Fundamental characteristics of the expressed immunoglobulin VH and VL repertoire in different canine breeds in comparison with those of humans and mice

Steiniger, Sebastian C.J.; Dunkle, William; Bammert, Gary F.; Wilson, Thomas L.; Krishnan, Abhiram; Dunham, Stephen; Ippolito, Gregory C.; Bainbridge, G.R.

doi:10.1016/j.molimm.2014.01.010

Cited by 22 publications

(25 citation statements)

References 49 publications

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“…As evidenced in prior studies, we observed the heavy chain CDR3 amino acid length is longest with shorter kappa and lambda chain CDR3 lengths (28 -32). The heavy chain CDR3 amino acid usage distribution in our data is also like that previously reported (29,33). Importantly, our study highlights additional novel aspects of CDR3 biology.…”

Section: Discussionsupporting

confidence: 88%

Deep Characterization of the Human Antibody Response to Natural Infection Using Longitudinal Immune Repertoire Sequencing

Mitsunaga

Snyder

2020

Molecular & Cellular Proteomics

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Human antibody response studies are largely restricted to periods of high immune activity (e.g. vaccination). To comprehensively understand the healthy B cell immune repertoire and how this changes over time and through natural infection, we conducted immune repertoire RNA sequencing on flow cytometry-sorted B cell subsets to profile a single individual's antibodies over 11 months through two periods of natural viral infection. We found that 1) a baseline of healthy variable (V) gene usage in antibodies exists and is stable over time, but antibodies in memory cells consistently have a different usage profile relative to earlier B cell stages; 2) a single complementarity-determining region 3 (CDR3) is potentially generated from more than one VJ gene combination; and 3) IgG and IgA antibody transcripts are found at low levels in early human B cell development, suggesting that class switching may occur earlier than previously realized. These findings provide insight into immune repertoire stability, response to natural infections, and human B cell development.

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Section: Discussionsupporting

confidence: 88%

Deep Characterization of the Human Antibody Response to Natural Infection Using Longitudinal Immune Repertoire Sequencing

Mitsunaga

Snyder

2020

Molecular & Cellular Proteomics

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“…The light chains were less variable, with CDRL3s of 8–9 amino acids. Both CDR3 length ranges are consistent with antibodies generated in other model systems (34, 35). Each of the six heavy and light chain pairs were co-transfected into HEK293 cells to express recombinant antibodies.…”

Section: Resultssupporting

confidence: 85%

A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning

Carbonetti

Oliver

Вигдорович

et al. 2017

Journal of Immunological Methods

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Monoclonal antibody technologies have enabled dramatic advances in immunology, the study of infectious disease, and modern medicine over the past 40 years. However, many monoclonal antibody discovery procedures are labor- and time-intensive, low efficiency, and expensive. Here we describe an optimized mAb discovery platform for the rapid and efficient isolation, cloning and characterization of monoclonal antibodies in murine systems. In this platform, antigen-binding splenic B cells from immunized mice are isolated by FACS and cocultured with CD40L positive cells to induce proliferation and mAb production. After 12 days of coculture, cell culture supernatants are screened for antigen, and IgG positivity and RNA is isolated for reverse-transcription. Positive-well cDNA is then amplified by PCR and the resulting amplicons can be cloned into ligation-independent expression vectors, which are then used directly to transfect HEK293 cells for recombinant antibody production. After 4 days of growth, conditioned medium can be screened using biolayer interferometry for antigen binding and affinity measurements. Using this method, we were able to isolate six unique, functional monoclonal antibodies against an antigen of the human malaria parasite Plasmodium falciparum. Importantly, this method incorporates several important advances that circumvent the need for single-cell PCR, restriction cloning, and large scale protein production, and can be applied to a wide array of protein antigens.

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“…We observed expression from 3 IGHV subgroups, with the majority (72%) of these sequences belonging to the IGHV1 subgroup. These findings indicates that feline IGHV diversity is derived from the use of more germline subgroups genes than the canine IGHV repertoire, in which over 90% of the sequences are derived from a single canine IGHV subgroup (IGHV1), and the remainder from IGHV2 40 .…”

Section: Discussionmentioning

confidence: 95%

Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus

Tallmadge

Callaway

et al. 2017

Sci Rep

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In response to immunization, B-cells generate a repertoire of antigen-specific antibodies. Antibody-based immunotherapies hold great promise for treating a variety of diseases in humans. Application of antibody-based immunotherapy in cats is limited by the lack of species-specific complete sequences for mRNAs encoding rearranged heavy and light chain immunoglobulins in B cells. To address this barrier, we isolated mRNAs from feline peripheral blood mononuclear cells (PBMCs), and used available immunoglobulin sequences and 5′ and 3′ RACE to clone and sequence heavy and light chain immunoglobulin mRNAs. We recovered mRNA from PBMCs from two cats, cloned and sequenced the variable and constant domains of the feline heavy chains of IgG1a (IGHG1a), IgG2 (IGHG2), and IgA (IGHA), and the light chains (lambda and kappa). Using these sequences, we prepared two bicistronic vectors for mammalian expression of a representative feline heavy (IGHG1a) together with a light (lambda or kappa) chain. Here we report novel feline Ig sequences, a technique to express antigen-specific felinized monoclonal antibodies, and the initial characterization of a functional felinized monoclonal antibody against feline panleukopenia virus.

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Fundamental characteristics of the expressed immunoglobulin VH and VL repertoire in different canine breeds in comparison with those of humans and mice

Cited by 22 publications

References 49 publications

Deep Characterization of the Human Antibody Response to Natural Infection Using Longitudinal Immune Repertoire Sequencing

Deep Characterization of the Human Antibody Response to Natural Infection Using Longitudinal Immune Repertoire Sequencing

A method for the isolation and characterization of functional murine monoclonal antibodies by single B cell cloning

Sequence analysis of feline immunoglobulin mRNAs and the development of a felinized monoclonal antibody specific to feline panleukopenia virus

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