Objective: To study the relationship between the onco-immunological and morphologic characteristics of lymphoepithelioma-like carcinoma (LELC) and peripheral blood lymphocyte subtypes and its clinical significance. Methods: The pathologic and clinical data of 117 LELC patients who were admitted to the Tumor Hospital of the University of Chinese Academy of Sciences from 2006 to 2018 were collected. The histological classification was based on previously reported morphological classification method. The onco-immunological and morphologic characteristics of the tumors such as lymphoid follicle formation and interstitial fibrous hyperplasia, patient's peripheral blood lymphocyte subtypes and prognosis data were collected. The relationship between various factors and their impact on prognosis were analyzed. Results: There were 117 patients, including 61 females and 56 males. The male to female ratio was 0.9∶1.0. The age of onset was 24-89 years (median 52 years). Primary sites included head and neck (68 cases), lungs (26 cases), stomach (15 cases), and others (eight cases). Morphologically, 54 cases were type Ⅰ, 62 cases were type Ⅱ, and one case could not be classified. The onco-immunological and morphologic features of the LELC tumors showed a continuous spectrum. Interstitial TILs were noted from focally to diffuse, and the interstitial fibrous tissues were from hardly visible to obvious sclerotic. Formation of lymphoid follicles was seen in 42 patients; obvious fibrosis was seen in 31 cases. Data of peripheral blood lymphocyte subtyping by flow cytometry were available in 73 cases. These data included CD3⁺total T cells, CD3⁺CD4⁺helper T cells, CD3⁺CD8⁺cytotoxic T cells, CD3^-CD56⁺natural killer (NK) cells, CD3^-CD19⁺B cells, CD4⁺CD45RA^-T helper induction subgroup, CD4⁺CD45RA⁺ T suppression induction subgroup, CD4⁺CD45RO⁺memory T cell subgroup, CD45RA⁺CD45RO⁺activated T cell subgroup, CD8⁺CD38⁺activated cytotoxic T cell, and CD25⁺lymphocytes and CD44⁺lymphocyte. The proportion of lymphocytes of each subtype was normal in most patients, but the proportion of CD44⁺lymphocytes in 61 cases (83.6%) was increased; the proportion of T cell suppression induced subgroups was decreased in 53 cases (72.6%). Correlation analysis found a significant correlation between clinical stage and NK cells (P=0.023); tumor histologic type and cytotoxic T cells were significantly positively correlated (P=0.012); while tumor cell morphologic differentiation was significantly related to total T cells (P=0.003) and NK cells (P=0.026); Formation of interstitial lymphoid follicles was positively correlated with memory T cell subsets (P=0.025); Tumor interstitial fibrosis was significantly positively correlated with T suppression-induced subpopulations (P=0.004), and was significantly negatively correlated with total T cells (P=0.023) and with the expression of CD44 adhesion molecules (P=0.003). Survival analysis found that lymphoid follicle formation was a favorable prognostic factor for LELC (P=0.001). Conclusions: The onco-immunological and morphologic features in LELC show a continuous spectrum; the tumor clinicopathological characteristics and onco-immunological morphology are closely related to peripheral blood T lymphocyte subtypes, and the formation of interstitial lymphoid follicles is a favorable prognostic factor for LELC.
Adult ; Aged ; Aged, 80 and over ; Carcinoma/metabolism* ; Female ; Fibrosis ; Flow Cytometry ; Humans ; Killer Cells, Natural/metabolism* ; Leukocyte Common Antigens/metabolism* ; Male ; Middle Aged ; Young Adult

OBJECTIVE: To investigate the predictive value of CD45CD117 phenotype-abnormal cells (hereinafter referred to as "abnormal cells") for relapse and prognosis in adult patients with acute myeloid leukemia (AML) within 2 weeks after the first complete remission (CR1). METHODS: The clinical data of patients with newly diagnosed AML (non-acute promyelocytic leukemia) admitted in our department from July 1, 2014 to June 30, 2017 were analyzed retrospectively, and the relationship between clinical features at the initial diagnosis and the abnormal phenotype cells of CD45CD117 within 2 weeks after CR1 with the prognosis were analyzed. RESULTS: A total of 91 patients with CD45CD117 abnormal cells were detected. The median age was 51 years old, the median WBC count was 11.60×109/L, and the median ratio of bone marrow blast cells was 0.35 at initial diagnosis. According to the FAB classification, 1 (1.1%), 7 (7.7%), 38 (41.7%), 20 (22.0%), 21 (23.1%) and 4 (4.4%) patients were classifice as M0, M1, M2, M4, M5, and M6, respectively. According to the NCCN risk stratification, 30 (33.0%), 51 (56.0%), and 10 (11.0%) patients were determined as good, moderate, and poor prognosis, respectively. The median ratio of abnormal cells within 2 weeks after CR1 was 1.8500 (0.0236-8.0000)%. The median time from initiation of induction therapy to the acquisition of CR was 46 days, median recurrence-free survival time was 319 days, and median overall survival time was 352 days. A total of 45 patients relapsed, of which 14 died; 46 patients did not relapse, of which 3 died. The cutoff of abnormal cells by receiver operating characteristic curve (ROC) analysis was 2.055% (Se=0.733，Sp=0.761). The abnormal cell ratio was＞2.055% in 44 patients, the median ratio of abnormal cells was 3.075%, among which 33 patients relapsed and 12 patients died; the abnormal cell ratio was ＜2.055% in 47 patients, the median ratio of abnormal cells was 1.150%, 12 patients relapsed and 5 patients died. Regression analysis showed that WBC count＞50×10/L and abnormal cell ratio＞2.055% were independent risk factors for recurrence. The abnormal cell ratio＞2.055% group had a 2-year RFS rate of 54.3% and a 2-year OS rate of 52.8%. The abnormal cell ratio＜2.055% group had a 2-year RFS rate of 86.6% (P=0.018), and a 2-year OS rate of 85.3% (P＜0.05). CONCLUSION For adult AML patients, CD45CD117 phenotypical abnormal cells ratio＞2.055% within 2 weeks after CR1 is an independent risk factor for recurrence, which also is an dverse factor for RFS and OS.
Humans ; Leukemia, Myeloid, Acute ; Leukocyte Common Antigens ; Leukocyte Count ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-kit ; Remission Induction ; Retrospective Studies

Objective: To explore the expression of CD45 in newly diagnosed multiple myeloma (MM) and its relationship with clinical efficacy and prognosis. Methods: This study retrospectively analyzed expression and distribution of CD45 in 130 cases of newly diagnosed MM, comparing clinical efficacy and prognosis in CD45(+)/CD45(-) groups. Results: ①The CD45(+) group was 33 cases (25.38%) , and CD45(-) group was 97 cases (74.62%) . ②The objective remission rate (ORR) of CD45(+) and CD45(-)group was 33.33% and 64.95%, respectively. The difference was statistically significant (P=0.002) . For patients in Bortezomib regimen, the ORR of CD45(+) and CD45(-) group was 35.71% and 66.25%, respectively. The difference was statistically significant (P=0.005) . ③The median progress free survival (PFS) of CD45(+) group and CD45(-) group was 29.8 (95%CI 10.0-59.0) months vs 34.5 (95%CI 6.0-69.0) months (χ(2)=14.59, P<0.001) and the median overall survival (OS) was 32.5 (95%CI 10.0-68.0) months vs 37.6 (95%CI 6.0-78.0) months (χ(2)=11.42, P=0.001) , respectively. Among the patients in bortezomib regimen, The median PFS and median OS of CD45 (+) group and CD45(-) group were 30.3 (95%CI 10.0-59.0) months vs 36.3 (95%CI 6.0-69.0) months (χ(2)=14.75, P=0.001) and 34.0 (95%CI 10.0-68.0) months vs 39.5 (95%CI 6.0-78.0) months (χ(2)=10.62, P=0.001) . ④Cox risk regression model analysis showed that serum creatinine≥176.8 μmol/L (HR=5.078, 95%CI 1.744-14.723, P=0.001) , CD45 positive (HR=14.504, 95%CI 0.168-0.42, P=0.001) , LDH≥220 IU/L (HR=1.308, 95%CI 1.16-2.417, P=0.015) were independent risk prognostic factors. Conclusion: CD45 expression is a risk prognostic factor of MM patients. Bortezomib did not improve the poor prognosis of CD45(+) MM patients.
Antineoplastic Combined Chemotherapy Protocols ; Disease-Free Survival ; Humans ; Leukocyte Common Antigens/metabolism* ; Multiple Myeloma/diagnosis* ; Prognosis ; Retrospective Studies

The patient-derived tumor xenografts (PDX) model is an animal model established by directly engrafting fresh tumor tissue of patients into immunodeficiency mice after surgery or biopsy, which plays an important role in the study of tumor biology. However, the transformation of PDX into lymphoma limits the application of this model. The characters of this transformation include that epithelial tumors origin, predorminance of B-cell lymphomas, lost of architectural feature of primary tumor, absence of epithelial tumor markers, and CD45 and CD20 expression. That were characteristics of human B lymphocytes, and possible infection of Epstein-Barr virus(EBV). The biology of primary tumor, EBV infection, inflammation infiltration in primary tumors and the host immune status are the main related factors in this transformation. Therefore, selective xenograft by the detection of EBV infection and inflammation infiltration in primary tumors may be effective methods to prevent lymphomagenesis.
Animals ; B-Lymphocytes ; Disease Models, Animal ; Epstein-Barr Virus Infections ; Herpesvirus 4, Human ; Heterografts ; Humans ; Leukocyte Common Antigens ; Lymphoma ; Lymphoma, B-Cell ; Mice ; Mice, SCID ; Neoplasms ; Transplantation, Heterologous

OBJECTIVETo investigate the immunophenotype of leukemia promyelocytes (LP) in bone marrow of patients with acute promyelocytic leukemia (APL) and to explore their characteristics and significance.

METHODSThe immunophenotypes of leukemia cells in 43 patients with APL were analyzed by means of 4 color immunophenotypes; the cell population in which CD45 strength localized at 10(2) and the SSC strength locatized at 10(2) was defined as R3, the cell population in which CD45 strength localized at 10(3) and the SSC strength localized at 10(2) was defined as R5, moreover the ratio of positive cells >80% was defined as strong positive expression, the ratio of positive cells between 20%-80% was difined as weak positive expression, the ratio of positive cells <20% was difined as negative by gating method of CD45/SSC.

RESULTSThere was a abnormal cell population (R3) in 79.07% cases; the immunophenotypes of R3 was cheracteried by high SSC, weaker expression of CD45, the rate of CD38, CD9 and CD13 all was 100%, moreover their bright expression (>80%) was 86.05%, 90.70% and 86.05%, respectively; the positive expression rate of CD33, CD117 and CD64 was 97.67%, 95.35% and 83.80% respectively, moreover thier bright expression was 84.04%, 69.77% and 30.23% respectively; the CD15 was weakly expressed in 39.53% cases, the CD34 and HLA-DR were weakly expression in 16.28% and 6.98% cases respectively. All the cases did not express CD116. There were 2 cell populations (R3 and R5) in 20.93% cases, the immunophenotypic features of R3 were cosistant with above mentioning, while the immunophenotypes of R5 were lower than those of R3 SSC; the fluorescence intensity of CD45 was higher, but lower than that in normal lymphycytes, the positive rate of CD9, CD13, MPO was 100%, moreover thier fluorescence intensity was high; they did not expressed CD123, CD25, CD22, CD4, CD64 and CD14. Thereby it can be concluded that the typical immunophenotypes is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-) in APL. There was a special immunophenotype in the APL with basophilic granules. Conclusoin: APL has a characteristic immunophenotypic profile, whose typical immunophenotype is characterized by CD13(+) CD9(+) CD38(+) CD33(+) CD117(+) CD64(+) CD11b(-) CD34(-) HLA-DR(-). The special immunophenotype exists in the APL with basophilic granules. Flow cytometric immunophenotyping may be a useful for rapid recognition of APL and has significant for prognosis.

Antigens, CD ; metabolism ; Cell Count ; Flow Cytometry ; Granulocyte Precursor Cells ; classification ; HLA-DR Antigens ; metabolism ; Humans ; Immunophenotyping ; Leukemia, Promyelocytic, Acute ; classification ; immunology ; Leukocyte Common Antigens ; metabolism ; Prognosis

OBJECTIVETo investigate the clinical significance of memory T cells (CD45RO⁺ T) and the initial T cells (CD45RA⁺ T) distribution in peripheral blood of patients with peripheral T-cell lymphoma (PTCL).

METHODSA total of 27 patients diagnosed as PTCL in our hospital from February 2010 to February 2014 were collected in this study; whereas 30 healthy people were enrolled as control. The distribution of CD45RO⁺ T and CD45RA⁺ T cells were detected seperately in each group, and the results were analysed further.

RESULTSThe expression of T cell antigens in lymphnodes of PTCL patients were diverse: the CD4⁺ T cells were the main immune phenotype, while the B cell-related antigen was not expressed. The CD4⁺/CD8⁺, CD4⁺ CD45RO⁺ T in the peripheral blood of PTCL patients were significantly lower than that in normal group (P < 0.05); while the CD4, CD45RA⁺ T, CD8⁺ CD45RA⁺ T and CD8⁺ CD45RO⁺ T were significantly higher than that in normal group (P < 0.05). The patients in stage I/II had higher CD4⁺/CD8⁺, CD4⁺ CD45RO⁺ T than those in stage III/IV (P < 0.05), whereas the CD4⁺ CD45RA⁺ T, CD8⁺ CD45RA⁺ T and CD8⁺ CD45RO⁺ T were significantly lower than those in the stage III/IV patients (P < 0.05).

CONCLUSIONThe distributions of CD45RA⁺ T and CD45RO⁺ T in PTCL patients are quite different, and the corresponding treatment might be adopted according to the different distribution of these cells, so as to improve the diagnosis and prognosis of PTCL.

CD4-Positive T-Lymphocytes ; Humans ; Leukocyte Common Antigens ; Lymphoma, T-Cell, Peripheral ; Phenotype ; Prognosis

OBJECTIVETo explore the clinical and laboratory characteristics of mutiple myeloma patients with CD20 expression.

METHODSReview the data of mutiple myeloma patients and analyze the clinical and laboratory characteristics of CD20 positive patients, compared with CD20 negative patients.

RESULTS(1)Totally 465 cases of newly-diagnosed MM were collected with CD20 expression status detected by multi-color flow cytometry. Sixty two patients (13.3%) were CD20 positive and the others were negative. (2)No statistical differences were found between CD20 positive and negative groups about the sex ratio, age predominance, D-S staging, ISS staging, renal insufficiency rate, platelet count, LDH level and classifications by paraprotein(all P value>0.05). (3)Compared with those of CD20 negative patients, the hemoglobulin value(74.5 g/L vs 83.5 g/L, P=0.021), extramedullary involvement rate (3.5% vs 13.7%, P=0.029), CD56-positive rate(36.7% vs 68.8%,P=0.000), t(4;14)translocation rate(2.4% vs 24.0%, P=0.001) in CD20 positive patients were lower statistically. (4)Compared with those of CD20 negative patients, the percentage of plasma cells (0.400 vs 0.295, P=0.045) by marrow smear differential counting, the percentage of myeloma cells(20.0% vs 6.8%, P=0.000) by multi-color flow cytometry analysis, CD45-positive rate(12.1% vs 4.5%, P=0.018), CD79a-positive rate(9.8% vs 1.5%, P=0.013) and t(11;14) translocation rate(60.5% vs 14.4%, P=0.000)in CD20 positive patients were higher statistically. (5)There was no statistical differences about the overall response rate (ORR), complete response rate (CRR), TTP(time to progression), PFS(progression free survival) and overall survival (OS) between CD20 positive and negative groups.

CONCLUSIONCD20 positive rate is 13.3% in multiple myeloma pateints according to our data. CD20 poaitive myeloma were prone to residing in bone marrow and affecting erythropoiesis. Atypical immunophenotypes were more common, and the incidence of t(11;14) were increased markedly while that of t(4;14)were rare for CD20 positive multiple myeloma.

Antigens, CD20 ; Bone Marrow ; Disease Progression ; Disease-Free Survival ; Flow Cytometry ; Humans ; Immunophenotyping ; Leukocyte Common Antigens ; Multiple Myeloma ; Myeloma Proteins ; Plasma Cells ; Remission Induction ; Translocation, Genetic

OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.

RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).

CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.

Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism

OBJECTIVETo clone human CD45 gene PTPRC and establish Hela cells overexpressing recombinant human CD45 protein.

METHODSThe intact cDNA encoding human CD45 amplified using RT-PCR from the total RNA extracted from peripheral blood mononuclear cells (PBMCs) of a healthy donor was cloned into pMD-18T vector. The CD45 cDNA fragment amplified from the pMD-18T-CD45 by PCR was inserted to the coding region of the PcDNA3.1-3xflag vector, and the resultant recombinant expression vector PcDNA3.1-3xflag-CD45 was transfected into Hela cells. The expression of CD45 in Hela cells was detected by flow cytometry and Western blotting, and the phosphastase activity of CD45 was quantified using an alkaline phosphatase assay kit.

RESULTSThe cDNA fragment of about 3 900 bp was amplified from human PBMCs and cloned into pMD-18T vector. The recombinant expression vector PcDNA3.1-3xflag-CD45 was constructed, whose restriction maps and sequence were consistent with those expected. The expression of CD45 in transfected Hela cells was detected by flow cytometry and Western blotting, and the expressed recombinant CD45 protein in Hela cells showed a phosphastase activity.

CONCLUSIONThe cDNA of human CD45 was successfully cloned and effectively expressed in Hela cells, which provides a basis for further exploration of the functions of CD45.

Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; Genetic Vectors ; HeLa Cells ; Humans ; Leukocyte Common Antigens ; genetics ; Leukocytes, Mononuclear ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; Transfection

OBJECTIVETo study the C-myc gene and protein in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) and its relationship to prognosis.

METHODS60 cases of T-LBL/ALL with follow-up data were studied by using immunohistochemical EnVision method for CD1a, CD3, εCD3, CD7, CD10, CD34, CD43, CD45RO, CD99, TDT, CD20, CD23, MPO, Ki-67 and C-myc. 20 cases of reactive lymph nodes were selected as normal control group of C-myc gene and protein. Fluorescence in-situ hybridization (FISH) for C-myc gene (located on chromosome8q24) was performed to detect its breakage and gain.

RESULTSAmong the 60 cases of T-LBL/ALL, immunohistochemistry results showed:the percentages of tumor cells expression of CD1a, CD3, εCD3, CD7, CD10, CD34, CD43, CD45RO, CD99 and TDT were 38.3% (23/60), 75.0% (45/60), 45.0% (27/60), 95.0% (57/60), 36.7% (22/60), 23.3% (14/60), 60.0% (36/60), 41.7% (25/60), 96.7% (58/60) and 93.3% (56/60). Separately, while CD20, CD23 and MPO were all negative. A figure of Ki-67 expression ≤ 80% was found in 36 cases and > 80% was found in 24 cases. The positive rate of C-myc protein was 66.7% (40/60) in 60 cases of T-LBL/ALL, was 0% (0/20) in 20 cases of reactivated lymphoid tissue (χ² = 26.67, P < 0.05). C-myc protein expression was positively correlated with the mediatinal width and Ki-67 index (P < 0.05). Fluorescence in-situ hybridization results showed that among the 60 cases of T-LBL/ALL, C-myc gene with breakage of 8q24 was detected in 6 cases (10.0%), and gains in 11 cases (18.3%). 20 cases of reactive lymph nodes were not occurred breakage and gains of C-myc gene. It is not significant between C-myc gene and protein expression (P > 0.05). In addition, in 60 cases of T-LBL/ALL, 12(20.0%) cases of C-myc protein and genetic abnormalities coexist. Log-rank analysis results: The prognosis of C-myc protein positive group was worse than negative group (P < 0.05). The relationship of C-myc gene and prognosis was not significant (P > 0.05). C-myc protein and genetic abnormality coexist is related with worse prognosis (P < 0.05). COX analysis results show that the C-myc protein positive group may be a independent poor prognosis factors (P < 0.05).

CONCLUSIONSC-myc may play an important role on the development of T-LBL/ALL. It may be a independent prognosis factors.

Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Leukocyte Common Antigens ; Lymph Nodes ; pathology ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-myc ; metabolism