HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Takahashi, M. Articles by Pressman, D. Search for Related Content PubMed Articles by Takahashi, M. Articles by Pressman, D.

Presence of Two Different Immunoglobulin Heavy Chains in Individual Cells of Established Human Hematopoietic Cell Lines¹

From the Departments of Biochemistry Research and Surgery, Roswell Park Memorial Institute, Buffalo, New York 14203³

Abstract

Presence of -, - and µ-heavy chains of immunoglobulin was examined in cultured cells of established human hematopoietic cell lines. Combinations of a fluorescein-conjugated antibody of one specificity and a tetramethylrhodamine-conjugated antibody of another specificity were used to see whether individual cells contain heavy chains of more than one class.

Studies were concentrated on two cell lines which produced immunoglobulins of more than one class.

Essentially all the immunoglobulin-containing cells in cell line LK1D (RPMI no. 4292) were found to contain both - and -heavy chains. Individual cells containing both - and -heavy chains were also found in high proportions in cell line RPMI no. 3157 along with the cells containing only -, only - or only µ-chains. There were a few cells containing both - and µ-chains, but no cells were found to contain both - and µ-chains.

In addition to the usual controls for fluorescent antibody techniques, specificity of antibody reagents was shown by staining bone marrows from myeloma patients and a mixture of cells from two cell lines, one producing only IgA and the other producing only IgG. Even when the cells of these two cell lines were grown together in the same culture, no cells were found to contain two heavy chains. This strongly suggests that the chains found in individual cells are being synthesized in the cells and are not derived from other cells in the culture.

Antibody reagents were specifically purified and were made monospecific to each heavy chain of immunoglobulin by the use of insoluble immunoadsorbents conjugated with homologous or heterologous antigens.

Footnotes

This work was supported in part by Grant no. AI-5158 from the National Institute of Allergy and Infectious Diseases.

² Work was done during the leave of absence from the Institute of Cancer Immunopathology, Hokkaido University, Sapporo, Japan. Present address: Cancer Research Institute, Kanazawa University, Kanazawa, Japan.

³ A unit of the New York State Department of Health.