Neoplastic angioendotheliosis: demonstration of immunoglobulin gene rearrangements by the Southern blot hybridization technique

Using the Southern blot hybridization technique, two cases of 'neoplastic angioendotheliosis' (NAE) were examined for immunoglobulin gene rearrangements. For DNA analysis, frozen kidney (Case 1) and lung (Case 2) tissue, in which intravascular neoplastic cells were abundant, were used. In each case rearrangements were observed in both heavy and light chain genes. On examination of the heavy chain gene, the JH DNA probe detected one rearranged band in Case 1 and two in Case 2 after EcoRI digestion. On examination of the light chain genes, the C kappa DNA probe revealed one rearranged band in Case 1 and two in Case 2 after BamHI digestion, while the C lambda 2 DNA probe revealed only germline configurations after EcoRI digestion in each case. Thus, B-lymphocytic differentiation at the gene level was demonstrated in neoplastic cells in both cases. Since no more than two rearrangements were detected in each immunoglobulin gene, the intravascular neoplastic cells were considered to be monoclonal in nature.     

DNA polymerase chain reaction using fine needle aspiration biopsy smears to evaluate non-Hodgkin's lymphoma.

OBJECTIVE: To apply polymerase chain reaction (PCR) analysis to the fine needle aspiration biopsy (FNAB) evaluation of lymphoid proliferations. STUDY DESIGN: We analyzed 37 consecutive archived FNAB malignant lymphoma specimens. Immunophenotypic data from the fine needle aspiration biopsy and excisional biopsy material was available for all specimens. PCR to identify monoclonal rearrangements of the immunoglobulin heavy chain gene, T-cell receptor and translocations involving the bcl-1 and bcl-2 genes was performed. RESULTS: Seventy-eight percent of cases were detected by at least one of these assays. Where DNA analysis was performed on excisional biopsy material, 70% of the cases had identical results; no discordant results for the immunoglobulin heavy chain gene or T-cell receptor were found. In 23% of cases, after review of all available data, a discordant result was thought to be a consequence of a false negative result in DNA analysis of excisional biopsy material. CONCLUSION: These findings indicate that PCR analysis of archived FNAB material, when necessary, provides useful information for diagnosis and staging of malignant non-Hodgkin's lymphomas.     

The immunoglobulin heavy chain class switch

Conclusions While much has been learned concerning the molecular structural basis for the heavy chain class switch, many questions relating to the regulation of the switch remain unanswered, or at least controversial. Identification of the enzyme system which mediates the class switch, as well as other regulatory, possibly X-linked, genes should provide the necessary key to our understanding of this unique process.AbbreviationsB cell lymphocyte derived from the bone marrow in adult mammals or the bursa of Fabricius in chickens - bp base pair - C immunoglobulin constant region - CDR complementarity-determining region of the immunoglobulin variable region - D diversity gene segment of the immunoglobulin heavy chain variable region gene - H immunoglobulin heavy chain - Ig immunoglobulin - J joining region gene segment of the immunoglobulin variable region gene - kb kilobase - L immunoglobulin light chain - LPS lipopolysaccharide - Pyr pyrimidine - S-, s-site, s-region switch rearrangement site - SCE sister chromatid exchange - sIg surface immunoglobulin - T cell lymphocyte derived from the thymus - USCE unequal sister chromatid exchange - V immunoglobulin variable region     

石蜡包埋组织的DNA提取及其在免疫球蛋白基因重排分析中的应用

基因重排分析在淋巴瘤诊断中具有重要意义.文章应用改良DNA提取方法,从30例淋巴增生性病变石蜡包埋组织获得的DNA虽有不同程度的降解,但适于PCR扩增Ig重链基因重排分析;约1/3病例提出高分子量DNA,可用于DNA印迹杂交.因此,石蜡包埋组织同样可为某些疾患,如淋巴瘤疑难和罕见病例的回顾性分子病理学研究提供基因诊断的DNA来源.     

甲型H1N1流感病毒血凝素单克隆抗体轻链和重链可变区基因的巢式PCR扩增及序列分析

目的：采用巢式PCR对甲型H1N1流感病毒血凝素单克隆抗体的轻链和重链基因进行扩增,对获得的基因进行序列分析,并找出克隆鼠Igκ轻链和重链可变区基因的通用方法。方法：设计22对扩增鼠Igκ轻链可变区和重链可变区基因的引物,对6株鼠抗人甲型H1N1流感病毒血凝素单克隆抗体的轻链和重链可变区基因进行克隆并测序,与NCBI公布的鼠免疫球蛋白序列比对分析。结果：巢式PCR方法可以有效避免单克隆抗体克隆过程的假基因,并且得到的单克隆抗体的氨基酸序列均符合鼠免疫球蛋白可变区特征。结论：建立了克隆鼠免疫球蛋白轻链和重链可变区基因的通用方法,为后期克隆鼠源性单克隆抗体的可变区基因提供了基础,并为研究甲型H1N1流感病毒血凝素与抗体的结合位点提供了实验数据。     

The immunoglobulin M heavy chain constant region gene of the channel catfish, Ictalurus punctatus: an unusual mRNA splice pattern produces the membrane form of the molecule.

The immunoglobulin (IgM) heavy chain constant region gene of the channel catfish, Ictalurus punctatus, has been cloned and characterized. The gene contains four constant region domain-encoding exons (CH1 to CH4) expressed in the secreted form of the immunoglobulin, and two exons encoding the transmembrane (TM) domain utilized in the lymphocyte membrane receptor form of the immunoglobulin. The sequence of a cDNA clone encoding the 3' region of the message for the membrane receptor form of the mu chain indicates that the TM1 exon is spliced directly to the CH3 exon, and not into a site within the CH4 exon, as occurs in the mammals, a shark and an amphibian. This unusual pattern of splicing, which produces a membrane heavy chain that is characteristically smaller than the secreted heavy chain, may be common to all teleost fish.     

Nucleotide sequences of immunoglobulin mu heavy chain deletion mutants.

Mutants of an IgM producing hybridoma cell line were isolated which produce mu heavy chain fragments. Two such mutants were found to have internal deletions in the mu gene and the nucleotide sequence of the deletion endpoints was determined. No evidence was found for a role of the heavy chain switch region in the formation of these deletions. The implications of these mutants in defining the requirements of immunoglobulin gene expression are discussed.     

The human T cell antigen receptor is encoded by variable, diversity, and joining gene segments that rearrange to generate a complete V gene

A cDNA clone YT35 , synthesized from poly(A)+ RNA of the human T cell tumor Molt 3, exhibits homology to the variable (V), joining (J), and constant (C) regions of immunoglobulin genes. We have isolated and sequenced the germ-line V and J gene segment counterparts to YT35 from a human cosmid library, and these failed to encode 14 nucleotides of the cDNA clone between the V and J regions. We postulate that these 14 nucleotides are encoded by a third gene segment analogous to the diversity (D) gene segments of immunoglobulin heavy chain genes. This T cell antigen receptor V gene appears to be assembled from three gene segments, V, D, and J, and accordingly most closely resembles immunoglobulin heavy chain V genes.     

BiP表达下调改善双载体转断裂FVIII基因的功效

双载体转凝血Ⅷ因子（FⅧ）基因可作为一种转基因策略克服腺相关病毒(AAV)载体容量限制,但重链分泌的低效性影响转基因功效. 为提高重链分泌,本文用RNA干扰技术下调内质网内蛋白伴侣分子免疫球蛋白重链结合蛋白(BiP)的表达,观察对HEK293细胞双载体共转FⅧ基因分泌重链和生物活性的影响. 结果显示,RNA干扰可明显下调BiP表达,但不影响细胞生长; ELISA检测BiP下调细胞单独转重链基因时的重链分泌量为98±38 ng/mL,与轻链共转基因时显著升高到157±32 ng/mL,明显高于对照细胞单独转重链基因和共转重链和轻链基因的重链分泌量(分别为29±8 ng/mL和79±19 ng/mL);Cotest法检测显示,BiP下调细胞共转重链和轻链基因细胞分泌的凝血生物活性为0.73±0.23 IU/mL,明显高于对照细胞共转重链和轻链基因(0.39±0.07 IU/mL). 结果表明, BiP表达下调通过促进重链分泌,可提高双载体共转FⅧ基因的功效,为进一步动物体内双AAV载体转FⅧ基因的甲型血友病基因治疗研究提供了实验依据.     

Duplications and deletions of Vh genes in inbred strains of mice

evolution of variable region (Vh) gene family copy number and polymorphism was investigated by the analysis of the immunoglobulin heavy chain variable region (Igh-V) locus in 74 inbred strains and substrains of mice. Several strains were found to have slight differences from Igh-V haplotypes previously identified, usually of a single Vh gene family. These results indicate that the evolution of copy number in the mouse Igh-V locus proceeds largely by the accumulation of incremental changes, reflecting the clustered organization of the mouse Igh-V locus. We have found no evidence of very large or frequent duplication or deletion events indicative of rapid expansion or contraction processes. The existence of one or more particularly large Vh gene families most likely reflects random copy number variation, rather than selection for the amplification of their members. The identification of strains with recombinant Vh gene arrays demonstrates that recombination, both within and between haplotypes, appears to be the predominant mechanism generating the high restriction fragment length polymorphism in the Igh-V locus.Abbreviations used in this paper Igh-V immunoglobulin heavy chain variable region locus - Vh heavy chain variable region gene - Dh heavy chain diversity region gene - V immunoglobulin kappa light chain variable region gene - V T-cell receptor beta chain variable region gene     

Retention and loss of immunoglobulin heavy chain alleles in helper T cell hybridoma clones

Retention or loss of immunoglobulin heavy chain genes was studied in 20 functional T cell hybridoma clones. DNA probes representing C mu, C alpha and JH genes, as well as VH subgroups II and III were hybridized with restriction enzyme fragments of hybridoma DNA by the Southern filter hybridization technique. Parental alleles of the hybridoma cells were distinguished on the basis of polymorphism of the lengths of restriction enzyme fragments. All clones retained the alleles of the lymphoma parent cell BW-5147 at all four loci. Thirteen clones lost both CH and VH alleles of the immune partner cell, whereas seven retained both VH alleles, and at least C alpha of the antigen-specific partner. Hence, T cell function in these cells is compatible with the loss of most immunoglobulin heavy chain alleles. This is interpreted to indicate either gene rearrangement and deletion, or chromosome loss. Accordingly, the T cell receptor is either controlled by two split gene loci in chromosome 12, at the two respective (5' and 3') ends of the mouse heavy chain gene family, or by a gene(s) outside chromosome 12.     

VPREB3: cDNA characterization and expression in human and chromosome mapping in human and mouse

The pre-B cell receptor (pre-BCR) regulates pre-B cell expansion and allelic exclusion at the immunoglobulin (Ig) heavy chain locus and mediates the selection of Ig heavy chain variable gene segments. During the early phase of pre-BCR assembly in the mouse, the membrane Ig mu heavy chain transiently associates with the VPREB3 protein in the endoplasmic reticulum. Here, we present the human VPREB3 cDNA sequence and its B cell-specific expression in hematopoietic cell lines. We have localized this gene to chromosome 22q11 close to IGLL genes in human and to chromosome 10C in mouse.