Description

Strain Information

Former Names B6.129P2-Ptprca Igh-Vtm1Mnz/J    (Changed: 11-JUL-07 ) Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Species laboratory mouse   Donating Investigator Michel Nussenzweig,   The Rockefeller University

Description
Mice homozygous for this targeted mutation are viable and fertile and do not display any gross physical or behavioral abnormalities. In combination with immunoglobulin lambda light chain, this heavy chain variable (V) region produces antibody that binds hapten NP (4-hydroxy-3-nitrophenylacetyl). A Trp to Leu point mutation at codon 33 results in a 40-fold increase in antigen binding affinity (B1-8^hi). Despite the significantly enhanced affinity for NP, there is only a 2-3 fold difference in the T-independent proliferative response of B cells that carry high or low affinity receptors. In T cell-dependent responses there is no difference in the ability of B1-8 high vs. B1-8 low affinity cells to enter germinal center reactions. However, high affinity B cells were recruited preferentially in response to both T cell-dependent and T cell-independent antigens. Expression has been verified by immunostaining cells that carry the mutation.

Development
PCR was used to introduce a point mutation conferring higher affinity 4-hydroxy-3-nitrophenyl)acetyl (NP)-binding to Igh (V_HB1-8): TGG->TTG (codon 33 Trp->Leu). A targeting vector incorporating a 5' floxed neomycin cassette in addition to the point mutation was transfected into 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. Positive clones were injected into C57BL/6 blastocysts, and the resulting chimeric males were bred to C57BL/6 EIIA-Cre females to delete the neomycin gene, and leave a loxP site. Mice were backcrossed to C57BL/6 congenic background CD45.1 (Ptprc^a) mice for at least 12 generations by the donating laboratory.

Related Strains

Strains carrying   Igh^tm1Mnz allele

007775

CBy.129P2(B6)-Ightm1Mnz/J

View Strains carrying   Igh^tm1Mnz     (1 strain)

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

Igh^tm1Mnz/Igh^tm1Mnz

        B6.129P2-Igh^tm1Mnz

• immune system phenotype
• abnormal B cell physiology (MGI Ref ID J:75653)
  • some mutant B cells that express Ig lambda differentiate into short-lived plasmacytes in red pulp of lymphoid follicles; plasmacyte response is greater than in Igh^tm2Mnz cells or wild-type
  • fraction of mutant B cells differentiate into short lived germinal center (GC) cells; GC response is greater than in Igh^tm2Mnz cells
  • when equal numbers of Igh^tm1Mnz and Igh^tm2Mnz B cells are transferred to wild-type recipient mice, after immunization with NP-Ficoll, Igh^tm1Mnz B cells preferentially expand and proliferation to make up ~46% of Ig lambda B cells whereas Igh^tm2Mnz B cells are not expanded
  • after 5 days post immunization, 85% of plasmacytes and 60% of germinal center cells are Igh^tm1Mnz B cells (high affinity)
  • mutant B cells with high affinity for antigen are preferentially selected during early phase of T1-2 immune response, over mutant B cells with low affinity for antigen
• increased B cell proliferation (MGI Ref ID J:75653)
  • mutant B cells in mice challenged with a T1-2 antigen (NP-Ficoll) undergo rapid proliferative expansion (from 3 to 25% of lymphocytes by 5 days after immunization
  • in immunized mice, on day 2 there is a much higher percentage of high-affinity B cells (58%) compared to wild-type cells (~3%)
• increased IgG level (MGI Ref ID J:75653)
  • 4-hydroxy-3-nitrophenyl)acetyl-specific (NP) IgM antibody production is greater than in Igh^tm2Mnz B cells or wild-type
• increased IgM level (MGI Ref ID J:75653)
  • NP-specific IgM antibody production is greater than in Igh^tm2Mnz B cells or wild-type
• increased B cell apoptosis (MGI Ref ID J:75653)
  • there are higher numbers of apoptotic mutant B cells than in wild-type or Igh^tm2Mnz B cells in spleens 3 and 5 days after immunization
• cellular phenotype
• abnormal cell cycle (MGI Ref ID J:75653)
  • absolute number of cells in cell cycle is always higher than in Igh^tm2Mnz or wild-type mice
• hematopoietic system phenotype
• increased B cell apoptosis (MGI Ref ID J:75653)
  • there are higher numbers of apoptotic mutant B cells than in wild-type or Igh^tm2Mnz B cells in spleens 3 and 5 days after immunization
• increased B cell proliferation (MGI Ref ID J:75653)
  • mutant B cells in mice challenged with a T1-2 antigen (NP-Ficoll) undergo rapid proliferative expansion (from 3 to 25% of lymphocytes by 5 days after immunization
  • in immunized mice, on day 2 there is a much higher percentage of high-affinity B cells (58%) compared to wild-type cells (~3%)

View Research Applications

Research Applications

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

Immunology and Inflammation Research

Igh related

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

Genes & Alleles

Gene & Allele Information

Allele Symbol		Ightm1Mnz
Allele Name		targeted mutation 1, Michel C Nussenzweig
Allele Type		Targeted (knock-in)
Common Name(s)		B1-8hi;
Mutation Made By		Michel Nussenzweig,   The Rockefeller University
Strain of Origin		129P2/OlaHsd
ES Cell Line Name		E14
ES Cell Line Strain		129P2/OlaHsd
Gene Symbol and Name		Igh, immunoglobulin heavy chain complex
Chromosome		12
Molecular Note		PCR was used to introduce a point mutation conferring higher affinity 4-hydroxy-3-nitrophenyl)acetyl (NP)-binding to Igh-V (VHB1-8): TGG->TTG (codon 33 Trp->Leu). A targeting vector incorporating a 5' floxed neomycin cassette in addition to the point mutation was transfected into 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. [MGI Ref ID J:75653]
Gene Symbol and Name		Ptprc, protein tyrosine phosphatase, receptor type, C
Chromosome		1
Gene Common Name(s)		B220; CD45; CD45 antigen; CD45R; Cd45; GP180; L-CA; LCA; LY5; Ly-5; Lyt-4; RT7; T-lymphocyte antigen 4; T200; lymphocyte antigen 5;

Genotyping

Genotyping Information

This strain will not have a genotyping protocol or one is not currently available.

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Shih TA; Roederer M; Nussenzweig MC. 2002. Role of antigen receptor affinity in T cell-independent antibody responses in vivo. Nat Immunol 3(4):399-406. [PubMed: 11896394]  [MGI Ref ID J:75653]

Additional References

Igh^tm1Mnz related

Reina-San-Martin B; Difilippantonio S; Hanitsch L; Masilamani RF; Nussenzweig A; Nussenzweig MC. 2003. H2AX is required for recombination between immunoglobulin switch regions but not for intra-switch region recombination or somatic hypermutation. J Exp Med 197(12):1767-78. [PubMed: 12810694]  [MGI Ref ID J:120658]

Health & husbandry

Health & Colony Maintenance Information

Currently there no information available for this strain. This may be due to the supply level of this strain.

Purchasing information

Pricing, Supply Level & Notes, Controls, General Terms & Conditions

Pricing

Supply Details

Standard Supply

Repository-Cryopreserved. Must Be Recovered. Please refer to pricing and supply notes for further information.

Supply Notes

Cryorecovery - Standard. The recovery process begins when a signed agreement form is returned to the Customer Service Department after order placement. Although results vary by strain, at least two males and two females (two pairs) will be provided, typically within 15 weeks of our receipt of the signed agreement form. If the first recovery attempt is unsuccessful or only one pair is recovered, a second recovery will be done, extending the delivery time to approximately 25 weeks. At least one member of each pair will be of known genotype and will carry the mutation if it is a mutant strain. Please note that pairs may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation of the strain. Mating schemes are sometimes modified for successful cryopreservation. Price represents a repository maintenance fee, which includes the cost of recovery of the strain from the cryopreservation resource and the periodic replacement of the frozen embryos used for recovery. Cryorecovery to establish a Dedicated Supply for greater quantities of mice. One to two pairs will be recovered to establish a Dedicated Supply of mice. Price by quotation. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 or 1-207-288-5845. This strain is included in the Induced Mutant Resource Colony collection.

General Terms and Conditions

See Terms of Use

The Jackson Laboratory's Genotype Promise

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.

---

Ordering and Purchasing Information

      Purchasing Information
      JAX^® Mice Orders
      Surgical Services

Contact Information
Orders & Technical Support
Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
Technical Support Email Form

Terms of Use

Terms of Use

General Terms and Conditions

Use restrictions apply, see Policy on Licensing and Use Restrictions.

Contact information

General inquiries

Contracts Administration

phone:	207-288-6470
fax:	207-288-6655

JAX^® Mice & Services Conditions of Use

“Each recipient institution, including its employees and other researchers under its control (RECIPIENT), of mice or services using mice from The Jackson Laboratory (TJL) agrees that such mice, descendants of those mice derived by inbreeding or crossbreeding, including unmodified derivatives of those mice or their descendants (“MICE”) shall not be: (i) used for any purpose other than the internal research of the RECIPIENT, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services with respect to MICE. Acceptance of MICE from TJL shall be deemed agreement by RECIPIENT to these conditions, and departure from these conditions requires The Jackson Laboratory’s prior written authorization.”

In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of MICE, products or services, The Jackson Laboratory will, at its option, provide credit or replacement for the MICE or product received or the services provided.

No Liability

In no event shall The Jackson Laboratory, its trustees, directors, officers, employees, and affiliates be liable for any causes of action or damages, including any direct, indirect, special, or consequential damages, arising out of the provision of MICE, products or services, including economic damage or injury to property and lost profits, and including any damage arising from acts or negligence on the part of The Jackson Laboratory, its agents or employees. In purchasing or receiving MICE, products or services from The Jackson Laboratory, purchaser or recipient, or any party claiming by or through them, expressly releases and discharges The Jackson Laboratory from all such causes of action or damages, and further agrees to defend and indemnify The Jackson Laboratory from any costs or damages arising out of any third party claims.

MICE and biological materials are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.

The foregoing represents the General Terms and Conditions applicable to The Jackson Laboratory’s MICE, products and services. In addition, special terms and conditions of sale of certain MICE, products and services may be set forth separately in The Jackson Laboratory web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, products and services by The Jackson Laboratory, and by its licensees and distributors.

Acceptance of delivery of MICE, products or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on The Jackson Laboratory, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, products services by The Jackson Laboratory.