白藜芦醇提高CD34+CD38-KG1a白血病细胞对人外周血单个核细胞的杀伤敏感性

目的:研究白藜芦醇(Resveratrol)作用前后CD34+CD38-KG1a白血病细胞对IL-15介导的人外周血单个核细胞(PBMC)杀伤敏感性的变化及机制的初步探讨.方法:应用CCK-8法检测白藜芦醇对白血病细胞的IC50值,以此量的白藜芦醇作用于白血病细胞.,并用LDH释放法检测白藜芦醇作用前后,效靶比分别为5∶1、10∶1、20∶1的IL-15介导的PBMC对CD34+CD38-KG1a细胞杀伤能力.流式细胞仪(FACS)检测IL-15介导前后PBMC表面NKG2D的表达.流式细胞仪(FACS)检测作用前后CD34+CD38-KG1a细胞表面NKG2D配体(MICA、MICB、ULBP1、ULBP2、ULBP3)表达.结果:白藜芦醇作用前后在不同效靶比时对IL-15介导的PBMC杀伤敏感性差异有统计学意义(P＜0.05).IL-15介导PBMC前后表面NKG2D的表达有显著变化,差异有统计学意义.白藜芦醇作用前后CD34+CD38-KG1a细胞表面NKG2D各配体中MICA、MICB无显著变化(P＞0.05),ULBP1、ULBP2、ULBP3有显著变化,差异有统计学意义(P＜0.05).结论:白藜芦醇可以提高CD34+CD38-KG1a细胞对IL-15介导PBMC的杀伤敏感性,其机制可能与白藜芦醇提高CD34+CD38-KG1a细胞表面NKG2D配体的表达有关.     

CD147 contains different bioactive epitopes involving the regulation of cell adhesion and lymphocyte activation

CD147 is a leukocyte surface molecule which belongs to the immunoglobulin superfamily. It is broadly expressed on various cell types and is a lymphocyte activation-associated molecule. In order to study the function of CD147, five CD147 monoclonal antibodies (mAbs) were generated: M6-2F9; M6-1D4; M6-1F3; M6-1B9; and M6-1E9. Biochemical characterizations and cross-blocking experiments indicated that M6-1B9 and M6-1E9 recognize the same or contiguous epitopes on CD147. By employing COS transfectants expressing CD147 membrane-distal domain (domain 1) and membrane-proximal domain (domain 2), mAbs M6-2F9, M6-1D4, M6-1B9, and M6-1E9 were shown to recognize epitopes located on domain 1 of the molecule. Functional studies indicated that engagement of CD147 by mAbs M6-1B9 and M6-1E9 strongly inhibited lymphocyte proliferation induced by a CD3 mAb. In contrast, mAbs M6-2F9, M6-1D4, and M6-1F3 induced U937 homotypic cell aggregation. The results indicate that CD147 contains at least two bioactive domains. Epitopes responsible for induction of cell aggregation are different from those regulating lymphocyte activation.     

CD23-mediated homotypic cell adhesion: The role of proteolysis

CD23 is a multifunctional molecule expressed by cells of lymphoid, myeloid and hematopoietic lineages. As a cell surface molecule CD23 acts both as a low-affinity receptor for IgE (FcεRII) and as a cell adhesion molecule. CD23 can undergo autoproteolysis to release soluble 37–25-kDa CD23 (s-CD23) molecules with a range of cytokine activities. Here we show a causal link between the two apparently disparate functions of autoproteolysis and cell adhesion. The Epstein-Barr virus-transformed B cell line RPMI-8866 formed macroscopic cell clusters solely via CD23. Cell adhesion was inhibited by mAb to CD23 and by IgE. Cell adhesion was also dependent on serum as cells grown in serum-free media failed to form clusters. In serum-free conditions cell adhesion could be induced by the addition of not only 10 % FCS but also s-CD23. As s-CD23 is reported to possess proteolytic activity we screened a range of proteases to determine whether they also could induce cell adhesion in serum-free medium. It was found that chymotrypsin and elastase induced cell: cell adhesion in RPMI-8866 cells. The same panel of proteases were screened against a range of CD23-positive (Jijoye, AF-10, T2, U937, ICH-1) and CD23-negative (RPMI-8226, U266, MOLT-4, Ramos) cell lines. It was found that chymotrypsin and elastase induce cell adhesion only in cells expressing CD23. Peptide mapping studies showed that chymotrypsin and elastase cleaved immunoprecipitated CD23 near the same site by which 37-kDa S-CD23 is released (Ala 80). Serum demonstrated no proteolytic activity towards CD23. However, it was found that cells grown in serum-free medium released 25-kDa S-CD23 without the need for prior cleavage at the 37-kDa cleavage site. To confirm the role of proteolysis in CD23-mediated cell adhesion we screened a range of protease inhibitors for their ability to antagonize this process. It was found that tosyl-lysine chloromethyl ketone inhibited CD23-mediated cell adhesion. Lactoperoxidase treatment, which inhibits CD23 cleavage, also inhibited cell adhesion. Addition of chymotrypsin and elastase to lactoperoxidase-treated cells induced cell adhesion. From these data we propose that intact CD23 has no demonstrable role in cell adhesion; instead, the portion of CD23 remaining on the cell surface following cleavage appears to mediate cell adhesion.     

A monoclonal antibody, 3/22, to rabbit CD11c which induces homotypic T cell aggregation: evidence that ICAM-1 is a ligand for CD11c/CD18

The rabbit CD11c molecule has been characterized by use of a new monoclonal antibody, mAb 3/22. Expression of the p150,95 integrin (CD11c/CD18) has been shown by flow cytometry and immunohistochemistry to be restricted to monocytes, macrophages, dendritic cells and a small population of lymphocytes in peripheral blood. No expression on neutrophils could be demonstrated. Incubation of the newly derived CD8⁺ T cell line, BJ/873, with mAb 3/22 causes homotypic aggregation, which has been shown to be a cell surface event that is not dependent on intracellular signaling or on receptor cross-linking. Inhibition studies show that the ligands responsible for this aggregation are CD11c/CD 18 and ICAM-1, both of which are expressed on BJ/873. One other rabbit T cell line, K34, that also expresses p150,95 and ICAM-1, shows a similar aggregation response when stimulated with 3/22. Cell lines that express p150,95 but not ICAM-1 do not aggregate. These observations suggest that ICAM-1 is a ligand for activated p150,95.     

Long-term treatment of pigs with low doses of monoclonal antibodies against porcine CD4 and CD8 antigens

In vivo depletion of lymphocyte subsets allows investigation of the role of specific subsets in protective immunity. In the present study we evaluated the effects of long-term, low-dose treatment with murine monoclonal antibodies (mAbs) against porcine CD4 and CD8 surface antigens on lymphocyte subsets in pigs. Four-week-old pigs were treated by intramuscular injections of hybridoma cell culture supernatants containing anti-CD mAbs twice a week for a period of 5 weeks. The immunomodulatory effects of the treatments were assessed by flow cytometry (FCM) analysis of peripheral blood lymphocytes. Treatment with the anti-CD4 mAb almost completely eliminated the CD4(+) T-cell subset from the circulation after 2 weeks of therapy. This depletion persisted until the end of the experimental period 5 weeks after initiated therapy. Treatment with the anti-CD8 mAb was less effective, reducing the CD8(+) T-cell subset in peripheral blood by approximately 50% of the initial level after 3 weeks of therapy. Further, the anti-CD8 mAb-treated pigs showed a parallel increase in the CD4(+) T-cell subset from day 7. Two-colour FCM analysis indicated that a shift in phenotype from single-positive CD4(+)/CD8(-) to double-positive CD4(+)/CD8(+) T-cells might have occurred in these pigs. In the present experiment we demonstrated specific modulation of the peripheral blood T-lymphocyte population in pigs with continuous low-dose injections of specific mAb. The ability to modulate individual T-cell subsets should provide a method to elucidate their functionality in protection against infectious disease.     

CD34免疫亲合柱在脐血造血干/祖细胞分离与扩增中的应用

目的 :观察CD34免疫亲合柱对脐血造血干 祖细胞分离纯化的效果及纯化后CD34 细胞的增殖分化特性。方法 :采用CD34免疫亲合柱分离脐血单个核细胞 (MNC)中的CD34 细胞 ,流式细胞技术 (FACS)进行细胞表面标志测定。将分离前后的细胞加入造血生长因子进行液态扩增培养和多向祖细胞集落 (CFU GEMM)培养。结果 :经CD34免疫亲合柱分离后脐血中CD34 细胞为 49 6 2 %± 17 6 9% ,明显高于脐血MNC(1 17%± 0 6 8% ) ,细胞回收率为 5 4 38%± 11 91%。分离后CD34 细胞和脐血MNC经造血生长因子刺激培养 2 0d分别扩增 5 6 1 0 0倍和 44 44倍。培养至 12d时 ,免疫亲合柱分离组CD34 细胞阳性率为 5 3 38% ,对照组为 7 91%。分离组CFU GEMM产率明显高于对照组 (P <0 0 0 1)。结论 :CD34免疫亲合柱应用于脐血造血干 祖细胞的分离可充分富集CD34 细胞 ,且分离后的CD34 细胞具有明显的增殖效应和CFU GEMM形成能力。     

Properties of mouse CD40: the role of homotypic adhesion in the activation of B cells via CD40

Stimulation of human B cells via CD40 is known to induce their homotypic aggregation. We show here that anti-mouse CD40 monoclonal antibodies (mAb) also induce B cells to form large, spherical, extremely stable clusters. This clustering is markedly enhanced by co-stimulation with either interleukin-4 (IL-4) or anti-immunoglobulin (Ig). The aggregation is slow in onset, and is largely (but not completely) abrogated by anti-LFA-1 mAb, but not by mAb directed against other potentially important adhesion molecules on B cells. Anti-LFA-1 mAb also partially suppressed DNA synthesis induced by anti-CD40, but not by other B cell mitogens, suggesting that clustering is an important component of B cell activation via CD40. This concept is supported by analyses of the phenotype of clustered B cells: the cells within clusters express higher levels of various activation markers, and also more of them are in cell cycle than non-clustered cells. These results therefore suggest that CD40 stimulation may either induce B cells to secrete soluble factors which act in an autocrine way to promote Bcell activation, or that clustering generates cell contact-mediated signals which are important in the activation cascade.     

A CD44 monoclonal antibody differentially regulates CD11a/CD18 binding to intercellular adhesion molecules CD54, CD102 and CD50

We have made a monoclonal anti-CD44 antibody which is able to activate the leukocyte integrin CD11a/CD18. Activated T cells strongly aggregated, and the aggregation was shown to be intercellular adhesion molecule (ICAM)-1 (CD54) and ICAM-2 (CD102) dependent. Using purified ICAM coated on plastic, only binding to ICAM-1 was increased by the CD44 antibody, whereas activation by phorbol ester increased binding to both ICAM-1 and ICAM-3. The binding to ICAM-2 was not affected by either treatment. These findings show that the CD11a/CD18 integrin can be activated in a ligand-specific manner by engagement of CD44.     

CD34抗体包被支架预防兔损伤动脉支架内狭窄

目的： 观察CD34抗体包被支架能否减少和预防球囊损伤动脉支架内狭窄。方法：①用明胶作为包被底物对支架进行包被，然后再包被CD34抗体。②复制股动脉损伤家兔模型，损伤后即刻于右侧植入普通支架，左侧植入抗体支架；45 d后，观察支架内最狭窄处的百分率、支架内和支架两端的血管新生内膜增生。结果：45 d 时，抗体包被支架内的最狭窄百分数明显低于普通侧支架（16.13±3.93 vs 47.03±11.89，P<0.05）；抗体包被支架段动脉血管新生内膜百分数明显低于普通支架段动脉血管（49.64±6.40 vs 66.83±6.00，P<0.05）；抗体包被支架远心端血管的新生内膜增生比普通支架远心端血管明显减轻。但两者的近心端动脉无明显差别。结论：CD34抗体包被支架能够有效减轻受损动脉支架内狭窄的发生。     

A new CD43 monoclonal antibody induces homotypic aggregation of human leucocytes through a CD11a/CD18-dependent and -independent mechanism.

We describe a monoclonal antibody (mAb), designated 1.C1, that causes rapid and vigorous aggregation among normal leucocytes and among T and myeloid/monocytic cell lines. As shown by competitive binding and sequential immunoprecipitation experiments, the antigen recognized by mAb 1.C1 is a 115,000 MW sialoglycoprotein, that corresponds to the human CD43 antigen, also known as leukosialin or sialophorin. The aggregation process starts within minutes and reaches maximum level 6-18 hr after addition of the antibody. It is dependent on active cell metabolism (inhibited at low temperatures and by a mixture of the metabolic poisons azide and 2-deoxy-D-glucose), a fluid plasma membrane (inhibited by pretreatment of the cells with paraformaldehyde) and an intact cytoskeleton (inhibited by cytochalasin B). Two reference CD43 antibodies (MEM-59 and DF-T1), both binding the same or closely related sialic acid-dependent epitope as mAb 1.C1, are also capable of inducing cell clump formation. CD11a/CD18 mAb block the 1.C1-induced adhesion of resting peripheral blood leucocytes, but not of haematopoietic cell line cells. In addition, mAb 1.C1 induces homotypic aggregation of K-562 cells, which do not express members of the beta 2 integrin subfamily on their surface. These data suggest that triggering of the CD43 antigen promotes homotypic cell adhesion that is mediated by both CD11a/CD18-dependent and -independent pathways.     

5株鼠抗人CD28单克隆抗体的研制及生物学特性的研究

目的：制备鼠抗人CD28分子功能性单克隆抗体，研究其对T细胞活化、增殖及信号转导等方面的生物学效应。方法：以天然高表达CD28分子的人多发性骨髓瘤细胞株U266和小鼠淋巴瘤细胞转人CD28基因细胞株CD28-T分别作为免疫原和检测细胞株，采用B淋巴细胞杂交瘤技术进行单抗的研制；以快速定性试纸法鉴定单抗亚类；腹水诱生法和免疫亲和层析法进行单抗的制备和纯化；经间接免疫荧光法分析单抗对不同细胞膜表面CD28分子的识别；采用竞争抑制法分析单抗识别的抗原位点；利用^3H-TdR掺入法分析单抗对PBTC的刺激效应和免疫荧光法分析PBTC活化前后的表型变化。结果：成功获得5株鼠抗人CD28功能性单克隆抗体，分别命名为2D5、2F5、3136、3F8和8G8，其中2D5为IgG2a亚类，其余均为IgG1亚类；流式细胞仪分析结果显示，5株单抗均能良好识别CD28-T、U266、XGI和Jurkat细胞表面的CD28分子；竞争抑制试验表明，2D5和8G8能完全阻断标准抗人CD28单抗与U266膜CD28分子的结合，其余3株为部分阻断；^3H-TdR掺入法实验结果表明，单抗8G8联合激发型CD3单抗能明显促进PBTC的增殖，刺激指数为7．4，活化细胞CD4、CD25、ICOS、4IBB及OX40分子的表达上调。结论：5株单抗均为抗人CD28单克隆抗体，具有重要的基础研究及潜在的临床应用价值。     

两步法从慢性髓系白血病患者骨髓CD34+细胞体外扩增诱导树突状细胞

目的探讨从慢性髓系白血病(CML)患者骨髓CD34~ 细胞体外扩增诱导树突状细胞(DC)的可行性,并比较CML-DC和正常DC的生物学特性。方法免疫磁珠法从CML患者和正常供者骨髓纯化CD34~ 细胞,在有血清条件下应用两步法:干细胞生长因子(SCF) FLT3配体(FL) 促血小板生成素(TPO) 白细胞介素-3(IL-3)扩增2周,然后以粒细胞巨噬细胞集落刺激因子(GM-CSF) 白细胞介素-4(IL-4) 肿瘤坏死因子-α(TNF-α)(GI方案)诱导DC。通过相差显微镜、电子显微镜、流式细胞仪分析DC的生物学特性,荧光原位杂交(FISH)检测培养前后CML细胞的bcr/abl融合基因表达。结果诱导后细胞较0d或诱导前细胞高表达DC相关抗原(CD1a,CD80,CD86,CD40,CD54,HLA-DR)。CML患者和正常供者CD34 细胞经GI方案诱导10d,CD1a阳性率分别为36.90%±26.94%和54.35%±16.34%,CD1a~ DC数是0d接种细胞的(54±54)倍和(122±129)倍。两组相比,总细胞扩增倍数、DC扩增倍数、细胞表型均无明显差异,诱导后的DC具有相似的超微结构。CML患者CD34~ 细胞诱导10d后bcr/abl融合基因阳性率为43.67%±21.55%,具有典型DC形态的细胞bcr/abl融合基因阳性率为53.2%。结论两步法GI方案能诱导CMLCD34~ 细胞产生大量DC,诱导生成的DC不但具有正常DC的典型形态、表型,而且起源于CML细胞。     

CD23 and CD21 function as adhesion molecules in homotypic aggregation of human B lymphocytes

We have previously found that interleukin-4 and CD40 monoclonal antibodies (mAb) are strong potentiatiors of homotypic B cell aggregation which is dependent on LFA-1. We show here that CD23 mAb were also able to inhibit aggregation to a similar extent as LFA-1 antibodies. This inhibition was restricted to the MHM6 epitope of CD23 and antibodies to other epitopes [Epstein-Barr virus (EBV) CS-1, EBV CS-2, EBV CS-5 and mAb 25] or occupation of the Fc-binding site by IgE had no or a slightly enhancing effect on aggregation. When testing two antibodies to CD21, the recently defined ligand for CD23, one of these (BU32) was found to be inhibitory whereas the other (THB5) had no effect. By combining antibodies to LFA-1 and CD23, aggregation was often completely inhibited. These data suggest that LFA-1/ICAM-1 and CD23/CD21 are the major molecules involved in homotypic aggregation of human B cells.     

恶性肿瘤干细胞标记物CD34在鼻咽癌细胞系的表达

BACKGROUND:Previous research have confirmed that CD34 is closely related to oncogenesis, progress, recurrence, metastasis and drug-resistance of various cancers, but its role in nasopharyngeal carcinoma remains unclear. OBJECTIVE:To sort cells positive and negative for CD34 in nasopharyngeal carcinoma cell lines and to detect cell proliferation and migration. METHODS:Expressions of CD34 in nasopharyngeal carcinoma cell lines 5-8F, 6-10B, CNE1 and CNE2 were detected by flow cytometry. And CD34+ and CD34^- cells were sorted based on cell surface markers for purity identification. Afterwards, proliferation and migration of CD34⁺ and CD34^- cells were detected by MTT assay, colony-formation assay and scratch assay. RESULTS AND CONCLUSION:All four nasopharyngeal carcinoma cell lines expressed CD34 in 0.1%-0.2%, and the level of CD34 was closely related to the cell growth density. The purity of CD34⁺ cell was more than 98% in the sorted CD34⁺ cell populations, but no CD34⁺ cells were found in the sorted CD34^- cell populations. At 1, 3, 5 and 7 days the proliferation rate of CD34⁺ cell, populations was significantly higher than that of CD34^- cells (P < 0.05). Consistently, the colony-formation efficiency of CD34⁺ cell was significantly higher than that of CD34^- cells (P < 0.05). Moreover, CD34⁺ cells migrated significantly faster than CD34^- cells by scratch assay (P < 0.05). In conclusion, CD34⁺ cells cultured in vitro display higher proliferation and migration capacities, indicating that CD34⁺ cells have the potential of nasopharyngeal carcinoma stem cells.     

Characterisation of CD4 T cells in healthy and diseased koalas (Phascolarctos cinereus) using cell-type-specific monoclonal antibodies

The koala (Phascolarctos cinereus) is an arboreal herbivorous marsupial that is an Australian icon. Koalas in many parts of Australia are under multiple threats including habitat destruction, dog attacks, vehicular accidents, and infectious diseases such as Chlamydia spp. and the koala retrovirus (KoRV), which may contribute to the incidence of lymphoma and leukaemia in this species. Due to a lack of koala-specific immune reagents and assays there is currently no way to adequately analyse the immune response in healthy, diseased or vaccinated animals. This paper reports the production and characterisation of the first anti-koala CD4 monoclonal antibody (mAb). The koala CD4 gene was identified and used to develop recombinant proteins for mAb production. Fluorochrome-conjugated anti-CD4 mAb was used to measure the levels of CD4⁺ lymphocytes collected from koala spleens (41.1%, range 20–45.1%) lymph nodes (36.3%, range 19–55.9%) and peripheral blood (23.8%, range 17.3–35%) by flow cytometry. Biotin-conjugated anti-CD4 mAb was used for western blot to determine an approximate size of 52 kDa for the koala CD4 molecule and used in immunohistochemistry to identify CD4⁺ cells in the paracortical region and germinal centres of spleen and lymph nodes. Using the anti-CD4 mab we showed that CD4 cells from vaccinated, but not control, koalas proliferated following in vitro stimulation with UV-inactivated Chlamydia pecorum and recombinant chlamydial antigens. Since CD4⁺ T cells have been shown to play a pivotal role in clearing chlamydial infection in both human and mouse infections, using this novel antibody will help determine the role CD4⁺ T cells play in protection against chlamydial infection in koalas and also enhance our knowledge of how KoRV affects the koala immune system.     

Properties of mouse CD40: Cellular distribution of CD40 and B cell activation by monoclonal anti-mouse CD40 antibodies

We describe here the derivation of a rat monoclonal antibody (mAb) against mouse CD40 (designated 3/23), which stains 45–50% of spleen cells of adult mice, approximately 90% of which are B cells. Interestingly, some 5–10% of both CD4⁺ and CD8⁺ T cells in the spleens of (some, but not all) adult, unimmunized mice are also CD40⁺, whereas CD40⁺ cells were not detectable in the thymus, even following collagenase digestion. Some 35–40% of lymphoid cells in the bone marrow of adult mice are CD40⁺ and virtually all of these are B220⁺, and hence of the B cell lineage: triple-color flow cytometry showed that CD40 is expressed at low levels on some 30% of pre-B cells, at intermediate levels on 80% of immature B cells and on essentially all mature B cells in the bone marrow. These results, therefore, suggest that in the mouse CD40 is expressed relatively late during the process of B cell differentiation. The mAb induced marked up-regulation of major histocompatibility complex class II molecules, CD23 and B7.2 antigens on mature B cells. It also stimulated modest levels of DNA synthesis in mature B cells by itself: this was markedly enhanced by suboptimal concentrations of mitogenic (but not non-mitogenic) anti-μ and anti-δ mAb, and moderately enhanced by co-stimulation with interleukin-4. Hyper-cross-linking of CD40 (using biotinylated mAb and avidin) also enhanced the proliferative response to anti-CD40.     

分泌高效价抗碱性磷酸酶McAb杂交瘤系的建立及其在APAAP免疫组化研究中的应用

用碱性磷酸酶长程多途径免疫BALB/C小鼠,采用反向间接ELISA法筛选融合孔上清,获得2株稳定分泌抗碱性磷酸酶McAb杂交瘤细胞株,腹水效价比国外报道高15～30倍,在APAAP免疫组化检测CD抗原、病毒抗原、肿瘤抗原以及细菌菌毛抗原等研究中均获得满意结果。     

Preventive effect of monoclonal antibodies to intercellular adhesion molecule-1 and leukocyte function-associate antigen-1 on murine spontaneous fetal resorption

PROBLEM: Are cell adhesion molecules involved in the murine model of immunologically‐mediated spontaneous abortion?
METHOD OF STUDY: Pregnant CBA/J female mice mated with DBA/2 male mice were injected with monoclonal antibodies (MAbs) to intercellular adhesion molecule‐1 (ICAM‐1) and leukocyte function‐associate antigen‐1 (LFA‐1). On day 13 of gestation, viable and resorbed embryos were counted. Natural killer (NK) cell activity in the spleen, mixed lymphocyte reactions (MLR), mixed lymphocyte‐placenta reactions (MLPR), and levels of interferon (IFN)‐Γ were assayed.
RESULTS: Significant suppression of fetal resorption was observed by the injection of MAb to ICAM‐1 and LFA‐1. NK cell activity and the MLR anti‐(CBA/J×DBA/2)F1 were reduced in the antibody‐treated CBA/J spleen. Moreover, the level of IFN‐Γ was significantly lower in the MLPR supernatants from the antibody‐treated group than those of the control group.
CONCLUSIONS: One mechanism in the murine model of spontaneous abortion may be through the interaction of cell adhesion molecules, which may modulate NK cell activities and cytokine production.     

甲型肝炎病毒的纯化及其单抗的制备

应用不连续蔗糖／甘油密度梯度超速离心的方法，从培养细胞中纯化甲肝病毒（Ｈｅ－ｐａｔｉｔｉｓＡＶｉｒｕｓ，ＨＡＶ）。纯化的ＨＡＶ抗原经ＳＤＳ－ＰＡＧＥ电泳分析，获得三条带：ＶＰ１（３４０００），ＶＰ２（３００００）和ＶＰ３（２６０００）。用纯化的ＨＡＶ抗原免疫Ｂａｌｂ／ｃ小鼠，将已免疫小鼠的脾细胞与ＳＰ２／０细胞融合，获得６株分泌抗ＨＡＶ单克隆抗体的杂交瘤。它们的培养上清和腹水的抗体滴度分别是５０～１０００和１０００～４０００。Ｉｇ亚类５株为ＩｇＧ１，１株为ＩｇＧ２ａ。采用间接的ＥＬＩＳＡ法分析ＨＡＶ抗原的抗原位点，结果表明ＨＡＶ抗原至少存在４个抗原位点，两个存在于ＶＰ１蛋白带，另两个分别存在于ＶＰ２和ＶＰ３蛋白带     

Presence of circulating abnormal CD34+ progenitors in adult Langerhans cell histiocytosis.

Langerhans cell histiocytosis (LCH) is related to the proliferation of cells, which are similar to Langerhans cells (LC) but possess many abnormal characteristics. Lesions are widespread and this fact suggests that LCH cells or their precursors are present in the blood of patients. In five adult patients, we have isolated and cultured CD34+ blood progenitors of dendritic cells. We studied their phenotype by flow cytometry and their functional properties in mixed culture with heterologous lymphocytes and with autologous lymphocytes in the presence of tri-nitro-phenyl antigen (TNP). The amount of CD34+ precursors was dramatically higher than controls but a high mortality occurred during the in vitro differentiation. The phenotype of surviving cells was similar to LC phenotype (CD1a+, CD83+, Lag+) but some of them expressed CD2. These cells were able to induce T cell proliferation in mixed culture. They could not initiate primary response to TNP, except in a patient treated with thalidomide. In our hands, these CD34+ cells may be precursors of LCH cells.