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series of 90 cases of CLL for which both immunophenotypic
and FISH data were available. Heavy chain isotype expression
was determined by 4-color flow cytometry, and karyotypic
abnormalities were identified using the Vysis CLL FISH probe
kit (Abbott Laboratories). Cases of CLL were categorized
as IgM/IgD+ (54%), IgD+ (19%), and IgD- (10%); in 17% of
cases, heavy chain expression was absent or undetectable.
Karyotypic abnormalities were categorized as described
previously (Döhner H, et al. N Engl J Med 2000; 343:1910-
6). The distribution of karyotypic abnormalities did not
differ significantly among the heavy chain isotype-defined
immunophenotypic categories; neither adverse (11q or 17p)
nor favorable (isolated 13q) deletions were more common
among any single heavy chain category (p > 0.05, Fisher-
Exact test). These data provide no evidence for an association
between immunoglobulin heavy chain expression and
karyotypic abnormalities in CLL.
P15
INKT CELLS IN NASAL POLYPOSIS: DIFFERENT
METHODS, DIFFERENT RESULTS
Mohammad Fereidouni
1
, Farahnaz Jabari Azad
2
, Reza Farid
Hosseini
2
1
Birjand University of Medical Sciences,
2
Mahshad University
of Medical Sciences
Background:
Invariant Natural killer T cells (iNKT) are a small
subset of T lymphocytes which are involved in a wide variety
of immune responses. In spite of their importance in different
disorders, low number of these cells and technical difficulties
affects the validity of studies in this field. The aim of this study
was to compare frequency of iNKT cells in nasal polyp with
flow cytometry and Real time PCR.
Materials and methods:
16 Nasal polyp tissue was isolated during the polypectomy
and each sample divided into two parts, in one part cell
suspension was made by Medimachine and the frequency of
iNKT cells among CD3+ gate, was measured by flow cyotmery
using vα24 and Vβ11 antibodies and in the other part, iNKT
cells evaluated by RT-qPCR using primers for Vα24 and Vβ11
genes as well as CD3 and GAPDH genes.
Results:
In flow
cytometry, the numbers of iNKT cells varied from 0.5% to 11%
(mean 4.7%) in Polyp cell suspensions while we could not
detect iNKT cells in most of polyp samples by real time PCR
and in a few positive samples, the relative expression of iNKT
genes was significantly lower than whatever detected in flow
cytometry.
Conclusion:
The results of this study show that
the method of iNKT detection has profound influence on the
results and it is necessary to set up standard protocols for
preventing errors in this filed.
P16
CONTRIBUTION OF MULTIPARAMETER FLOW
CYTOMETRY IMMUNOPHENOTYPING TO THE
DIAGNOSTIC SCREENING AND CLASSIFICATION OF
PEDIATRIC CANCER
Cristiane S Ferreira-Facio
1
, Cristiane B Milito
2
, Vitor Botafogo
1
,
Marcela Fontana
1
, Leandro S Thiago
3
, Elen Oliveira
1
,
Ariovaldo S da Rocha-Filho
4
, Fernando Werneck
4
, Danielle
N Forny
1
, Samuel Dekermacher
4
, Ana Paula de Azambuja
5
,
Sima E Ferman
3
, Paulo Antonio S Faria
3
, Marcelo G P Land
1
,
Alberto Orfao
6
, Elaine S Costa
1
1
Pediatrics Institute
IPPMG, Universidade Federal do Rio de Janeiro - UFRJ,
2
Pathologic Anatomy Department, Medicine Faculty/ UFRJ,
3
Servidores do Estado Hospital- HSE,
4
National Cancer
Institute of Brazil – INCA, 5Clinics Hospital, Federal University
of Paraná - UFPR.,
6
Cytometry Service, Department of
Medicine and Cancer Research Center (IBMCC, University of
Salamanca-CSIC and IBSAL), University of Salamanca
Pediatric cancer is a heterogeneous group of tumors which
require multiple procedures, for its diagnostic screening and
classification. Until now, flow cytometry (FC) has not been
POSTER ABSTRACTS