Overview

These "kappa-macroself Ag#2" transgenic mice may be useful to study B cell receptor editing, B cell tolerance in polyclonal immune systems, and for the rapid screening of mutants of interest for potential defects in B cell tolerance.

Donating Investigator

David Nemazee, The Scripps Research Institute

Genetic overview

Genetic Background	Generation

Tg(UBC-scFv)2Nemz

Allele Type
Transgenic (Inserted expressed sequence)

Pepc^b

Allele Type	Gene Symbol	Gene Name
Not Applicable	Pepc	peptidase C

Ptprc^a

Allele Type	Gene Symbol	Gene Name
Not Applicable	Ptprc	protein tyrosine phosphatase, receptor type, C

View Genetics

Research Applications

Research Tools
Immunology, Inflammation and Autoimmunity Research

View All Research Applications

Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

"Kappa-macroself Ag#2" transgenic mice express a membrane-bound "macroself" superantigen with specificity for the mouse immunoglobulin kappa light chain (IgΚ). Hemizygous mice are viable and fertile. The transgene is expressed in virtually all cells tested, including lymphoid tissue and bone marrow. B cells expressing the IgΚ-macroself superantigen are absent from the peripheral lymphoid organs while Igl⁺ B cell numbers are substantially increased. Although bone marrow B cell numbers are unchanged, total peripheral B cell populations are reduced by approximately one half. These mice also express the CD45.1 (Ly5.1 or Ptprca) allele, which is atypical for this C57BL/6 (B6) congenic background, and this marker may be used to track donor cell populations in bone marrow transplantation studies with B6 (CD45.2: Ptprc^b) mice.
These "kappa-macroself Ag#2" transgenic mice may be useful to study B cell receptor editing and B cell tolerance in a polyclonal immune system, and to rapidly screen other mutations for potential defects in B cell tolerance.

Genotyping Note: As the transgene expresses a superantigen reactive to immunoglobulin kappa of mouse and may interfere with antibody specificity if using mouse anti-CD45 antibodies, a pre-blocking step using unlabeled mouse IgG:kappa (or equivalent blocking step) is recommended for FACS-typing of the Ptprc (CD45) allele.

Development

These mice are designed to express a membrane-tethered single-chain variable fragment antibody with specificity to mouse IgΚ light chain. The 4 kb Ulik transgene contains the 1.2 kb human ubiquitin C promoter, rheumatoid factor Ig light chain gene leader exon and first intron, the expressed variable genes from the rat anti-mouse Ig CΚ hybridoma 187 line, the hinge, CH2, and CH3 sequences from rat IgG1 heavy chain cDNA, and the transmembrane and intracytoplasmic sequences from the mouse H2-k^b gene. This construct was microinjected into (B6xDBA/2)F1 embryos and then implanted into pseudopregnant Igk-deficient foster dams. "Kappa-macroself" transgenic mice (founder line 2) were backcrossed to mice on a "B6.CD45.1" background (derived from Stock No. 002014, B6.SJL-Ptprc^a Pepc^b/BoyJ) for approximately 20 generations before arriving at The Jackson Laboratory.

Expression Data

Expressed Gene	scFv, single chain variable fragment,
Site of Expression
Site of Expression
Site of Expression	Widely expressed on all adaptive and innate immune cells.

Control Suggestions

Noncarrier

Approximate Controls

Additional Information

Considerations for Choosing Controls

Selected References

Ait-Azzouzene D; Verkoczy L; Peters J; Gavin A; Skog P; Vela JL; Nemazee D. 2005. An immunoglobulin C{kappa}-reactive single chain antibody fusion protein induces tolerance through receptor editing in a normal polyclonal immune system. J Exp Med 201(5):817-28PubMed: 15738053 MGI: J:96759

View All References

Genetics

Tg(UBC-scFv)2Nemz

Allele Symbol: Tg(UBC-scFv)2Nemz

Allele Name	transgene insertion 2, David Nemazee
Allele Type	Transgenic (Inserted expressed sequence)
Allele Synonym(s)	kappa-macroself; Ulik
Gene Symbol and Name	Tg(UBC-scFv)2Nemz, transgene insertion 2, David Nemazee
Gene Synonym(s)
Promoter	UBC, ubiquitin C, human
Expressed Gene	scFv, single chain variable fragment,
Strain of Origin	(C57BL/6 x DBA/2)F1
Chromosome	UN
Molecular Note	These mice are designed to express a membrane-tethered single chain variable fragment antibody with specificity to mouse Igk light chain. The 4 kb "Ulik" transgene contained the 1.2 kb human ubiquitin C promoter, rheumatoid factor Ig light chain gene leader exon and first intron, the expressed variable genes from the rat anti-k hybridoma 187 line, the hinge, CH2, and CH3 sequences from rat gamma chain 1 (CH1 domain deleted), and the transmembrane and intracytoplasmic sequences from the H2-kb gene.
Mutations Made By	Djemel Ait Azzouzene , The Scripps Research Institute

Pepc^b

Allele Symbol: Pepc^b

Allele Name	b variant
Allele Type	Not Applicable
Allele Synonym(s)	Pep3^b
Gene Symbol and Name	Pepc, peptidase C
Gene Synonym(s)
Strain of Origin	Not Applicable
Chromosome	1
Molecular Note	The Pepc gene controls electrophoretic variation of peptidase C (PEP-C; EC 3.4.11.-) of red blood cells. The allele Pepc^b determines a faster migrating band and is found in the CBA/J and A/He strains.

Ptprc^a

Allele Symbol: Ptprc^a

Allele Name	a variant
Allele Type	Not Applicable
Allele Synonym(s)	CD45.1; Ly5^a; Ptprc^SJL
Gene Symbol and Name	Ptprc, protein tyrosine phosphatase, receptor type, C
Gene Synonym(s)
Site of Expression	Widely expressed on all adaptive and innate immune cells.
Strain of Origin	Not Applicable
Chromosome	1
Molecular Note	Ptprc^a is found in strains SJL/J, STS/A, and DA. Ptprc^b is found in strains C57BL/6, C3H/An, DBA/2, AKR, and many others (J:13367, J:12054, J:12077, J:8603). Twelve nucleotide differences between the a and b alleles have been identified. These base substitutions correspond to five amino-acid changes within the extracellular domain of the encoded protein. These amino-acid differences are clustered in a region that also contains the greatest divergence between mouse and rat sequences (J:22485). Note that the allele designations originally described were reversed in 1987 (J:8603); all publications prior to 1987 show SJL/J, STS/A, and DA as having the b allele and the C57BL/6J group as having the a allele (J:22341).

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Research Tools
- Immunology, Inflammation and Autoimmunity Research
  - B cell deficiency
  - production of B cells and antibodies
Immunology, Inflammation and Autoimmunity Research
- Lymphoid Tissue Defects
- Autoimmunity
  - B cell deficiency
- Immunodeficiency
  - defects in humoral immune responses
  - B cell deficiency

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Tg(UBC-scFv)2Nemz/0
involves: C57BL/6 * DBA/2

hematopoietic system phenotype

abnormal B cell differentiation

abnormal B cell morphology

abnormal B cell receptor editing
- B220+ cells fail to express kappa light chain
- immature B cells expressing kappa light chain are forced to undergo receptor editing
- kappa light chain is detected in cytoplasm
- levels of lambda V and J excision products exhibit a three fold increase in bone marrow and spleen cells
- proliferation is not increased

abnormal B-1 B cell morphology
- kappa positive B1-b cells are absent in the peritoneal cavity

abnormal immunoglobulin level
- kappa serum IgM and IgG2a/b are decreased, however, lambda serum IgM and IgG2a/b are increased

abnormal mature B cell morphology
- kappa positive B cells are absent from peripheral lymphoid tissues
- percentage of lambda positive B cells in peripheral lymphoid tissues is increased seven fold relative to non-transgenic mice

decreased mature B cell number
- B220+ fraction of spleen cells is reduced 60%
- B220+ peripheral B cell numbers are reduced by approximately 50%

decreased splenocyte number
- total number of splenic cells is reduced 80% as compared to control

spleen hypoplasia
- total number of splenic cells is reduced 80% as compared to control

immune system phenotype

abnormal B cell differentiation

abnormal B cell morphology

abnormal B cell receptor editing
- B220+ cells fail to express kappa light chain
- immature B cells expressing kappa light chain are forced to undergo receptor editing
- kappa light chain is detected in cytoplasm
- levels of lambda V and J excision products exhibit a three fold increase in bone marrow and spleen cells
- proliferation is not increased

abnormal B-1 B cell morphology
- kappa positive B1-b cells are absent in the peritoneal cavity

abnormal immune tolerance
- mice are tolerized to anti-kappa light chain resulting in the absence of kappa expressing B cells

abnormal immunoglobulin level
- kappa serum IgM and IgG2a/b are decreased, however, lambda serum IgM and IgG2a/b are increased

abnormal mature B cell morphology
- kappa positive B cells are absent from peripheral lymphoid tissues
- percentage of lambda positive B cells in peripheral lymphoid tissues is increased seven fold relative to non-transgenic mice

decreased mature B cell number
- B220+ fraction of spleen cells is reduced 60%
- B220+ peripheral B cell numbers are reduced by approximately 50%

decreased splenocyte number
- total number of splenic cells is reduced 80% as compared to control

spleen hypoplasia
- total number of splenic cells is reduced 80% as compared to control

References

Ait-Azzouzene D; Verkoczy L; Peters J; Gavin A; Skog P; Vela JL; Nemazee D. 2005. An immunoglobulin C{kappa}-reactive single chain antibody fusion protein induces tolerance through receptor editing in a normal polyclonal immune system. J Exp Med 201(5):817-28PubMed: 15738053 MGI: J:96759

Additional - Pepc^b related

Additional - Ptprc^a related

Additional - Tg(UBC-scFv)2Nemz related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Probe:Ptprc End Point-Alternate 1

Breeding Considerations

When maintaining a live colony, hemizygous mice can be bred to wildtype siblings or to Stock No. 002014. The donating investigator has not attempted to breed hemizygous mice together.
Genotyping Note: As the transgene expresses a superantigen reactive to immunoglobulin kappa of mouse and may interfere with antibody specificity if using mouse anti-CD45 antibodies, a pre-blocking step using unlabeled mouse IgG:kappa (or equivalent blocking step) is recommended for FACS-typing of the Ptprc (CD45) allele.

Additional Breeding and Husbandry Support

Citation
When using the B6.Cg-Pepc^b Ptprc^a Tg(UBC-scFv)2Nemz/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #006259 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic

International

Live Mouse

Age Genotype Price

weeks

Cryorecovery - Pricing

Service/Product Description Price

Homozygous for Pepc<b>, Homozygous for Ptprc<a>, Hemizygous or Non carrier for Tg(UBC-scFv)2Nemz

Payment Terms and Conditions

Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.

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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.

Terms Of Use

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General Terms and Conditions

Questions about Terms of Use

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Licensing Information

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Tg(UBC-scFv)2Nemz

Allele Symbol: Tg(UBC-scFv)2Nemz

Pepcb

Allele Symbol: Pepcb

Ptprca

Allele Symbol: Ptprca

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Tg(UBC-scFv)2Nemz/0involves: C57BL/6 * DBA/2

hematopoietic system phenotype

immune system phenotype

References

Additional - Pepcb related

Additional - Ptprca related

Additional - Tg(UBC-scFv)2Nemz related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms Of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Pepc^b

Allele Symbol: Pepc^b

Ptprc^a

Allele Symbol: Ptprc^a

Genotype: Tg(UBC-scFv)2Nemz/0
involves: C57BL/6 * DBA/2

Additional - Pepc^b related

Additional - Ptprc^a related