12th Congress of the European Hematology ... - Haematologica

acquired mutation most prone to cause IM resistance. The heterozigosity
of p.I432I (c.1677T>C) in the oral sample, and the fact that it does not
cause an amino-acid change, led us to suppose that this should be a polymorphism.
We plan, also, to sequence the patient cDNA sample collected
at diagnosis. The absence of p.K404E in this sample will be other evidence
pointing out to an acquired mutation. Being so, this amino-acid
should be taken into account when screening BCR-ABL mutations resistant
to IM.
References
1. Hofmann WK, et al. Leukemia & Lymphoma 2004; 45:655-60.
2. Towatari M, et al. Blood 2004; 104:3507-12.
3. Talpaz M, et al. NEJM 2006; 354:2531-41.
4. van Dongen JJ et al. Leukemia 1999; 13:1901-28.
5. van der Velden V, et al. Leukemia 2003; 17:1013-34.
1460
ANTIBODIES IN THE PATIENTS WITH HAEMOPHILIA EXPOSED TO MULTIPLE
TRANSFUSIONS
V. Mineeva, 1 O. Zavarzina, 2 V. Soldatenkov, 1 N. Bodrova, 1 E. Elkhina1 1 Haematology&Transfusiology, SAINT-PETERSBURG; 2 Hematology&Transfusiology,
ST-PETERSBURG, Russian Federation
Although the indications for transfusion of fresh-frozen plasma and
cryoprecipitate are limited, the use of plasma continues to rise in Russia.
Residual platelets and RBCs in plasma can provide immunization of
the patients. Limited data are available concerning antibodies in multitransfused
haemophiliacs. The purpose of our work was the identification
of auto- and alloantibodies in the haemophilia patients exposed to
multiple transfusions. Auto- and alloantibodies to red cells were identified
by direct and indirect antiglobulin test (DiaMed AG, Switzerland).
For identification of granulocyte autoantibodies the modified method of
agglutination in gel (DiaMed) was used. Platelet auto- and alloantibodies
were tested by mixed passive haemagglutination test (Shibata et. al.,
1981), which has been modified to avoid HLA antibodies detection.
Specificity of HPA antibodies was confirmed by the use of panel HPA
genotyped donor platelets. We examined 162 blood samples of patients
with haemophilia. Fifteen samples (9,26%) had alloantibodies to red
cells (anti-D,-E,-K,-CW). Autoantibodies to red cells were not detected.
Blood of 139 patients was investigated for platelet autoantibodies, forty
eight (34,53%) of them were positive. Out of 97 analysed blood samples,
41 were positive for platelet alloantibodies (42,27%). The specificity of
antibodies was the following: anti- HPA-1a (6 samples), anti- HPA-1b (4
samples), anti- HPA- 2a (2 samples), anti- HPA- 2b (7 samples), anti-
HPA- 3a (8 samples), anti- HPA 5a (2 samples). Positive antibody test
without detection of specific antibodies was found in 12 patients. 136
patients were investigated for granulocyte auto-immunization, three of
them (2,21%) were positive. As a rule, patients with haemophilia receive
the transfusions repeatedly, that enables to observe the process of antibodies
formation in dynamics. 18 patients were repeatedly investigated
for platelet autoantibodies: 10 remained negative, 6 patients developed
the autoantibodies, platelet autoantibodies disappeared in 2 patients.
Thus, after multiple transfusion of fresh-frozen plasma and cryoprecipitate,
platelet auto- and allo-immunization (34,53% and 42,27% accordingly)
develops in patients with haemofilia in addition to RBCs alloimmunization.
1461
RADIOFREQUENCY ABLATION FOR RESIDUAL HEPATIC HODGKINS LYMPHOMA:
A CASE REPORT
A.M. Nosari, C. Rusconi, A. Rampoldi, L. Gargantini, E. Ravelli,
M. Turrini, C. Gabutti, E. Morra
Ospedale Niguarda Ca’Granda, MILAN, Italy
Percutaneous radiofrequency (RF) ablation is a well documented treatment
for primary and metastatic localized hepatic tumors. 1 Until now
few cases of RF in lymphoma patients have been reported. 2 In October
2003 a 33-year-old woman presented with cervical lymphoadenopathies,
fever, fatigue and abdominal pain. Hodgkin’s lymphoma (HL), scleronodular
variety, was diagnosed by lymphonode biopsy. CT scan showed
mediastinal and para-aortic not bulky lymphoadenopathies and a hepatic
lesion suggestive for a disease localization with a maximum diameter
of 23 mm. HL was staged as IVB and ABVD treatment was started. After
4 cycles patient achieved a partial response, with complete regression of
lymphoadenopathies but a stable size of the hepatic lesion. At the end
of 8 ABVD cycles restaging by CT and PET confirmed the complete
regression of nodal disease while the hepatic lesion, measuring 16 mm,
was still present and metabolically active. Two DHAP cycles were
administered and peripheral blood stem cells harvest after the first cycle
was successful (20.3×10 6 CD34 + /kg). After the second DHAP the hepatic
lesion exhibited further reduction, being the unique residual site of HL
with a maximum diameter of 14 mm. In January 2005 patient underwent
eco-guided percutaneous RF ablation of the hepatic lesion with no significant
toxicity. After RF PET was negative and CT still showed the
hepatic focal lesion, with a maximum diameter of 14 mm. Patient is
now alive and in complete remission after a 26 months follow-up. This
case shows that RF, a safe and well known treatment for solid tumors,
could be considered as a promising chance for residual HL in the liver, a
location not susceptible of external radiation therapy.
References
12 th Congress of the European Hematology Association
1. Image-guided Percutaneous Radiofrequency Ablation and Incidence of
Post-Radiofrequency Ablation Syndrome: Prospective Survey. T.M. Wah,
R.S. Arellano, D.A. Gervais et al. Radiology 2005; 237:1097-102
2. Radiofrequency ablation of lymphoma. D. Sudheendra, M.M. Barth, U.
Hegde et al. Blood 2006; 107:1624-6.
1462
ORGAN DERIVED MATRIX EXTRACTS INDUCE TISSUE SPECIFIC DIFFERENTIATION OF
HUMAN BONE MARROW DERIVED MESENCHYMAL STEM CELLS
A. Bazarbachi, 1 H. El-Khoury, 1 R. Hamdan-Khalil, 1 M. Springer, 2
A. Briest, 2 S. Sindet-Pedersen, 3 M. El-Sabban1 1 American University of Beirut, BEIRUT, Lebanon; 2 Ossacur AG, OBERSTEN-
FELD, Germany; 3 The London Research Company, LONDON, United Kingdom
Background. Bone marrow derived mesenchymal stem cells (BMMSC)
are rare multipotent cells that can differentiate into multiple lineages
such as bone, cartilage and muscle cells among others. Hence, these cells
are of potential clinical importance for the repair of damaged tissues. The
local microenvironment in vivo is critical to support the desired differentiation
of stem cells or to sustain the phenotype of the stem cell-derived
in vitro differentiated cells. This local microenvironment comprises a
physical support supplied by the organ matrix as well as tissue specific
cytokines. Colloss ® and Colloss ® E (Ossacur AG) are sterile acellular
lyophilizates extracted from bovine and equine bone matrix, respectively.
Method. We tested the effect of Colloss ® and Colloss ® E, and cartilage
acellular extracts of bovine origin on human BMMSC differentiation.
Results. BMMSC treated with either Colloss ® or Colloss ® E exhibited
characteristic morphological changes accompanied by the expression of
osteoblast specific markers such as alkaline phosphatase activity, osteopontin
secretion and calcium deposits, explicitly demonstrating that
these bone matrix extracts induce osteoblastic differentiation of BMM-
SC in vitro. Similarly, treatment of BMMSC with either meniscus or joint
derived cartilage extracts resulted in the formation of cell aggregates
with a central alignment containing pre-hypertrophic cells in the middle
suggestive of chondrocytic differentiation. This was accompanied by
the expression of chondrocyte specific markers such as Sox9, aggrecan
and glycoso-amino-glycans production, hence demonstrating that they
induce chondrocytic differentiation of BMMSC in vitro. Control cultures,
studied at all time points, maintained a spindle shaped morphology typical
of bone marrow-derived mesenchymal stem cells. Importantly, Colloss
® or Colloss ® E did not induce chondrocytic markers, and conversely,
cartilage extracts did not induce osteoblastic markers, hence demonstrating
organ specificity in BMMSC differentiation phenotype. Conclusions.
Colloss ® , Colloss ® E, and cartilage extracts seem to provide stem
cells with specific structural and soluble mediators that mimic the in vivo
microenvironment, and induce bone- or cartilage-specific differentiation
of BMMSC, respectively. These results support a novel tissue engineering-based
tissue repair, using autologous BMMSC pretreated with organ
derived matrix extracts.
haematologica/the hematology journal | 2007; 92(s1) | 521