The conjugation of immunoglobulins with tetramethylrhodamine isothiocyanate by utilization of dimethylsulfoxide (DMSO) as a solvent

Bergquist, N.R.; Nilsson, P. M.

doi:10.1016/0022-1759(74)90009-x

Cited by 30 publications

(9 citation statements)

References 8 publications

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“…The main difficulty in the analysis of the interaction of antibodies with lipid antigens stems from their insolubility in aqueous solutions. Bergquist and Nilsson (1974) havle shown that the presence of up to 30% DMSO in an aqueous solution containing antibody did not noticeably denature the antibody activity. This observation led us to include for each concentration of the standard range, as well as in the samples for assay, 1 I % DMSO as a solvent for the competitor GalC.…”

Section: Discussionmentioning

confidence: 99%

Immunological Determination of Galactosylceramide Level in Blood as a Serum Index of Active Demyelination

Thuillier¹,

Lubetzki²,

Goujet‐Zalc³

et al. 1988

Journal of Neurochemistry

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An enzyme-linked immunosorbent assay (ELISA) to determine the level of galactosylceramide (GalC) in biological fluids is described. The assay uses GalC-coated plastic microtiter plates, with binding of an antibody to GalC detected by a peroxidase-labeled second antibody. The GalC level was directly estimated in the biological samples, without prior extraction, by competition with the coated hapten. This method allows the detection of 62 pmol of GalC (1.2 nmol/ml). Results using this procedure revealed positive sera only among patients suffering a myelin-destructive process: either primary, as in multiple sclerosis, or secondary to brain damage, as during ischemic strokes.

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Section: Discussionmentioning

confidence: 99%

Immunological Determination of Galactosylceramide Level in Blood as a Serum Index of Active Demyelination

Thuillier¹,

Lubetzki²,

Goujet‐Zalc³

et al. 1988

Journal of Neurochemistry

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“…The anii-CD43 antibody B1B6 (IgGl) [3] and the anti-CD40 antibody S2C6 (IgGl) [18] were generated as fusion products of Sp 2,0 AgI4 myeloma cells and spleen cells taken from BALB/c mice previously immunized with PBL from a patient with chronic lymphocylic leukaemia oC T-cell type and cells from tninsiiional cell carcinoma of ihe human urinary bladder, respectively. Conjugation ol" MoAb S2C6 wilh fluorescein isothiocyanate (FITC) was performed as described previously [5]. Anii-CD23-FITC antibody wiis obtained from Immunotech (Marseille, France) and anti-CD7I antibody (OKT9) from American Type Culture Collection (Rockville.…”

Section: Methodsmentioning

confidence: 99%

Expression of CD40 and CD43 during Activation of Human B Lymphocytes

1991

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CD40 and CD43 are two cell-surface glycoproteins that appear to be functionally involved in the growth stimulation of human B cells. Whereas CD40 is structurally similar to the NGF receptor and is present on all resting B cells, CD43 displays no homology to other known proteins and is expressed only on a subpopulation of these cells. To further understand the extra- and intracellular signals regulating these molecules and in which stage of activation they may play a role, we used various activation strategies and studied their expression on tonsillar B cells. As expected, activation of protein kinase C by TPA increased both CD40 and CD43. In contrast, a rise in intracellular Ca2+, e.g. by ionomycin, did not influence the expression of these antigens. However, in the presence of TPA, ionomycin further up-regulated CD43 but not CD40. Anti-IgM behaved similarly to ionomycin suggesting that the effect of this reagent was due primarily to its ability to increase intracellular Ca2+. Of three interleukins (IL-2, IL-4 and IL-6) only IL-4 had a significant effect when used alone in that it up-regulated CD40 but not CD43. However, in the presence of anti-IgM, both IL-2 and IL-4 synergistically up-regulated the two antigens. Complementation of antigen receptor stimulation with TPA or IL-4 increased CD40 during the first 24 h, whereas up-regulation of CD43 did not occur until 24 to 48 h after stimulation. With regard both to up-regulation in response to different stimuli and to kinetics, CD40 expression paralleled that of the early activation antigen CD23, whereas CD43 was induced in parallel with the transferrin receptor (CD71). Taken together, our results suggests that the expression of CD40 and CD43 is regulated by different intracellular signals and that CD40 may be important during early activation, whereas CD43 may have its major function during later stages of B-cell differentiation. These assumptions are in line with the observations that CD40 antibodies can directly activate resting B cells and that CD43 are retained on plasma cells.

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“…OVA was insolubilized either by precipitating with methylated bovine serum albumin (BSA) as described by Kapp et al [15] or by coupling to acroleine-activated latex beads as described by Kumakura et al [16]. Fluorescein isothiocyanate (F1TC)-OVA was prepared following the method of Bergquist and Nilsson [17]. OVA was labeled to gold particles according to the method of Horisberger "1.…”

Section: Antigens and Immunizationmentioning

confidence: 99%

Antigen‐presenting cell function of dendritic cells and macrophages in proliferative T cell responses to soluble and particulate antigens

Kapsenberg¹,

Teunissen²,

Stiekema³

et al. 1986

Eur J Immunol

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The capacity of dendritic cells (DC) and macrophages (M phi) to present soluble and particulate antigen was tested in an ovalbumin (OVA)-specific T cell proliferation assay. In a comparative investigation we found that both DC and M phi were able to present soluble OVA, but that only M phi could present insolubilized OVA to T cells. DC were found to be able to present OVA in collaboration with M phi. The failure of DC to present insolubilized OVA is probably caused by their inability to endocytose these antigens. DC appeared not to endocytose substantial amounts of soluble OVA either. In contrast to M phi, antigen presentation by DC is not blocked by lysosomotropic drugs. Taken together, these observations suggest that DC can present soluble protein antigens without intracellular degradation.

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The conjugation of immunoglobulins with tetramethylrhodamine isothiocyanate by utilization of dimethylsulfoxide (DMSO) as a solvent

Cited by 30 publications

References 8 publications

Immunological Determination of Galactosylceramide Level in Blood as a Serum Index of Active Demyelination

Immunological Determination of Galactosylceramide Level in Blood as a Serum Index of Active Demyelination

Expression of CD40 and CD43 during Activation of Human B Lymphocytes

Antigen‐presenting cell function of dendritic cells and macrophages in proliferative T cell responses to soluble and particulate antigens

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