Oral Presentations - Federation of Clinical Immunology Societies

S52 Abstracts
Based on H&E and direct IF studies, 34 cases were diagnosed as
MMP. The most common antibody deposited was IgG (90%)
followed by C3 (82%) and IgG4 (71%). Strikingly, IgG4 was the
sole antibody detected in two cases (6%). Conclusion: Our
results suggest that the use of monoclonal IgG4 is important in
the diagnosis of MMP. We suggest adding monoclonal IgG4 to the
routine panel of antibodies used in studies of cases suspected to
have MMP to avoid false-negatives.
doi:10.1016/j.clim.2007.03.324
F.111 Immuophenotyping of PBL and B.M Markers in
Patients with Leukemia by Flowcytometry
Fereshteh Alesahebfosoul, Academic Member of Isfahan
University, Isfahan University Medical School, Immunology
Department, Isfahan
Introduction: Peripheral blood lymphocytes (PBL) and bone
marrow (B.M.) immunophenotyping is widely used for diagnosis
and prognosis of leukemia. Flow cytometry technique provided
facilities to assess the frequency of malignant cell population
and the protein expression in each group of cells which have
been gated. Method and material: Fresh peripheral blood and
bone marrow of patient were collected and tested for following
examination: total white blood cells and leukocyte percentage
rate were determined by H1. Machine: The samples were
treatedwithconjugatedMoAbwithFITCand/orphycoErythrine
and then dual staining of cells was performed. The following
monoclonal antibodies were used: CD10, CD19, CD20, CD5,
CD3, CD22, CD7, CD33, anti-HLA DR, and CD13. After direct
staining of cell samples, 30,000 cells were analyzed by flow
cytometry. Result and discussion: The results of the first case of
the study were WBC=780 and LUC=2.2%, CD7=60%, CD=57%,
CD5=56%ofthelymphocytepopulationofperipheralblood.But
the same markers in bone marrow sample were determined for
D7=5%, CD3=5%, and CD5=6.5%. The results of the second case
of the study were CD19=90%, CD20=89%, CD22=82%, and
CD5=95% of the lymphocyte population of peripheral blood. In
addition the same markers in bone marrow samples were
determined for CD19=76%, CD20==85%, CD22=75.9%, and
CD5=93%. HLA-DR marker was shown to be over 80% in both
peripheral blood and bone marrow for this case. The result of
cell counting for the third case of the study showed WBC=4800
and LUC=6.3%, CD19=41.5%, CD20=29.5%, CD22=39% and
CD5=43.12%, CD3=40.3%, and CD7=41.1% in peripheral blood.
But the same markers from bone marrow sample were
determined for CD19=85.6%, CD22=81.2% and CD5=2.08%
and CD3=1.88%. The results of this study show that the
lymphocyte protein expression in leukemic samples is the value
in diagnostic and classification of leukemia.
doi:10.1016/j.clim.2007.03.325
F.113 An All-Human Culture Model Supporting
Human B Lymphopoesis
Mercy Kagoda, Graduate Research Assistant/Student, Loma
Linda University School of Medicine Department of Pathology
and Human Anatomy, Loma Linda, CA, Yasmin Parrish,
Research Specialist II, Childrens Hospital Los Angeles Research
Immunology Bone Marrow Transplant, Los Angeles, CA,
Jaqueline Rogerio, Post Doctoral Fellow, Childrens Hospital Los
Angeles Research Immunology Bone Marrow Transplant, Los
Angeles, CA, Ineavely Baez, Senior Research Assistant, Loma
Linda University School of Medicine Department of Pathology
and Human Anatomy, Loma Linda, CA, Qian-Lin Hao, Research
Specialist IV, Childrens Hospital Los Angeles Research
Immunology Bone Marrow Transplant, Los Angeles, CA, Lora
Barsky, Flow Technologist, Childrens Hospital Los Angeles
Research Immunology Bone Marrow Transplant, Los Angeles,
CA, Ewa Zielinska, Flow Technologist, Childrens Hospital Los
Angeles Research Immunology Bone Marrow Transplant, Los
Angeles, CA, Monika Smogorzewska, Flow Technologist,
Childrens Hospital Los Angeles Research Immunology Bone
Marrow Transplant, Los Angeles, CA, Kimberly Payne, Assistant
Professor, Loma Linda University School of Medicine
Department of Pathology and Human Anatomy, Loma Linda,
CA
Traditionally, nonfetal models of human B cell development
have been based on co-culturing hematopoietic stem cells
(HSCs) from cord blood (CB) and bone marrow (BM) on murine
stromal cell lines. Here we describe the progressive replacement
of murine components to generate a totally human
culture model that supports human hematopoiesis, including B
cell development. First, we replaced murine stromal cell lines
with primary human BM stroma, next human serum was
substituted for nonhuman sera. It has been reported that
human B cell precursors selectively interact with CD10+ stromal
cells during in vivo B cell development. In our cultures, human
stroma express CD10 and secrete IL-7 at levels comparable to
primary murine BM stroma. We found that human stroma bind
IL-7 independently of the IL-7 receptor thereby having the
potential for presenting IL-7 to developing B cell precursors.
This is important as we have recently shown that IL-7 expands
human B cell precursors by ∼50 fold and that IL-7 is essential for
production of B cells from HSCs in adult human BM. While
human stroma in our cultures express the human pre-BCR
ligand, galectin-1 (Gal-1), the presence of Gal-1 was not
required for IL-7-induced expansion of human B cell precursors.
HSCs in co-cultures with human stroma and serum showed a
generative capacity for CD19+ and CD19− progeny that was at
least equal to that seen on murine stroma with nonhuman sera.
Planned experiments will assess the ability of this culture model
to support primary B cell malignancies.
doi:10.1016/j.clim.2007.03.326
F.114 Expression of CD3 and T Cell Receptor (TCR)
on a Minor Population of Human CD14 Positive Cells
Daron Forman, Senior Scientist, Tolerx, Cambridge, MA,
Michael Rosenzweig, Senior Director of Preclinical
Research, Tolerx, Cambridge, MA, Lou Vaickus, Chief
Medical Officer, Tolerx, Cambridge, MA
The TCR associates with CD3 polypeptides including CD3
delta, gamma, epsilon, and zeta. It was commonly believed
that TCR and CD3 expression occur exclusively on T cells.
Recently however, TCR expression was reported on a minor
population of both mouse and human neutrophils indicating