Organization of the variable region of the immunoglobulin heavy-chain gene locus of the rat

Hendricks, Jacobus; Terpstra, Peter; Dammers, Peter M.; Somasundaram, Rajesh; Visser, Anita; Stoel, Maaike; Bos, Nicolaas A.; Kroese, Frans G. M.

doi:10.1007/s00251-010-0448-x

Cited by 13 publications

(18 citation statements)

References 34 publications

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“…56,57,67 Two possible explanations for this difference are that only one particular IGHV gene family (viz., the IGHV5 family, the homologue of PC7183 in the mouse) has been analyzed in the (PVG) rat and that this IGHV gene family is not representative for other IGHV genes, or IGHV gene families. By establishing the genomic germline IGHV gene repertoire of the BN rat 68 as illustrated in Fig. 2, it became possible to accurately analyze other IGHV gene families as well.…”

Section: Igm Expressing Marginal Zone Memory B Cellsmentioning

confidence: 99%

“…69 These three IGHV gene families have 4, 2, and 26 functional IGHV genes, respectively. 68 The IGHV3 and IGHV4 genes families were chosen to determine whether there is a difference in mutation frequencies among members of IGHV gene families that are relatively small and to compare this frequency to the second-largest IGHV gene family (IGHV5) in the rat, which had also been analyzed previously in the PVG rat. 53 The BN rat strain contains 26 functional IGHV5 (germline) genes compared to the 28 germline genes in the PVG rat.…”

Section: Igm Expressing Marginal Zone Memory B Cellsmentioning

confidence: 99%

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Heterogeneity of Memory Marginal Zone B Cells

2018

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The marginal zone (MZ) is largely composed of a unique subpopulation of B cells, the so-called MZ-B cells. At a molecular level, memory B cells are characterized by the presence of somatically mutated IGV genes. The earliest studies in the rat have documented the presence of hapten-specific MZ-B cells after immunization in the MZ. This work later received experimental support demonstrating that the IGHV-Cµ transcripts expressed by phenotypically defined splenic MZ-B cells (defined as CD90negIgMhighIgDlow B cells) can carry somatic hypermutation. However, only a minor fraction (< 10%–20%) of these MZ-B cells is mutated and is considered to represent memory B cells. Memory B cells can either be class-switched (IgG, IgA, IgE), or non–class-switched (IgM) B cells. B cells in the MZ are a heterogeneous population of cells and both naïve MZ-B cells; class switched and unswitched memory MZ-B cells are present at this unique site in the spleen. Naïve MZ-B cells carry unmutated Ig genes, produce low-affinity IgM molecules and constitute a first line of defense against invading pathogens. Memory MZ-B cells express high-affinity Ig molecules, directed to (microbial) antigens that have been encountered. In this review, we report on the memory compartment of splenic MZ-B cells in the rat to provide insights into the origin and function of these memory MZ-B cells.

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Section: Igm Expressing Marginal Zone Memory B Cellsmentioning

confidence: 99%

Section: Igm Expressing Marginal Zone Memory B Cellsmentioning

confidence: 99%

Heterogeneity of Memory Marginal Zone B Cells

2018

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“…Also the utilization of IGHD genes by the IGHV5-C␥ transcripts is similar for all three B cell fractions (Fisher's exact test, P = 0.794). Of the known functional IGHD genes (Hendricks et al, 2010) the IGHD genes 1-6 and 1-7 appear to be preferentially used to encode for IgG antibodies in all three fractions.…”

Section: The Repertoire Of Class-switched Ighv Region Genes Obtained mentioning

confidence: 99%

Class-switched marginal zone B cells in spleen have relatively low numbers of somatic mutations

Hendricks

Visser

Dammers

et al. 2011

Molecular Immunology

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The vast majority of rodent splenic marginal zone (MZ)-B cells are naive IgM(+) cells. A small fraction of these MZ-B cells carry mutated V-genes, and represent IgM(+) memory MZ-B cells. Here we reveal further heterogeneity of B cells with a MZ-B cell phenotype, by providing evidence for the existence of class-switched memory MZ-B cells in the rat. In essence, we observed IGHV5 encoded Cγ transcripts, among FACS-purified MZ-B cells, defined as HIS24(low)HIS57(bright) cells. Furthermore, we found that most IgG encoding transcripts are mutated. There is no significant difference in IGHV5 repertoire and subclass usage of these IgG encoding transcripts collected from B cells with a MZ-B cell phenotype and B cells with a follicular (FO) B cell phenotype. However, the IGHV5 genes encoding for IgG antibodies of MZ-B cells exhibited significantly fewer mutations, compared to those with a FO-B cell phenotype. In one rat we found a clonally related set of IgG encoding sequences, of which one was derived from the MZ-B cell fraction and the other from the FO-B cell fraction. We speculate that these two subpopulations of class-switched B cells are both descendants from naive FO-B cells and are generated in germinal centers. Class-switched memory cells with a MZ-B cell phenotype may provide the animal with a population of IgG memory cells that can respond rapidly to blood-borne pathogens.

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“…In 2008, Sepulveda and Shoemaker 6 compared rat EST sequences and mouse VL/VH coding sequences and designed a primer set which amplifies 65 heavy and 77 light chains-predicted variable domains. From the curated IMGT database, we currently know that in spite of the close similarity to mouse, rats have a very high number of functional variable genes in the genome (232 VH and 164VLK) with IGVH1, IGVH2 and IGVH5 33 being the most abundant families. Sepulveda's primer set only covers family 1 and 2, lacking coverage of the highly abundant family IGVH5 and other families such as IGVH8 and IGVH11.…”

Section: Discussionmentioning

confidence: 99%

A primer set for the rapid isolation of scFv fragments against cell surface antigens from immunised rats

Nannini

Parekh

Wawrzyniecka

et al. 2020

Sci Rep

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Antibody phage display is a powerful platform for discovery of clinically applicable high affinity monoclonal antibodies against a broad range of targets. Libraries generated from immunized animals offer the advantage of in vivo affinity-maturation of V regions prior to library generation. Despite advantages, few studies have described isolation of antibodies from rats using immune phage display. In our study, we describe a novel primer set, covering the full rat heavy chain variable and kappa light chain variable regions repertoire for the generation of an unbiased immune libraries. Since the immune repertoire of rats is poorly understood, we first performed a deep sequencing analysis of the V(D)J regions of VH and VLK genes, demonstrating the high abundance of IGVH2 and IGVH5 families for VH and IGVLK12 and IGVLK22 for VLK. The comparison of gene’s family usage in naïve rats have been used to validate the frequency’s distribution of the primer set, confirming the absence of PCR-based biases. The primers were used to generate and assemble a phage display library from human CD160-vaccinated rats. CD160 represents a valid therapeutic target as it has been shown to be expressed on chronic lymphocytic leukaemia cells and on the surface of newly formed vessels. We utilised a novel phage display panning strategy to isolate a high affinity pool (KD range: 0.399–233 nM) of CD160 targeting monoclonal antibodies. Subsequently, identified binders were tested for function as third generation Chimeric Antigen Receptors (CAR) T cells demonstrating specific cytolytic activity. Our novel primer set coupled with a streamlined strategy for phage display panning enable the rapid isolation and identification of high affinity antibodies from immunised rats. The therapeutic utility of these antibodies was demonstrated in CAR format.

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Organization of the variable region of the immunoglobulin heavy-chain gene locus of the rat

Cited by 13 publications

References 34 publications

Heterogeneity of Memory Marginal Zone B Cells

Heterogeneity of Memory Marginal Zone B Cells

Class-switched marginal zone B cells in spleen have relatively low numbers of somatic mutations

A primer set for the rapid isolation of scFv fragments against cell surface antigens from immunised rats

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