Surface immunoglobulin light chain restriction in B-cell non-Hodgkin's malignant lymphomas

Twenty-four control cases of reactive follicular hyperplasia (RFH) and 28 cases of B-cell non-Hodgkin's malignant lymphoma (ML) were evaluated for surface immunoglobulin (sIg) light chain expression with flow cytometry. Because there was no significant difference between the distribution of the ML kappa/lambda ratios for which kappa/lambda greater than 1 compared with those ML lambda/kappa ratios for which lambda/kappa greater than 1 using the Kolmogorov-Smirnov statistic, the authors treated kappa/lambda and lambda/kappa data symmetrically. By transforming all kappa/lambda ratios to maximum ratios, that is, Rmax = max (kappa/lambda, lambda/kappa), so that all resulting ratios were at least one, the authors defined a new statistic for evaluating sIg light chain restriction. There was a highly significant difference for equality of distribution of the data using the Mann-Whitney test (P less than 0.0001). By choosing cut-points based on the Rmax statistic, the authors were able to separate the RFH and ML cases with sensitivity = 0.96 +/- 0.07 (95% confidence) and with specificity = 0.91 +/- 0.11 (95% confidence). These point estimates of sensitivity and specificity appear to be promising for the clinical utility of the Rmax statistic, even for cases of B-cell ML with discordantly low expression of sIg light chains. The Rmax statistic will facilitate the comparison of data between different laboratories because it does not require prior assignment of cut-points and can be made using a simple nonparametric test.     

The correlation dimension identifies B-cell immunoglobulin light chain restriction in peripheral blood flow cytometry data

The correlation dimension (D2) is a mathematical tool that quantifies dimensional properties of fractals. We set out to determine whether immunoglobulin light chain restriction (LCR) in B-cell lymphoproliferative disorders (BCLPDs) could be identified using the D2. The D2 was calculated from flow cytometry data files (FCS 2.0) from a training set of peripheral blood specimens from 22 patients with and 23 patients without BCLPD. A cutoff value derived from the training set was applied to a validation set of 69 patients with and 64 patients without BCLPD. In the training set, all BCLPDs had a D2 of less than 0.72 (CD19-gated kappa-lambda data). When this cutoff was applied to the validation set, the D2 had sensitivity and specificity values of 88% and 98%, respectively. The D2 can identify LCR in peripheral blood flow cytometry data. It offers operator-independent data analysis and may be useful in objectively quantifying differences in data distribution.     

Excessive amounts of mu heavy chain block B-cell development

Antigen-independent B-cell development occurs in several stages that depend on the expression of Ig heavy and light chain. We identified a line of mice that lacked mature B cells in the spleen. This mouse line carried approximately 11 copies of a transgene of the murine heavy chain constant region locus, and B-lineage cells expressed excessive amounts of the intracellular μ heavy chain. B-cell development failed in the bone marrow at the pro/pre B-cell transition, and examination of other lines with various copy numbers of the same transgene suggested that deficiencies in B-cell development increased with increased transgene copy number. Expression of a transgenic (Tg) light chain along with the Tg μ heavy chain led to minimal rescue of B-cell development in the bone marrow and B cells in the spleen. There are several potential mechanisms for the death of pro/pre B cells as a consequence of excess heavy chain expression.     

Powered by pairing: the surrogate light chain amplifies immunoglobulin heavy chain signaling and pre-selects the antibody repertoire

Selective expansion of functional pre-B cells is accomplished by the assembly of a signaling-competent pre-B cell receptor (pre-BCR) consisting of immunoglobulin mu heavy chains (muHC), surrogate light chains (SLC) and Igalpha/Igbeta. Here, we review recent data showing that muHCs, in the absence of SLC, deliver autonomous differentiation signals. However, enhanced signaling necessary for pre-B cell expansion requires cross-linking of pre-BCRs via the non-immunoglobulin tail of SLC's subunit lambda5. We also discuss how SLC's ability to modulate the strength of pre-BCR signals is controlled by a muHC's idiotype and its affinity to the chaperone BiP. In this model, BiP in concert with SLC functions as a pre-selector of the antibody repertoire.     

The immunoglobulin light chain in poikilothermic vertebrates

Summary: The immunoglobulin light chains are classified as κ or δ in mammals and birds (homoeothermic vertebrates), but the traditional criteria for this classification are not applicable to the light chains found in poikilothermic vertebrates. Still it is possible to find some relationships between Ig light chain sequences in these animals and in those of the homoeothermic animals. It is generally accepted that the Ig light chains contribute to the antigen binding capacity of antibodies and the variability is approximately similar in all studied vertebrate species except the elasmobranchs. This might be explained by the organisation of the Ig light chain locus in these animals and the fact that the variable and joining DNA segments are joined in the genome. These conclusions are limited by the small number of species studied in this respect.     

B-cell development

The B cell population represents an extremely complex set of cells with respect to the existence of funclional and developmental cell subpopulations and their extremely diverse repertoire of antibody specificities. This article summarizes the key features of the ontogeny and developmental stages of murine B cells, the subjects of a recent extensive review¹, and considers several crucial unresolved issues central to the ultimate understanding of B cell function and expression.     

Immunocytochemical detection of kappa and lambda light chain V region subgroups in human B-cell malignancies.

We have used a sensitive immunoperoxidase method and highly specific anti-light chain antisera to determine the light chain variable region (VL) subgroup nature of cytoplasmic (c) and cell surface (s) Ig expressed by human monoclonal plasma cells and B lymphocytes. The immunocytochemical characterization of cIg and sIg used antisera specific for the established kappa light chain V kappa subgroups (V kappa I, V kappa II, V kappa III, and V kappa IV) and the lambda light chain V lambda subgroups (V lambda I, V lambda II/V, V lambda IV, and V lambda VI). Studies were performed using cytospin preparations of bone marrow-, peripheral blood-, and lymph node-derived cells from patients with multiple myeloma, amyloidosis AL, and Waldenström''s macroglobulinemia and with low-, mid-, and high-grade B-cell malignancies. The V kappa or V lambda subgroup of the cIg or sIg also could be identified after deparaffinization and enzyme treatment of formalin-fixed, paraffin-embedded specimens. For those patients who had monoclonal serum or urinary Igs, there was complete concordance between the VL subgroup of the secreted Ig and that of the cIg or sIg. The percentage distribution of V kappa or V lambda subgroups on the sIg of cells from patients with chronic lymphocytic leukemia (CLL) and other cytomorphologic types of B-cell malignancies differed from that found for kappa- or lambda-type Bence Jones proteins obtained from patients with multiple myeloma, amyloidosis AL, and Waldenström''s macroglobulinemia. In contrast to the plasma cell and lymphocytoid plasma cell diseases, a relative predominance of certain VL subgroups, ie, V kappa IV, V lambda III, and V lambda IV, and the absence of the amyloid-associated V lambda VI subgroup were found in CLL and related diseases. The immunocytochemical techniques used make possible a rapid means to demonstrate B-cell monoclonality and provide further evidence for the selective expression of certain VL genes in human B-cell neoplasia.     

Embryonic essential myosin light chain regulates fetal lung development in rats

Congenital diaphragmatic hernia (CDH) is currently the most life-threatening congenital anomaly the major finding of which is lung hypoplasia. Lung hypoplasia pathophysiology involves early developmental molecular insult in branching morphogenesis and a late mechanical insult by abdominal herniation in maturation and differentiation processes. Since early determinants of lung hypoplasia might appear as promising targets for prenatal therapy, proteomics analysis of normal and nitrofen-induced hypoplastic lungs was performed at 17.5 days after conception. The major differentially expressed protein was identified by mass spectrometry as myosin light chain 1a (MLC1a). Embryonic essential MLC1a and regulatory myosin light chain 2 (MLC2) were characterized throughout normal and abnormal lung development by immunohistochemistry and Western blot. Disruption of MLC1a expression was assessed in normal lung explant cultures by antisense oligodeoxynucleotides. Since early stages of normal lung development, MLC1a was expressed in vascular smooth muscle (VSM) cells of pulmonary artery, and MLC2 was present in parabronchial smooth muscle and VSM cells of pulmonary vessels. In addition, early smooth muscle differentiation delay was observed by immunohistochemistry of alpha-smooth muscle actin and transforming growth factor-beta1. Disruption of MLC1a expression during normal pulmonary development led to significant growth and branching impairment, suggesting a role in branching morphogenesis. Both MLC1a and MLC2 were absent from hypoplastic fetal lungs during pseudoglandular stage of lung development, whereas their expression partially recovered by prenatal treatment with vitamin A. Thus, a deficiency in contractile proteins MLC1a and MLC2 might have a role among the early molecular determinants of lung hypoplasia in the rat model of nitrofen-induced CDH.     

The chicken B-cell receptor complex and its role in avian B-cell development

Summary: The bursa of Fabricius is critical to normal B-lymphocyte development in birds. During embryonic life, B-cell precursors migrate to the bursal rudiment and those which have undergone productive V(D)J recombination colonize lymphoid follicles and undergo immunoglobulin V gene diversification by gene conversion. The chicken surface IgM complex appears structurally and functionally equivalent to its mammalian counterpart, with homologs to CD79a and CD79b. Expression of a truncated Ig chain is sufficient to drive the early stages of B-cell development in the embryo bursa. Bursal cells expressing the truncated receptor complex proliferate in bursal follicles, and those which contain V gene rearrangements undergo V gene diversification by gene conversion. The bursa is a gut-associated organ and antigen is focused to bursal lymphoid follicles after hatch. While expression of the truncated chain is sufficient to support B-cell development in the embryo, B cells expressing this receptor are rapidly eliminated after hatch. We suggest the possibility that B-cell development in the bursa after hatch is driven by encounter with antigen leading to redistribution of B cells within the lymphoid follicle, B-cell proliferation and V gene repertoire development by gene conversion.     

The mysterious immunoglobulin light chain.

The immunoglobulin light chain is mysterious from different points of view. In an evolutionary perspective there seems to be at least three major pathways but it is today impossible to say which of them is the most ancient one and which isotype belongs to which branch. It is also difficult to assess the contribution of the light chain to the binding capacity of the antibodies. It is possible that it differs from antibody to antibody but the fact that there are antibodies in camels and sharks without any light chain suggests that it is perhaps not necessary for good antigen binding.     

Flow cytometric analysis of surface light chain expression patterns in B-cell lymphomas using monoclonal and polyclonal antibodies

Light chain (LC) expression by flow cytometry (FC) in B cell non-Hodgkin lymphomas (B-NHLs) can occasionally be detected with one anti-LC antibody but not with another. We retrospectively analyzed 564 four-color FC files from B-NHLs, assessing LC staining with monoclonal antibodies (mAbs) and polyclonal antibodies (pAbs). Discrepancies in LC expression between mAbs and pAbs were present in 9.2% of cases, mainly in chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL; 11.1%), diffuse large B-cell lymphoma (DLBCL; 10.2%), follicular lymphoma (9.5%), and mantle cell lymphoma (11.1%) and most frequently in body fluids. Equal proportions of cases were LC+ only with pAbs (4.8%) or mAbs (4.4%). Negative LC expression with both antibodies was present in 7.5% of cases, most frequently in DLBCL (21.6%) and body fluids (27.6%). Evaluation with both mAbs and pAbs increases the sensitivity for LC detection, with no single reagent outperforming the other, although CLL/SLL preferentially showed LC expression with pAbs.     

Determination of light chain restriction in fine-needle aspiration-type preparations of B-cell lymphomas by mRNA in situ hybridization.

The purpose of this study was to investigate whether B-cell clonality could be demonstrated in fine-needle aspiration (FNA)-type preparations by using automated and manual in situ hybridization (ISH) systems. FNA-like preparations were made from 10 cases of B-cell lymphoma and 5 cases of reactive lymphoid hyperplasia. Kappa/lambda expression was determined using an automated mRNA ISH assay or a manual ISH system. Other variables tested included type and length of fixation, protease digestion, and time in chromogen. Clonality data were corroborated by either flow cytometry or tissue-based analysis. Optimal conditions required formalin fixation, strong protease digestion, and prolonged hybridization and chromogen times; under these conditions, monoclonality was demonstrated by in situ in 8 of 10 cases. Each of the five cases of reactive lymphoid hyperplasia showed polyclonal light chain expression by automated mRNA ISH. In situ kappa/lambda mRNA analysis of FNA-type specimens allows direct determination of monoclonality in cytologic preparations.     

Antigen-dependent B-cell development

There is growing evidence that the development of na?ve B cells depends on the interaction of self antigens with the BCR. A view that has emerged over the past year is that BCR signal output contributes in a large part to the developmental fate of peripheral B cells. Differences in antigen-receptor signal strength may determine whether B cells assume a marginal zone, follicular or B-1 phenotype.     

The role of microRNAs in B-cell development and function

MicroRNA (miRNA)-mediated gene silencing at the translational level has led to novel discoveries for numerous biological processes. Recently, there has been increasing evidence to indicate that miRNAs are involved in normal immune functions and inflammation. In this review, we focus on recent advances that have elucidated the role of miRNAs in B-cell development, differentiation, apoptosis and function. While the regulatory mechanisms of miRNAs in controlling and maintaining B-cell fate remain largely uncharacterized, further studies on miRNAs and their targets will increase our understanding of B-cell development and function. Such studies may be able to provide new therapeutic strategies for treating autoimmune diseases.     

Precursor B cells of mouse bone marrow express two different complexes with the surrogate light chain on the surface

Two monoclonal antibodies raised against the complex of μ heavy (H) chain and V_pre-B/λ5 surrogate light (L) chains recognize surrogate L chain in different conformations on normal pre-B cells. One, LM34 recognizes free λ5 protein and free λ5/V_pre-B surrogate L chains and binds to surrogate L chains on the surface of early, pro-B and pre-B-I cells where the surrogate L chain is associated with a gp130/gp35–65 complex of proteins. It also recognizes the surrogate L chain associated with the μH chain on pre-B-II cells. The other monoclonal antibody, SL156, does not recognize free surrogate L chain or its components, nor its complex with gp130/gp35–65 on pro-B and pre-B-I cells. However, it does bind to a conformational epitope on the surrogate light chain/μH chain complex on a subpopulation of pre-B-II cells and on μH chain-positive pre-B cell lines. On mouse precursor B cells prepared ex vivo on ice, expression of the surrogate L chain is very low and almost undetectable. Incubation of the precursor cells for 1 h at 37 °C up-regulates the surface expression of surrogate L chain associated with gp130/gp35–65 (early complex) as well as the μH chain/surrogate L chain complex. These results reconcile some of the apparently discrepant results on surface expression of the surrogate L chain obtained with human and mouse bone marrow pre-B cells, and show that a surrogate L chain/μH chain-containing pre-B cell receptor can be expressed also on the surface of mouse pre-B-II cells.