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B cell clonal expansion was identified in 3 of 15 OND. There were numerous somatic mutations, mainly in the CDRs, with a high replacement-to-silent ratio; the mutations were distributed in a way suggesting that these B cells had been positively selected through their antigen receptor.

Our results demonstrate that in MS CSF, there is a high frequency of clonally expanded B cells that have properties of postgerminal center memory or antibody-forming lymphocytes. The primary antibody repertoire is generated by mechanisms involving the assembly of the exons that encode the antigen-binding variable regions of immunoglobulin heavy Ig H and light Ig L chains during the early development of B lymphocytes.

After antigen-dependent activation, mature B lymphocytes can further alter their Ig H and Ig L variable region exons by the process of somatic hypermutation SHM , which allows the selection of B cells in which SHMs resulted in the production of antibodies with increased antigen affinity. In addition, during antigen-dependent activation, B cells can also change the constant region of their Ig H chain through a DNA double-strand-break DSB dependent process referred to as Ig H class switch recombination CSR , which generates B cell progeny that produce antibodies with different Ig H constant region effector functions that are best suited for a elimination of a particular pathogen or in a particular setting.

This review describes in depth the processes of SHM and CSR with a focus on mechanisms that direct AID cytidine deamination in activated B cells and mechanisms that promote the differential outcomes of such cytidine deamination. Although the primary function of the DNA mismatch repair MMR system is to identify and correct base mismatches that have been erroneously introduced during DNA replication, recent studies have further implicated several MMR components in somatic hypermutation of immunoglobulin Ig genes.

APOBEC3-mediated hypermutation of retroviral vectors produced from some retrovirus packaging cell lines. APOBEC3 proteins are packaged into retrovirus virions and can hypermutate retroviruses during reverse transcription. We found that HT human fibrosarcoma cells hypermutate retroviruses, and that the HT cell-derived FLYA13 retrovirus packaging cells also hypermutate a retrovirus vector produced using these cells.

We found no hypermutation of the same vector produced by the mouse cell-derived packaging line PT67 or by human cells transfected with the vector and retrovirus packaging plasmids. We expect that avoidance of vector hypermutation will be particularly important for vectors used in gene therapy, wherein mutant proteins might stimulate deleterious immune responses.

The rabbit is unique in having well-defined allotypes in the variable region of the heavy chain. Products of the VHa locus, with alleles a1, a2, and a3 , account for the majority of the serum immunoglobulins. A small percentage of the serum immunoglobulins are a-negative. From this animal the Alicia rabbit strain was developed and the mutation was termed ali. We previously showed, using Southern analysis and the transverse alternating field electrophoresis technique, that the difference between the ali rabbit and normal is a relatively small deletion including some of the most 3' VH genes.

The most JH proximal 3' VH1 genes in DNA from normal rabbits of a1, a2 and a3 haplotypes encode a1, a2 and a3 molecules respectively, and it has been suggested that these genes are responsible for allelic inheritance of VHa allotypes. The present study suggests that the 3' end of the VH locus probably plays a key role in regulation of VH gene expression in rabbits because VH gene s in this region are the target s of preferential VDJ rearrangements.

This raises the possibility that mechanisms such as somatic gene conversion and hypermutation are at work to generate the antibody repertoire in this species. Our data support the view that the 3' VH1 gene may be the preferred target for rearrangement in normal rabbits, and for the normal chromosome in heterozygous ali animals.

However, homozygous ali rabbits with a deletion that removed the a2-encoding VH1 on both chromosomes do survive, rearrange other VH genes and produce normal levels of immunoglobulins as well as a significant percentage of B cells which bear the a2 allotype. Combined hereditary and somatic mutations of replication error repair genes result in rapid onset of ultra- hypermutated cancers.

DNA replication-associated mutations are repaired by two components: polymerase proofreading and mismatch repair. The mutation consequences of disruption to both repair components in humans are not well studied. We sequenced cancer genomes from children with inherited biallelic mismatch repair deficiency bMMRD. We previously identified a candidate Ig kappa gene corresponding to this manuscript. We show now the gene referred to as: "sea star Ig kappa gene in its specificity".

Together with Southern blotting, our data suggest that multiple genes encoding both Ig Y forms are present in the turtle genome. Both of the Ig Y forms were detected in the serum using rabbit polyclonal Abs. Somatic immunoglobulin hypermutation. Immunoglobulin hypermutation provides the structural correlate for the affinity maturation of the antibody response.

Characteristic modalities of this mechanism include a preponderance of point-mutations with prevalence of transitions over transversions, and the mutational hotspot RGYW sequence. The nature of the targeting mechanism and the trans-factors effecting such breaks and their repair remain to be determined.

X-linked hyper-immunoglobulin M hyper- Ig M syndrome XHIM is a primary immunodeficiency due to mutations in CD40 ligand that affect immunoglobulin class-switch recombination and somatic hypermutation. The disease is amenable to gene therapy using retroviral vectors, but dysregulated gene expression results in abnormal lymphoproliferation in mouse models, highlighting the need for alternative strategies.

Site-specific insertion of the donor sequence downstream of the endogenous CD40L promoter maintained physiologic expression of CD40L while overriding all reported downstream mutations. Notably, gene -corrected HSCs engrafted in immunodeficient mice at clinically relevant frequencies.

These studies provide the foundation for a permanent curative therapy in XHIM. Published by Elsevier Inc. HIV1 V3 loop hypermutability is enhanced by the guanine usage bias in the part of env gene coding for it. Distribution of guanine between three codon positions should influence the probability for G to A mutation to be nonsynonymous to occur in first or second codon position.

We discovered that nucleotide sequences of env genes coding for third variable regions V3 loops of gp from HIV1 and HIV2 have different kinds of guanine usage biases. According to the HIV1 and HIV2 V3 alignment, insertion of the sequence enriched with 2G 21 codons in length occurred during the evolution of HIV1 predecessor, while insertion of the different sequence enriched with 3G 19 codons in length occurred during the evolution of HIV2 predecessor.

The higher is the level of 3G in the V3 coding region, the lower should be the immune escaping mutation occurrence rates. This hypothesis was tested in this study by comparing the guanine usage in V3 loop coding regions from HIV1 fast and slow progressors. A hypermutated subtype of advanced prostate cancer was recently described, but prevalence and mechanisms have not been well-characterized.

Our findings identify parallels and differences in the mechanisms of hypermutation in prostate cancer compared with other MSI-associated cancers. Somatic hypermutation SHM mediated by activation-induced cytidine deaminase AID is a transcription-coupled mechanism most responsible for generating high affinity antibodies. By modelling the dynamical behaviour of IGHV DNA, a commonly used human variable gene segment, we observed that hypermutable bases on the non-transcribed strand are paired whereas those on transcribed strand are mostly unpaired.

Hypermutable bases both paired and unpaired are made accessible to AID in stabilised secondary structures formed with increasing transcription levels. This observation provides a rationale for the hypermutable bases on both the strands of DNA being targeted to a similar extent despite having differences in unpairedness. Hypermutation In Pancreatic Cancer. Pancreatic cancer is molecularly diverse, with few effective therapies. Increased mutation burden and defective DNA repair are associated with response to immune checkpoint inhibitors in several other cancer types.

We interrogated pancreatic cancer genomes to define hypermutation and its causes. Mutational signatures inferring defects in DNA repair were enriched in those with the highest mutation burdens. Defining mutation load in individual pancreatic cancers and the optimal assay for patient selection may inform clinical trial design for immunotherapy in pancreatic cancer.

Ig W is present in the lungfish, a bony fish sharing a common ancestor with sharks million years ago, implying that the Ig W VH family is as old as the Ig M VH family. This nurse shark study examined the Ig M and Ig W VH repertoire from birth through adult life, and analyzed the phylogenetic relationships of these gene families.

Despite the clear presence of N-region additions, newborn CDR3 were significantly shorter than those of adults. The Ig M clones are all included in a conventional VH family that can be classified into five discrete groups, none of which is orthologous to Ig M VH genes in other elasmobranchs.

In addition, a novel divergent VH family was orthologous to a published monotypic VH horn shark family. Ig W VH genes have diverged sufficiently to form three families. Phylogenetic analysis of cartilaginous fish and lungfish Ig M and Ig W demonstrated they form two major ancient gene groups; furthermore, these VH genes generally diversify duplicate and diverge within a species. As in ratfish, sandbar and horn sharks, most nurse shark Ig M VH.

Conclusion As in ratfish, sandbar and horn. What do somatic hypermutation and class switch recombination teach us about chronic lymphocytic leukaemia pathogenesis? Interestingly, the presence or absence of somatic hypermutation SHM process is associated with the use of particular VH genes.

Particular alleles of the VH gene and the VH gene are preferentially expressed in an unmutated form, while VH or the majority of VH3 family genes frequently contain somatic mutations. The mutational profile of Ig genes has been shown to be associated with disease prognosis. These results could favour the idea that CLL could correspond to two different diseases that look alike in morphologic and phenotypic terms.

Donehower, Lawrence A. These tumors, unlike the majority of colorectal carcinomas, are often diploid, exhibit frequent epigenetic silencing of the MLH1 DNA mismatch repair gene , and have a better clinical prognosis. As an adjunct study to The Cancer Genome Atlas consortium that recently analyzed colorectal cancers by whole exome sequencing, we compared the 35 CRC These patterns were confirmed in an independent validation set of exome-sequenced CRC. Our findings suggest that hypermutated CRC include one subgroup with fundamentally different pathways to malignancy than the majority of CRC.

Detection of hypermutated human papillomavirus type 16 genome by Next-Generation Sequencing. Human papillomavirus type 16 HPV16 is a major cause of cervical cancer. To investigate in vivo characteristics of hypermutation , differential DNA denaturation-PCR 3D-PCR was performed using three clinical specimens obtained from HPVpositive cervical dysplasia, and detected hypermutation from two out of three specimens.

One sample accumulating hypermutations in both E2 and the long control region LCR was further subjected to Next-Generation Sequencing, revealing that hypermutations spread across the LCR and all early genes. Notably, hypermutation was more frequently observed in the LCR, which contains a viral replication origin and the early promoter.

To analyse the restriction of salivary Ig G subclasses, we used unstimulated whole saliva and sera collected at the same time from individuals with homozygous gene deletions, two with G1 deletion, one with G4 deletion, six with both G2 and G4 deletions and from eight individuals without IGHG gene deletions and expressing all four Ig G subclasses.

The median values of salivary Ig G from individuals with homozygous G1, or G4, or both G2 and G4 deletions, and from individuals expressing all four subclasses were The median values of serum Ig G were Our results show that the salivary and serum Ig G levels were both within the normal range in individuals with homozygous gene deletions of either G1, or G4, or both G2 and G4. Structure and antigenicity analysis of the Ig G gene for Nyctereutes procyonoides.

Nyctereutes procyonoides immunoglobulin G Ig G gene is partially cloned. And the serum antibody of Nyctereutes procyonoides had obviously cross-reaction with HRP conjugated rabbit anti-dog Ig G, compared with those of canine, oryctolagus cuniculus, mus musculus, mink, gallus. There is the closest ties of consanguinity of Ig G exist between Nyctereutes procyonoides and canine among the mammal through the genetic evolution.

The detection and treament of canine distemper can be used on Nyctereutes procyonoides. The remnant of the European rabbit Oryctolagus cuniculus Ig D gene. Although Ig D first appeared, along with Ig M, in the cartilaginous fishes and has been retained throughout subsequent vertebrate evolution, it has been lost in a diverse group of vertebrate species. Chicken immune responses to human proteins are often more robust than rodent responses because of the phylogenetic relationship between the different species.

For discovery of a diverse panel of unique therapeutic antibody candidates, chickens therefore represent an attractive host for human-derived targets. Recent advances in monoclonal antibody technology, specifically new methods for the molecular cloning of antibody genes directly from primary B cells, has ushered in a new era of generating monoclonal antibodies from non-traditional host animals that were previously inaccessible through hybridoma technology.

However, such monoclonals still require post-discovery humanization in order to be developed as therapeutics. To obviate the need for humanization, a modified strain of chickens could be engineered to express a human-sequence immunoglobulin variable region repertoire. Here, human variable genes introduced into the chicken immunoglobulin loci through gene targeting were evaluated for their ability to be recognized and diversified by the native chicken recombination machinery that is present in the B-lineage cell line DT After expansion in culture the DT40 population accumulated genetic mutants that were detected via deep sequencing.

Bioinformatic analysis revealed that the human targeted constructs are performing as expected in the cell culture system, and provide a measure of confidence that they will be functional in transgenic animals. Characterization of a gene coding for a type IIo bacterial Ig G-binding protein. Two antigenic classes of non-immune Ig G-binding proteins can be expressed by group A streptococci. One antigenic group of proteins is recognized by an antibody prepared against the product of a cloned fcrA gene anti-FcRA.

In this study, the immunogen used to prepare the antibody that defines the second antigenic class was shown to be the product of the emm-like emmL gene of M serotype 55 group A isolate, A The emmL55 gene expressed in E. These properties are characteristic of the previously described type IIo Ig G-binding protein isolated from this strain. In addition, the recombinant protein was reactive with human serum albumin and fibrinogen.

The emmL 55 gene sequence was analysed and found to have the organization and sequence characteristics of a typical class I emm-like gene. A key event in this differentiation is the extensive cell loss due to apoptosis. Because the proto-oncogene bcl-2 can promote cell survival, we established pre-B cell lines from E mu-bcl-2 transgenic mice.

These pre-B cells have the same properties as those derived from non-bcl-2 transgenic mice except that they do not die by apoptosis. This allowed us to study the fate of newly formed B cells in vitro for a longer period of time. Here we show that early during the differentiation of pre-B cells, upregulation of RAG-1 and RAG-2 expression go hand in hand with rearrangements of the Ig gene loci. Thus, deposition of a complete Ig molecule on the surface of a B cell does not automatically stop the Ig -rearrangement machinery.

Somatic hypermutation and antigen-driven selection of B cells are altered in autoimmune diseases. B cells have been found to play a critical role in the pathogenesis of several autoimmune AI diseases. A common feature amongst many AI diseases is the formation of ectopic germinal centers GC within the afflicted tissue or organ, in which activated B cells expand and undergo somatic hypermutation SHM and antigen-driven selection on their immunoglobulin variable region Ig V genes.

However, it is not yet clear whether these processes occurring in ectopic GCs are identical to those in normal GCs. The analysis of Ig V mutations has aided in revealing many aspects concerning B cell expansion, mutation and selection in GC reactions. We have found that RA and MS sequences exhibited normal mutation spectra and targeting motifs, but a stricter selection compared to normal controls, which was more apparent in RA.

SS sequence analysis results deviated from normal controls in both mutation spectra and indications of selection, also showing differences between light and heavy chain Ig V and between different tissues. The differences revealed between AI diseases and normal control mutation patterns may result from the different microenvironmental influences to which ectopic GCs are exposed, relative to those in normal secondary lymphoid tissues.

Identification of a cofactor responsible for the fact that Ig V genes are much more sensitive to AID-induced mutagenesis than other genes is a key question in immunology. SRSF1 not only acts as an essential splicing factor but also regulates diverse aspects of mRNA metabolism and maintains genome stability.

Our findings, thus, define an unexpected and important role for SRSF1, particularly for its splice variant, in enabling AID to function specifically on its natural substrate during SHM. Compared with mammals, the bird Ig genetic system relies on gene conversion to create an Ab repertoire, with inversion of the Ig A-encoding gene and very few cases of Ig subclass diversification.

Although gene conversion has been studied intensively, class-switch recombination, a mechanism by which the Ig H C region is exchanged, has rarely been investigated in birds. In this article, we show that both DNA deletion and inversion may result from Ig A and Ig Y class switching, and similar junction patterns were observed for both types of class-switch recombination.

The data obtained in this study provide new insight into class-switch recombination and Ig gene evolution in birds. Ig VH gene analysis suggests that peritoneal B cells do not contribute to the gut immune system in man.

The contribution of peritoneal B cells to the intestinal lamina propria plasma cell population is well documented in mice, but unknown in humans. We have analyzed immunoglobulin Ig genes of human peritoneal B cells, because such genes show distinctive characteristics in mucosal B cells, particularly highly mutated variable regions. We focused on the properties of Ig V H to allow comparisons of like-with-like between different isotypes and cells from different immune compartments.

We observed that the Ig M genes were mostly unmutated, and that the mutated subset had less mutations than would be expected in a mucosal B cell population. Likewise, the Ig V H genes used by Ig A and Ig G from peritoneal B cells had significantly lower numbers of mutations than observed in the mucosal counterparts. Other trends observed, while not reaching statistical significance, followed the trend of peripheral B cells.

The peritoneal B cell population had more Ig A1 than Ig A2 sequences, and there was no dominance of J H 4 in the Ig A from peritoneum or spleen, in contrast to the mucosal sequences. Overall, this study suggested that human peritoneal B cell are either peripheral or mixed in origin; they are unlikely to represent an inductive compartment for the mucosal B cell system. Antibody repertoire diversification through VH gene replacement in mice cloned from an Ig A plasma cell.

In mammals, VDJ recombination is responsible for the establishment of a highly diversified preimmune antibody repertoire. Acquisition of a functional Ig heavy H chain variable V gene rearrangement is thought to prevent further recombination at the Ig H locus. VH replacement occurred in early pro-B cells, was independent of pre-B-cell receptor signaling, and involved predominantly one adjacent VH germ-line gene. Fanconi anemia is a rare genetic disorder that can lead to bone marrow failure, congenital abnormalities, and increased risk for leukemia and cancer.

Cells with loss-of-function mutations in the FANC pathway are characterized by chromosome fragility, altered mutability, and abnormal regulation of the nonhomologous end-joining NHEJ pathway. Both processes are initiated after the generation of dG:dU mismatches by activation-induced cytidine deaminase. Enhancement of hypermutation frequency in the chicken B cell line DT40 for efficient diversification of the antibody repertoire.

Chicken B cell line DT40 continuously accumulates mutations in the immunoglobulin variable region Ig V gene by gene conversion and point mutation, both of which are mediated by activation-induced cytidine deaminase AID , thereby producing an antibody Ab library that is useful for screening monoclonal Abs mAbs in vitro. To efficiently create an Ab library with sufficient diversity, higher hypermutation frequency is advantageous.

However, the protein level of the truncated AID mutant was less than that of the wild type. Chronic lymphocytic leukemia CLL patients exhibit a variable clinical course. To investigate the association between clinicobiologic features and responsiveness of CLL cells to anti- Ig M stimulation, we evaluated gene expression changes and modifications in cell-cycle distribution, proliferation, and apoptosis of Ig V H mutated M and unmutated UM samples upon BCR cross-linking.

Supervised analysis identified several genes modulated exclusively in the UM cases upon BCR cross-linking. Functional gene groups, including signal transduction, transcription, cell-cycle regulation, and cytoskeleton organization, were up-regulated upon stimulation in UM cases.

Cell-cycle and proliferation analyses confirmed that Ig M cross-linking induced a significant progression into the G 1 phase and a moderate increase of proliferative activity exclusively in UM patients. These results document that a differential genotypic and functional response to BCR ligation between Ig V H M and UM cases is operational in CLL, indicating that response to antigenic stimulation plays a pivotal role in disease progression.

Human Ig G2- and Ig G4-expressing memory B cells display enhanced molecular and phenotypic signs of maturity and accumulate with age. The mechanisms involved in sequential immunoglobulin G Ig G class switching are still largely unknown. Sequential IG class switching is linked to higher levels of somatic hypermutation SHM in vivo, but it remains unclear if these are generated temporally during an immune response or upon activation in a secondary response.

We here aimed to uncouple these processes and to distinguish memory B cells from primary and secondary immune responses. SHM levels and Ig G subclasses were studied with pyrosequencing on blood mononuclear cells from young children and adults as models for primary and secondary immunological memory.

We conclude that secondary immunological memory accumulates with age and these memory B cells express CD27, high levels of activation markers, and carry high SHM levels and frequent usage of IGHG2. These new insights contribute to our understanding of sequential Ig G subclass switching and show a potential relevance of using serum Ig G2 levels or numbers of Ig G2-expressing B cells as markers for efficient generation of memory responses.

Aberrant O-glycosylation of serum immunoglobulin A1 Ig A1 represents a heritable pathogenic defect in Ig A nephropathy, the most common form of glomerulonephritis worldwide, but specific genetic factors involved in its determination are not known.

These genes encode molecular partners essential for enzymatic O-glycosylation of Ig A1. Our findings provide novel insights into the genetic regulation of O-glycosylation and are relevant not only to Ig A nephropathy, but also to other complex traits associated with O-glycosylation defects, including inflammatory bowel disease, hematologic disease, and cancer. Integrating multiple immunogenetic data sources for feature extraction and mining somatic hypermutation patterns: the case of "towards analysis" in chronic lymphocytic leukaemia.

The analysis of SHM has offered critical insight into the physiology and pathology of B cells, leading to strong prognostication markers for clinical outcome in chronic lymphocytic leukaemia CLL , the most frequent adult B-cell malignancy.

In this paper we present a methodology for integrating multiple immunogenetic and clinocobiological data sources in order to extract features and create high quality datasets for SHM analysis in IG receptors of CLL patients. This dataset is used as the basis for a higher level integration procedure, inspired form social choice theory. This is applied in the Towards Analysis, our attempt to investigate the potential ontogenetic transformation of genes belonging to specific stereotyped CLL subsets towards other genes or gene families, through SHM.

The data integration process, followed by feature extraction, resulted in the generation of a dataset containing information about mutations occurring through SHM. The Towards analysis performed on the integrated dataset applying voting techniques, revealed the distinct behaviour of subset compared to other subsets, as regards SHM related movements among gene clans, both in allele-conserved and non-conserved gene areas.

With respect to movement between genes , a high percentage movement towards pseudo genes was found in all CLL subsets. This data integration and feature extraction process can set the basis for exploratory analysis or a fully automated computational data mining approach on many as yet unanswered, clinically relevant biological questions. Ig G1 memory B cells keep the memory of Ig E responses. The unique differentiation of Ig E cells suggests unconventional mechanisms of Ig E memory.

By identifying the cellular origin of high affinity Ig E and the clonal selection of high affinity memory B cells into the plasma cell fate, our findings provide fundamental insights into the pathogenesis of allergies, and on the mechanisms of antibody production in memory B cell responses. Ig E is an important mediator of protective immunity as well as allergic reaction, but how high affinity Ig E antibodies are produced in memory responses is not clear.

Here the authors show that Ig E can be generated via class-switch recombination in Ig G1 memory B cells without additional somatic hypermutation. Genes encoding Xenopus laevis Ig L chains: Implications for the evolution of [kappa] and [lambda] chains.

Xenopus laevis Ig contain two distinct types of L chains, designated [rho] or L1 and [sigma] or L2. Many fragments hybridized to the V probe, but only one or two fragments hybridized to the C probe. Corresponding C, J, and V gene segments were identified on clones isolated from a genomic library prepared from the same DNA. One clone contains a C gene segment separated from a J gene segment by an intron of 3.

The C segment is somewhat more similar and the J segment considerably more similar in sequence to the corresponding segments of mammalian [kappa] chains than to those of mammalian [lambda] chains. Upstream of the J segment is a typical recombination signal sequence with a spacer of 23 bp, as in J[kappa].

A second clone from the library contains four V gene segments, separated by 2. Two of these, V1 and V3, have the expected structural and regulatory features of V genes , and are very similar in sequence to each other and to mammalian V[kappa].

A third gene segment, V2, resembles V1 and V3 in its coding region and nearby 5[prime]-flanking region, but diverges in sequence 5[prime] to position [minus]95 with loss of the octamer promoter element. The fourth V-like segment is similar to the others at the 3[prime]-end, but upstream of codon 64 bears no resemblance in sequence to any Ig V region.

All four V segments have typical recombination signal sequences with bp spacers at their 3[prime]-ends, as in V[kappa]. Taken together, the data suggest that Xenopus L1 L chain genes are members of the [kappa] gene family. Somatic hypermutation SHM , coupled with Ag selection, provides a mechanism for generating Abs with high affinity for invading pathogens. Class-switch recombination CSR ensures that these Abs attain pathogen-appropriate effector functions. Although the enzyme critical to both processes, activation-induced cytidine deaminase, has been identified, it remains unclear which cis-elements within the Ig loci are responsible for recruiting activation-induced cytidine deaminase and promoting its activity.

Hyper- Ig M syndrome type 4 with a B lymphocyte—intrinsic selective deficiency in Ig class-switch recombination. We investigated the clinical and immunological characteristics of 15 patients with an unidentified form of HIGM. We found that B cell CSR was intrinsically impaired. However, the generation of somatic hypermutations was observed in the variable region of the Ig heavy chain gene , as in control B lymphocytes. Polymorphisms in positions 46, 79 and were not associated with fenoterol- or budesonide-modulated Ig E synthesis.

No differences in the distribution of Ig E synthesis was seen between atopic and non-atopic individuals carrying the same alleles. Published by Elsevier Espana. Anti-h Ig E gene therapy of peanut-induced anaphylaxis in a humanized murine model of peanut allergy. Peanuts are the most common food to provoke fatal or near-fatal anaphylactic reactions.

Treatment with an anti-h Ig E mAb is efficacious but requires frequent parenteral administration. Based on the knowledge that peanut allergy is mediated by peanut-specific Ig E, we hypothesized that a single administration of an adeno-associated virus AAV gene transfer vector encoding for anti-h Ig E would protect against repeated peanut exposure in the host with peanut allergy.

We developed a novel humanized murine model of peanut allergy that recapitulates the human anaphylactic response to peanuts in NOD-scid IL2Rgamma null mice transferred with blood mononuclear cells from donors with peanut allergy and then sensitized with peanut extract. As therapy, we constructed an adeno-associated rh. In the reconstituted mice peanut-specific Ig E was induced by peanut sensitization and hypersensitivity, and reactions were provoked by feeding peanuts to mice with symptoms similar to those of human subjects with peanut allergy.

A single administration of AAVrh. The anti-h Ig E vector, administered either before sensitization or after peanut sensitization and manifestation of the peanut-induced phenotype, blocked Ig E-mediated alterations in peanut-induced histamine release, anaphylaxis scores, locomotor activity, and free Ig E levels and protected animals from death caused by anaphylaxis.

If this degree of persistent efficacy translates to human subjects, AAVrh. AID attacks differ from those in cCSR because they are not accompanied by extensive somatic hypermutation SHM of targeted regions and because repaired junctions exhibit features of the alternative end-joining A-EJ pathway.

Histone H3. Immunoglobulin diversification is driven by activation-induced deaminase AID , which converts cytidine to uracil within the Ig variable Ig V regions. Loss of H3. Published under the terms of the CC BY 4. T cell-dependent B cell responses typically develop in germinal centers.

Abs generated during such responses are isotype switched and have a high affinity to the Ag because of somatic hypermutation of Ab genes. B cell responses to purified polysaccharides are T cell independent and do not result in the formation of bona fide germinal centers, and the dominant Ab isotype produced during such responses is Ig M with very few or no somatic mutations.

Activation-induced cytidine deaminase AID is required for both somatic hypermutation and Ig isotype switching in humans and mice. Typhi or purified Vi polysaccharide ViPS. However, the differences in the Ab-dependent killing of S.

Typhi mediated by the classical pathway of complement activation were not statistically significant. Typhi bactericidal Ab titers at 7 but not at 28 d postimmunization were also comparable. To test the protective immunity conferred by these immunizations, mice were challenged with a chimeric S. Typhimurium strain expressing ViPS. With the recent discovery of mutations in the STAT3 gene in the majority of patients with classic Hyper- Ig E syndrome, it is now possible to make a molecular diagnosis in most of these cases.

We scanned for mutations in 10 unrelated pedigrees, which include 16 patients with classic Hyper- Ig E syndrome. These pedigrees include both sporadic and familial cases and their relatives, and we have found STAT3 mutations in all affected individuals. High-resolution melting analysis allows a single day turn-around time for mutation scanning and targeted sequencing of the STAT3 gene , which will greatly facilitate the rapid diagnosis of the Hyper- Ig E syndrome, allowing prompt and appropriate therapy, prophylaxis, improved clinical outcome, and accurate genetic counseling.

Extensive plasma cell infiltration with crystal Ig G inclusions and mutated Ig V H gene in an osteoarthritis patient with lymphoplasmacellular synovitis. A case report. The presence of immunoglobulin crystal inclusions in plasma cells from plasmacytomas and B-NHLs linked to overstimulation and overproduction has been frequently reported.

Our case describes a lymphoplasmacellular synovitis in a patient with osteoarthritis OA showing an unusually high plasma cell infiltration and for the first time crystals in plasma cells. Using immunohistochemistry. Since no germinal centers could be detected and the analyzed B cells showed antigen selectivity, it may be concluded that already antigenically activated B cells migrated into the synovium and locally differentiated into plasma cells, leading to the extensive infiltration observed.

Rheumatoid fibroblasts were shown to support terminal B cell differentiation. Our data suggests that the ability of fibroblasts to activate B cells is not only restricted to RA, but also occurs in OA. The intense plasma cell infiltration contributed to further cartilage damage by altering the microenvironment of the nourishing synovial tissue or by the local production of pathogenic autoantibodies. Hypermutation in Burkholderia cepacia complex is mediated by DNA mismatch repair inactivation and is highly prevalent in cystic fibrosis chronic respiratory infection.

The Burkholderia cepacia complex Bcc represents an important group of pathogens involved in long-term lung infection in cystic fibrosis CF patients. A positive selection of hypermutators , linked to antimicrobial resistance development, has been previously reported for Pseudomonas aeruginosa in this chronic infection setting.

Hypermutability , however, has not yet been systematically evaluated in Bcc species. A total of well characterized Bcc isolates recovered from 48 CF patients, 10 non-CF patients and 15 environmental samples were analyzed. In order to determine the prevalence of mutators their spontaneous mutation rates to rifampicin resistance were determined.

In addition, the genetic basis of the mutator phenotypes was investigated by sequencing the mutS and mutL genes , the main components of the mismatch repair system MRS. The overall prevalence of hypermutators in the collection analyzed was Thirteen Finally, searching for a possible association between antimicrobial resistance and hypermutability , the resistance-profiles to 17 antimicrobial agents was evaluated. High antimicrobial resistance rates were documented for all the Bcc species recovered from CF patients, but, except for ciprofloxacin, a significant association with hypermutation was not detected.

In conclusion, in the present study we demonstrate for the first time that, MRS-deficient Bcc species mutators are highly prevalent and positively selected in CF chronic lung infections. Hypermutation therefore, might be playing a key role in increasing bacterial adaptability to the CF-airway environment, facilitating the persistence of chronic lung infections. Presence of Streptococcus mutans strains harbouring the cnm gene correlates with dental caries status and Ig A nephropathy conditions.

Streptococcus mutans is a major pathogen of human dental caries. Strains harbouring the cnm gene , which encodes Cnm, a collagen-binding protein, contribute to the development of several systemic diseases. In this study, we analysed S. Saliva specimens were collected from Ig AN patients and the cnm status of isolated S. Berghof, Tom V. Natural antibodies NAb are antigen binding antibodies present in individuals without a previous exposure to this antigen.

Keyhole limpet hemocyanin KLH -binding NAb levels were previously associated with survival in chickens. This suggests that selective breeding for KLH-binding NAb may increase survival by means of improved general disease resistance. The region of interest was fine mapped by imputing the region of the study population to whole genome sequence, and subsequently performing an association study using the imputed sequence variants.

SNP located in coding regions of the candidate genes were checked for predicted changes in protein functioning. One SNP at 69,, base pairs received the maximum impact score from two independent prediction tools, which makes this SNP the most likely causal variant. This study contributes to increased understanding of genetic regulation of KLH-binding NAb levels, and total antibody concentrations.

A locus control region LCR containing four transcriptional enhancers lies downstream of the Ig H chain locus. We also compared transgenes that were or were not flanked by two copies of the beta-globin HS4 insulator, an element defined by its ability to protect transgenes from the influences of surrounding genes at the insertion site.

Flow cytometry experiments reported a strictly B cell specific GFP expression from pre-B cells in bone marrow to mature B cells in spleen. Despite addition of 5 ' HS4 insulators to the GFP-3 ' LCR construct, complete transgene silencing occurred in some transgenic lines and was systematically observed in ageing animals from all lines.

Analysis of gene expression and Ig transcription in PU. A number of presumptive target genes for the Ets-family transcription factor PU. However, the precise function of PU. In this study, we take advantage of recently developed PU. We show that contrary to previously published expectations, PU.

In fact, PU. In addition, high levels of Igkappa transcription are induced after IL-7 withdrawal of wild-type or PU. In contrast, we found that Iglambda transcription is reduced in PU. The PU. Taken together, our observations suggest that reduction of PU. Here, we show that certain pairwise combinations of base analog mutagens display synergy or suppression.

In particular, the combination of 2-aminopurine and zebularine, analogs of adenine and cytidine, respectively, shows a fold increased mutation frequency compared with that of either mutagen alone. Understanding the mechanism of synergy can lead to increased understanding of mutagenic processes. As combinations of base analogs are used in certain chemotherapy regimens, including those involving ZEB and 5AZ, these results indicate that testing the mutagenicity of all drug combinations is prudent.

Overlapping activation-induced cytidine deaminase hotspot motifs in Ig class-switch recombination. Ig class-switch recombination CSR is directed by the long and repetitive switch regions and requires activation-induced cytidine deaminase AID. By using somatic gene targeting and recombinase-mediated cassette exchange, we established a cell line-based CSR assay that allows manipulation of switch sequences at the endogenous locus.

This finding leads to a much clearer model for the difference between CSR and somatic hypermutation. VH—expressing clones are common in the naive B cell repertoire but are rarely found in Ig G memory B cells from healthy individuals. However, this paradoxical self-reactivity correlated with these VH—encoded Ig G clones binding commensal bacteria antigens.

Hence, B cells expressing germline-encoded self-reactive VH antibodies may represent an innate-like B cell population specialized in the containment of commensal bacteria when gut barriers are breached. A role for Msh6 but not Msh3 in somatic hypermutation and class switch recombination. Somatic hypermutation is initiated by activation-induced cytidine deaminase AID , and occurs in several kilobases of DNA around rearranged immunoglobulin variable V genes and switch S sites before constant genes.

AID deaminates cytosine to uracil, which can produce mutations of C:G nucleotide pairs, and the mismatch repair protein Msh2 participates in generating substitutions of downstream A:T pairs. Msh2 is always found as a heterodimer with either Msh3 or Msh6, so it is important to know which one is involved. Therefore, we sequenced V and S regions from Msh3- and Msh6-deficient mice and compared mutations to those from wild-type mice.

Msh6-deficient mice had fewer substitutions of A and T bases in both regions and reduced heavy chain class switching, whereas Msh3-deficient mice had normal antibody responses. This establishes a role for the Msh2-Msh6 heterodimer in hypermutation and switch recombination. An Ig M monoclonal gammopathy points to a diagnosis of Waldenstrom's Macroglobulinemia. Other B lymphoproliferatives disorders should be ruled out but the limits are sometimes difficult to define.

For the 49 who had a monoclonal peak, we analysed the underlying pathology in termes of general, clinical and biological characteristics. We checked if the MYD88 mutation had been detected. The overall survival rate was studied. The MW group was compared to the other groups. Regarding biological characteristics, the Ig M level upon diagnosis was statistically higher in the MW group with a median level at A precise diagnosis requires collating clinical, histological, immunophenotypical and genetical data.

The murine Cd48 gene : allelic polymorphism in the Ig V-like region. The murine CD48 molecule is a member of the immunoglobulin superfamily which regulates the activation of T lymphocytes. To resolve this issue, we have directly sequenced genomic DNA of 10 laboratory strains and two inbred strains of wild origin.

The results of our analysis reveal an allelic polymorphism within the Ig V-like domain of murine CD A double-strand break can trigger immunoglobulin gene conversion. All three B cell-specific activities of the immunoglobulin Ig gene re-modeling system— gene conversion, somatic hypermutation and class switch recombination—require activation-induced deaminase AID. The pattern and pseudogene usage of DSB-induced gene conversion were comparable to those of AID-induced gene conversion; surprisingly, sometimes a single DSB induced multiple gene conversion events.

These constitute direct evidence that a DSB in the V region can be an intermediate for gene conversion. Humanized mouse models present an important tool for preclinical evaluation of new vaccines and therapeutics. Published by Elsevier B. Immunoglobulin affinity maturation depends on somatic hypermutation SHM in immunoglobulin variable Ig V regions initiated by activation-induced cytidine deaminase AID. Abstract Immunoglobulin affinity maturation depends on somatic hypermutation SHM in immunoglobulin variable Ig V regions initiated by activation-induced cytidine deaminase AID.

Recurrence of base decamers, nonomers, and octamers within mouse C mu gene of Ig heavy chain and its primordial building block. Within the published 2,base-long mouse C mu gene of Ig heavy chain consisting of four coding and four noncoding segments, 2 base decamers, 8 nonomers, and 39 octamers recurred.

Recurring base heptamers about and hexamers about were simply too numerous to merit individual identification. In spite of extensive overlaps between these recurring base decamers to hexamers, they occupied nearly the entire length of mouse Ig C mu gene. As with other genes of the beta-sheet-forming beta 2-microglobulin family, the Ig C mu gene flanking and intervening noncoding sequences included is not a unique sequence but rather it is degenerate repeats of the base-long primordial building-block sequence uniquely its own.

This primordial building block must originally have specified the amino-acid-residue-long primordial arm of beta-sheet-forming loops, the characteristics of the beta 2-microglobulin family of polypeptides. HIDS is an autosomal recessive disorder characterized by recurrent fever attacks in early childhood. During febrile episodes, urinary mevalonate concentrations were found to be significantly elevated in patients.

Diagnosis of HIDS was retrieving gene or measurement of the enzyme activity in peripheral blood lymphocyte in general. The relation between the fever and inflammation of mevalonate or isoprenoid products are uncertain. The therapy attempt with statins, which is inhibited the next enzyme after HMG-CoA reductase, or inhibit the proinflammatory cytokines.

In vitro studies showed that single enzyme-DNA encounters result in distributive deamination of adjacent cytidines. Increased ssDNA concentrations abolished enzyme processivity in the case of short, but not long, DNA substrates, emphasizing the key role of rapid intersegmental transfer in targeting the deaminase. Selection of genes that have not been horizontally transferred for prokaryote phylogenetic inferences is regarded as a challenging task.

The markers internal transcribed spacer of ribosomal genes 16SS ITS and phycocyanin intergenic spacer PC- IGS , based on the operons of ribosomal and phycocyanin genes respectively, are among the most used markers in cyanobacteria.

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Cryptocurrency 2021 nfl One advantage of Ramos cells is that they have a built-in convenient semi-quantitative measure of SHM. Hence, B cells expressing germline-encoded self-reactive VH antibodies may represent an innate-like B cell population specialized in the containment of commensal bacteria when gut barriers are breached. Open demo account. Sale, Julian E. However, its re-expression in disease states, including cancer and sepsis, results in destabilization of blood vessels contributing to the pathology of these conditions. The receptor functions as a second line of antibacterial defense involving serum Ig A rather than secretory Ig A. Published by Elsevier Espana.
Betting assistant windows 8 Ig spread betting apic, we determined the effects of hypermutation on the HIV-1 recombination rate and its contribution to genetic variation through recombination to team totals sports betting progeny genomes containing portions of hypermutated genomes without lethal mutations. Characteristic modalities of this mechanism include a preponderance of point-mutations with prevalence of transitions over transversions, and the mutational hotspot RGYW sequence. Our case describes a lymphoplasmacellular synovitis in a patient with osteoarthritis OA showing an unusually high plasma cell infiltration and for the first time crystals in plasma cells. So go as huge as potential along with your qualifying deposits in order to maximize this nice supply. The Ig M clones are all included in a conventional VH family that can be classified into five discrete groups, none of which is orthologous to Ig M VH genes in other elasmobranchs. Since no germinal centers could be detected and the analyzed B cells showed antigen selectivity, it may be concluded that already antigenically activated B cells migrated into the synovium and locally differentiated into plasma cells, leading to the extensive infiltration observed.
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