Double fluorescence technique for measurement of complement-fixing antibody to lymphocyte subsets.

Abstract:

:A double fluorescent antibody method for quantitating human complement-fixing antibody to lymphocyte subclasses has been developed. The indicators in this system are a C6-deficient serum as a non-lytic source of complement, rhodamine-labeled anti-C3 and fluorescein-labeled murine monoclonal antibodies to human lymphocyte subsets. The basic procedure is to incubate lymphocytes with the unknown serum and then to add C6-deficient serum. The binding of C3 is indicated by staining with rhodamine-labeled anti-C3 and the subset class of the lymphocyte so stained is determined by binding of fluorescein-tagged anti-OKT4 or -OKT8 antibodies. The occurrence of both red and green cell surface fluorescence denotes the presence of a complement-binding antibody to the lymphocyte subset defined by the monoclonal antibody. In addition to defining the specificity of complement-fixing anti-lymphocyte antibodies, this technique is more sensitive than the microcytotoxicity assay.

journal_name

J Immunol Methods

authors

Ozturk GE,Hayward AR,Weil R 3rd,Kohler PF

doi

10.1016/0022-1759(85)90284-4

subject

Has Abstract

pub_date

1985-12-17 00:00:00

pages

163-8

issue

1

eissn

0022-1759

issn

1872-7905

pii

0022-1759(85)90284-4

journal_volume

85

pub_type

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