ISSN 1866-8836
Клеточная терапия и трансплантация
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Gorchakova, Ekaterina B. Rusanova, Konstantin U. Slobodnyuk, Irina J. Saburova, Elena A. Stadnik, Michail I. Zaraiski, Yekaterina E. Zueva</p>" ["TYPE"]=> string(4) "HTML" } ["DESCRIPTION"]=> string(0) "" ["VALUE_ENUM"]=> NULL ["VALUE_XML_ID"]=> NULL ["VALUE_SORT"]=> NULL ["~VALUE"]=> array(2) { ["TEXT"]=> string(158) "

Margarita V. Gorchakova, Ekaterina B. Rusanova, Konstantin U. Slobodnyuk, Irina J. Saburova, Elena A. Stadnik, Michail I. Zaraiski, Yekaterina E. Zueva

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Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia


Correspondence
Margarita V. Gorchakova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: Rita.gorchakova@gmail.com 

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Aims

We present a rare clinical case of combined myeloid and lymphoid lineage disorders in patients with chronic eosinophilic leukemia and T-cell lymphoma.

Materials

The patient (female, 51) complained of general weakness, a permanent hyperthermia up to 38.5–39ºC, constant painful dry cough, and also she expressed redness, peeling and itchy skin on her face. An external inspection showed a significant peripheral lymphadenopathy. A clinical blood analysis revealed the following: leukocytes – 21.9 *10^9 /L, segmented neutrophils – 47%, eosinophilia – 17% (3.7 *10^9 /L), lymphocytes – 28% (6.1 *10^9 /L), monocytes – 8%, hemoglobin – 96 g /L, and ESR – 32 mm /h. Up to 44% of eosinophils were found in the myelogram.

Results

The bulk of lymphoid bone marrow cells were represented by a homogeneous population of CD2+CD3+CD4+CD5+ T-cells. A significant portion of the T-cells had an early thymic phenotype CD4+CD8+CD3+. The eosinophils phenotype was represented by markers CD13+CD15+CD33+ with a lack of CD117 and CD23. Molecular biological studies identified chimerical gene transcripts FIP1L1/PDGRFα and monoclonal T-cell receptors. The level of mRNA expression of the interleukin–5 gene was negative.

Summary

T-lymphocytes represented a clone of transformed cells confirmed by the molecular genetics data (receptor clonality) and flow cytometry (aberrant phenotype). The presence of mutations of the gene FIP1L1/PDGRFα showed the clonal nature of hypereosinophilia. A lack of increase in IL5 proved the lymphoproliferative variant of HES to be incorrect. Thus, using molecular genetics and immunological techniques we verified the presence of two diseases: chronic eosinophilic leukemia and T-cell lymphoma unspecified IVA.

Keywords

hypereosinophilic syndrome, T-cell lymphoma, case report, chronic eosinophilic leukemia

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Margarita V. Gorchakova, Ekaterina B. Rusanova, Konstantin U. Slobodnyuk, Irina J. Saburova, Elena A. Stadnik, Michail I. Zaraiski, Yekaterina E. Zueva

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Margarita V. Gorchakova, Ekaterina B. Rusanova, Konstantin U. Slobodnyuk, Irina J. Saburova, Elena A. Stadnik, Michail I. Zaraiski, Yekaterina E. Zueva

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Aims

We present a rare clinical case of combined myeloid and lymphoid lineage disorders in patients with chronic eosinophilic leukemia and T-cell lymphoma.

Materials

The patient (female, 51) complained of general weakness, a permanent hyperthermia up to 38.5–39ºC, constant painful dry cough, and also she expressed redness, peeling and itchy skin on her face. An external inspection showed a significant peripheral lymphadenopathy. A clinical blood analysis revealed the following: leukocytes – 21.9 *10^9 /L, segmented neutrophils – 47%, eosinophilia – 17% (3.7 *10^9 /L), lymphocytes – 28% (6.1 *10^9 /L), monocytes – 8%, hemoglobin – 96 g /L, and ESR – 32 mm /h. Up to 44% of eosinophils were found in the myelogram.

Results

The bulk of lymphoid bone marrow cells were represented by a homogeneous population of CD2+CD3+CD4+CD5+ T-cells. A significant portion of the T-cells had an early thymic phenotype CD4+CD8+CD3+. The eosinophils phenotype was represented by markers CD13+CD15+CD33+ with a lack of CD117 and CD23. Molecular biological studies identified chimerical gene transcripts FIP1L1/PDGRFα and monoclonal T-cell receptors. The level of mRNA expression of the interleukin–5 gene was negative.

Summary

T-lymphocytes represented a clone of transformed cells confirmed by the molecular genetics data (receptor clonality) and flow cytometry (aberrant phenotype). The presence of mutations of the gene FIP1L1/PDGRFα showed the clonal nature of hypereosinophilia. A lack of increase in IL5 proved the lymphoproliferative variant of HES to be incorrect. Thus, using molecular genetics and immunological techniques we verified the presence of two diseases: chronic eosinophilic leukemia and T-cell lymphoma unspecified IVA.

Keywords

hypereosinophilic syndrome, T-cell lymphoma, case report, chronic eosinophilic leukemia

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Aims

We present a rare clinical case of combined myeloid and lymphoid lineage disorders in patients with chronic eosinophilic leukemia and T-cell lymphoma.

Materials

The patient (female, 51) complained of general weakness, a permanent hyperthermia up to 38.5–39ºC, constant painful dry cough, and also she expressed redness, peeling and itchy skin on her face. An external inspection showed a significant peripheral lymphadenopathy. A clinical blood analysis revealed the following: leukocytes – 21.9 *10^9 /L, segmented neutrophils – 47%, eosinophilia – 17% (3.7 *10^9 /L), lymphocytes – 28% (6.1 *10^9 /L), monocytes – 8%, hemoglobin – 96 g /L, and ESR – 32 mm /h. Up to 44% of eosinophils were found in the myelogram.

Results

The bulk of lymphoid bone marrow cells were represented by a homogeneous population of CD2+CD3+CD4+CD5+ T-cells. A significant portion of the T-cells had an early thymic phenotype CD4+CD8+CD3+. The eosinophils phenotype was represented by markers CD13+CD15+CD33+ with a lack of CD117 and CD23. Molecular biological studies identified chimerical gene transcripts FIP1L1/PDGRFα and monoclonal T-cell receptors. The level of mRNA expression of the interleukin–5 gene was negative.

Summary

T-lymphocytes represented a clone of transformed cells confirmed by the molecular genetics data (receptor clonality) and flow cytometry (aberrant phenotype). The presence of mutations of the gene FIP1L1/PDGRFα showed the clonal nature of hypereosinophilia. A lack of increase in IL5 proved the lymphoproliferative variant of HES to be incorrect. Thus, using molecular genetics and immunological techniques we verified the presence of two diseases: chronic eosinophilic leukemia and T-cell lymphoma unspecified IVA.

Keywords

hypereosinophilic syndrome, T-cell lymphoma, case report, chronic eosinophilic leukemia

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Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia


Correspondence
Margarita V. Gorchakova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: Rita.gorchakova@gmail.com 

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Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia


Correspondence
Margarita V. Gorchakova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: Rita.gorchakova@gmail.com 

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Rudnitskaya, Elena V. Morozova, Boris V. Afanasyev</p>" ["TYPE"]=> string(4) "HTML" } ["DESCRIPTION"]=> string(0) "" ["VALUE_ENUM"]=> NULL ["VALUE_XML_ID"]=> NULL ["VALUE_SORT"]=> NULL ["~VALUE"]=> array(2) { ["TEXT"]=> string(66) "

Yulia V. Rudnitskaya, Elena V. Morozova, Boris V. Afanasyev

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Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia

Correspondence
Elena V. Morozova, Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: dr_morozova@mail.ru

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Introduction

The only curative treatment available for myelodysplastic syndromes (MDS) is hematopoietic stem cell transplantation (HSCT). There are two types of conditioning regimens followed by HSCT: reduced-intensity conditioning (RIC) and standard (myeloablative) conditioning. RIC regimens were designed to perform allogeneic HSCT in patients who had a high risk of death due to toxicity of standard preparative chemotherapy regimens (elder patients, patients with severe comorbid conditions, or heavy pretreated patients). However, the use of RIC can increase the risk of relapse after HSCT. We report the single case of treatment with decitabine of relapsed MDS after RIC matched related allograft stem cell transplantation.

Patient characteristics

The patient was a 49-year-old female, who had been diagnosed with MDS, refractory anemia with an excess of blasts II, and IPSS-3, which was verified in September 2007 based on anemia, leucopenia, blasts in bone marrow >10%, multiple dysplastic signs, and multiple chromosomes’ disorders (46,хх,del(2)(p22),add(3)(q29)[15] / 46,хх,del(2)(p22) [3] / 46,xx [2]).

SCT and follow up

Due to high risk MDS (IPSS-3), it was decided to perform allograft HSCT from an HLA identical sibling as the first line treatment (day 0, 18 January 2008). The conditioning regimen was RIC (Fludarabin+Busulfan). GVHD prophylaxis was CSA + MTX.

The engraftment was carried out on day +16. On day +30 the patient had mixed chimerism (50%); 46,xy; and complete disappearance of chromosomes’ disorders and no evidence of acute GVHD.

However, disease relapse was revealed on day +68 (15% of blasts in bone marrow, cytopenia, multiple chromosomes’ disorders). On day +72 CSA was stopped. On day +82 a donor lymphocytes infusion (DLI) №1 was performed. Nevertheless, on day +93 there were still signs of disease progression (22% of blasts in bone marrow, mixed chimerism 30%).

Decitabine 20 mg/m2/day, 1-hour IV infusion for 5 days, was started with the main goal to reduce the blasts quantity in the bone marrow. DLI №2 and DLI №3 were performed on days +110 and +136, respectively. Since day +154 patient has had chronic GVHD of the skin.

We achieved complete remission: normal blood counts, normal account of blasts in bone marrow, full donor’s chimerism, and 46,xy.

Conclusion

Combined treatment (Decitabine and DLI) showed clinical activity and can be used for treatment of relapsed myelodysplastic syndrome after allogeneic HSCT.

Keywords

myelodysplastic syndrome, reduced-intensity conditioning, allogeneic HSCT, relapsed myelodysplastic syndrome, donor lymphocytes infusion, decitabine

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Rudnitskaya, Elena V. Morozova, Boris V. Afanasyev</p>" ["TYPE"]=> string(4) "HTML" } ["DESCRIPTION"]=> string(0) "" ["VALUE_ENUM"]=> NULL ["VALUE_XML_ID"]=> NULL ["VALUE_SORT"]=> NULL ["~VALUE"]=> array(2) { ["TEXT"]=> string(66) "

Yulia V. Rudnitskaya, Elena V. Morozova, Boris V. Afanasyev

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Yulia V. Rudnitskaya, Elena V. Morozova, Boris V. Afanasyev

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Introduction

The only curative treatment available for myelodysplastic syndromes (MDS) is hematopoietic stem cell transplantation (HSCT). There are two types of conditioning regimens followed by HSCT: reduced-intensity conditioning (RIC) and standard (myeloablative) conditioning. RIC regimens were designed to perform allogeneic HSCT in patients who had a high risk of death due to toxicity of standard preparative chemotherapy regimens (elder patients, patients with severe comorbid conditions, or heavy pretreated patients). However, the use of RIC can increase the risk of relapse after HSCT. We report the single case of treatment with decitabine of relapsed MDS after RIC matched related allograft stem cell transplantation.

Patient characteristics

The patient was a 49-year-old female, who had been diagnosed with MDS, refractory anemia with an excess of blasts II, and IPSS-3, which was verified in September 2007 based on anemia, leucopenia, blasts in bone marrow >10%, multiple dysplastic signs, and multiple chromosomes’ disorders (46,хх,del(2)(p22),add(3)(q29)[15] / 46,хх,del(2)(p22) [3] / 46,xx [2]).

SCT and follow up

Due to high risk MDS (IPSS-3), it was decided to perform allograft HSCT from an HLA identical sibling as the first line treatment (day 0, 18 January 2008). The conditioning regimen was RIC (Fludarabin+Busulfan). GVHD prophylaxis was CSA + MTX.

The engraftment was carried out on day +16. On day +30 the patient had mixed chimerism (50%); 46,xy; and complete disappearance of chromosomes’ disorders and no evidence of acute GVHD.

However, disease relapse was revealed on day +68 (15% of blasts in bone marrow, cytopenia, multiple chromosomes’ disorders). On day +72 CSA was stopped. On day +82 a donor lymphocytes infusion (DLI) №1 was performed. Nevertheless, on day +93 there were still signs of disease progression (22% of blasts in bone marrow, mixed chimerism 30%).

Decitabine 20 mg/m2/day, 1-hour IV infusion for 5 days, was started with the main goal to reduce the blasts quantity in the bone marrow. DLI №2 and DLI №3 were performed on days +110 and +136, respectively. Since day +154 patient has had chronic GVHD of the skin.

We achieved complete remission: normal blood counts, normal account of blasts in bone marrow, full donor’s chimerism, and 46,xy.

Conclusion

Combined treatment (Decitabine and DLI) showed clinical activity and can be used for treatment of relapsed myelodysplastic syndrome after allogeneic HSCT.

Keywords

myelodysplastic syndrome, reduced-intensity conditioning, allogeneic HSCT, relapsed myelodysplastic syndrome, donor lymphocytes infusion, decitabine

" ["TYPE"]=> string(4) "HTML" } ["~DESCRIPTION"]=> string(0) "" ["~NAME"]=> string(21) "Description / Summary" ["~DEFAULT_VALUE"]=> array(2) { ["TEXT"]=> string(0) "" ["TYPE"]=> string(4) "HTML" } ["DISPLAY_VALUE"]=> string(2788) "

Introduction

The only curative treatment available for myelodysplastic syndromes (MDS) is hematopoietic stem cell transplantation (HSCT). There are two types of conditioning regimens followed by HSCT: reduced-intensity conditioning (RIC) and standard (myeloablative) conditioning. RIC regimens were designed to perform allogeneic HSCT in patients who had a high risk of death due to toxicity of standard preparative chemotherapy regimens (elder patients, patients with severe comorbid conditions, or heavy pretreated patients). However, the use of RIC can increase the risk of relapse after HSCT. We report the single case of treatment with decitabine of relapsed MDS after RIC matched related allograft stem cell transplantation.

Patient characteristics

The patient was a 49-year-old female, who had been diagnosed with MDS, refractory anemia with an excess of blasts II, and IPSS-3, which was verified in September 2007 based on anemia, leucopenia, blasts in bone marrow >10%, multiple dysplastic signs, and multiple chromosomes’ disorders (46,хх,del(2)(p22),add(3)(q29)[15] / 46,хх,del(2)(p22) [3] / 46,xx [2]).

SCT and follow up

Due to high risk MDS (IPSS-3), it was decided to perform allograft HSCT from an HLA identical sibling as the first line treatment (day 0, 18 January 2008). The conditioning regimen was RIC (Fludarabin+Busulfan). GVHD prophylaxis was CSA + MTX.

The engraftment was carried out on day +16. On day +30 the patient had mixed chimerism (50%); 46,xy; and complete disappearance of chromosomes’ disorders and no evidence of acute GVHD.

However, disease relapse was revealed on day +68 (15% of blasts in bone marrow, cytopenia, multiple chromosomes’ disorders). On day +72 CSA was stopped. On day +82 a donor lymphocytes infusion (DLI) №1 was performed. Nevertheless, on day +93 there were still signs of disease progression (22% of blasts in bone marrow, mixed chimerism 30%).

Decitabine 20 mg/m2/day, 1-hour IV infusion for 5 days, was started with the main goal to reduce the blasts quantity in the bone marrow. DLI №2 and DLI №3 were performed on days +110 and +136, respectively. Since day +154 patient has had chronic GVHD of the skin.

We achieved complete remission: normal blood counts, normal account of blasts in bone marrow, full donor’s chimerism, and 46,xy.

Conclusion

Combined treatment (Decitabine and DLI) showed clinical activity and can be used for treatment of relapsed myelodysplastic syndrome after allogeneic HSCT.

Keywords

myelodysplastic syndrome, reduced-intensity conditioning, allogeneic HSCT, relapsed myelodysplastic syndrome, donor lymphocytes infusion, decitabine

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Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia

Correspondence
Elena V. Morozova, Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: dr_morozova@mail.ru

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Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia

Correspondence
Elena V. Morozova, Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: dr_morozova@mail.ru

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Rusanova<sup>1</sup>, Konstantin U. Slobodnyuk<sup>1</sup>, Margarita V.Gorchakova<sup>1</sup>, Yekaterina E. Zueva<sup>1</sup>, Ramon Simon-Lopez<sup>2</sup></p>" ["TYPE"]=> string(4) "HTML" } ["DESCRIPTION"]=> string(0) "" ["VALUE_ENUM"]=> NULL ["VALUE_XML_ID"]=> NULL ["VALUE_SORT"]=> NULL ["~VALUE"]=> array(2) { ["TEXT"]=> string(178) "

Ekaterina B. Rusanova1, Konstantin U. Slobodnyuk1, Margarita V.Gorchakova1, Yekaterina E. Zueva1, Ramon Simon-Lopez2

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1Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia; 2Ramon Simon-Lopez, Beckman Coulter, Nyon, Switzerland


Correspondence
Ekaterina B. Rusanova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
Phone/Fax: +7 (812) 233 9726, E-mail: katerina.rusanova@gmail.com

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Purpose

Myelodysplastic syndrome (MDS) is a group of clonal bone marrow disorders. The list of features used commonly for diagnosis of MDS includes several morphological cell characteristics, such as the presence of hypogranulation in the neutrophils. We have observed in our daily experience that the NE Mean scatter (SS) was lower in MDS than in normal cases and the scatter of lymphocyte SS is quite constant in normal cases and in the majority of MDS.

Methods

Twenty-four diagnosed with MDS and 16 normal blood donors as a control group were enrolled into this study. The protocol of study included evaluation of blood cell populations with the GenS (BC) hematological analyzer, smears microscopy, and flow cytometry for identifying neutrophils granularity levels (FC500, BC). We created a single tube protocol for immunophenotyping of blood cells with multiparametric flow cytometry using a panel of monoclonal antibodies: CD14-FITC, CD16-PE, CD33-PC5, and CD45-PC7 (BC). In order to standardize the NE mean SS we also used an index granularity of neutrophils (IGN), which was calculated by dividing the neutrophil mean SS by lymphocyte mean SS.

Results

The results show that IGN was significantly lower in MDS than in the normal group (p<0.001). We obtained AUC=0.874 (significance level 0.0001) with a cut-off of <=8.34, a sensitivity of 87.5% with a specificity of 81.5% using the IGN to detect MDS.

Conclusions

The information about hypogranularity of the neutrophils is a well-known feature of MDS. Using IGN may be a useful tool to detect or flag neutrophil hypogranularity and could facilitate the differential diagnosis of MDS.

Keywords

myelodysplastic syndrome, dysplasia, hypogranularity, flow cytometry, screening, peripheral blood

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Ekaterina B. Rusanova1, Konstantin U. Slobodnyuk1, Margarita V.Gorchakova1, Yekaterina E. Zueva1, Ramon Simon-Lopez2

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Purpose

Myelodysplastic syndrome (MDS) is a group of clonal bone marrow disorders. The list of features used commonly for diagnosis of MDS includes several morphological cell characteristics, such as the presence of hypogranulation in the neutrophils. We have observed in our daily experience that the NE Mean scatter (SS) was lower in MDS than in normal cases and the scatter of lymphocyte SS is quite constant in normal cases and in the majority of MDS.

Methods

Twenty-four diagnosed with MDS and 16 normal blood donors as a control group were enrolled into this study. The protocol of study included evaluation of blood cell populations with the GenS (BC) hematological analyzer, smears microscopy, and flow cytometry for identifying neutrophils granularity levels (FC500, BC). We created a single tube protocol for immunophenotyping of blood cells with multiparametric flow cytometry using a panel of monoclonal antibodies: CD14-FITC, CD16-PE, CD33-PC5, and CD45-PC7 (BC). In order to standardize the NE mean SS we also used an index granularity of neutrophils (IGN), which was calculated by dividing the neutrophil mean SS by lymphocyte mean SS.

Results

The results show that IGN was significantly lower in MDS than in the normal group (p<0.001). We obtained AUC=0.874 (significance level 0.0001) with a cut-off of <=8.34, a sensitivity of 87.5% with a specificity of 81.5% using the IGN to detect MDS.

Conclusions

The information about hypogranularity of the neutrophils is a well-known feature of MDS. Using IGN may be a useful tool to detect or flag neutrophil hypogranularity and could facilitate the differential diagnosis of MDS.

Keywords

myelodysplastic syndrome, dysplasia, hypogranularity, flow cytometry, screening, peripheral blood

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Purpose

Myelodysplastic syndrome (MDS) is a group of clonal bone marrow disorders. The list of features used commonly for diagnosis of MDS includes several morphological cell characteristics, such as the presence of hypogranulation in the neutrophils. We have observed in our daily experience that the NE Mean scatter (SS) was lower in MDS than in normal cases and the scatter of lymphocyte SS is quite constant in normal cases and in the majority of MDS.

Methods

Twenty-four diagnosed with MDS and 16 normal blood donors as a control group were enrolled into this study. The protocol of study included evaluation of blood cell populations with the GenS (BC) hematological analyzer, smears microscopy, and flow cytometry for identifying neutrophils granularity levels (FC500, BC). We created a single tube protocol for immunophenotyping of blood cells with multiparametric flow cytometry using a panel of monoclonal antibodies: CD14-FITC, CD16-PE, CD33-PC5, and CD45-PC7 (BC). In order to standardize the NE mean SS we also used an index granularity of neutrophils (IGN), which was calculated by dividing the neutrophil mean SS by lymphocyte mean SS.

Results

The results show that IGN was significantly lower in MDS than in the normal group (p<0.001). We obtained AUC=0.874 (significance level 0.0001) with a cut-off of <=8.34, a sensitivity of 87.5% with a specificity of 81.5% using the IGN to detect MDS.

Conclusions

The information about hypogranularity of the neutrophils is a well-known feature of MDS. Using IGN may be a useful tool to detect or flag neutrophil hypogranularity and could facilitate the differential diagnosis of MDS.

Keywords

myelodysplastic syndrome, dysplasia, hypogranularity, flow cytometry, screening, peripheral blood

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1Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia; 2Ramon Simon-Lopez, Beckman Coulter, Nyon, Switzerland


Correspondence
Ekaterina B. Rusanova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
Phone/Fax: +7 (812) 233 9726, E-mail: katerina.rusanova@gmail.com

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1Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia; 2Ramon Simon-Lopez, Beckman Coulter, Nyon, Switzerland


Correspondence
Ekaterina B. Rusanova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
Phone/Fax: +7 (812) 233 9726, E-mail: katerina.rusanova@gmail.com

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J. Myeloproliferative diseases

Case report: combined myeloid and lymphoid lineage disorders in patient with chronic eosinophilic leukemia and T-cell lymphoma
Константин Слободнюк, Маргарита Горчакова, Екатерина Русанова, Михаил Зарайский, Екатерина Зуева, Ирина Сабурова, Елена Стадник
Decitabine for treatment of relapsed myelodysplastic syndrome after allogeneic stem cell transplantation: Case report
Борис Афанасьев, Елена В. Морозова, Юлия Рудницкая
Index granularity of neutrophils in peripheral blood as a new application for screening myelodysplastic syndromes using flow cytometry
Константин Слободнюк, Маргарита Горчакова, Екатерина Русанова, Екатерина Зуева, Ramon Simon-Lopez

J. Myeloproliferative diseases

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Margarita V. Gorchakova, Ekaterina B. Rusanova, Konstantin U. Slobodnyuk, Irina J. Saburova, Elena A. Stadnik, Michail I. Zaraiski, Yekaterina E. Zueva

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Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia


Correspondence
Margarita V. Gorchakova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: Rita.gorchakova@gmail.com 

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Aims

We present a rare clinical case of combined myeloid and lymphoid lineage disorders in patients with chronic eosinophilic leukemia and T-cell lymphoma.

Materials

The patient (female, 51) complained of general weakness, a permanent hyperthermia up to 38.5–39ºC, constant painful dry cough, and also she expressed redness, peeling and itchy skin on her face. An external inspection showed a significant peripheral lymphadenopathy. A clinical blood analysis revealed the following: leukocytes – 21.9 *10^9 /L, segmented neutrophils – 47%, eosinophilia – 17% (3.7 *10^9 /L), lymphocytes – 28% (6.1 *10^9 /L), monocytes – 8%, hemoglobin – 96 g /L, and ESR – 32 mm /h. Up to 44% of eosinophils were found in the myelogram.

Results

The bulk of lymphoid bone marrow cells were represented by a homogeneous population of CD2+CD3+CD4+CD5+ T-cells. A significant portion of the T-cells had an early thymic phenotype CD4+CD8+CD3+. The eosinophils phenotype was represented by markers CD13+CD15+CD33+ with a lack of CD117 and CD23. Molecular biological studies identified chimerical gene transcripts FIP1L1/PDGRFα and monoclonal T-cell receptors. The level of mRNA expression of the interleukin–5 gene was negative.

Summary

T-lymphocytes represented a clone of transformed cells confirmed by the molecular genetics data (receptor clonality) and flow cytometry (aberrant phenotype). The presence of mutations of the gene FIP1L1/PDGRFα showed the clonal nature of hypereosinophilia. A lack of increase in IL5 proved the lymphoproliferative variant of HES to be incorrect. Thus, using molecular genetics and immunological techniques we verified the presence of two diseases: chronic eosinophilic leukemia and T-cell lymphoma unspecified IVA.

Keywords

hypereosinophilic syndrome, T-cell lymphoma, case report, chronic eosinophilic leukemia

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Case report: combined myeloid and lymphoid lineage disorders in patient with chronic eosinophilic leukemia and T-cell lymphoma

J. Myeloproliferative diseases

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Yulia V. Rudnitskaya, Elena V. Morozova, Boris V. Afanasyev

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Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia

Correspondence
Elena V. Morozova, Memorial R.M. Gorbacheva Institute of Children Hematology and Transplantation, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
E-mail: dr_morozova@mail.ru

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Introduction

The only curative treatment available for myelodysplastic syndromes (MDS) is hematopoietic stem cell transplantation (HSCT). There are two types of conditioning regimens followed by HSCT: reduced-intensity conditioning (RIC) and standard (myeloablative) conditioning. RIC regimens were designed to perform allogeneic HSCT in patients who had a high risk of death due to toxicity of standard preparative chemotherapy regimens (elder patients, patients with severe comorbid conditions, or heavy pretreated patients). However, the use of RIC can increase the risk of relapse after HSCT. We report the single case of treatment with decitabine of relapsed MDS after RIC matched related allograft stem cell transplantation.

Patient characteristics

The patient was a 49-year-old female, who had been diagnosed with MDS, refractory anemia with an excess of blasts II, and IPSS-3, which was verified in September 2007 based on anemia, leucopenia, blasts in bone marrow >10%, multiple dysplastic signs, and multiple chromosomes’ disorders (46,хх,del(2)(p22),add(3)(q29)[15] / 46,хх,del(2)(p22) [3] / 46,xx [2]).

SCT and follow up

Due to high risk MDS (IPSS-3), it was decided to perform allograft HSCT from an HLA identical sibling as the first line treatment (day 0, 18 January 2008). The conditioning regimen was RIC (Fludarabin+Busulfan). GVHD prophylaxis was CSA + MTX.

The engraftment was carried out on day +16. On day +30 the patient had mixed chimerism (50%); 46,xy; and complete disappearance of chromosomes’ disorders and no evidence of acute GVHD.

However, disease relapse was revealed on day +68 (15% of blasts in bone marrow, cytopenia, multiple chromosomes’ disorders). On day +72 CSA was stopped. On day +82 a donor lymphocytes infusion (DLI) №1 was performed. Nevertheless, on day +93 there were still signs of disease progression (22% of blasts in bone marrow, mixed chimerism 30%).

Decitabine 20 mg/m2/day, 1-hour IV infusion for 5 days, was started with the main goal to reduce the blasts quantity in the bone marrow. DLI №2 and DLI №3 were performed on days +110 and +136, respectively. Since day +154 patient has had chronic GVHD of the skin.

We achieved complete remission: normal blood counts, normal account of blasts in bone marrow, full donor’s chimerism, and 46,xy.

Conclusion

Combined treatment (Decitabine and DLI) showed clinical activity and can be used for treatment of relapsed myelodysplastic syndrome after allogeneic HSCT.

Keywords

myelodysplastic syndrome, reduced-intensity conditioning, allogeneic HSCT, relapsed myelodysplastic syndrome, donor lymphocytes infusion, decitabine

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Decitabine for treatment of relapsed myelodysplastic syndrome after allogeneic stem cell transplantation: Case report

J. Myeloproliferative diseases

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Ekaterina B. Rusanova1, Konstantin U. Slobodnyuk1, Margarita V.Gorchakova1, Yekaterina E. Zueva1, Ramon Simon-Lopez2

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1Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University,
St. Petersburg, Russia; 2Ramon Simon-Lopez, Beckman Coulter, Nyon, Switzerland


Correspondence
Ekaterina B. Rusanova, Department of Clinical Laboratory Diagnostics, Center for Laboratory Diagnostics, St. Petersburg Pavlov State Medical University, 6/8, Tolstoy str., St. Petersburg, 199044, Russia
Phone/Fax: +7 (812) 233 9726, E-mail: katerina.rusanova@gmail.com

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Purpose

Myelodysplastic syndrome (MDS) is a group of clonal bone marrow disorders. The list of features used commonly for diagnosis of MDS includes several morphological cell characteristics, such as the presence of hypogranulation in the neutrophils. We have observed in our daily experience that the NE Mean scatter (SS) was lower in MDS than in normal cases and the scatter of lymphocyte SS is quite constant in normal cases and in the majority of MDS.

Methods

Twenty-four diagnosed with MDS and 16 normal blood donors as a control group were enrolled into this study. The protocol of study included evaluation of blood cell populations with the GenS (BC) hematological analyzer, smears microscopy, and flow cytometry for identifying neutrophils granularity levels (FC500, BC). We created a single tube protocol for immunophenotyping of blood cells with multiparametric flow cytometry using a panel of monoclonal antibodies: CD14-FITC, CD16-PE, CD33-PC5, and CD45-PC7 (BC). In order to standardize the NE mean SS we also used an index granularity of neutrophils (IGN), which was calculated by dividing the neutrophil mean SS by lymphocyte mean SS.

Results

The results show that IGN was significantly lower in MDS than in the normal group (p<0.001). We obtained AUC=0.874 (significance level 0.0001) with a cut-off of <=8.34, a sensitivity of 87.5% with a specificity of 81.5% using the IGN to detect MDS.

Conclusions

The information about hypogranularity of the neutrophils is a well-known feature of MDS. Using IGN may be a useful tool to detect or flag neutrophil hypogranularity and could facilitate the differential diagnosis of MDS.

Keywords

myelodysplastic syndrome, dysplasia, hypogranularity, flow cytometry, screening, peripheral blood

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Index granularity of neutrophils in peripheral blood as a new application for screening myelodysplastic syndromes using flow cytometry