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| 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 (B1-8hi). 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. | |||||||||||
Type Congenic; Mutant Strain; Targeted Mutation; 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 (B1-8hi). 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. The phenotype of this mutation on the BALB/cBy background is not known to differ from that on the C57BL/6 background (Stock No. 007594).Development
PCR was used to introduce a point mutation conferring higher affinity 4-hydroxy-3-nitrophenyl)acetyl (NP)-binding to Igh (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. 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 (Ptprca) mice for at least 12 generations by the donating laboratory, then backcrossed to BALB/cBy for 10 generations. This strain does not carry the Ptprca allele.
Strains carrying Ightm1Mnz allele
007594 B6.129P2-Ptrpca Ightm1Mnz/J View Strains carrying Ightm1Mnz (1 strain)
Strains carrying other alleles of Igh
008332 C.129S1-Ightm1Janz/J 002595 C57BL/6-Tg(IghelMD4)4Ccg/J 003395 CD1-Tg(Igh-HOX11)11Idd/J 006345 NOD.B6-Tg(IghelMD4)4Ccg/DvsJ 006608 NOD.Cg-Igh-6tm1Cgn Tg(IghelMD4)4Ccg/DvsJ View Strains carrying other alleles of Igh (5 strains)
Congenic Nomenclature
Mammalian Phenotype Terms assigned by genotype
The following phenotype information may relate to a genetic background differing from this JAX® Mice strain.
Ightm1Mnz/Ightm1Mnz
B6.129P2-Ightm1Mnz
- 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 Ightm2Mnz cells or wild-type
- fraction of mutant B cells differentiate into short lived germinal center (GC) cells; GC response is greater than in Ightm2Mnz cells
- when equal numbers of Ightm1Mnz and Ightm2Mnz B cells are transferred to wild-type recipient mice, after immunization with NP-Ficoll, Ightm1Mnz B cells preferentially expand and proliferation to make up ~46% of Ig lambda B cells whereas Ightm2Mnz B cells are not expanded
- after 5 days post immunization, 85% of plasmacytes and 60% of germinal center cells are Ightm1Mnz 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 Ightm2Mnz B cells or wild-type
- increased IgM level (MGI Ref ID J:75653)
- NP-specific IgM antibody production is greater than in Ightm2Mnz 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 Ightm2Mnz 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 Ightm2Mnz 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 Ightm2Mnz 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%)
Research Applications
This mouse can be used to support research in many areas including:
Igh relatedImmunology and Inflammation Research
Immunology and Inflammation Research
CD Antigens, Antigen Receptors, and Histocompatibility Markers
| Allele Symbol | Ightm1Mnz | ||
|---|---|---|---|
| Allele Name | targeted mutation 1, Michel C Nussenzweig | ||
| 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] | ||
Genotyping Protocols
Ightm1Mnz, STD PCR, vers. 1
Helpful Links
Optimizing PCR Protocols
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]
Ightm1Mnz 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]
Currently there no information available for this strain. This may be due to the supply level of this strain.
*Price(s) in US dollars ($)
Weeks of Age Price* Gender Cryorecovery Fee $1900.00
| Standard Supply | Repository-Cryopreserved. Must Be Recovered. Please refer to pricing and supply notes for further information. |
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| Supply Notes |
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*Price(s) in US dollars ($)
Weeks of Age Price* Gender Cryorecovery Fee $2470.00
| Standard Supply | Repository-Cryopreserved. Must Be Recovered. Please refer to pricing and supply notes for further information. |
|---|---|
| Supply Notes |
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