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 language | English |
|---|---|
| Pages (from-to) | 1248-1252 |
| Number of pages | 5 |
| Journal | Clinical and Diagnostic Laboratory Immunology |
| Volume | 9 |
| Issue number | 6 |
| DOIs | |
| State | Published - Nov 1 2002 |
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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 journal › Article
}
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.
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U2 - 10.1128/CDLI.9.6.1248-1252.2002
DO - 10.1128/CDLI.9.6.1248-1252.2002
M3 - Article
VL - 9
SP - 1248
EP - 1252
JO - Clinical and Vaccine Immunology
JF - Clinical and Vaccine Immunology
SN - 1556-6811
IS - 6
ER -