Overview

Also Known As: CD28^-

These Cd28 knock-out mice exhibit rapid onset Type 1 diabetes in both males and females.

Donating Investigator

Dr. Jeffrey A. Bluestone, University of California, San Francisco

Genetic overview

Genetic Background	Generation

Cd28^tm1Mak

Allele Type	Gene Symbol	Gene Name
Targeted (Null/Knockout)	Cd28	CD28 antigen

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Research Applications

Immunology, Inflammation and Autoimmunity Research
Diabetes and Obesity Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

NOD mice heterozygous for the Cd28^tm1Mak mutation are viable, fertile; exhibiting diabetes onset and incidence similar to NOD. NOD mice homozygous for Cd28^tm1Mak experience rapid onset Type 1 diabetes in both males and females. When stimulated with anti-Cd3 mAB, T cells from CD28 deficient NOD mice proliferate poorly when compared with wildtype controls. Nor do T cells proliferate in culture after the mice have been immunized with ovalbumin (OVA) in complete Freunds adjuvant. The IL-2 level of T-cells from mice that have been stimulated with either anti-Cd3 or OVA was significantly reduced. However, when challenged with GAD the CD28 deficient mice produced normal levels of IL-2. RT-PCR indicates a dramatic increase in the amount of IFNgamma and decreased IL4 compared to heterozygous controls. 1.5% of CD4 T cells from homozygous mice express CD25, while 6.5% of the CD4 T cells express CD25 in their wildtype cohorts.

This model is useful for studying the CTLA4/CD28:CD80/CD86 pathways in the regulation of self tolerance and susceptibility to autoimmune diseases.

Development

Cd28, located on Chr. 1, 30.1cM, is involved in costimulatory signaling important for T cell activation.
A construct containing a neomycin expression cassette replacing part of exon 2 of Cd28 was transfected into D3 (129S2/SvPas derived) embryonic stem cells (ES cells). These ES cells were injected into C57BL/6 blastocysts. As reported by Lenschow et al 1996, chimeric founders were initially mated to C57BL/6 and intercrossed to generate Cd28 deficient mice. B6 congenic mice homozygous for the mutation were subsequently mated to NOD for 4 generations, Lenschow et al 1996, prior to making homozygous. In 2006, the T1DR received mice heterozygous for the mutation at generation N11.

Control Suggestions

Wild-type from the colony
001976 NOD/ShiLtJ

Additional Information

Considerations for Choosing Controls

Genetics

Cd28^tm1Mak

Allele Symbol: Cd28^tm1Mak

Allele Name	targeted mutation 1, Tak Mak
Allele Type	Targeted (Null/Knockout)
Allele Synonym(s)	targeted mutation 1, Tak Mak; Cd28^tm1Mak
Gene Symbol and Name	Cd28, CD28 antigen
Gene Synonym(s)	Tp44; CD28RNA
Strain of Origin	129S2/SvPas
Chromosome	1
Molecular Note	A genomic fragment containing part of exon 2 was replaced by a neomycin resistance cassette. Flow-cytometry analysis on peripheral blood lymphocytes demonstrated that the protein was not expressed in these cells in homozygous mice.
Mutations Made By	Dr. Tak Mak, University Health Network/Un of Toronto

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Genotype: Cd28^tm1Mak related

Immunology, Inflammation and Autoimmunity Research
- CD Antigens, Antigen Receptors, and Histocompatibility Markers

Diabetes and Obesity Research
- Type 1 Diabetes (IDDM)

Mammalian Phenotype Terms by Genotype

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

Genotype: Cd28^tm1Mak/Cd28^tm1Mak
involves: 129S2/SvPas * NOD

cellular phenotype

decreased T cell proliferation
- T cells also produce less IL-2 during these in vitro stimulations
- T cells fail to proliferate when restimulated with OVA peptide
- T cells stimulated in vitro proliferate much less than controls to anti-CD3

immune system phenotype

abnormal T-helper 1 physiology
- splenic T cells secrete high levels of IFN-gamma while expressing very little to no IL-4 upon stimulation
- Th1 responses are enhanced in these mice

abnormal T-helper 2 physiology
- serum contains significantly less of the Th2-dependent anti-GAD IgG2a antibodies
- splenic T cells secrete high levels of IFN-gamma while expressing very little to no IL-4 upon stimulation
- Th2 responses are dampened in these mice

decreased IgG2a level
- serum contains significantly less of the Th2-dependent anti-GAD IgG2a antibodies

decreased interleukin-2 secretion
- T cells also produce less IL-2 during stimulation with anti-CD3 antibody or upon restimulation with OVA peptide
- Normal - T cells secrete the same amout of IL-2 as controls when stimulated with the diabetic autoantigen GAD

decreased interleukin-4 secretion
- T cell secrete little to no IL-4 upon activation in vitro

decreased T cell proliferation
- T cells also produce less IL-2 during these in vitro stimulations
- T cells fail to proliferate when restimulated with OVA peptide
- T cells stimulated in vitro proliferate much less than controls to anti-CD3

increased interferon-gamma secretion
- splenic T cells secrete high levels of IFN-gamma, sometimes as much as 4-fold compared to controls

increased susceptibility to autoimmune diabetes
- 70-80% of female mice in the fourth backcross onto the NOD strain are hyperglycemic by 24 weeks of age compared to 30% for non-transgenic littermate controls
- nearly 90% of male mice in this backcross are hyperglycemic by 24 weeks of age compared to 10% of non-transgenic littermate controls

hematopoietic system phenotype

abnormal T-helper 1 physiology
- splenic T cells secrete high levels of IFN-gamma while expressing very little to no IL-4 upon stimulation
- Th1 responses are enhanced in these mice

abnormal T-helper 2 physiology
- serum contains significantly less of the Th2-dependent anti-GAD IgG2a antibodies
- splenic T cells secrete high levels of IFN-gamma while expressing very little to no IL-4 upon stimulation
- Th2 responses are dampened in these mice

decreased IgG2a level
- serum contains significantly less of the Th2-dependent anti-GAD IgG2a antibodies

decreased T cell proliferation
- T cells also produce less IL-2 during these in vitro stimulations
- T cells fail to proliferate when restimulated with OVA peptide
- T cells stimulated in vitro proliferate much less than controls to anti-CD3

The following phenotype relates to a compound genotype created using this strain

Genotype: Cd28^tm1Mak/Cd28^tm1Mak
NOD.129S2-Cd28

cellular phenotype

increased T cell proliferation
- expansion of transferred CD25-depleted T cells from Tg(TcraBDC2.5)1Doi Tg(TcrbBDC2.5)2Doi is dramatically accelerated compared to NOD controls; there are 5-fold more autoreactive transferred T cells in pancreatic lymph nodes compared to NOD mice

hematopoietic system phenotype

abnormal CD4-positive, alpha-beta T cell physiology
- only 1.5% of CD4 T cells in homozygotes express CD25 compared to 6.5% in wild-type

increased T cell proliferation
- expansion of transferred CD25-depleted T cells from Tg(TcraBDC2.5)1Doi Tg(TcrbBDC2.5)2Doi is dramatically accelerated compared to NOD controls; there are 5-fold more autoreactive transferred T cells in pancreatic lymph nodes compared to NOD mice

immune system phenotype

abnormal CD4-positive, alpha-beta T cell physiology
- only 1.5% of CD4 T cells in homozygotes express CD25 compared to 6.5% in wild-type

abnormal response to transplant
- CD28-deficient NOD mice injected with CD4+CD25+ NOD T cells showed significant delay in the development of diabetes with no disease apparent through 15 weeks of age, whereas transfer of CD4+CD25+ T cells resulted in diabetes development by 11 weeks of age

increased susceptibility to autoimmune diabetes
- mice show higher incidence of diabetes compared to Cd40lg, Cd28 double null mice

increased T cell proliferation
- expansion of transferred CD25-depleted T cells from Tg(TcraBDC2.5)1Doi Tg(TcrbBDC2.5)2Doi is dramatically accelerated compared to NOD controls; there are 5-fold more autoreactive transferred T cells in pancreatic lymph nodes compared to NOD mice

References

Additional References

Shahinian A; Pfeffer K; Lee KP; Kundig TM; Kishihara K; Wakeham A; Kawai K; Ohashi PS; Thompson CB; Mak TW. 1993. Differential T cell costimulatory requirements in CD28-deficient mice. Science 261(5121):609-12PubMed: 7688139 MGI: J:14194
Salomon B; Lenschow DJ; Rhee L; Ashourian N; Singh B; Sharpe A; Bluestone JA. 2000. B7/CD28 costimulation is essential for the homeostasis of the CD4+CD25+ immunoregulatory T cells that control autoimmune diabetes. Immunity 12(4):431-40PubMed: 10795741 MGI: J:61905

Additional - Cd28^tm1Mak related

Technical Support

Contact Technical Support

Genotyping Protocols
- Standard PCR:Cd28^tm1Mak
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred together.
- Additional Breeding and Husbandry Support
Appearance
- albino, pink-eyed
  Related Genotype: A/A Tyr^c/Tyr^c
Citation
When using the CD28^- mouse strain in a publication, please cite the originating article(s) and include JAX stock #004761 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 - Not-For-Profit & Academic Pricing

Service	Genotype	Price
	Heterozygous or Wild-type for Cd28<tm1Mak>

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.

Related Products and Services

Frozen Mouse Embryo	NOD.129S2(B6)-Cd28<tm1Mak>/JbsJ Frozen Embryo	$2595.00
Frozen Mouse Embryo	NOD.129S2(B6)-Cd28<tm1Mak>/JbsJ Frozen Embryo	$2595.00
Frozen Mouse Embryo	NOD.129S2(B6)-Cd28<tm1Mak>/JbsJ Frozen Embryo	$3373.50
Frozen Mouse Embryo	NOD.129S2(B6)-Cd28<tm1Mak>/JbsJ Frozen Embryo	$3373.50

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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.

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Also Known As: CD28-

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Control Suggestions

Additional Information

Cd28tm1Mak

Allele Symbol: Cd28tm1Mak

Disease Terms

Research Areas By Genotype

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

Genotype: Cd28tm1Mak related

Mammalian Phenotype Terms by Genotype

Genotype: Cd28tm1Mak/Cd28tm1Makinvolves: 129S2/SvPas * NOD

cellular phenotype

immune system phenotype

hematopoietic system phenotype

Genotype: Cd28tm1Mak/Cd28tm1MakNOD.129S2-Cd28

cellular phenotype

hematopoietic system phenotype

immune system phenotype

References

Additional References

Additional - Cd28tm1Mak related

Genotyping Protocols

Breeding Considerations

Appearance

Citation

Animal Health Reports

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

Live Mouse

Cryorecovery - Not-For-Profit & Academic Pricing

Related Products and Services

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

Also Known As: CD28^-

Cd28^tm1Mak

Allele Symbol: Cd28^tm1Mak

Genotype: Cd28^tm1Mak related

Genotype: Cd28^tm1Mak/Cd28^tm1Mak
involves: 129S2/SvPas * NOD

Genotype: Cd28^tm1Mak/Cd28^tm1Mak
NOD.129S2-Cd28

Additional - Cd28^tm1Mak related