健康人C3/Ig双特异性免疫复合物与体液免疫的相关性

采用直线相关分析法,较系统地研究了健康人 Ｃ３／ Ｉｇ 双特异性循环免疫复合物（ Ｃ３／ Ｉｇ Ｍ  Ｔ Ｃ Ｉ Ｃ、 Ｃ３／ Ｉｇ Ｇ Ｔ Ｃ Ｉ Ｃ 和 Ｃ３／ Ｉｇ Ａ Ｔ Ｃ Ｉ Ｃ）与 Ｉｇ Ｍ 、 Ｉｇ Ｇ、 Ｉｇ Ａ 和 Ｃ３ 的相关性．结果发现, Ｃ３／ Ｉｇ Ｍ  Ｔ Ｃ Ｉ Ｃ 和 Ｃ３／ Ｉｇ Ｇ Ｔ Ｃ Ｉ Ｃ 含量主要取决于血清总 Ｉｇ Ｍ 和 Ｉｇ Ｇ 量; Ｃ３／ Ｉｇ Ｇ Ｔ Ｃ Ｉ Ｃ 与复合的 Ｉｇ Ｍ 、复合的 Ｉｇ Ｇ、总的 Ｃ３ 呈正相关,而与复合的 Ｃ３ 呈负相关．这些结果提示, Ｉｇ Ｇ 在健康人体液免疫中的重要作用．同时说明,健康人机体免疫反应是复杂多样的,深入研究 Ｃ３／ Ｉｇ Ｔ Ｃ Ｉ Ｃ 与体液免疫的相关性及其形成机理,将为揭示健康人的体液免疫机制提供重要参考．     

健康人Ig/C3双特异性免疫复合物与体液免疫的相关性

采用直线相关分析法,较系统地研究了健康人Ｉｇ／Ｃ３双特异性循环免疫复合物（ＩｇＭ／Ｃ３－ＴＣＩＣ、ＩｇＧ／Ｃ３－ＴＣＩＣ和ＩｇＡ／Ｃ３－ＴＣＩＣ）与ＩｇＭ、ＩｇＧ、ＩｇＡ和Ｃ３的相关性,比较了Ｉｇ／Ｃ３－ＴＣＩＣ和Ｃ３／Ｉｇ－ＴＣＩＣ分别与有关体液免疫指标关系的异同。结果发现,ＩｇＭ／Ｃ３－ＴＣＩＣ与总ＩｇＭ和总ＩｇＡ／Ｃ３－ＴＣＩＣ与总ＩｇＭ、总ＩｇＭ、总ＩｇＧ、总ＩｇＡ、和复合ＩｇＧ以及总Ｃ３     

HMC14单克隆抗体杂交瘤株的建立

以北京鸭红细胞提取的ＨＭＧｓ免疫Ｂａｌｂ／ｃ小鼠，取其脏压细胞与Ｓｐ２／０（小鼠骨髓瘤细胞）融合，经选择培养，阳性筛选，克隆化以及冻存复苏，获得了三株稳定分泌抗ＨＭＧ１４的杂交瘤细胞株，经ＷｗｓｔｅｒｎＢｌｏｔ鉴定，与其它几种ＨＭＧ无反应，抗体为ＩｇＧ１亚型。腹水ＨＭＧ１４单隆抗体经ＳｅｐｈａｄｘＣＬ－４Ｂ亲和柱得到了纯化。     

HMG＿（14）单克隆抗体杂交瘤株的建立

以北京鸭红细胞提取的ＨＭＧｓ（ＨＭＧ＿１，ＨＭＧ＿（２ａ），ＨＭＧ＿（２ｂ），ＨＭＧ＿（１４），ＨＭＧ＿（１７））免疫Ｂａｌｂ／ｃ小鼠，取其脏压细胞与Ｓｐ２／０（小鼠骨髓瘤细胞）融合，经选择培养，阳性筛选，克隆化以及冻存复苏，获得了三株稳定分泌抗ＨＭＧ＿（１４）的杂交瘤细胞株（１Ｃ＿（１２），１Ｏ＿３和２Ｆ＿２），经ＷｅｓｔｅｒｎＢｌｏｔ鉴定，与其它几种ＨＭＧ无反应，抗体为ＩｇＧ＿１亚型。腹水ＨＭＧ＿（１４）单克隆抗体经ＳｅｐｈａｄａｘＣＬ－４Ｂ亲和柱得到了纯化。     

鸭乙肝病毒表面抗原特异性单抗的研制及应用

以纯化的鸭乙型肝炎病毒表面抗原（ＤＨＢｓＡｇ）免疫ＢＡＬＢ／ｃ小鼠,取免疫鼠脾细胞与小鼠骨髓瘤ＳＰ２／Ｏ－Ａｇ１４融合,应用ＥＬＩＳＡ筛选出５Ｂ８和８Ｇ１１ ２株抗体分泌滴度较高的杂交瘤细胞,二者所分泌的单抗仅与ＤＨＢｓＡｇ发生反应,而与其它９种禽类常见的病毒均不发生反应,其亚类鉴定分别属于ＩｇＭ和ＩｇＧ２ａ。用鸭乙型肝炎病毒（ＤＨＢＶ）阳性鸭血清中粗提的病毒悬液进行５Ｂ８株单抗介导的ＳＰＡ胶体金     

冠心病患者血清抗心磷脂抗体检测的临床研究

为探讨ＡＣＡ对ＣＨＤ的临床意义,分别用ＥＬＩＳＡ和免疫比浊法对３４例住院心病和１４８名健康体检人员的ＡＣＡ－ＩｇＧ,ＡＣＡ－ＩｇＭ,ＡＣＡ－ＩｇＡ和ＴＣ,ＴＧ,ＨＤＬ进行了检测。结果ＣＨＤ组与正常组比较ＨＤＬ,ＴＣ,ＴＧ均具显著统计学差异;３类ＡＣＡ水平与正常组无明显差异,但ＡＣＡ－ＩｇＧ的消长与血清ＴＧ呈正相关;冠心病组的ＡＣＡ－ＩｇＧ阳性检出经也明显高于正常组,提示血清ＡＣＡ可能通过影响脂代谢     

静脉注射免疫丙种蛋白柱层析分离的研究

用凝胶渗透谱色法（ＧＰＣ）检测了柱层析分离纯化后所得的免疫丙种球蛋白（ＩｇＧ）组分。结果表明，使用ＤＥＡＥ阴闻子交换纤维素－葡聚糖凝胶Ａ－５０柱层析法能从人血浆蛋白中分离含聚集体的纯ＩｇＧ单体。对肌注ＩｇＧ制剂使用葡聚糖凝胶Ｓｅｐｈａｃｒｙ１ Ｓ－２００凝胶过滤法能除动ＩｇＧ中的聚集体，但却不能分离出存在于其中的小量血清白蛋白；但使用ＤＥＡＥ阴离子交换纤维素层析法效果却恰好相反。     

PVG膜剂量计线性响应范围的估计

对辐射致色薄膜剂量计（ＰＶＧ）的辐照响应过程进行了研究，发现其剂量响应曲线上有一极大点，考查了隐色孔雀绿（ＬＭＧ）的极大转化率Ｃｅｘｔ与隐色孔雀绿（ＬＭＧ）和有机齿代物（ＲＸ）量含量的关系，进而讨论了ＰＶＧ膜剂量计线性响应范围的上限与膜中ＬＭＧ是含量的关系。在该体系中当ＬＭＧ量含量在２０～１００μｍｏｌ·ｇ＾－１范围内时，其线性响应范围时上限估计在１４０～１１０ｋＧｙ范围内。     

甲亢患者尿β2微球蛋白,白蛋白和IgG水平的检测

采用放射免疫分析法测定了７１例甲亢患者尿β２微球蛋白（β２－Ｍ）、白蛋白（ＡＬｂ）和ＩｇＧ水平，以观察甲亢患者肾脏功能的变化。结果表明甲亢患者尿β２－Ｍ、ＡＬｂ和ＩｇＧ水平明显高于正常对照组，肾脏损害检出率６７．６％，肾小球或肾小管同时受损率为２６．８％。提示部分甲亢患者存在肾脏功能异常     

淋巴细胞性脉络丛胞膜炎病毒单克隆抗体的研制及初步鉴定

用提纯的ＬＣＭ病毒抗原（主要为ＮＰ６３）免疫ＢＡＬＢ／Ｃ小鼠，应用鼠—鼠杂交瘤技术获得了三珠（１Ｅ２、１Ｃ６和２Ｄ２）分泌抗ＬＣＭＶ单克隆抗体的杂交瘤细胞系。经检测，它们所分泌的抗体亚类分别为ＩｇＧ１（１Ｅ２）和ＩｇＭ（１Ｃ６）。亲和力为７５μｇ（１Ｅ２）和５μｇ（１Ｃ６）。腹水效价为１０５。免疫荧光阻断法和ＥＬＩＳＡ阻断试验测定结果一致，１Ｅ２的标记物能阻断２Ｄ２，但不能阻断１Ｃ６。三株ＭｃＡｂ对７种鼠源性病毒抗原（ＲｅＯ３、Ｓｅｎｄａｉ、ＭＨＶ、ＧＤⅦ、ＥＨＦＰＶＭ和Ｅｃｔｒｏ）均无反应。     

The immunoglobulin mu constant region gene is expressed in mouse thymocytes

It has been a matter of controversy whether the functional capacity of T cells to discriminate between antigens is mediated via immunoglobulin, an immunoglobulin-like molecule, or by the product(s) of unrelated genes. The progenitors of immunoglobulin-secreting cells, B cells, express membrane-bound immunoglobulin as the antigen-specific receptor on their surface. For T cells, although products of immunoglobulin heavy chain variable region genes are implicated as receptor components, there has been no compelling immunochemical evidence for participation of either immunoglobulin light chains or heavy chain constant regions (see refs 2-6 for the disparate views). Recently, using cloned immunoglobulin DNA sequences as hybridization probes, we have demonstrated that the immunoglobulin Cmu gene, but not the Cmu gene, is expressed as polyadenylated RNA in some T cell tumour (T lymphoma) cell lines. Individual T lymphoma lines yielded up to three discrete mu RNA species of different sizes (1.9, 2.2 and 3.0 kilobases), each species being different in size from the major mu RNA species present in B lymphoma cells (2.4 and 2.7 kilobases). We show here that cells from the normal mouse thymus contain mu RNA species, indistinguishable in size from those in T lymphoma cells, but contain little if any kappa RNA.     

Diversity of immunoglobulin expression in leukaemic cells resembling B-lymphocyte precursors

Approximately 20% of patients with acute lymphocytic leukaemia (ALL) have leukaemic blasts with features of pre-B cells which are the recently characterized precursors of B lymphocytes in normal development (for a review, see ref. 2). Pre-B cells isolated from normal bone marrow or fetal liver, and malignant cells from patients with pre-B cell leukaemia, are rapidly dividing lymphoid cells that contain cytoplasmic immunoglobulin mu heavy chains, but have no detectable surface immunoglobulin. The resemblance of immunoglobulin-containing ALL cells to normal precursors of B lymphocytes and their availability in relatively pure preparations allowed us to explore them as models of early stages in the differentiation of the B-lymphocyte line. We report here observations on the occurrence of intermediate pre-B/B-cell phenotypes, immunoglobulin isotype switching and the asynchrony of immunoglobulin heavy and light chain expression in 30 cases of ALL and 3 cases of chronic myelogenous leukaemia in lymphoblastic crisis (CML-BC).     

Transfer of a cloned immunoglobulin light-chain gene to mutant hybridoma cells restores specific antibody production

The expression of immunoglobulin (Ig) genes is regulated at several levels. For example, although kappa-chain production requires a DNA rearrangement that juxtaposes variable and joining segments, this rearrangement is not sufficient for kappa-chain gene expression; that is, some cell types do not permit immunoglobulin production. The mechanisms responsible for the regulation of the expression of rearranged immunoglobulin genes are poorly understood. The technique of modifying cloned genes in vitro and transferring the modified genes to cells in culture provides a tool for identifying the structural features required for gene expression. To analyse immunoglobulin genes in this manner, however, it is first necessary to use, as recipients, cells that normally permit immunoglobulin production. We report here that a cloned kappa-chain gene is expressed in immunoglobulin-producing hybridoma cells. Furthermore, the product of the transferred kappa-chain gene is capable of restoring specific antibody production to the transformed cells.     

Epstein-Barr virus transforms precursor B cells even before immunoglobulin gene rearrangements

The very early stages of the human B-cell differentiation pathway are poorly understood, primarily because of the lack of appropriate permanent cell lines. Epstein-Barr virus (EBV) is a putative human oncogenic virus which transforms human B cells in vitro into continuously proliferating cells. It has been believed that EBV transforms mature B cells, but recently, transformation of immature pre-B-cell lines has been reported, suggesting that EBV might also transform cells much earlier in the B-cell lineage. We report here the establishment of cell lines transformed by EBV at various stages of the B-cell differentiation pathway. Interestingly, two lines showed the complete absence of immunoglobulin synthesis and the lack of immunoglobulin gene rearrangement despite containing EBV genome and surface markers of B cells. Our results indicate that EBV can infect and transform cells of the B lymphocyte lineage even before immunoglobulin gene rearrangement.     

A novel cell surface molecule on early B-lineage cells

B cells and their antibody-secreting progeny represent one of several differentiation pathways that haematopoietic stem cells (HSC) may enter. Cells representing intermediate stages between HSC and B cells have been identified in mammalian haematopoietic tissues and studied intensively over the past decade. This population of early B-lineage cells, termed pre-B, is characterized by cellular proliferation and an orderly cascade of immunoglobulin gene rearrangements, a combination of events leading to the generation of clonally diverse B cells which then migrate to peripheral lymphoid tissues. It remains to be determined what elements determine the polyclonal growth of pre-B cells, how immunoglobulin gene rearrangements are regulated, and what happens to pre-B cells undergoing 'non-productive' immunoglobulin gene rearrangements. These issues could be resolved more easily if early B-lineage cells could be identified precisely and isolated. Here, we describe a cell surface glycoprotein that is selectively expressed by pre-B and newly formed B cells in murine haematopoietic tissues. The molecule, a homodimer formed by disulphide-linked chains of relative molecular mass (Mr) 140,000, is identified by a mouse monoclonal alloantibody called BP-1.     

Functional V region formation during in vitro culture of a murine immature B precursor cell line

B lymphocytes originate from pluripotential haematopoietic stem cells and differentiate into immunoglobulin (Ig)-producing cells. B-cell lineage differentiation is accompanied by two types of immunoglobulin gene rearrangements--rearrangement of V, D and J gene segments to create a functional V gene and rearrangement of CH genes for heavy-chain switching. These results, however, have been obtained mainly by analysis of immunoglobulin gene organization of myeloma cells. Baltimore and his colleagues have established Abelson murine leukaemia virus (A-MuLV)-transformed cell lines and found a few lines capable of carrying out kappa-gene rearrangement or undergoing isotype switching during in vitro culture. To study early B-cell lineage differentiation events, we have now also established A-MuLV-transformed cell lines which are capable of differentiating from mu- to mu+ and of undergoing continuing rearrangement of heavy-chain genes in culture. Analysis of immunoglobulin gene organization of these transformed cells revealed that mu- cells have already undergone DNA rearrangements involving JH segments but an additional rearrangement of JH segments is required for initiation of mu-chain synthesis. Southern blot analysis of the DNA and two-dimensional gel electrophoresis of intracytoplasmic mu-chain show that mu-chain diversity with respect to antigen specificity may be generated during this second rearrangement process. As no rearrangement of light-chain genes takes place in these cells, this implies that light-chain gene rearrangement requires some further change, or a different enzyme.     

Production of functional chimaeric mouse/human antibody

The availability of monoclonal antibodies has revived interest in immunotherapy. The ability to influence an individual's immune state by administering immunoglobulin of the appropriate specificity may provide a powerful approach to disease control and prevention. Compared with immunoglobulin from other species, human immunoglobulin (Ig) might be best for such therapeutic intervention; it might function better with the recipient's effector cells and should itself be less immunogenic. The success of the mouse hybridoma system suggests that immunoglobulin of virtually any specificity can be obtained from a properly immunized animal. In the human system, however, immunization protocols are restricted by ethical considerations, and it is not yet clear whether human antibody-producing cell lines of the required specificity can be obtained from adventitiously immunized individuals or from in vitro immunized cells. A method which might circumvent these difficulties is to produce antibodies consisting of mouse variable regions joined to human constant regions. Therefore, we have constructed immunoglobulin genes in which the DNA segments encoding mouse variable regions specific for the hapten trinitrophenyl (TNP) are joined to segments encoding human mu and kappa constant regions. These 'chimaeric' genes are expressed as functional TNP-binding chimaeric IgM. We report here some of the properties of this novel IgM.