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.
Diagnosis ; Genes, T-Cell Receptor ; Genes, T-Cell Receptor gamma ; Humans ; Leukemia ; Leukemia, Myeloid, Acute ; Precursor Cell Lymphoblastic Leukemia-Lymphoma* ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; T-Lymphocytes

Genes, T-Cell Receptor gamma ; Humans ; Lymphomatoid Papulosis ; classification ; diagnosis ; Skin Neoplasms ; classification ; diagnosis

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.
Diagnosis ; Genes, T-Cell Receptor gamma ; Herpesvirus 4, Human ; Incidence ; Korea ; Lymphoma* ; Lymphoma, Large-Cell, Anaplastic ; Lymphoma, T-Cell, Cutaneous ; Lymphomatoid Papulosis ; Lymphoproliferative Disorders ; Mycosis Fungoides

OBJECTIVETo investigate the distribution and clonality of T cell receptor (TCR) Vγ and Vδ subfamily in peripheral blood of patients with allergic rhinitis before and after 1 year treatment with immunotherapy.

METHODSThe specific IgE and the complementary determinant region 3 (CDR3) of TCR V γ (I-III) and Vδ(1-8) subfamily genes in mononuclear cells were amplified from 10 effective cases of allergic rhinitis before and after 1 year treatment with immunotherapy, to observe the distribution and utilization of TCR Vγ and Vδ repertoire. The positive PCR products were further labeled with RT-PCR and analyzed by gene scan technique to determine the CDR3 size and evaluate the clonality of the detectable TCR Vγ and Vδ T cells. Peripheral blood of 10 healthy adults served as controls.

RESULTSAll symptoms were significantly improved after 1 year specific immunotherapy, but no changes were seen in specific IgE [(22.89 ± 9.60) kU/L before treatment, (19.62 ± 7.63) kU/L after treatment, Z = 1.051, P > 0.05]. No statistically significant differences of expression levels of the TCR V&gamma; I-III subfamily genes were found between patients with allergic rhinitis normal control group (t value were -0.679, -0.516, -0.808, all P > 0.05), but significantly decreased after 1 year treatment. There were statistically significant differences of expression levels of the TCR V&gamma;I-II subfamily genes before and after treatment (t value were -2.904, -2.217, all P < 0.05). 5.30 ± 0.82, 4.90 ± 0.57 and 5.20 ± 1.40 out of TCR Vδ (1-8) subfamilies were selectively expressed in T cells in patients with allergic rhinitis before and after 1 year treatment and normal control group, predominantly for TCR Vδ 1, 2, 3 and 6. The TCR Vδ 6 subfamily was found to have statistically significant differences in these groups (Fisher's Exact Test, P < 0.05). Compared with the normal control group and the allergic rhinitis group before treatment, a significant higher frequency of Vδ 6 oligoclonal was identified in T cells in patients with allergic rhinitis after 1 year treatment.

CONCLUSIONSThere was difference in the expression levels of the TCR V&gamma; I-III subfamily genes and distribution and clonality of TCR V&gamma; and Vδ subfamily T cells in peripheral blood of patients with allergic rhinitis before and after 1 year treatment. Specific immunotherapy can be effective in alleviation of the symptom in patients with allergic rhinitis during the early stage, possibly by inducing TCR &gamma;δ T cells, especially the TCR Vδ6 subfamily, and possibly no significant relativity between symptom and specific IgE.

Adolescent ; Adult ; Case-Control Studies ; Female ; Genes, T-Cell Receptor ; Humans ; Immunoglobulin E ; blood ; Immunotherapy ; Male ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; immunology ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; immunology ; Rhinitis ; genetics ; immunology ; therapy ; Young Adult

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.
Cell Proliferation ; Dermatitis, Allergic Contact ; Diagnosis* ; Diagnosis, Differential ; Gene Rearrangement ; Genes, T-Cell Receptor gamma* ; Humans ; Lichen Planus ; Mycosis Fungoides* ; Neurodermatitis ; Paraffin ; Polymerase Chain Reaction* ; Psoriasis ; Skin ; Skin Diseases ; T-Lymphocytes

OBJECTIVETo 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.

METHODSBronchoalveolar 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.

RESULTSThe 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.

CONCLUSIONSThe 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.

Adult ; Asthma ; genetics ; immunology ; Bronchoalveolar Lavage Fluid ; cytology ; Case-Control Studies ; Cell Proliferation ; Cytokines ; metabolism ; Female ; Genes, T-Cell Receptor delta ; genetics ; Genes, T-Cell Receptor gamma ; genetics ; Humans ; Immunoglobulin Variable Region ; genetics ; Lung ; immunology ; Male ; Middle Aged ; Polymorphism, Genetic ; T-Lymphocyte Subsets ; immunology ; metabolism ; Th1-Th2 Balance

OBJECTIVETo investigate the clinical and laboratory features of chronic eosinophilic leukemias (CEL) and hypereosinophilic syndrome (HES).

METHODSThe 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.

RESULTSOf 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.

CONCLUSIONS(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.

Adult ; Female ; Gene Rearrangement ; Genes, T-Cell Receptor gamma ; genetics ; Humans ; Hypereosinophilic Syndrome ; diagnosis ; genetics ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Mutation ; Receptor, Platelet-Derived Growth Factor alpha ; genetics ; Retrospective Studies ; Young Adult ; mRNA Cleavage and Polyadenylation Factors ; genetics

OBJECTIVETo investigate the rate of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphomas (NHL) and its pathogenesis and pathologic significance.

METHODSPCR 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.

RESULTSCombination 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.

CONCLUSIONDual rearrangements in NHL are not rare and have no relationship with proliferation index.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carrier Proteins ; genetics ; Cell Proliferation ; Child ; Female ; Gene Rearrangement ; Genes, T-Cell Receptor gamma ; genetics ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Lymphoma, B-Cell ; genetics ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; genetics ; metabolism ; pathology ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Polymerase Chain Reaction ; Receptors, Antigen ; genetics ; Young Adult

OBJECTIVETo investigate effect of tumor-specific T cell receptor gene transfection on memory T cell differentiation in vitro.

METHODSTCRVbeta7.1 gene was transferred into peripheral blood mononuclear cells (PBMCs) obtained from healthy adults, and the expression of Vbeta7.1 was detected by flow cytometry before and after the transfection. Memory T cell differentiation was induced by stimulation with the hepatocarcinoma cell line BEL-7402 in vitro. The expression of surface molecules CD45RO, CD45RA and CCR7 was analyzed by flow cytometry to identify the phenotype and subsets of the memory T cells. Fluorescence-activated cell sorting was performed to detect the apoptosis of the tumor cells, and enzyme-linked immunoabsorbent assay was used to determine the production of interferon-gamma (IFN-gamma) for assessing the immune function of the memory T cells.

RESULTSFlow cytometry showed that TCRVbeta7.1 gene was efficiently expressed after transfection. After stimulation by the tumor cells in vitro, the expression of CD45RO in TCRVbeta7.1 gene-modified T cells increased gradually, and analysis of the coexpression of CD45RA and CCR7 revealed that the effector memory T cells constituted the majority of the differentiated memory T cells. The apoptotic rate of the tumor cells induced by the T cells increased significantly with also obviously increased INF-gamma secretion in the memory T cells.

CONCLUSIONTumor-specific TCRVbeta7.1 gene transfection can promote the differentiation of the memory T cells, the majority of which belongs to effector memory T cells that perform immune functions by inducing apoptosis and cytokine secretion.

Adult ; Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Genes, T-Cell Receptor alpha ; genetics ; Humans ; Immunologic Memory ; immunology ; Interferon-gamma ; metabolism ; Leukocyte Common Antigens ; metabolism ; Liver Neoplasms ; genetics ; metabolism ; pathology ; T-Lymphocytes ; cytology ; immunology ; metabolism ; Transfection

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.
Abdomen ; Aged ; Arthralgia ; Biopsy ; Blood Vessels ; Brain ; Carcinogenesis ; Central Nervous System* ; Dermis ; Diagnosis ; Dysarthria ; Epstein-Barr Virus Infections ; Female ; Fever ; Genes, T-Cell Receptor gamma ; Herpesvirus 4, Human ; Humans ; In Situ Hybridization ; Lymphatic Diseases ; Lymphocytes ; Lymphohistiocytosis, Hemophagocytic ; Lymphoma ; Lymphoma, T-Cell* ; Lymphoma, T-Cell, Peripheral ; Magnetic Resonance Imaging ; Nausea ; Panniculitis ; Phenotype ; Polymerase Chain Reaction ; Prednisolone ; Sepsis ; Skin* ; Subcutaneous Fat ; Subcutaneous Tissue ; T-Lymphocytes* ; Thigh ; Ulcer ; Upper Extremity ; Urinary Tract Infections ; Vincristine ; Vomiting ; Weight Loss