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october 7-9, 2012 • NEW ORLEANS, LA
we were able to identify a severe decrease in memory B
cells number (relative and absolute) in six patients. Age,
sex, anamnesis morbi and clinical manifestation were highly
variable in accordance with heterogeneity of a disease.
Overall, the severity of the course was more dependent on
memory B cell count, than on serum immunoglobulin levels.
In some patients we also found association with neutropenia
that should be later characterized in details.
P20
COMPARISON OF BD™ STEM CELL ENUMERATION
KIT (BD-SCE) WITH CURRENT LABORATORY ISHAGE
PROTOCOL STEM CELL ENUMERATION ASSAY
Abigail S Kelliher, Michelle E Delelys, Laura J Dillon,
Frederic I Preffer
Massachusetts General Hospital
In February of 2012, BD released the Stem Cell Enumeration
Kit (SCE) for absolute counting of CD34+ stem cells.
Recently, new requirements by the Foundation for the
Accreditation of Cellular Therapies (FACT) necessitate the
presence of a simultaneous viability marker to determine
CD34+ cell viability. BD’s new single platform kit satisfies
these requirements. We performed a comparison with our
current dual platform method that is based upon the ISHAGE
Stem Cell Enumeration guidelines. Percentages and
absolute counts of viable CD34+ cells were compared. We
tested staining the cells in the dark and in light, determined
how long cells remained viable after staining and whether
or not cells needed to be left on ice. There was excellent
correlation between the two assays. In comparing viable
absolute counts and percentages of CD34+ cells, the
r-squared values were 0.977 and 0.895, respectively. If
two samples are removed from the study that were stained
the following day (all others were stained the same day),
the r-squared value increased to 0.942. Since the absolute
value of viable CD34+ cells ultimately determines the
potential engraftment of the transplant, that value is of
greatest importance. The BD™ SCE Kit and the automated
FACSCanto™ software require that 7- Color Setup Beads
as well as BD™ Stem Cell controls be utilized prior to the
analysis of patient samples. The new BD assay is 8 times
more expensive than our current assay, although it is more
streamlined and because the software is automated, easier
to ensure accurate gating between users.
P21
FLOW CYTOMETRY REVEALS IMMUNE SIGNATURES
ASSOCIATED TO HUMAN AGING
Anis Larbi
1
, Tze Pin Ng
2
, Tamas Fulop
3
1
Singapore
Immunology Network, Biopolis, A*STAR,
2
NUHS, Singapore,
3
Research Center on Aging
The proportion of elderly individuals (over 65 years old)
is increasing worldwide. The problem with the elderly
population is that a significant fraction is not healthy.
Although the number of centenarians is increasing, the
number of clinical conditions affecting mortality and morbidity
is high. Most of elderly individuals display at least one
chronic condition that overall affects health. Based on this,
we aimed at identifying in a very well-characterized elderly
population the changes at the immunological levels that
may be associated to one or several conditions. For this
we recruited apparently healthy home-dwelling individuals
via the Singapore Longitudinal Aging Study (SLAS). Using
state-of-the-art flow cytometry we tested the hypothesis
that T cell subset distribution was different in healthy elderly
vs. individual suffering from diabetes, frailty, dementia and
compared this with various clinical parameters including
inflammation-related markers, also assessed by flow
POSTER ABSTRACTS