of human B cell responses in both polyclonal and antigen-specific assay systems. Unfortunately, there have been conflicting data among sev-. eral of these systems (1-10). Of particular note has been the divergence of results with regard to the radiosensitivity of T cell help and the relative contributions of allogeneic effects in the mitogen-induced vs. antigen-induced human B cell systems. Disagreement with regard to this latter point has led to difficulty in delineating the precise nature of genetic restrictions in T cell-B cell collaboration in antigen-specific responses as well in appreciating the scope of allogeneic effects in co-culture experiments now commonly used in the study of patients with immunemediated or immunodeficiency diseases.Recently, in vitro systems have been developed for the study of antigen-induced, antigen-specific antibody production and secretion into culture supernatants by human B cells (11)(12)(13)(14). The use of enzymelinked immunosorbent assays (ELISA)l has proven to be extremely useful in these studies because of their simplicity, sensitivity, and specificity. These advances have allowed for a more precise study of the mechanisms involved in...

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Section: Discussionmentioning

confidence: 99%

Antigen-induced human T cell help. Precursor frequency, radiation sensitivity, and allogeneic effects.

Lane¹,

Whalen²,

Fauci³

1983

J. Clin. Invest.

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“…After exposure to trypsin, the adherent cells were removed from the plastic with a rubber policeman; >95% of these cells showed positive staining for nonspecific esterase. The nonadherent population was further subdivided into T cells and B cells by double rosetting with fresh sheep erythrocytes treated with 2-amino-ethylisothiouranium bromide (31). To obtain a sufficient mass ofendothelial cells for these studies, primary endothelial cell cultures from -10 donors were subcultured through four passages.…”

Section: Methodsmentioning

confidence: 99%

Presence of complement-fixing anti-endothelial cell antibodies in systemic lupus erythematosus.

Cines¹,

Lyss²,

Reeber³

et al. 1984

J. Clin. Invest.

244

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“…E rosette assay. For the demonstration of spontaneous E rosette formation, one volume of washed and packed E was pretreated with four volumes of the sulfidic compound, 2-aminoethylisothio-uronium bromide hydrobromide (0.28 M, pH 8.5), for 30 min at 37°C according to the method described by Pellegrino et al (18). For the rosette assay, 100 ,ul of 1% treated E and 100 ,ul of lymphocytes (4 x 106 cells/ml), both in minimum essential Eagle's medium containing 2.5% heat-inactivated fetal calf serum (MEM-FCS), were mixed and incubated 10 min at 37°C before centrifuging the cells at 200 g for 5 min at 40C and incubating them for 60 min at 4°C.…”

Section: Methodsmentioning

confidence: 99%

A subpopulation of normal human peripheral B lymphcytes that bind IgE.

González-Molina¹,

Spiegelberg²

1977

J. Clin. Invest.

143

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Enhancement of sheep red blood cell human lymphocyte rosette formation by the sulfhydryl compound 2-amino ethylisothiouronium bromide

Cited by 386 publications

References 16 publications

Antigen-induced human T cell help. Precursor frequency, radiation sensitivity, and allogeneic effects.

Antigen-induced human T cell help. Precursor frequency, radiation sensitivity, and allogeneic effects.

Presence of complement-fixing anti-endothelial cell antibodies in systemic lupus erythematosus.

A subpopulation of normal human peripheral B lymphcytes that bind IgE.

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