Quantitative fluorescence measures for determination of intracellular perforin content

Kevin J. Maher, Nancy G. Klimas, Barry Hurwitz, Richard Schiff, Mary Ann Fletcher

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We present methodologic details and operating characteristics of a procedure with whole blood for the quantitative assessment of intracellular perforin within distinct lymphocyte subsets. Using this method, we analyzed 20 healthy controls and 2 individuals with an inherited deficiency of perforin. The mean ± standard deviation perforin contents of natural killer (NK) cells and cytotoxic T cells of healthy controls were 3,561 ± 1,157 and 500 ± 779 relative number of molecules (rMol) of antiperforin antibody bound per cell, respectively. The NK cell perforin contents of individuals with heterozygous and homozygous perforin deficiency (familial hemophagocytic lymphohistiocytosis) were 2,260 and 212 rMol of antiperforin antibodies per NK cell. While the homozygous deficiency was found to be associated with negligible antiperforin binding, the heterozygous condition was associated with a level of perforin binding that was below the 15th percentile for healthy individuals. Because 83% of this subject's NK cells were shown to bind to antiperforin antibodies by conventional flow cytometry (relative to the normal range of 81% ± 25%), quantitative cytometry may be more sensitive than conventional cytometric methods in identifying cytolytic defects.

Original languageEnglish
Pages (from-to)1248-1252
Number of pages5
JournalClinical and Diagnostic Laboratory Immunology
Volume9
Issue number6
DOIs
StatePublished - Nov 1 2002

Fingerprint

Perforin
Fluorescence
Natural Killer Cells
Antibodies
Molecules
T-cells
Lymphocytes
Flow cytometry
Lymphocyte Subsets
Flow Cytometry
Reference Values
Blood
T-Lymphocytes
Defects

ASJC Scopus subject areas

  • Microbiology (medical)
  • Immunology and Allergy
  • Clinical Biochemistry
  • Immunology

Cite this

Quantitative fluorescence measures for determination of intracellular perforin content. / Maher, Kevin J.; Klimas, Nancy G.; Hurwitz, Barry; Schiff, Richard; Fletcher, Mary Ann.

In: Clinical and Diagnostic Laboratory Immunology, Vol. 9, No. 6, 01.11.2002, p. 1248-1252.

Research output: Contribution to journalArticle

Maher, Kevin J. ; Klimas, Nancy G. ; Hurwitz, Barry ; Schiff, Richard ; Fletcher, Mary Ann. / Quantitative fluorescence measures for determination of intracellular perforin content. In: Clinical and Diagnostic Laboratory Immunology. 2002 ; Vol. 9, No. 6. pp. 1248-1252.
@article{8436e595bbe94ac2a2fec9e3453306e3,
title = "Quantitative fluorescence measures for determination of intracellular perforin content",
abstract = "We present methodologic details and operating characteristics of a procedure with whole blood for the quantitative assessment of intracellular perforin within distinct lymphocyte subsets. Using this method, we analyzed 20 healthy controls and 2 individuals with an inherited deficiency of perforin. The mean ± standard deviation perforin contents of natural killer (NK) cells and cytotoxic T cells of healthy controls were 3,561 ± 1,157 and 500 ± 779 relative number of molecules (rMol) of antiperforin antibody bound per cell, respectively. The NK cell perforin contents of individuals with heterozygous and homozygous perforin deficiency (familial hemophagocytic lymphohistiocytosis) were 2,260 and 212 rMol of antiperforin antibodies per NK cell. While the homozygous deficiency was found to be associated with negligible antiperforin binding, the heterozygous condition was associated with a level of perforin binding that was below the 15th percentile for healthy individuals. Because 83{\%} of this subject's NK cells were shown to bind to antiperforin antibodies by conventional flow cytometry (relative to the normal range of 81{\%} ± 25{\%}), quantitative cytometry may be more sensitive than conventional cytometric methods in identifying cytolytic defects.",
author = "Maher, {Kevin J.} and Klimas, {Nancy G.} and Barry Hurwitz and Richard Schiff and Fletcher, {Mary Ann}",
year = "2002",
month = "11",
day = "1",
doi = "10.1128/CDLI.9.6.1248-1252.2002",
language = "English",
volume = "9",
pages = "1248--1252",
journal = "Clinical and Vaccine Immunology",
issn = "1556-6811",
publisher = "American Society for Microbiology",
number = "6",

}

TY - JOUR

T1 - Quantitative fluorescence measures for determination of intracellular perforin content

AU - Maher, Kevin J.

AU - Klimas, Nancy G.

AU - Hurwitz, Barry

AU - Schiff, Richard

AU - Fletcher, Mary Ann

PY - 2002/11/1

Y1 - 2002/11/1

N2 - We present methodologic details and operating characteristics of a procedure with whole blood for the quantitative assessment of intracellular perforin within distinct lymphocyte subsets. Using this method, we analyzed 20 healthy controls and 2 individuals with an inherited deficiency of perforin. The mean ± standard deviation perforin contents of natural killer (NK) cells and cytotoxic T cells of healthy controls were 3,561 ± 1,157 and 500 ± 779 relative number of molecules (rMol) of antiperforin antibody bound per cell, respectively. The NK cell perforin contents of individuals with heterozygous and homozygous perforin deficiency (familial hemophagocytic lymphohistiocytosis) were 2,260 and 212 rMol of antiperforin antibodies per NK cell. While the homozygous deficiency was found to be associated with negligible antiperforin binding, the heterozygous condition was associated with a level of perforin binding that was below the 15th percentile for healthy individuals. Because 83% of this subject's NK cells were shown to bind to antiperforin antibodies by conventional flow cytometry (relative to the normal range of 81% ± 25%), quantitative cytometry may be more sensitive than conventional cytometric methods in identifying cytolytic defects.

AB - We present methodologic details and operating characteristics of a procedure with whole blood for the quantitative assessment of intracellular perforin within distinct lymphocyte subsets. Using this method, we analyzed 20 healthy controls and 2 individuals with an inherited deficiency of perforin. The mean ± standard deviation perforin contents of natural killer (NK) cells and cytotoxic T cells of healthy controls were 3,561 ± 1,157 and 500 ± 779 relative number of molecules (rMol) of antiperforin antibody bound per cell, respectively. The NK cell perforin contents of individuals with heterozygous and homozygous perforin deficiency (familial hemophagocytic lymphohistiocytosis) were 2,260 and 212 rMol of antiperforin antibodies per NK cell. While the homozygous deficiency was found to be associated with negligible antiperforin binding, the heterozygous condition was associated with a level of perforin binding that was below the 15th percentile for healthy individuals. Because 83% of this subject's NK cells were shown to bind to antiperforin antibodies by conventional flow cytometry (relative to the normal range of 81% ± 25%), quantitative cytometry may be more sensitive than conventional cytometric methods in identifying cytolytic defects.

UR - http://www.scopus.com/inward/record.url?scp=0036844619&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036844619&partnerID=8YFLogxK

U2 - 10.1128/CDLI.9.6.1248-1252.2002

DO - 10.1128/CDLI.9.6.1248-1252.2002

M3 - Article

C2 - 12414757

AN - SCOPUS:0036844619

VL - 9

SP - 1248

EP - 1252

JO - Clinical and Vaccine Immunology

JF - Clinical and Vaccine Immunology

SN - 1556-6811

IS - 6

ER -