Isolation of MUC1-primed B lymphocytes from tumour-draining lymph nodes by immunomagnetic beads |
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Authors: | Claudia Petrarca Beniamino Casalino Silvia von Mensdorff-Pouilly Aurelia Rughetti Hassan Rahimi Giovanni Scambia Joseph Hilgers Luigi Frati Marianna Nuti |
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Institution: | (1) Department of Experimental Medicine and Pathology, University of Rome, Viale Regina Elena 324, I-00161 Rome, Italy e-mail: mnuti@axrma.uniroma1.it Tel.: +39-6-494-05-40; Fax: +39-6-445-48-20, IT;(2) Division of Gynecology and Obstetrics, Catholic University of Rome, Largo Gemelli 1, 00168 Rome, Italy, IT;(3) Department of Obsterics and Gynaecology, Academic Hospital Vrije Universiteit, Amsterdam, The Netherlands, NL |
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Abstract: | The humoral immune response against a tumour-associated antigen, polymorphic epithelial mucin (PEM, MUC1) in cancer patients
was studied by isolating specific B cells primed for the antigen. Human B lymphocytes from tumour-draining lymph nodes, obtained
from 12 patients with epithelial cancers, were immunoselected with magnetic beads coated with a 60mer synthetic peptide corresponding
to three tandem repeats of the protein core of the MUC1 antigen. Short-term cultures of B cells were established utilizing
interleukin-10 (IL-10), IL-4 and monoclonal antibody anti-CD40, and were maintained for a maximum of 3␣weeks. B cell culture
supernatants contained human anti-MUC1 antibodies, as detected by enzyme-linked immunosorbent assay, in 6/12 of the patients
tested. Five of these patients, all with early-stage cancer, also had high levels of circulating anti-MUC1 IgM antibodies
in the serum. A significant correlation was found (two-tailed P = 0.041) between the presence of circulating anti-MUC1 antibodies and the ability to isolate PEM-specific B cells from tumour-draining
lymph nodes. The technique proposed provides a useful method for the analysis of natural immunity against defined tumour antigens.
Received: 30 June 1998 / Accepted: 5 October 1998 |
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Keywords: | Immune response Polymorphic epithelial mucin MUC1 Tumour-associated antigen B lymphocytes Lymph nodes |
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