FcγRIIa and Fcγ RIIIa genes polymorphism in Egyptian children with primary immune thrombocytopenia

Abstract Introduction Phagocytosis of autoantibody-sensitized coated platelets through Fc gamma receptors on phagocytic cells is an important mechanism of thrombocytopenia in primary immune thrombocytopenia (ITP). Objective We aimed to investigate the contribution of the FcγRIIa and FcγRIIIa genes polymorphism to the risk of ITP and their association with disease characteristics in Egyptian children. Methods A case control study was conducted on eighty children with primary ITP and eighty age and sex healthy matched subjects as a control group. The FcγRIIa and FcγRIIIa genes polymorphism was detected using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results We found that the FcγRIIa‐131H and ‐131R allele frequencies were 51.3 % and 48.7%, respectively, in children with ITP, versus 75% and 25%, respectively, in controls (p= 0.002). The compound heterozygous HR genotype was significantly higher in ITP patients (p < 0.05). The FcγRIIIa-158F and ‐158V allele frequencies were 46.3% and 53.7%, respectively, in children with ITP, versus 70% and 30%, respectively, in controls (p= 0.002). The compound heterozygous VF genotype was significantly higher in ITP patients (p < 0.05). The combined HR/FV genotype was 47.5% in ITP patients, versus 10% in controls (p < 0.001). No significant difference was found between children with newly diagnosed ITP and those who developed chronic ITP, regarding the frequency distribution of the FcγRIIa and FcγRIIIa alleles and genotypes (p > 0.05). Conclusion There is a possible association of the FcγRIIa and FcγRIIIa genes polymorphism with the risk for, and genetic susceptibility to ITP in Egyptian children, but large-scale studies are still needed to support our findings.

Mechanism of Qihuang Yiqi Shexue Prescription in Treatment of ITP Model Mice Based on Autophagy Mediated by AMPK/mTOR/ULK1 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Qihuang Yiqi Shexue prescription （QHYQSX） in the treatment of immune thrombocytopenia （ITP） model mice based on the autophagy mediated by the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase 1 （ULK1） signaling pathway. MethodFifty BALB/c mice were randomly divided into normal group， model group， high- and low-dose QHYQSX groups， and prednisone group， with 10 mice in each group. The ITP model was induced by intraperitoneal injection of anti-platelet serum （APS） of guinea pig. On the 8th day of the APS injection， drugs were administered by gavage for 14 days. Peripheral blood platelet （PLT） count and hemoglobin （Hb） concentration were detected. Spleen and thymus were separated， weighed， and the organ index was calculated. Sternum was sampled for bone marrow smear， and bone marrow megakaryocytes were classified under a microscope. Thrombopoietin （TPO）， interleukin-6 （IL-6）， IL-10， tumor necrosis factor-α （TNF-α）， transforming growth factor-β1 （TGF-β1）， and interferon-γ （IFN-γ） in the serum were detected by enzyme-linked immunosorbent assay（ELISA）. AMPK， mTOR， ULK1， microtubule-associated protein light chain 3 （LC3）， Beclin1， and p62 mRNA expression levels in the spleen were detected by Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR）. The protein expression of AMPK， p-AMPK， p-mTOR， p-ULK1， LC3Ⅱ/LC3Ⅰ， Beclin1， and p62 in the spleen was detected by Western blot. ResultCompared with the normal group， the model group showed reduced peripheral blood PLT count， Hb， and TPO levels （P<0.05，P<0.01）， increased spleen and thymus indexes （P<0.01）， decreased number of bone marrow megakaryocytes （P<0.01）， elevated serum levels of IL-6， TNF-α， and IFN-γ （P<0.01）， and reduced IL-10 and TGF-β1 levels （P<0.01）. Compared with the model group， the groups with drug intervention showed increased PLT counts and TPO levels （P<0.01）， decreased spleen and thymus indexes （P<0.05， P<0.01）， elevated number of bone marrow megakaryocytes （P<0.05， P<0.01）， reduced serum levels of IL-6， TNF-α， and IFN-γ （P<0.05， P<0.01）， and up-regulated IL-10 and TGF-β1 levels （P<0.05，P<0.01）. Compared with the low-dose QHYQSX group， the high-dose QHYQSX group and the prednisone group showed different degrees of significant differences in improving PLT counts and levels of cellular inflammatory factors （P<0.05， P<0.01）. Real-time PCR and Western blot results showed that compared with the normal group， the model group showed up-regulated mRNA expression of AMPK， LC3， and Beclin1 and protein expression of p-AMPK/AMPK， LC3Ⅱ/LC3Ⅰ， and Beclin1 in the spleen （P<0.05， P<0.01）， and down-regulated mRNA expression of mTOR， ULK1， and p62 and protein expression of p-mTOR， p-ULK1， and p62 （P<0.05， P<0.01）. Compared with the results in the model group， high- and low-dose QHYQSX and prednisone could down-regulate the mRNA expression of AMPK， LC3， and Beclin1 and protein expression of p-AMPK/AMPK， LC3Ⅱ/LC3Ⅰ， and Beclin1 in the spleen （P<0.05， P<0.01）， and up-regulate the mRNA expression of mTOR， ULK1， and p62 and protein expression of p-mTOR， p-ULK1， and p62 （P<0.05， P<0.01）. ConclusionQHYQSX may inhibit excessive autophagy by regulating the AMPK/mTOR/ULK1 signaling pathway， thereby regulating immune intolerance and playing a role in the treatment of ITP.

Effect of anti-platelet antibodies on the efficacy of immunoglobulin therapy and Treg/Th17 ratio in children with immune thrombocytopenia / 中国输血杂志

【Objective】 To investigate the relationship between anti-platelet antibodies, therapeutic effect of intravenous immunoglobulin (IVIG) and Treg/Th17 cells imbalance in children with immune thrombocytopenia (ITP). 【Methods】 The changes and correlation of platelet count and Treg/Th17 ratio before and after IVIG treatment in 60 newly diagnosed ITP children with anti-platelet antibodies and 60 children with primary ITP without anti-platelet antibodies were analyzed. 【Results】 1) Compared with the control group, the efficacy of IVIG treatment was better in children with ITP in the case group(CR+ R cases: 50 vs 32) (P<0.01). 2) After IVIG treatment, platelet count(×109/L)(case: 4.5±2.9 vs 327.4±69.5, control: 4.1±3.2 vs 304.7±75.9), Treg cell level(%)(case: 2.15±1.08 vs 5.09±1.37, control: 2.41±0.92 vs 4.98±1.10), Treg/Th17 ratio(case: 1.10±0.19 vs 7.75±1.11, control: 1.27±0.21 vs 4.69±0.81)significantly increased while Th17 cell level(%) significantly decreased in the 2 groups of children(case: 2.07±1.31 vs 1.37±0.92, control: 2.13±1.18 vs 1.48±1.01); compared with the control group, there was no significant change in Treg, Th17 and Treg/Th17 ratio before and after treatment in the case group (P>0.05), but platelet count increased more significantly (P<0.05). 3) There were 3 positive cases in the control group and 12 negative cases in the case group after IVIG treatment, and IVIG treatment probably had no effect on the positive rate of anti-platelet antibodies in children with ITP (P>0.05). 4) The change in platelet count after IVIG treatment was significantly positively correlated with Treg levels (r=0.49 in the case group and r=0.441 in the control group) and negatively correlated with Th17 cell levels (r=-0.390 in the case group and r=-0.364 in the control group). 【Conclusion】 Anti-platelet antibodies can be used as a predictor of the efficacy of IVIG therapy in children with ITP, but they are not associated with changes in the Treg/Th17 ratio.

A Study On Etiology And Clinical Profile Of Patients With Thrombocytopenia

BACKGROUND: Thrombocytopenia refers to a reduction in platelet count below 1.5 lakh/microliter. The presence of thrombocytopenia in a hemogram should alert the physician to identify the underlying etiology for the prompt management of the patient. Timely identification and treatment prevent bleeding manifestations, requirement of platelet transfusions/steroids and overall impact on mortality of the patients. AIM OFSTUDY:Analysis to study the etiology, bleeding manifestation, percentage of patients requiring platelet transfusion, length of hospital stay in patients with thrombocytopenia. METHODOLOGY: 100 cases thrombocytopenia both male and female were included in the study. The diagnosis was made on peripheral smear and Hemogram. RESULTS: Dengue fever was the most common cause of thrombocytopenia with 43 cases. Sepsis with 23 cases was the second commonest. Bleeding manifestations were seen in 23% of the study population.100% of the patients with platelet count less than 10,000/microlitre had bleeding manifestations. 26 patients (26%) received platelet transfusion out of which 23 were therapeutic and 3 were prophylactic transfusions. Steroid therapy was given in 11% of patients. Mortality was highest in patients with sepsis induced thrombocytopenia. CONCLUSION:This study shows that Dengue fever is the commonest diagnosis made in patients who are detected to have thrombocytopenia. One fifth of patients with platelet count less than 1,00,000/microlitre tend to have bleeding manifestation, commonest being GI bleed, petechial rash and epistaxis. Majority of the bleeding occurs with platelet count less than 10,000. The proportion of patients receiving therapeutic platelet transfusion was higher compared to prophylactic transfusion.

Clinicopathological Profile of Thrombocytopenia in Sitapur and Shahjahanpur Districts of Uttar Pradesh

Introduction: Thrombocytopenia is common haematologicalfinding with different specific etiologies. The aim ofpresent study was to find clinicopathological correlation ofthrombocytopenia in adults by causes, severity and clinicalpresentation.Material and Methods: This was a cross sectional studydone for a period of 6 months. After clinical profile laboratorydata and complications of patients with a platelet count of lessthan 1,50,000 were analyzed and tabulated.Results: The total sample size was 135 with 95(70.37%) malesand 40 (29.63%) females. Out of 135 patients, 32(23.70%)were diagnosed dengue. Malaria was found in 20% patients.Most cases were presented with fever(56%) followed bybodyache (40.74%), joint pain(33.33%), bleeding (17.03%)and hepatosplenomegaly(15.55%).Conclusion: Infectious diseases were the most common causeof thrombocytopenia out of which dengue was commonestfollowed by malaria and typhoid. Early recognition anddiagnosis of cause of thrombocytopenia can avoid bleedingmanifestations and serious complications.

Effects of bone marrow mesenchymal cells from immune thrombocytopenia patients on the biological behaviors of megakaryocytes / 上海交通大学学报（医学版）

Objective • To investigate changes of immune thrombocytopenia (ITP) patients-derived bone marrow mesenchymal cells (BMCs) in cells survival, cytokines expression as well as the effects of BMCs on the biological behaviors of megakaryocytes. Methods • BMCs were collected from 7 ITP patients and 5 normal controls (NC), and cultivated by the whole marrow adherent method. Surface markers and basal apoptosis rate of BMCs were analyzed by flow cytometry (FCM). Proliferation of BMCs was assessed by CCK-8 method. Phorbol 12-myristate 13-acetate (PMA) was used to stimulate differentiation of HEL cells. The induced HEL cells (inHEL) were divided into 3 groups: inHEL cultured alone (group a), inHEL co-cultured with BMCs derived from ITP patients (group b), inHEL co-cultured with BMCs derived from NC (group c). After 72 h incubation, the expression of cell surface proteins (CD41a, CD42b) and cell apoptosis rate were analyzed by FCM. The mRNA and proteins expression levels of cytokines IL6, IL11, TPO, SCF were detected by real-time fluorescent quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), respectively. Results • Compared with NC, BMCs from ITP patients grew progressively slowly (Day 4, P=0.039; Day 6, 10, P=0.009; Day 8, P=0.007), cell basal apoptosis rates were increased [AV+PI- (early apoptosis rate), P=0.036; AV+PI+ (late apoptosis rate), P=0.003; AV+PI-/+ (total apoptosis rate), P=0.004]. Compared with group a, the expression of CD41a in group c was much higher (P=0.000). The expression of CD41a in group b was higher than that in group a (P=0.015), but still much less than that in group c (P=0.000). Compared with group a, the early and total apoptosis rate in group b, c and the late apoptosis rate in group c were decreased obviously (all P=0.000), whereas there was no obvious change of the late apoptosis rate in group b. However, compared with group c, the late and total apoptosis rate in group b were significantly increased (both P=0.000). The expression levels of IL6, SCF mRNA and IL6 protein were significantly decreased in ITP BMCs (all P=0.000), but there was no obvious difference in the expression levels of IL11 and TPO between ITP BMCs and NC BMCs. Conclusion • BMCs from ITP patients show some defects in supporting megakaryocytic differentiation and survival under co-culture conditions, which mechanisms are related to the reduction of IL6 and SCF expression.

Effects of bone marrow mesenchymal cells from immune thrombocytopenia patients on the biological behaviors of megakaryocytes / 上海交通大学学报（医学版）

Objective·To investigate changes of immune thrombocytopenia (ITP) patients-derived bone marrow mesenchymal cells (BMCs) in cells survival, cytokines expression as well as the effects of BMCs on the biological behaviors of megakaryocytes. Methods?·?BMCs were collected from 7 ITP patients and 5 normal controls (NC), and cultivated by the whole marrow adherent method. Surface markers and basal apoptosis rate of BMCs were analyzed by flow cytometry (FCM). Proliferation of BMCs was assessed by CCK-8 method. Phorbol 12-myristate 13-acetate (PMA) was used to stimulate differentiation of HEL cells. The induced HEL cells (inHEL) were divided into 3 groups: inHEL cultured alone (group a), inHEL co-cultured with BMCs derived from ITP patients (group b), inHEL co-cultured with BMCs derived from NC (group c). After 72 h incubation, the expression of cell surface proteins (CD41a, CD42b) and cell apoptosis rate were analyzed by FCM. The mRNA and proteins expression levels of cytokines IL6, IL11, TPO, SCF were detected by real-time fluorescent quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), respectively. Results?·?Compared with NC, BMCs from ITP patients grew progressively slowly (Day 4, P=0.039; Day 6, 10, P=0.009; Day 8, P=0.007), cell basal apoptosis rates were increased [AV+PI- (early apoptosis rate), P=0.036; AV+PI+(late apoptosis rate), P=0.003; AV+PI-/+(total apoptosis rate), P=0.004]. Compared with group a, the expression of CD41a in group c was much higher (P=0.000). The expression of CD41a in group b was higher than that in group a (P=0.015), but still much less than that in group c (P=0.000). Compared with group a, the early and total apoptosis rate in group b, c and the late apoptosis rate in group c were decreased obviously (all P=0.000), whereas there was no obvious change of the late apoptosis rate in group b. However, compared with group c, the late and total apoptosis rate in group b were significantly increased (both P=0.000). The expression levels of IL6, SCF mRNA and IL6 protein were significantly decreased in ITP BMCs (all P=0.000), but there was no obvious difference in the expression levels of IL11 and TPO between ITP BMCs and NC BMCs. Conclusion?·?BMCs from ITP patients show some defects in supporting megakaryocytic differentiation and survival under co-culture conditions, which mechanisms are related to the reduction of IL6 and SCF expression.

Progress on management of primary immune thrombocytopenia in pregnancy / 中华全科医师杂志

Primary immune thrombocytopenia (ITP) in pregnancy is a special type of ITP,its impact on the mother and fetus cannot be ignored.The correct diagnosis and effective treatment of ITP in pregnancy are the focus of the clinical practice and medical research.This article reviews the progress on the management of the primary immune thrombocytopenia in pregnancy.

Diagnostic Approach of Childhood Immune Thrombocytopenia / 임상소아혈액종양

Immune thrombocytopenia (ITP) is the most common cause of thrombocytopenia in children and can be defined as an autoimmune disorder of isolated thrombocytopenia without other causes of thrombocytopenia. This review will focus on the diagnostic approach of ITP, especially regarding the differential diagnosis. The practice of differential diagnosis has the goal of distinguishing primary ITP from secondary ITP and nonimmune thrombocytopenia requiring different treatments and showing different prognoses.

Management of immune thrombocytopenia: Korean experts recommendation in 2017

Management options for patients with immune thrombocytopenia (ITP) have evolved substantially over the past decades. The American Society of Hematology published a treatment guideline for clinicians referring to the management of ITP in 2011. This evidence-based practice guideline for ITP enables the appropriate treatment of a larger proportion of patients and the maintenance of normal platelet counts. Korean authority operates a unified mandatory national health insurance system. Even though we have a uniform standard guideline enforced by insurance reimbursement, there are several unsolved issues in real practice in ITP treatment. To optimize the management of Korean ITP patients, the Korean Society of Hematology Aplastic Anemia Working Party (KSHAAWP) reviewed the consensus and the Korean data on the clinical practices of ITP therapy. Here, we report a Korean expert recommendation guide for the management of ITP.

A four-point clinical criteria distinguishes immune thrombocytopenia from acute lymphoblastic leukaemia

Immune thrombocytopenia is the most common diagnosis of isolated thrombocytopenia. The dilemma encountered by paediatricians is missing diagnosis of acute leukaemia in children with isolated thrombocytopenia. We demonstrated childhood ITP could be diagnosed using a four point clinical criteria without missing a diagnosis of acute leukaemia. Hence, bone marrow examination is not necessary in children with typical features compatible with ITP prior to steroid therapy. This can encourage paediatricians to choose steroid therapy, which is cheaper and non-blood product, as first line platelet elevating therapy in children with significant haemorrhage.

Prognostic Significance of IL-10 in Childhood Chronic Immune Thrombocytopenic Purpura / 임상소아혈액종양

BACKGROUND: The proposal of the present study was to assess the cytokine imbalances such as serum levels of IL-2, IL-10 and TGF-β1 in childhood chronic ITP during the thrombocytopenic and spontaneous remission phases. The findings of this study are anticipated to offer new insights into the pathogenesis of childhood chronic ITP and the prognostic factors involved in spontaneous remission. METHODS: Ten children with chronic ITP, 6 thrombocytopenic and 4 recovered cases, were enrolled. Five healthy children and eight healthy adults were included in this study as controls. The serum levels of IL-2, IL-10 and TGF-β1 were measured by ELISA technique. RESULTS: During this study, four patients achieved spontaneous remission. The serum level of IL-10 was higher in the patients that recovered spontaneously compared to the patients with persistent chronic thrombocytopenia. The serum level of IL-2 was not able to be detected as the levels were too low for analysis. TGF-β1 showed no significant differences among the groups in this study. CONCLUSION: These data suggest that increased serum level of IL-10 could have prognostic significance in the natural course and long-term outcome of childhood chronic ITP.

Effect of short-term, high-dose methylprednisolone on oxidative stress in children with acute immune thrombocytopenia

BACKGROUND: Immune thrombocytopenia (ITP) is the most common cause of acquired childhood thrombocytopenia and is characterized by increased immune-mediated destruction of circulating thrombocytes. Oxidative damage may be involved in ITP pathogenesis; paraoxonase (PON) and arylesterase (ARE) enzymes are closely associated with the cellular antioxidant system. We investigated the effect of short-term high-dose methylprednisolone (HDMP) treatment on the total oxidant status (TOS), total antioxidant capacity (TAC), oxidative stress index (OSI), and PON and ARE enzymatic activity in children with acute ITP. METHODS: Thirty children with acute ITP constituted the study group and 30 healthy children constituted the control group. Children with acute ITP were treated with HDMP: 30 mg/kg for 3 days, then 20 mg/kg for 4 days. The TOS, TAC, OSI, PON, and ARE levels were determined before and after 7 days of HDMP treatment. RESULTS: The TAC level (P<0.001), and PON (P<0.001) and ARE (P=0.001) activities were lower and the TOS (P=0.003) and OSI (P<0.001) levels were higher in children with acute ITP than those in healthy children in the control group. We also observed statistically significant increases in the TAC (P<0.01), PON (P<0.001) and ARE levels (P=0.001) and decreases in the TOS (P<0.05) and OSI levels (P<0.05) with 7 days of HDMP treatment compared to their values before treatment. CONCLUSION: Our study demonstrated increased oxidative stress (OSI and TOC) and decreased antioxidant capacity (TAC), PON, and ARE in ITP patients and that steroid treatment could be effective in reducing the oxidative stress.

Prognostic Significance of IL-10 in Childhood Chronic Immune Thrombocytopenic Purpura / 임상소아혈액종양

BACKGROUND: The proposal of the present study was to assess the cytokine imbalances such as serum levels of IL-2, IL-10 and TGF-β1 in childhood chronic ITP during the thrombocytopenic and spontaneous remission phases. The findings of this study are anticipated to offer new insights into the pathogenesis of childhood chronic ITP and the prognostic factors involved in spontaneous remission.METHODS: Ten children with chronic ITP, 6 thrombocytopenic and 4 recovered cases, were enrolled. Five healthy children and eight healthy adults were included in this study as controls. The serum levels of IL-2, IL-10 and TGF-β1 were measured by ELISA technique.RESULTS: During this study, four patients achieved spontaneous remission. The serum level of IL-10 was higher in the patients that recovered spontaneously compared to the patients with persistent chronic thrombocytopenia. The serum level of IL-2 was not able to be detected as the levels were too low for analysis. TGF-β1 showed no significant differences among the groups in this study.CONCLUSION: These data suggest that increased serum level of IL-10 could have prognostic significance in the natural course and long-term outcome of childhood chronic ITP.

A Case of Type A Acute Aortic Dissection in an Elderly Woman with Immune Thrombocytopenia Who Underwent Replacement of the Ascending Aorta and Aortic Arch and Later Required Aortic Root Replacement for Redissection of the Aortic Root / 日本心臓血管外科学会雑誌

We report a case of type A acute aortic dissection in an elderly woman with immune thrombocytopenia (ITP) who underwent replacement of the ascending aorta and aortic arch and later required aortic root replacement for redissection of the aortic root one month after her initial surgery. She was an 86-year-old woman with severe mitral regurgitation, and surgery was contraindicated because of her age and ITP. In October 2014, the patient presented with back pain. Computed tomography confirmed the diagnosis of her condition as type A acute aortic dissection, and she was immediately transferred to our hospital. Because echocardiography showed severe aortic regurgitation, severe mitral regurgitation, and moderate tricuspid regurgitation, we performed replacement of the ascending aorta and aortic arch, mitral valve repair, and tricuspid annuloplasty. We used Bioglue to fuse the false lumen of the type A acute aortic dissection and used a Teflon felt sandwich for the proximal anastomosis technique. Respiratory support was discontinued 91 h after her first operation ; however, 30 days after surgery, she developed a to-and-fro murmur-a sign of the progression of heart failure. Echocardiography showed aggravation of aortic regurgitation, and computed tomography showed aortic root redissection ; therefore, 39 days after the initial surgery, we performed aortic root replacement. During the operation, we found the entry under the proximal anastomosis with an almost semicircle form at the right coronary cusp to the noncoronary cusp, and the dissection extended close to the right coronary artery ; thus, we performed bypass to the right coronary artery. Pathologic findings did not establish a causal association between the redissection and Bioglue, and we believed the fragility of the tissue and the selection of the surgical procedure to be the cause of redissection. The patient was transferred to another hospital when she was able to walk and eat, which was 121 days after her first operation. The patient required 50 units of platelet transfusion during her first and second operations, but her bleeding was easily controlled during surgery. She needed two procedures of pericardium drainage for pericardiac effusion and cardiac tamponade, which may relate to ITP. The diagnosis of redissection of the aortic root was made 30 days after the patient's first operation, on the basis of exacerbation of the to-and-fro murmur. Here, we emphasize the clinical importance of basic observations over time, such as auscultation, that are liable to be overlooked in the intensive care unit.

Mechanisms of loss of immune tolerance in the production of antiplatelet autoantibodies in primary immune thrombocytopenia / 中华微生物学和免疫学杂志

Primary immune thrombocytopenia ( ITP) is an organ-specific autoimmune hemorrhagic disease. The etiology of ITP is still unclear, but loss of immune tolerance to platelet surface antigens is con-sidered as a fundamental cause of ITP. Therapeutic strategies that prevent the activation and proliferation of autoreactive cells have been suggested, which includes clearance of autoreactive cells ( apoptosis) , receptor editing, induction of anergy and extrinsic cellular suppression. Failure at any of these steps may lead to the production of autoantibodies against platelet and megakaryocyte glycoproteins. An improved understanding of the mechanisms for autoantibody production will provide theoretical basis for optimal diagnosis and treatment of ITP.

Evaluation of 303 Cases of Thrombocytopenia

Thrombocytopenia is a common clinical problem with many etiological causes. Although transient bone marrow suppression and marrow infiltration by malignancies are important causes, certain non-malignant conditions such as nutritional causes and infections are equally important as the treatment is simple and cure is possible. Depending on the aetiology, the clinical presentation may vary. Knowing the exact aetiology is important for specific treatment and prognostication. A total of 303 cases of thrombocytopenia were studied out of which males were 44% and females were 56%. The patients’ age ranged from 5 months to 84 yrs. The commonest presenting symptom was fever with bleeding manifestations and jaundice. 14% of cases are of Grade 1, 20% of the cases of Grade 2, 5% of cases of Grade 3 and 31% of cases had counts less than 25000/cu.mm i.e. of Grade 4. 50% of cases in Grade 4 had a decreased number of megakaryocytes in the bone marrow. The most common cause of thrombocytopenia in our study was megaloblastic anaemia (48.6%), ITP (20%), post-viral (10.9%) followed by leukaemia, aplastic anaemia and others. Thrombocytopenia has a spectrum of causes which can be diagnosed by detailed history and peripheral smear examination supported by bone marrow examination. Megaloblastic anaemia was the commonest cause of thrombocytopenia followed by immune thrombocytopenia. Unlike in the western India megaloblastic anemia is highly prevalent and is the leading correctable cause of thrombocytopenia. Most of the patients with Grade 4 thrombocytopenia had a decreased number of megakaryocytes in the bone marrow suggesting a production defect.

Roles of CD5+, CD19+, CD41a+, CD55+ and CD59+ in Chronic Immune Thrombocytopenic Purpura (ITP).

Aims: The aim of this study was to investigate of the roles of CD5+ and CD19+ on lymphocytes, CD5+ on B lymphocytes, CD41a+ on platelets and CD55+ and CD59+ on erythrocytes in platelet destruction; and evaluate them according to the patient response status to steroid therapy and platelet counts in chronic immune thrombocytopenic purpura (ITP). Study Design: This study included 20 chronic ITP patients and 20 healthy controls. We investigated the roles of CD5+ and CD19+ expression on lymphocytes, CD5+ expression on B lymphocytes, CD41a+ expression on platelets, and CD55+ and CD59+ expression on erythrocytes, as well as the platelet counts in healthy and chronic ITP patients. Additionally, these markers were evaluated according to the patient response status to steroid therapy and platelet counts. Place and Duration of Study: This study took place at the Department of Internal Medicine and Haematology, Meram Medical Faculty at Selçuk University in Turkey, between November, 2008 and July, 2009. Methodology: A total of 40 patients (26 women, 14 men, age range: 19-79 years) were studied. The study group included 20 chronic ITP patients (12 women and 8 men, age range: 19-78 years) and the control group included 20 healthy volunteers (14 women and 6 men, age range: 22-79 years). The platelet counts and expressions of CD5+ and CD19+ on lymphocytes, CD5+ on B lymphocytes, CD41a+ on platelets, and CD55+ and CD59+ on erythrocytes were analysed in the patients and control subjects. The chronic ITP patients were evaluated according to their requirements of treatment. Five patients whose platelet counts were above 50,000 mm–3 were observed without treatment. The other 15 patients whose platelet counts were under 50.000 mm–3 and had bleeding, or whose platelet counts were under 20,000 mm–3, were given methylprednisolone treatments (1 mg/kg/day orally). Three of the 15 patients discontinued treatment for various reasons. The twelve patients who continued the methylprednisolone treatment were divided into two subgroups according to their responder status of steroid treatment. The patients whose platelet counts slowly increased above 30,000 mm–3 within three months included the steroid treatment responder subgroups. The chronic ITP patients were also divided into two subgroups according to the severity of their thrombocytopenia. The limit of the platelet count was 30,000 mm–3 for severe thrombocytopenia. These parameters were analysed according to the response status of the steroid treatment and platelet counts. The platelet counts, and the expressions of these markers, were compared between the subgroups. Results: The level of CD5+ on B lymphocyte expression (2.19 ± 1.65) in peripheral blood lymphocytes was significantly higher in the immune thrombocytopenic purpura patients than in the controls (P = .05). The CD55+ + CD59+ expression on erythrocytes (98.03 ± 1.77) was significantly higher in the ITP patients than in the controls (P = .05). There was no significant relationship between the expression of CD5+, CD19+ or CD5+ on B lymphocytes, CD41a+ expression on platelets or CD55+ and CD59+ expression on erythrocytes, according to the response status to steroid therapy in the patient group (P > 0.05). Additionally, the patients were evaluated according to platelet counts, and there was a significantly positive correlation between the level of CD41a+ expression on the platelets and the platelet count (P = .05). Conclusion: The level of CD5+ on B lymphocytes was significantly higher in the ITP patients than in the controls. A relationship between CD55+ plus CD59+ expression on erythrocytes and immune destruction of platelets was not observed in the chronic ITP patients.

Clinical Significance of Antibodies Against Platelet HLA Class I in Children with Idiopathic Thrombocytopenic Purpura / 대한수혈학회지

BACKGROUND: A previous history of transfusion has been known to be associated with production of anti-HLA class I antibodies. However, platelet glycoproteins are the main target of idiopathic thrombocytopenic purpura (ITP). The mechanism of antibody production is known to differ significantly between glycoproteins and anti-HLA class I. The aim of this study was to evaluate the clinical significance of anti-HLA class I antibodies in childhood ITP. METHODS: Enrollment for the normal control group targeted 48 people who visited Gyeongsang National University Hospital from 1990 to 2010, and 48 young children with ITP. Anti-glycoproteins and anti-HLA class I antibodies were tested using the Modified Antigen Capture Enzyme-linked immunosorbent assay (MACE) kit. RESULTS: The positive rate of anti-HLA antibodies was significantly different [36/39 (92.3%) vs 29/46 (63%)] [ITP group vs normal control group] (P=0.002). The mean positive S/C ratio of anti-HLA antibodies was also significantly different (3.55 vs 1.51) [ITP group vs normal control group] (P=0.0000). The positive rate of anti-HLA did not differ significantly between the transfused group and the non-transfused group [12/12 (100%) vs 24/27 (88%)] [transfused ITP vs non-transfused ITP]. The mean positive S/C ratio of anti-HLA antibodies did not differ significantly between the transfused ITP group and the non-transfused ITP group (4.30 vs 3.25) [transfused ITP vs non-transfused ITP]. Consecutive testing showed that positive rate and positive S/C ratio of anti-HLA antibodies did not change significantly between sampling times in both groups [transfused ITP vs non-transfused ITP] (P=1.00 and P=0.15). CONCLUSION: Anti-HLA class I antibodies may be involved in childhood ITP. Transfusion did not affect the course of childhood ITP.