γδ T lymphocyte function and the polymorphism of T cell receptor V δ chain in lungs of asthmatic patients / 中国医学科学院学报

<p><b>OBJECTIVE</b>To observe the function of gamma delta T lymphocytes and the polymorphism of T cell receptor V delta chain in the lungs of asthmatic patients and explore the role of gamma delta T cells in airway inflammation.</p><p><b>METHODS</b>Bronchoalveolar lavage fluid BALF was obtained from 7 asthmatic patients and 7 healthy control individuals. The percentage of gamma delta T cell in BALF was measured by flow cytometry. The gamma delta T cell in BALF was purified by magnetic labeled beads. Proliferous activity was examined by MTT assay. Cytokines secreted by gamma delta T cells in medium was assessed by enzyme-linked immunosorbent assay. Polymorphism of T cell receptor V delta chain was detected by RT-PCR and gene scan analysis.</p><p><b>RESULTS</b>The proportion of gamma delta T cell in the BALF of asthmatic patients [(6.39+/-0.71)%] was significantly higher than that in control subjects [(2.62+/-0.37)%] (P<0.01). The proportion of macrophage in the BALF of asthmatic patients [(81+/-4)] was significantly lower than that in control subjects [(86+/-2)] (P<0.05). The proliferation rate of asthmatic patients [(284.2+/-43.6)%] was significantly higher than that of control subjects [(217.5+/-59.5)%] (P<0.05). Interleukin-4 secreted by gamma delta T cells of asthmatic patients [(18.9+/-3.1) pg/ml)] significantly increased when compared with the control subjects [(14.1+/-3.0) pg/ml] (P<0.05). The polymorphism of T cell receptor V delta chain was not significantly different between these two groups.</p><p><b>CONCLUSIONS</b>The increase of gamma delta T cells in the lung of asthmatic patients further exacerbates Th1/Th2 disturbance and airway inflammation. Antigen recognition by gamma delta T cells is non-specific.</p>

Studying of clinical and laboratory features of chronic eosinophilic leukemias /hypereosinophilic syndrome / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical and laboratory features of chronic eosinophilic leukemias (CEL) and hypereosinophilic syndrome (HES).</p><p><b>METHODS</b>The clinical manifestations, laboratory parameters were retrospectively analyzed in 20 patients with HES/CEL. Detection of the FIP1L1-PDGFRA fusion gene was performed by nested RT-PCR. JAK2 V617F mutation screening was processed through allele-specific PCR combined with sequence analysis. PCR-RFLP was used to discriminate homozygous from heterozygous mutation patterns. TCR gamma rearrangement was detected by PCR.</p><p><b>RESULTS</b>Of the 20 patients, 19 were males and one female, with a median age of 33 (20 to 57) years. The FIP1L1-PDGFRA fusion gene positivity in bone marrow mononuclear cells in 12 cases was identified. All the breakpoints were identified by direct sequencing of cloned RT-PCR products in FIP1L1 intron 10 - 12 and in PDGFRA exon 12. In CEL the most common involved organs were lungs, heart and nervous system. Splenomegaly was significantly more frequent in CEL than in HES (92.5% vs 42.5%, P = 0.031). Anemia and myelofibrosis were common in CEL. There was no significant difference in circulating absolute eosinophil, leukocyte, platelet counts, hemoglobin level and percentages of eosinophil and blast cell in bone marrow between CEL and HES. The morphological abnormalities of eosinophils on bone marrow smear were easily found in CEL, including hypogranularity, and cytoplasmic vacuolization, increased basophilic granule. One patient with HES was found to have heterozygous JAK2 V617F mutation. Six patients had TCR gamma rearrangement, including 4 CEL and 2 HES.</p><p><b>CONCLUSIONS</b>(1) There is a male predominance in HES/CEL, and the median age was in the thirties. (2) The most common involved organs in CEL were lung, heart and nervous system. Bone marrow morphology might be of a little help in diagnosis of CEL. (3) JAK2 V617F may be involved in the pathogenesis of HES. (4) Patients with CEL carried the FIP1L1-PDGFRA fusion gene and TCR gamma rearrangement concurrently, their relationship warrants further study.</p>

Anemia falciforme em Salvador - Bahia: caracterização fenotípica, molecular e de sequências gênicas potencialmente importantes na expressão dos genes gama da hemoglobina fetal / Sickle cell anemia in Salvador - Bahia: phenotype and molecular characterization and search of specific gene sequences potentially important to gama gennes expression of the fetal hemoglobin

(...) O estudo investigou as características fenopíticas e os marcadores moleculares presentes em portadores da anemia falciforme de Salvador-BA, identificando seqüências gênicas potencialmente importantes para a expressão dos genes gama. O perfil de hemoglobinas e o nível de HbF foram determinados por cromatografia líquida de alta performance (HPLC). Informações sobre o perfil clínico dos pacientes foram obtidas através da análise de prontuários. A talassemia alfa2 3.7kb foi investigada pela reação em cadeia da polimerase (PCR) e os haplótipos ligados ao grupo de genes da globina betaS foram investigados por PCR e análise de sítios polimórficos utilizando endonucleases de restrição (RFLP) (...) Foram analisados 131 pacientes, dos quais 125 tiveram identificado o genótipo BetaS, tendo sido encontrado 64 (51,2 por cento) CAR/Ben; 36 (28,8 por cento) Ben/Ben; 18 (14,4 por cento) CAR/CAR; dois (1,6 por cento) Ben/Cam; um (0,8 por cento) CAR/Cam; um (0,8 por cento) Car/Arábia-India e um (0,8 por cento) Sen/Aty. A talassemia alfa2 3.7kb foi estudada em 110 pacientes, onde 30 (27,3 por cento) foram heterozigotos e dois (1,8 por cento) homozigotos. O uso de transfusão sanguínea foi maior em pacientes com HbF menor ou igual 10,0 por cento (p=0,009). Pacientes com genótipos alfa diferentes apresentaram diferenças para os valores de Hb (p=0,018); Ht (p=0,019); VCM (p=0,0004) e HCM (p=0,039). Os níveis de HbF foram maiores entre os pacientes Ben/Ben que entre os CAR/CAR (p=0,007) e CAR/Ben (p=0,013). A análise das seqüências do HS2-LCR de dez indivíduos demonstrou a substituição G-A na posição -10.677, presente apenas entre os portadores do haplótipo Ben com nível elevado de HbF, sugerindo uma possível associação entre este polimorfismo, a expressão dos genes y e a síntese da HbF. A análise da região promotora do gene yG demonstrou a substituição T-C na posição -157, que parece ser uma seqüência característica entre os pacientes estudados. Também foi encontrada a deleção de 4 pb na posição -222 a -225 no gene yG e relacionado ao haplótipo Cam. Os dados demonstraram um novo polimorfismo localizado no HS2-LCR e na região promotora do gene yG da globina, justificando a realização de estudos adicionais, associando os níveis de HbF, marcadores biológicos e mecanismos relacionados, visando esclarecer um possível papel no desenvolvimento do fenótipo da doença

Study of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphoma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the rate of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphomas (NHL) and its pathogenesis and pathologic significance.</p><p><b>METHODS</b>PCR analysis of monoclonal, polyclonal and dual rearrangements of IgH and TCR gamma, TCR beta genes was carried out in 125 cases of NHL to evaluate the rate of dual rearrangements, immunohistochemistry was performed for a Ki67 protein expression in 117 cases and the proliferation index was calculated. The relationship between antigen receptor gene rearrangements and proliferation index was analyzed.</p><p><b>RESULTS</b>Combination of the two pairs of IgH gene primers with the multiplex PCR for TCR gamma and TCR beta gene revealed dual rearrangements in 8% (8/96) of B-NHL, 17% (5/29) of T-NHL. In B cell NHL, IgH gene monoclonal, dural and polyclonal rearrangements were identified in 65, 8 and 15 cases respectively, while in T-cell NHL, they were in 15, 5 and 9 cases, respectively. There was no significant difference between proliferation index and monoclonal, dual, polyclonal rearrangements in both B-NHL and T-NHL by One-way test.</p><p><b>CONCLUSION</b>Dual rearrangements in NHL are not rare and have no relationship with proliferation index.</p>

A Case of Subcutaneous Panniculitic T-cell Lymphoma Involved in Skin and Central Nervous System / 대한피부과학회지

Subcutaneous panniculitic T-cell lymphoma(SPTCL) is categorized as a rare subtype of peripheral T-cell lymphoma. It is characterized by primary involvement of the subcutaneous fat in a manner mimicking panniculitis with/without hemophagocytic syndrome. They share a generally aggressive course and are highly associated with Epstein-Barr virus infection. EBV infection plays an important role in the tumorigenesis and may be related to cutaneous lymphoma with hemophagocytic manifestations. We have seen a patient, a 67-year-old woman with tender erythematous nodules on both upper extremities and abdomen. With time, the skin lesion showed ulcerative change on her right thigh. She has also suffered from fever, weight loss, arthralgia, and general weakness without hepatosplenomegaly or lymphadenopathy for 4 months. During the admission, she complained of nausea, vomiting and dysarthria. On the MRI examination, we found a multi-focal solid lesion on her brain. The histopathological findings of the biopsy from her abdominal skin lesion showed a septal and lobular, histiocytic panniculitis with bean bag cells and atypical lymphoid cells identified as NK like T-cells and also dense diffuse infiltrates localized in the lower dermis and subcutaneous tissue, with minimal epidermal and upper dermal infiltrates without destructive change of blood vessels. The infiltrating atypical lymphoid cells expressed the phenotype of LCA, CD45RO, CD3, CD8, CD56 and also positive for EBV by in situ hybridization. Our case showed a clonal TCR gamma gene rearrangement by polymerase chain reaction. She was subjected to a course of treatment(cyclophosphamide, vincristine, prednisolone and radiation therapy) under the diagnosis of SPTCL. but died of sepsis due to urinary tract infections after 2 months.

A Case of CD7+, CD4-, CD8-, CD3-acute T cell lymphoblastic leukemia / 대한임상병리학회지

A CD7 positive acute leukemia, lacking CD4, CD8, CD3, CD13 and CD33 expression may include 4 categories; acute T-cell leukemia, mixed lineage leukemia, acute undifferentiated leukemia and CD7 positive acute myeloid leukemia. Therefore, the expression of cyCD3 or the presence of TCR gene rearrangement can make the diagnosis of acute T-cell leukemia. We report a patient with acute T-cell lymphoblastic leukemia, showing CD7+, CD4-CD8-, and CD3-expression and TCR gamma gene rearrangement.

The Value of PCR Detection of TCR gamma Gene Rearrangement in the Diagnosis of Early Mycosis Fungoides / 대한피부과학회지

BACKGROUND: The diagnosis and differential diagnosis of mycosis fungoides(MF) is often difficult, clinically and histologically. A variety of inflammatory dermatoses may be seen with similar clinical and histological features. Given the limitation of histologic diagnosis in the early MF, an ancillary method to demonstrate a clonal T cell proliferation would be helpful. Recent attempts to enhance diagnostic sensitivity have involved T-cell receptor(TCR) gene rearrangement studies, using polymerase chain reaction(PCR) technique. OBJECTIVE: The purpose of this study was the value of PCR detection of TCR gamma gene rearrangement in the diagnosis of early MF. MATERIALS AND METHODS: Thirty-two cases of paraffin embedded tissue(PET) from patients of early MF were investigated for the presence of TCR gamma gene rearrangement using a nested PCR technique and analyzed by polyacrylamide gel electrophoresis(PAGE). As a control, PET from patients of 6 psoriasis, 3 lichen planus, 1 neurodermatitis, and 1 allergic contact dermatitis were tested. RESULTS: Monoclonal TCR gamma gene rearrangement was detected in 28 out of 32 (88%) patients with early MF and in none out of 11 patients with inflammatory skin diseases. CONCLUSION: TCR gamma gene rearrangement studies on lesional skin using PCR may be helpful as an adjunct to the histopathologic diagnosis of early MF.

Histopathologic Features and Immunophenotype of 19 Primary Cutaneous Lymphomas

The diagnosis of primary cutaneous lymphoma is based on a combination of clinical, histological, immunophenotypic and genetic criteria. Nineteen cases of primary cutaneous lymphomas were studied for clinicopathologic, immunophenotypic, and genetic features. Seventeen (89%) cases were T cell origin and two cases (11%) were B cell origin. CD30-positive cutaneous lymphoproliferative disorder was the most frequent subtype, occupying 42% (8 cases) of the cases. CD8 was positive in 5 cases consisting of 3 cutaneous T cell lymphomas and 2 anaplastic large cell lymphomas. CD4 was positive in 2 cases of mycosis fungoides and 3 cases of lymphomatoid papulosis. Six (67%) of 9 cases of cutaneous T cell lymphoma were positive for TIA-1. Ten (83%) out of 12 cases showed clonal rearrangements of TCR gamma genes, however, one T/NK cell lymphoma and one anaplastic large cell lymphoma did not. EBV association was detected only in T/NK cell lymphomas among 10 cases examined. In conclusion, our study showed higher proportion of CD30-positive lymphoproliferative disorders and less frequent mycosis fungoides in Korea compared to the incidences in Western countries. Our immunostaining results suggested that mycosis fungoides and lymphomatoid papulosis are CD4-positive T cell origin, however, the remaining primary cutaneous T cell lymphoma is predominantly CD8-positive cytotoxic T cell origin.