OBJECTIVES: Multiple myeloma (MM) is a hematologically malignant clonal plasma cell disease. This study aims to explore the association between immunophenotypes and prognosis in patients with MM, to determine whether the expression of CD45 and CD200 is related to the prognosis of newly diagnosed MM (NDMM) patients, and to evaluate the significance of the combined expression of CD45 and CD200 in NDMM. METHODS: A total of 123 NDMM patients admitted to Shengjing Hospital of China Medical University from July 2015 to August 2019 were enrolled. Five key immunophenotypic markers (including CD38, CD138, CD45, CD56, and CD200) were screened through flow cytometry and identified using random forest analysis and univariate Cox regression analysis. Patients were divided into 3 groups: Group A, CD45 and CD200 double-positive; Group B, CD45 or CD200 single-positive; Group C, CD45 and CD200 double-negative. Kaplan-Meier curves were used to analyze overall survival (OS) and progression-free survival (PFS) across groups. Multivariate Cox regression was performed to evaluate prognostic factors, and a nomogram was constructed based on these results. RESULTS: The OS and PFS of single-positive groups for CD38, CD138, CD45, CD56, and CD200 were all shorter than those of their respective single-negative groups (all P<0.05). Significant differences were observed in OS (P<0.001) and PFS (P=0.001) among Groups A, B, and C. Group A had shorter OS and PFS (all P=0.001) compared to the Group B+C (cases from Group B and Group C were combined). CD45 and CD200 double-positive was an independent prognostic factor for NDMM [hazard ratio (HR)=2.178, 95% confidence interval (CI) 1.048 to 4.529; P=0.037]. The nomogram and calibration curves constructed from multivariate Cox regression analysis demonstrated good concordance (concordance index=0.706; 95% CI 0.661 to 0.751). CONCLUSIONS NDMM patients with double-positive expression of CD45 and CD200 have significantly shorter OS and PFS. Compared with the use of either marker alone, the combined assessment of CD45 and CD200 may provide better prognostic stratification for MM patients.
Humans ; Multiple Myeloma/metabolism* ; Male ; Female ; Middle Aged ; Antigens, CD/metabolism* ; Prognosis ; Leukocyte Common Antigens/metabolism* ; Aged ; Adult ; Immunophenotyping ; Nomograms ; Biomarkers, Tumor ; Clinical Relevance

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 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

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 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

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 analyze the clinicopathologic and immunohistochemical features of nodular histiocytic/mesothelial hyperplasia (NHMH) and to improve the knowledge of this disease.

METHODSSeven cases of NHMH were collected and the clinicopathologic and immunohistochemical data were analyzed with review of the literature.

RESULTSSeven male patients aged from 1.5 to 5.0 years (mean 2.8). The main clinical symptom was an inguinal mass.Grossly, main pathological changes were the mural nodule or free nodule in lumen, with diameter of 0.1-0.5 cm.Histologically, the tumor cell morphology was relatively single, cohesive polygonal or oval cells which were arranged in solid sheets or nests, usually with ovoid or deeply grooved nuclei and a moderate amount of pale pink cytoplasm in the nodular collection area. The nuclei had delicate chromatin and no obvious atypia, and mitosis was incidentally found. A few scattered lymphocytes were found in the stroma. The cyst wall was lined by a single layer of mesothelial cells.Immunohistochemically, the most cells in nodular lesion were strongly positive for the histiocytic marker CD68, vimentin and α1-antichymotrypsin, while lining mesothelial cells on the wall were positive for calretinin, MC, WT1, CK5/6, CKpan and EMA.

CONCLUSIONSNHMH is a rare and benign tumor-like lesion, and easy to be misdiagnozed, which should be distinguished from neuroendocrine tumors, Langerhans cell histiocytosis, seminoma, mesothelioma and so on. The correct diagnosis of this lesion depends on the clinical characteristics, morphology and immunohistochemistry.

Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Calbindin 2 ; metabolism ; Child, Preschool ; Diagnosis, Differential ; Epithelium ; metabolism ; pathology ; surgery ; Histiocytes ; metabolism ; pathology ; Histiocytosis, Langerhans-Cell ; metabolism ; pathology ; Humans ; Hyperplasia ; metabolism ; pathology ; surgery ; Infant ; Leukocyte Common Antigens ; metabolism ; Male ; Mesothelioma ; metabolism ; pathology ; Mucin-1 ; metabolism ; Neuroendocrine Tumors ; metabolism ; pathology ; Seminoma ; metabolism ; pathology ; Vimentin ; metabolism ; WT1 Proteins ; metabolism ; alpha 1-Antichymotrypsin ; metabolism

BACKGROUNDPrimary systemic light chain amyloidosis (AL) is a rare plasma cell disease, our purpose was to analyze the immunophenotypic characteristics of the plasma cells in bone marrow in AL patients, and explore whether the detection of abnormal plasma cell clones in bone marrow by flow cytometry (FCM) could be used as an important indicator of AL diagnosis.

METHODSFresh bone marrow samples were collected from 51 AL, 21 multiple myeloma (MM), and 5 Waldenström's macroglobulinemia (WM) patients. The immunophenotype of bone marrow cells were analyzed and compared by FCM using a panel of antibodies including CD45, CD38, CD138, CD117, CD56, and CD19.

RESULTSIn AL, light chain restriction could be identified in 31 cases (60.9%), in which the λ light chain restriction was found in 24 cases (77.4%). In MM, κ light chain restriction was found in 13 cases (61.9%), and λ light chain restriction in eight cases. CD45 on abnormal plasma cells was negative to weakly positive in both AL and MM, but was positive to strongly positive in WM. In the bone marrow plasma cells of the 51 AL, 78.4% were CD56+, 68.6% were CD117+, and 88.2% were CD19-. While in the 21 MM cases, 66.7% were CD56+, 38.1% were CD117+, and 90.4% were CD19-. The plasmacytoid lymphocytes in the five WM patients were CD19+ and CD56-, CD117-.

CONCLUSIONDetection of abnormal plasma cell clones in bone marrow by FCM is valuable for the diagnosis of AL.

Adult ; Aged ; Aged, 80 and over ; Amyloidosis ; immunology ; metabolism ; CD56 Antigen ; metabolism ; Female ; Flow Cytometry ; Humans ; Immunoglobulin Light-chain Amyloidosis ; Immunoglobulin lambda-Chains ; metabolism ; Immunophenotyping ; Leukocyte Common Antigens ; metabolism ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; metabolism ; Proto-Oncogene Proteins c-kit ; metabolism ; Waldenstrom Macroglobulinemia ; immunology ; metabolism

Antibodies, Monoclonal, Murine-Derived ; therapeutic use ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD5 Antigens ; metabolism ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Follow-Up Studies ; Heart Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Heart Ventricles ; Humans ; Interferon Regulatory Factors ; metabolism ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; PAX5 Transcription Factor ; metabolism ; Prednisone ; therapeutic use ; Vincristine ; therapeutic use

OBJECTIVETo observe the Immune No. 2 (2) on the immune reconstitution in patients with human immunodeficiency virus or acquired immune deficiency syndrome (HIV/AIDS) after highly active antiretroviral therapy (HAART).

METHODSA randomized, double-blind, placebo-controlled clinical trial was designed. 233 patients failing immune reconstitution after HAART were randomly divided into treatment group (116 cases) and control group (117 cases), respectively using Immune No. 2 plus HAART and placebo combined with HAART for 6 months. CD4, CD45RA, CD45RO cell numbers, as well as the symptoms, signs and integral improvement rates were observed in order to evaluate the immune reconstitution efficiency.

RESULTSafter the intervention for 1 month, the effective rate of the treatment group (18.97%, 22/116) was significantly higher than that of the control group (9.40%, 11/117) (P=0.02); 3 months after treatment, the effective rate of the treatment group (27.59%, 32/116) was no difference from that of the control group (22.22%, 26/117) (P=0.31); 6 months after treatment, the effective rate of the treatment group (34.48%, 40/116) was significantly superior to the control group (21.37%, 25/117) (P=0.02). CD4, CD45RA, CD45RO count of the treatment group was significantly higher than that of the control group (P<0.05). The total score of symptoms and signs in the treatment group was significantly lowered compared with the control group (P=0.02), and the improvement of fatigue, muscle and joint pain, pruritus and shortness of breath in the treatment group was better than the control group (P<0.05).

CONCLUSIONImmune No. 2 can effectively improve the numbers of CD4 cells and its subgroups, as well as the main clinical symptoms and signs of patients after HAART, thereby promoting the immune reconstitution.

Acquired Immunodeficiency Syndrome ; drug therapy ; immunology ; Adult ; Antiretroviral Therapy, Highly Active ; CD4 Lymphocyte Count ; Double-Blind Method ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Leukocyte Common Antigens ; metabolism ; Male ; Placebos ; Viral Load