An immunofluorescent method for identifying individual IFN-γ-producing lymphocytes

Tamás Laskay; Ulf Andersson; Jan Andersson; Rolf Kiessling; Marc DeLey

doi:10.1016/0022-1759(86)90310-8

An immunofluorescent method for identifying individual IFN-γ-producing lymphocytes

Tamás Laskay, Ulf Andersson^*, Jan Andersson, Rolf Kiessling, Marc DeLey

^*Corresponding author for this work

Institute of Medical Microbiology

24 Citations (Scopus)

Abstract

An immunofluorescent staining method was developed for detecting human IFN-γ-producing cells in single cell suspensions. Mononuclear leukocytes, stimulated in vitro to produce IFN-γ, were fixed and made permeable. The cytoplasmic presence of IFN-γ was visualized by indirect immunofluorescence using IFN-γ-specific mouse monoclonal antibodies. The staining was found to be specific for IFN-γ and allowed the detection of newly synthesized rather than internalized IFN-γ molecules. The cytoplasmic fluorescence appeared locally in a polar, juxtanuclear position, which overlapped the Golgi apparatus, probably reflecting the glycosylation site of the newly formed IFN-γ molecules. Two-colour staining experiments showed that the method is useful not only for the detection and enumeration but also for the phenotypic characterization of IFN-γ-producing cells.

Original language	English
Journal	Journal of Immunological Methods
Volume	95
Issue number	1
Pages (from-to)	1-7
Number of pages	7
ISSN	0022-1759
DOIs	https://doi.org/10.1016/0022-1759(86)90310-8
Publication status	Published - 04.12.1986

Funding

This work was supported by PHS Grant no. 2RO1 CA 26782-06 awarded by the National Cancer Institute, DHEW, the Swedish Cancer Society, the Swedish Medical Research Council and the L~ikars~illskapet. We gratefully acknowledge the advice of Dr. Hans Wigzell.

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

Access to Document

10.1016/0022-1759(86)90310-8

Cite this

@article{c806f961ebd94befa7851dc1d870aa62,

title = "An immunofluorescent method for identifying individual IFN-γ-producing lymphocytes",

abstract = "An immunofluorescent staining method was developed for detecting human IFN-γ-producing cells in single cell suspensions. Mononuclear leukocytes, stimulated in vitro to produce IFN-γ, were fixed and made permeable. The cytoplasmic presence of IFN-γ was visualized by indirect immunofluorescence using IFN-γ-specific mouse monoclonal antibodies. The staining was found to be specific for IFN-γ and allowed the detection of newly synthesized rather than internalized IFN-γ molecules. The cytoplasmic fluorescence appeared locally in a polar, juxtanuclear position, which overlapped the Golgi apparatus, probably reflecting the glycosylation site of the newly formed IFN-γ molecules. Two-colour staining experiments showed that the method is useful not only for the detection and enumeration but also for the phenotypic characterization of IFN-γ-producing cells.",

author = "Tam{\'a}s Laskay and Ulf Andersson and Jan Andersson and Rolf Kiessling and Marc DeLey",

year = "1986",

month = dec,

day = "4",

doi = "10.1016/0022-1759(86)90310-8",

language = "English",

volume = "95",

pages = "1--7",

journal = "Journal of Immunological Methods",

issn = "0022-1759",

publisher = "Elsevier B.V.",

number = "1",

}

TY - JOUR

T1 - An immunofluorescent method for identifying individual IFN-γ-producing lymphocytes

AU - Laskay, Tamás

AU - Andersson, Ulf

AU - Andersson, Jan

AU - Kiessling, Rolf

AU - DeLey, Marc

PY - 1986/12/4

Y1 - 1986/12/4

N2 - An immunofluorescent staining method was developed for detecting human IFN-γ-producing cells in single cell suspensions. Mononuclear leukocytes, stimulated in vitro to produce IFN-γ, were fixed and made permeable. The cytoplasmic presence of IFN-γ was visualized by indirect immunofluorescence using IFN-γ-specific mouse monoclonal antibodies. The staining was found to be specific for IFN-γ and allowed the detection of newly synthesized rather than internalized IFN-γ molecules. The cytoplasmic fluorescence appeared locally in a polar, juxtanuclear position, which overlapped the Golgi apparatus, probably reflecting the glycosylation site of the newly formed IFN-γ molecules. Two-colour staining experiments showed that the method is useful not only for the detection and enumeration but also for the phenotypic characterization of IFN-γ-producing cells.

AB - An immunofluorescent staining method was developed for detecting human IFN-γ-producing cells in single cell suspensions. Mononuclear leukocytes, stimulated in vitro to produce IFN-γ, were fixed and made permeable. The cytoplasmic presence of IFN-γ was visualized by indirect immunofluorescence using IFN-γ-specific mouse monoclonal antibodies. The staining was found to be specific for IFN-γ and allowed the detection of newly synthesized rather than internalized IFN-γ molecules. The cytoplasmic fluorescence appeared locally in a polar, juxtanuclear position, which overlapped the Golgi apparatus, probably reflecting the glycosylation site of the newly formed IFN-γ molecules. Two-colour staining experiments showed that the method is useful not only for the detection and enumeration but also for the phenotypic characterization of IFN-γ-producing cells.

UR - https://www.scopus.com/pages/publications/0022981675

U2 - 10.1016/0022-1759(86)90310-8

DO - 10.1016/0022-1759(86)90310-8

M3 - Journal articles

C2 - 2431063

AN - SCOPUS:0022981675

SN - 0022-1759

VL - 95

SP - 1

EP - 7

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

IS - 1

ER -

An immunofluorescent method for identifying individual IFN-γ-producing lymphocytes

Abstract

Funding

UN SDGs

Research Areas and Centers

Access to Document

Other files and links

Fingerprint

Cite this