T-Cell Membrane CD45RA (2H4) and CD45RO (UCHL1) Determinants: I, Diverse Patterns of Expression in Mature (Post-Thymic) T-Cell Proliferations

By simultaneous two- and three-colour flow cytometry, this study analysed the expression of membrane CD45RA (2H4) and CD45RO (UCHL1) determinants by normal thymocytes (n = 5) and peripheral blood lymphocyte subpopulations (CD4⁺, n = 21; CD8⁺, n = 12; CD8^dim+, n = 12) and compared these patterns with those of T-cells from representative CD4⁺CD8^- (n = 8), CD4⁺CD8⁺ (n = 2), CD4^-CD8⁺ (n = 10) and CD4^-CD8^- (n = 1) proliferations. These comprised cases of prolymphocytic leukaemia (T-PLL, n = 5), adult T-cell leukaemia-lymphoma (ATLL, n = 2), Sezary Syndrome (SS, n = 4), chronic lymphocytic leukaemia (T-CLL, n = 4), and lymphoproliferative disease of granular lymphocytes (LDGL, n = 5). Normal thymocyte fractions, of which a mean of 85% cells co-expressed membrane CD4 and CD8, were predominantly (mean 89%) 2H4^-UCHL1⁺ with the remaining cells consisting of 2H4^intUCHL1⁺ and 2H4⁺UCHL1^- components. Further analysis showed that virtually all CDla⁺ thymocytes were UCHL1⁺ whereas the CD1a^- fraction comprised similar proportions of both UCHL1^- and UCHL1⁺ subpopulations. Similarly, normal blood CD4⁺, CD8⁺ and CD8^dim+ lymphocytes showed reciprocal CD45RA/CD45RO expression and could be phenotypically grouped into 2H4⁺UCHL1^- 2H4^intUCHL1⁺ and 2H4^-UCHL1⁺ subpopulations. Mean proportions of 48% and 68%, for CD4⁺ and CD8⁺ lymphocytes respectively, showed a composite 2H4⁺UCHL1^- phenotype, whereas the percentage of NK-associated CD8^dim+ cells with this phenotypic pattern was considerably higher (mean, 85%). Normal lymphocyte subpopulations lacking both determinants (2H4^-UCHL1^-) were only rarely noted. Comparing normal patterns of CD45RA/CD45RO expression with those of the T-cell proliferations revealed diverse and abnormal patterns of staining for 3/6 of the CD4⁺CD8^- SS and ATLL, and for 5/5 of the T-PLL (CD4⁺CD8^-, n = 2; CD4⁺CD8⁺, n = 2; and CD4^-CD8⁺, n = 1) cases studied. In contrast, the nine cases of CD4^-CD8⁺ T-CLL and LDGL all showed CD45RA/CD45RO staining patterns similar to that of normal CD8⁺/CD8^dim+ blood lymphocytes (i.e. a predominance of 2H4⁺UCHL1^- cells). Although the variant CD45RA/CD45RO pattern types of the CD4⁺ proliferations did not appear to be related to either the diagnostic category or other phenotypic characteristics, the high proportion of abnormal patterns within this case group suggests that recognition of these abnormalities may be potentially relevant to the differentiation of benign and malignant CD4⁺ proliferations and, in addition, may be of aetiological importance with respect to the diverse acquired defects in immunity commonly seen in patients with such disorders.     

Homing-Associated Cell Adhesion Molecule (H-CAM/CD44) on Human CD34+ Hematopoietic Progenitor Cells

Human CD34⁺ hematopoietic progenitor cells (HPCs) express CD44 and can directly adhere to hyaluronate (HA) via CD44. Furthermore, CD44 may also be involved in the regulation of CD34⁺ HPC proliferation and development. The expression of CD44 molecules on CD34⁺ hematopoietic progenitor cells is significantly lower on bone marrow (BM) CD34⁺ cells compared with circulating CD34⁺ cells in cord blood and peripheral blood. Myeloid and erythroid progenitor cells are found predominantly in CD34⁺CD44^- cell fractions. More interestingly, CD34⁺CD44⁺ cells expressing B-lymphocyte-associated CD10 and CD19 would represent unique B-lymphocyte committed precursors in the BM, which might undergo apoptotic cell death in the early steps of B-cell differentiation.     

Multi-drug resistance mediated by P-glycoprotein overexpression is not correlated with ZAP-70/CD38 expression in B-cell chronic lymphocytic leukemia

ZAP-70 and CD38 expression can identify B-cell chronic lymphocytic leukemia with an inferior clinical outcome. Many groups have investigated the meaning of the expression of these two proteins and the correlation with the bad prognosis in B-CLL. But nobody has investigated the relation between the multidrug resistance mediated by Pgp overexpression (MDR1) and ZAP-70/CD38 coexpression. Forty-one untreated and stage A patients, either ZAP-70⁺CD38⁺ or ZAP-70^-CD38^-, were tested to determine the MDR1 status. MDR1 was observed in 41% of CLL ZAP-70⁺CD38⁺ and in 37% of CLL ZAP-70^-CD38^-. The difference was not significant (p = 0.745). Patients with ZAP-70 and CD38 positive CLL can not be candidates for MDR1 antagonists.     

Clinicopathological correlates of plasma cell CD56 (NCAM) expression in multiple myeloma

The aim of this prospective, long-term study was to define the flow cytometric characteristics of plasma cell CD56 expression as well as determine the clinical characteristics of 204 multiple myeloma (MM) patients and 26 plasma cell leukemia (PCL) patients with regard to CD56 expression. CD56 expression intensity was determined by measurement of antigen molecules on the cell defined as Antibodies Bound per Cell (ABC) and calculation of Relative Fluorescence Intensity (RFI). CD56 expression was found in 66% of MM and 54% of PCL cases. The RFI values for individual MM patients ranged from 7.6 to 27.4 while ABC values on MM plasma cells from 2255 to 58469. There was a correlation between the proportion of all bone marrow CD38⁺⁺/CD138⁺ cells with CD56 expression and ABC and RFI indices. With regard to CD56 expression positive patients, the CD56^- MM patients presented lower frequency of osteolysis (p = 0.01). The median survival was 48 months in CD56⁺ patients and 43 months (p = 0.84) in CD56^- cases. In conclusion, CD56 expression carries no distinct adverse prognosis and the lack of CD56 expression does not define a unique clinicopathological or prognostic entity in MM. A remarkable heterogeneity of CD56 expression intensity in CD56⁺ patients imposes the necessity of determining CD56 expression intensity in candidates to antibody-based therapy.     

Ecto-5'-Nucleotidase (E5'-NT, 27.2, CD73), 27.1, Leu13, CD28 and LAM-1(Leu8) Antigens in Mycosis Fungoides (MF)

Two monoclonal antibodies (MoAbs) B121 and B124 to the human lymphocyte differentiation antigen E5'-NT (CD73), and the MoAbs 27.2 and 27.1 that were raised to HTLV-1⁺CD4⁺CD25⁺ leukemic T cells were tested immunohistochemically on frozen sections of 13 cases of MF together with Leu13, CD28, LAM-1(Leu8) and other standard T-cell markers. Controls included 7(5T, 2B) non-MF cutaneous non-Hodgkin's lymphomas (NHL), 2 cutaneous T lymphoid leukemias (T-ALL), 13 miscellaneous non-neoplastic dermatoses, 11(5T, 6B) extracutaneous NHL, and 5 splenic B-hairy cell leukemias. Previous studies suggest that 27.2 also recognizes the CD73 antigen and is present in high density in some cases of HTLV-1⁺ adult thymic leukemia/lymphoma (ATL) and HTLV-1^- Sezary syndrome (Y. Fukunaga et al., Blood 74:2486-2492, 1989). In the studies reported here B121, B124 and 27.2 reacted similarly with the MF and control samples tested. The CD73 antigen was present in the majority of lymphoid cells from 8 (62%) of 13 cases of MF. In one of these cases the CD73 antigen was detected only in the deeper and more pleomorphic MF dermal infiltrate. Although CD73 expression was noted in varying numbers of lymphocytes from many of the control benign and malignant lymphoproliferative lesions of the skin, none of the extracutaneous NHL or HCL cases tested showed significant numbers of CD73⁺ cells. Ten, 11 and 11 of the 13 MF cases also showed moderate to high numbers of cells reactive with 27.1, Leu13 and CD28, respectively. Most of the MF cases also showed moderate to high co-expression of βF1 (TCRβ), CD3 and CD4, and 9 cases showed moderate numbers of CD8+ cells as well. In contrast to the control dermatoses, where significant co-expression of the majority T-cell markers LAM-1(Leu8), CD5 and CD7 was the rule, 11 of the 13 MF cases showed significant loss of one or more of these antigens. One or more of these antigens was notably also lost with CD73 in 3 patients who had, or developed, systemic MF. The above results indicate that MF cases often co-express the CD73/27.2 antigen with 27.1, Leu13, CD28, βF1 (TCRβ), CD3 and CD4, and may show a concurrent absence of CD5, CD7 or LAM-1(Leu8) antigens with or without loss of CD73. The possible roles of CD73, Leu 13, CD28 and LAM-1(Leu8) in controlling cell proliferation, adhesion and migration in MF are discussed.     

Expression and Production of Interleukin 4 in B-Cell Chronic Lymphocytic Leukaemia

Interleukin (IL) 4 is a T-cell derived pleiotropic cytokine whose properties include alterations of B-cell function. In B-cell chronic lymphocytic leukaemia (B-CLL), IL4 is involved in the mechanism of survival of the leukaemic B-cells. The present study examines the expression and production of IL4 by B- and T-lymphocytes derived from patients with B-CLL and provides evidence that IL4 is not an autocrine factor in B-CLL. Freshly isolated B-CLL cells enriched for B- and T-cells did not express mRNA for IL4 but expressed mRNA for IL4 receptor (IL4R). Activation of B-cells with phorbol ester and calcium ionophore and of T-cells with phytohaemaglutinin (PHA) upregulated IL4 mRNA expression. However phorbol ester and calcium ionophore did not affect the mean level of IL4 production by either B-CLL or normal B-cells. Furthermore, in the presence or absence of activation, the amount of IL4 synthesised by B-CLL B-cells was not significantly different than that observed with peripheral blood B-cells isolated from normal individuals (with activation: P=0.239; without activation: P=0.565). Also, there was no significant difference between normal and B-CLL B-cells in the level of cytoplasmic IL4 (P=0.47).

PHA-activated enriched B-CLL T-cells produced significantly higher levels of IL4 compared to normal control T-cells (P=0.0136). In addition, in 47% of cases with B-CLL T-cells, a significant higher level of intracellular IL4 was observed (P=0.0027). The levels of production of IL4 by the T-cell-enriched preparations correlated positively with the intensity of cytoplasmic IL4 in CD4⁺ and CD8⁺ cells in tested samples (r=0.49 and r=0.76, respectively).

The significant differences observed in the production of IL4 by B-CLL B- and T-lymphocytes may suggest a paracrine function of IL4 in B-CLL.     

Interleukin 3 Stimulation Enhances Tyrosine Phosphorylation and Proliferative Activity of Human Fetal Thymocytes and Leukemic T-cell Precursors at Multiple Developmental Stages of T-Cell Ontogeny

The purposes of this study were to elucidate the biologic effects of recombinant human interleukin 3 (rhIL-3) on leukemia T-cell precursors and fetal thymocytes corresponding to discrete and sequential developmental stages of human T-cell ontogeny, and to examine the biochemical nature of the IL-3 receptor linked transmembrane signal in human T-cell precursor populations. The specific binding of biosynthetically labeled ³⁵S-rhIL-3 to leukemic T-cell precursors from T-lineage ALL patients was initially investigated. In 5 of 9 cases, the binding of ³⁵S-rhIL-3 was significantly blocked by excess cold rhIL-3, and the percentage of inhibitable binding ranged from 40% to 64% (mean ± SE = 53 ± 4%). In these cases, 5-13 femtomols (mean + SE = 7.0 ± 1.5 fms) of ³⁵S-rhIL-3/10⁷ cells were specifically bound. rhIL-3 stimulated in a dose dependent fashion the in vitro clonal proliferation of leukemic T-cell precursors with composite immunophenotypes corresponding to the developmental stages of prothymocytes/double negative immature thymocytes (3 of 5 cases) and corticothymocytes/double positive CD4⁺CD8⁺ immature thymocytes (4 of 4 cases). By contrast, leukemic T-cell precursors from 2 T-lineage ALL cases with a composite immunophenotype of medullary thymocytes/single positive CD4⁺CD8^-/CD4^-CD8⁺ mature thymocytes did not show an enhanced proliferative activity after stimulation with increasing concentrations of rhIL-3. All of the 5 cases with significant ³⁵S-rhIL-3 binding and none of the 4 cases with no ³⁵S-rhIL-3 binding showed a proliferative response to rhIL-3. Thus, there was a high correlation between ³⁵S-rhIL-3 binding and proliferative response of leukemic T-cell precursors in colony assays, indicating that functional IL-3 receptors were detected in ligand binding assays. rhIL-3 also stimulated the proliferation of immature double positive CD4⁺CD8⁺ thymocytes from 9 of 10 fetal thymuses without inducing differentiation and with no selective advantage for the development of CD4⁺CD8^- or CD4^- or CD4^-CD8⁺ single positive thymocytes. The observed differences in IL-3 responsiveness among T-cell precursor populations at distinct developmental stages indicates that IL-3 receptors may be expressed only during the early stages of human T-cell ontogeny preceding the negative or positive selection events within the thymic microenvironment. Stimulation of fetal thymocytes with rhIL-3 resulted in enhanced tyrosine phosphorylation of 8 distinct cellular substrates with molecular weights of 44 kDa, 55 kDa, 60 kDa, 69 kDa, 98 kDa, 123 kDa, 150 kDa and 190 kDa, but it did not result in stimulation of phosphoinositide turnover and increased Ins-1,4,5-P₃ production. The induction of tyrosine phosphorylation by rhIL-3 was augmented by further crosslinking surface bound IL-3 molecules with an anti-IL-3 antibody and it was abrogated by the tyrosine kinase inhibitor genistein. The ligation of the CD45 protein tyrosine phosphatase markedly suppressed the tyrosine phosphorylation of specific substrates induced by IL-3 stimulation. Thus, the mitogenic transmembrane signal triggered by the engagement of the IL-3 R on human T-cell precursors is linked to a functional protein tyrosine kinase (PTK)/protein tyrosine phosphatase (PTP) regulatory pathway. Taken together, these results indicate that IL-3 may have an important growth regulatory role in early stages of human T-cell ontogeny.     

Enhanced antitumor effects by the coculture of allotumor RNA-pulsed dendritic cells with autologous cytokine-induced killer cells on hormone-refractory prostate cancer

In this study, we evaluated antitumor effects of allotumour RNA-transfected dendritic cells (DCs) cocultured with autologous cytokine-induced killer cells (CIKs) on hormone-refractory prostate cancer. The cocultured cells enhanced prostate cancer cytolysis from 26% (CIKs-induced cytolysis) to 80.8%. They also increased the productions of CD4⁺ Th1 (IFN-γ⁺IL-4^-, 55.52%) and CD8⁺ T (IFN-γ⁺, 69.59%) cells determined by intracellular cytokines IFN-γ /IL-4 staining and reduced the rate of CD4⁺ CD25⁺ cells from 18.72% (in CIKs) to 9.72%. The cocultured cells significantly inhibited tumor growth in SCID mouse and induced cancer cells necrosis and apoptosis. Our study indicates that tumor RNA-pulsed DCs cocultured with autologous CIKs significantly enhance antitumor immunity, which can be induced by increased CD4⁺ Th1 and CD8⁺ T cells and decreased CD4⁺CD25⁺ regulatory T (T_reg) cells. This provides a potential immunotherapy strategy for HRPC.     

不同乳腺癌干细胞亚群的自我更新和致瘤能力差异

目的 分析原代培养乳腺癌细胞中不同乳腺癌干细胞亚群的自我更新、致瘤能力差异。方法 采用流式细胞术和免疫荧光实验,从乳腺癌原代培养细胞中分选、鉴定CD44⁺CD24^-/low、ALDH1⁺和ALDH1⁺CD44⁺CD24^-/low细胞亚群,分析各分选细胞占总肿瘤细胞的比例;通过克隆形成实验观察各组细胞的自我更新能力;通过裸鼠致瘤实验观察各组细胞的致瘤能力。结果 CD44+CD24^-/low细胞占总细胞比例的7.2%,ALDH1⁺细胞所占比例为4.6%,ALDH1⁺CD44⁺CD24^-/low细胞所占比例为1.5%。三组细胞自我更新和致瘤能力由强至弱依次为ALDH1⁺CD44⁺CD24^-/low、ALDH1⁺、CD44⁺CD24^-/low细胞,各组细胞间比较差异均存在统计学意义（P<0.05）。 结论 乳腺癌组织中CD44+CD24-/low、ALDH1⁺和ALDH1⁺CD44⁺CD24^-/low三组乳腺癌干细胞亚群之间的自我更新和致瘤能力有明显差异,ALDH1⁺CD44⁺CD24^-/low亚群细胞具有最强的自我更新和致瘤能力,提示ALDH1⁺CD44⁺CD24^-/low可能是更具有特异性的乳腺癌干细胞标志物。     

Proportion of CD4+CD25+ regulatory T cell is increased in the patients with ovarian carcinoma

Objective: To study the frequency of the CD4⁺CD25⁺ regulatory T cells (Tregs) in the patients with ovarian carcinoma and its possible mechanism. Methods: The percentages of CD4⁺CD25⁺ Tregs in the peripheral blood lymphocytes (PBLs), tumor infiltrating lymphocytes (TILs) and tumor associated lymphocytes (TALs) from 13 patients with ovarian carcinoma and in the PBLs from 14 healthy women were determined by flow cytometry. The expression of CD69 on CD4⁺PBLs from the patients was detected. PBLs from healthy women were cultured in complete RPMI 1640 containing the supernatant from SKOV3 cell line with or without PHA (phytohemagglutinin) stimulation for 72 hours, then the percentage of CD4⁺CD25⁺ T cells was detected. Results: CD4⁺CD25⁺ Tregs in the PBLs from patients with ovarian carcinoma were significantly increased compared with those from the control. The percentage of CD4⁺CD25⁺ Tregs in TILs was higher than that in PBLs and TALs from the patients, but not significantly. The expression of CD69 on CD4⁺PBLs from the patients was negative. The percentages of CD4⁺CD25⁺ T cells in PBLs cultured with SKOV3 supernatant elevated significantly compared with those without supernatant whether PHA was added or not (P = 0.001 and 0.001, respectively). Conclusion: There is an increasing of the proportion of CD4⁺CD25⁺ Tregs in PBLs, TILs and TALs of the patients with ovarian carcinoma, which probably results from up-regulation of soluble factor secreted by ovarian carcinoma cells.     

结直肠癌及腺瘤患者CD4+、CD8+和CD28+T淋巴细胞的亚群变化及临床意义

背景与目的：结直肠癌（colorectal cancer,CRC）严重影响患者生存。探讨肿瘤微环境（tumor microenvironment,TME）T细胞亚群在CRC和腺瘤中的表达及意义。方法：用免疫组织化学法和流式细胞术对51例健康人（对照组）、46例结直肠腺瘤（腺瘤组）、100例CRC（癌症组）和15例CRC术后（癌术后组）患者进行T细胞亚群检测。结果：① 对照组、腺瘤组及癌症组3组中CD4⁺T细胞的阳性率分别是90.00%、43.75%及32.65%,CD8⁺T淋巴细胞的阳性率分别是30.00%、56.25%及75.51%,CD28⁺T淋巴细胞的阳性率分别是42.86%、30.00%及20.00%。② 对照组、腺瘤组及癌症组3组中CD4⁺、CD4⁺/CD8⁺、CD28⁺、CD8⁺CD28⁺和CD8^－CD28⁺逐渐降低,CD8⁺、CD8⁺CD28^－逐渐增加（P＜0.05）;癌术前术后T细胞亚群差异有统计学意义（P＜0.05）。结论：① CRC微环境T细胞亚群中CD4⁺、CD4⁺/CD8⁺、CD28⁺、CD8⁺CD28⁺和CD8^－CD28⁺呈递减趋势,CD8⁺、CD8⁺CD28^－呈递增趋势,且在癌前病变腺瘤中已逐步出现上述趋势变化。② CRC患者行肿瘤切除术后,其T细胞亚群有所恢复,故在一定程度上,CRC中T细胞亚群的变化可以早期预测结直肠疾病的发展。     

CD44+/CD24+表型的宫颈癌Siha细胞对顺铂的耐药及机制研究

背景与目的：肿瘤干细胞的存在是肿瘤细胞抗拒化疗的原因之一。该研究探讨CD44⁺/CD24⁺宫颈癌Siha细胞对顺铂的耐药性及相关机制。方法：体外培养Siha细胞,流式荧光激活细胞分选仪分选CD44⁺/CD24⁺Siha细胞,噻唑蓝(thiazolyl blue,MTT)法检测不同浓度顺铂对细胞的体外抑制情况,采用流式细胞仪检测10 μg/mL顺铂作用于CD44⁺/CD24⁺ Siha细胞24、48和72 h时的细胞凋亡率,实时定量聚合酶链式反应(quantitutive real-time polymerase chain reaction,qRT-PCR)和蛋白［质］印迹法(Western blot)检测CD44⁺/CD24⁺ Siha细胞中Oct-4、ABCG2、Bcl-2的表达,同时设立亲代Siha细胞作为对照。结果：不同浓度顺铂(0.1、1、5、10、15和20 μg/mL)对CD44⁺/CD24⁺ Siha细胞的增殖抑制作用较亲代Siha细胞小［(88.4±1.5)% vs (92.9±1.5)%,(79.9±1.0)% vs (84.7±1.1)%,(69. 8±0.8)% vs (75.1±2.9)%,(59.0±0.7)% vs (65.8±2.7)%,(49.6±0.9)% vs(52.1±0.5)%,(45.1±0.7)% vs (48.8±1.0)%,P<0.05］;与亲代细胞相比,当10 μg/mL顺铂作用于CD44⁺/CD24⁺Siha细胞时,发现在24、48和72 h时的细胞凋亡率均较小［(3.05±0.16)% vs (5.17±0.27)%,(17.94±2.02)%vs (32.60±4.28)%和(40.14±3.01)% vs (56.62±5.32)%,P<0.05］。qRT-PCR和Western blot实验均显示CD44⁺/CD24⁺ Siha细胞高表达Oct-4、ABCG2和Bcl-2,与亲代Siha细胞比较差异有统计学意义(P=0.015)。结论：CD44⁺/CD24⁺ Siha细胞可以抵抗顺铂诱导的细胞凋亡,具有化疗抵抗性,并且高表达肿瘤干细胞表面标志物如Oct-4和ABCG2,该结果在宫颈癌肿瘤干细胞的有效分选以及肿瘤靶向治疗方面有深远意义。     

Immunologic, morphologic and chromosomal characterization of a cell line (TC78) established from a child with acute lymphoblastic leukemia

We characterized a cell line established from bone marrow cells from a child with acute lymphoblastic leukemia. This cell line, TC78, had lymphoblastic morphology and was cytoplasmic peroxidase and esterase negative. The cells did not have T- or B-cell properties such as E- or EAC-rosette forming ability, reactivity with monoclonal T-cell or B2 antibodies, or immunoglobulin synthesis. We concluded that TC78 was a pre-pre B-cell line based on the following monoclonal antibody staining pattern: BA-I⁺, BA-2⁺, cALLa⁺, Ia⁺, 2H7⁺ and OKB2⁺. Growth in ‘Dickie’ culture and reactivity with 1G10 myeloid antibody suggested coexpression of lymphoid and myeloid characteristics. However, 1G10 expression proved dependent on culture conditions, illustrating one caveat in application of monoclonal antibodies in lineage determination.     

The clonal hierachy in multiple myeloma

In this report we evaluated the number and phenotype of blood circulating B-cell subsets at different stages of differentiation in 26 patients with newly diagnosed multiple myeloma (MM). In all patients, plasma cells and/or plasma blasts could be identified by flow cytometry with a mean frequency of 1.20% and 0.07%, respectively. In 76.9% of the patients these cells showed aberrant expression mainly of CD56, CD28 and CD117, none of these markers were found on the earlier B-lymphocytes. Clonal B-cells preceding the plasma blast stage were identified by patient specific IgH RT-PCR on sorted B-cell subsets. The clonal cells included the less differentiated CD38⁺/CD19⁺ and CD38^-/CD19⁺ subsets, illustrating that the clonal cells are part of an ongoing differentiation process. Further, the presence of CD38^-/CD19⁺     

Expression of MHC class I and class I-like gene products on the cell membrane of mature and immature T cells

Beta 2-microglobulin (beta 2m) constitutes the 12 kD light chain of the MHC-encoded 43-kD glycoprotein HLA-ABC molecules, which are expressed on most nucleated cells. The T6 (CD1a) antigen is expressed on thymocytes in reciprocal manner to HLA-ABC, having a similar structure to the HLA-ABC molecule. We have determined the expression of beta 2m, HLA-ABC and the T6 antigen on the cell-surface of CD1a+ and CD1a- thymocytes (defined by NA1/34 expression), and have shown that despite immunophenotypic differences between these two stages of thymic maturation, the quantity of beta 2m-associated molecules expressed was not significantly different. The nature of the heavy (alpha) chain associated with beta 2m, however, differed since HLA-ABC had largely replaced T6 on the surface of CD1a- thymocytes. We also determined the expression of beta 2m and HLA-ABC on the surface of peripheral blood CD4+ and CD8+ T-cells, and showed that the CD8+ population expressed higher levels of these antigens than CD4+ T-cells, with no detectable excess beta 2m over HLA-ABC for either subpopulation. In addition, thymocytes expressed fewer beta 2m-associated determinants than peripheral blood T-cells. These results indicate an increase in the expression of beta 2m-associated molecules with differentiation from thymic to peripheral T-cell, with a further increase in such molecules expressed on the CD8+ compared to the CD4+ peripheral T-cell subpopulation.     

CD44+CD24-/low在基底样乳腺癌中过表达与其恶性预后的相关性研究

目的探讨乳腺癌干细胞样标志物CD44⁺CD24^-/low在基底样乳腺癌(basal-like breast carcinoma, BLBC)中过表达与BLBC恶性预后的相关性。方法 在乳腺癌基因表达分型的基础上, 根据雌激素受体(ER)、孕激素受体(PR)、人类表皮生长因子受体2(Her-2)免疫表型的表达选取乳腺癌组织四组:管腔A组、管腔B/C组、Her-2高表达组及三阴性组;对三阴性组检测CK5/6、EGFR, 分为正常乳腺样型和BLBC型两组;对上述5组进行免疫组化Envision法染色, 选用抗体为CD44、CD24, 观察CD44⁺CD24^-/low表型表达并比较BLBC组与其它各组的差异。结果 (1)三阴性组共60例, CK5/6和或EGFR阳性者41例(68.3%), 确定为BLBC;CK5/6、EGFR阴性者19例(31.7%), 确定为正常乳腺样组;(2)CD44⁺CD24^-/low表型在BLBC组中占78.0%(32/41), 相对于管腔A组37.9%(11/29)、管腔B组25.9%(7/27)、Her-2高表达组17.2%(5/29)、正常乳腺样组26.3%(5/21), 表达增高并具有显著性(P＜0.05);(3)所有150例乳腺癌中(可评价145例)具有CD44⁺CD24^-/low免疫表型者其Ki-67指数增高相对于非CD44⁺CD24^-/low表型具有统计学差异(P＜0.001)。结论 BLBC型乳腺癌表达乳腺癌干细胞样标志物CD44⁺CD24^-/low显著高于其它各型乳腺癌, CD44⁺CD24^-/low与BLBC独特的恶性生物学行为相关。     

CD15及CD133在高、低级别胶质瘤中的表达及意义

目的探讨CD15在胶质瘤的表达及分布特征,与目前公认的脑肿瘤干细胞标志物CD133作对照,并探讨两者相关性。方法采用免疫组织化学法检测80例胶质瘤组织（高级别39例,低级别41例）CD15和CD133的表达。用Image-Pro Plus 6.0图像分析处理软件计算CD15和CD133阳性细胞积分光密度值（IOD）。结果CD15与CD133在高、低级别的胶质瘤均有表达。CD15及CD133阳性细胞数以及壁龛随胶质瘤病理级别增高而增多。在高、低级别胶质瘤中CD15⁺细胞与CD133⁺细胞IOD值差异均有统计学意义（z=－3.432,P=0.001;z=－4.192,P=0.000）;CD15⁺细胞与CD133⁺细胞IOD值呈显著正相关（r=0.535,P=0.000）;CD133还表达于含有肥胖细胞成分的胶质瘤肥胖细胞中,并可见CD133+血管。结论CD15及CD133阳性细胞数以及壁龛随胶质瘤病理级别增高而增多。CD15及CD133表达成正相关关系。含有肥胖细胞成分的胶质瘤中大量肥胖细胞CD133阳性,与其不良临床预后相符。关键词：     

CD56+表达对急性单核细胞白血病儿童的临床特征、疗效及预后的影响

目的探讨CD56⁺表达对急性单核细胞白血病(AML-M5)儿童临床特征、疗效及预后的影响。方法对84例14岁以下AML-M5患儿临床资料进行回顾性分析,分析CD56的表达与临床特征、疗效及预后的关系。结果CD56⁺表达率为32.14%,白细胞计数(73.21±12.93)×10⁹/l,高白细胞血症占51.85%,髓外浸润占59.26%;CD56^-表达率为67.86%,白细胞计数(54.21±10.64)×10⁹/l,高白细胞血症占15.79%,髓外浸润占29.82%,比较差异具有统计学意义(P<0.05)。CD56+患儿治疗完全缓解率为62.96%、死亡率为7.41%,CD56^-患儿完全缓解率66.67%、死亡率8.77%,比较差异无统计学意义(P>0.05);CD56⁺达完全缓解的患儿随访期间复发率76.47%、持续缓解率为23.53%,平均生存时间(10.92±4.71)个月,CD56^-患儿复发率28.95%、持续缓解率为71.05%、平均生存时间(15.15±8.12)个月,比较差异具有统计学意义(P<0.05)。结论CD56⁺表达的14岁以下AML-M5患儿高白细胞血症、髓外浸润发生及复发率较高,平均生存时间较短。CD56表达能够为患儿的治疗及预后提供指导信息。     

CD4+/CD8+ T淋巴细胞在肝细胞癌组织中的浸润及与预后的相关性研究

目的 检测CD4⁺/CD8⁺ T淋巴细胞在肝细胞癌（hepatocellular carcinoma,HCC）组织中的浸润程度,并分析其与预后的相关性。方法 收集行肝切除术的HCC患者215例,采用免疫组化技术检测CD4⁺/CD8⁺ T淋巴细胞在HCC癌组织中的浸润程度,根据浸润情况比较患者肝切除术后无瘤生存率和总生存率。结果 CD4⁺ T淋巴细胞高浸润和低浸润比例分别为60.9%和39.1%。CD4⁺ T淋巴细胞高浸润组患者总生存率和无瘤生存率均显著高于低浸润组（P=0.015,P=0.038）。CD8⁺ T淋巴细胞高浸润和低浸润比例分别为34.9%和65.1%。CD8⁺ T淋巴细胞高浸润组患者的总生存率和无瘤生存率亦显著高于低浸润组患者（P=0.033,P=0.047）。结论 CD4⁺或CD8⁺ T淋巴细胞低浸润可能与HCC患者术后不良预后相关。