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A novel CD93 polymorphism in non-obese diabetic (NOD) and NZB/W F1 mice is linked to a CD4+ iNKT cell deficient state.

Zekavat G, Mozaffari R, Arias VJ, Rostami SY, Badkerhanian A, Tenner AJ, Nichols KE, Naji A, Noorchashm H - Immunogenetics (2010)

Bottom Line: This allele carries a coding polymorphism in the first epidermal growth factor-like domain of CD93, which results in an amino acid substitution from Asn-->His at position 264.This polymorphism does not appear to influence protein translation or ecto-domain cleavage, as CD93 is detectable in bone-marrow-derived macrophage and B-cell precursor lysates and in soluble form in the serum.These data suggest that Cd93 may be an autoimmune susceptibility gene residing within the Idd13 locus, which plays a role in regulating absolute numbers of CD4(+) NKT cells.

View Article: PubMed Central - PubMed

Affiliation: Harrison Department of Surgical Research, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.

ABSTRACT
In the present study, we characterize a polymorphism in the CD93 molecule, originally identified as the receptor for the C1q complement component (i.e., C1qRp, or AA4.1) in non-obese diabetic (NOD) mice. This allele carries a coding polymorphism in the first epidermal growth factor-like domain of CD93, which results in an amino acid substitution from Asn-->His at position 264. This polymorphism does not appear to influence protein translation or ecto-domain cleavage, as CD93 is detectable in bone-marrow-derived macrophage and B-cell precursor lysates and in soluble form in the serum. The NOD CD93 isoform causes a phenotypic aberrancy in the early B-cell developmental stages (i.e., pro-, pre-, immature, and transitional), likely related to a conformational variation. Interestingly, the NZB/W F1 strain, which serves as a murine model of Lupus, also expresses an identical CD93 sequence polymorphism. Cd93 is located within the NOD Idd13 locus and is also tightly linked to the NZB/W F1 Wbw1 and Nkt2 disease susceptibility loci, which are thought to regulate natural killer T (NKT) cell homeostasis. Consistent with this genetic linkage, we found B6 CD93(-/-) and B6.NOD(Idd13) mice to be susceptible to a profound CD4(+) NKT cell deficient state. These data suggest that Cd93 may be an autoimmune susceptibility gene residing within the Idd13 locus, which plays a role in regulating absolute numbers of CD4(+) NKT cells.

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Identification of a coding polymorphism in the first EGF-like domain of Cd93 in NOD and NZB/W F1 mice. a The Cd93 gene was PCR amplified from genomic DNA. Sequencing revealed a point mutation at cDNA nucleotide position 790. The AAC→CAC codon change in NOD and NZB/W F1 mice led to an amino acid substitution in the first EGF-like domain of Cd93 at position 264. The first EGF-like domain in which the defined polymorphism resides is stippled. b Bone marrow macrophage lysates (left) were separated on an 8% SDS-PAGE under reducing conditions, transferred to PVDF, and probed with 5 μg/ml polyclonal anti-CD93 cytoplasmic tail Ab 1150 (black arrow). Isolated bone marrow B-cell lysate immunoblots (right) were probed with 0.1 μg/ml polyclonal anti-CD93 (anti-C1qR1). The blots were stripped and re-probed for β-actin or β-tubulin to control for similar protein loading. The blots are from one experiment, representative of two, using different animals as a source of bone-marrow-derived macrophages or B cells. c sCD93 is detected in sera from NOD and NZB/W F1 mice. Sera were diluted (1:50) and tested for the presence of sCD93 by sandwich ELISA. Shown is the average of triplicate wells (± SD) from one experiment, representative of two, using different animals as a source of sera
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Fig1: Identification of a coding polymorphism in the first EGF-like domain of Cd93 in NOD and NZB/W F1 mice. a The Cd93 gene was PCR amplified from genomic DNA. Sequencing revealed a point mutation at cDNA nucleotide position 790. The AAC→CAC codon change in NOD and NZB/W F1 mice led to an amino acid substitution in the first EGF-like domain of Cd93 at position 264. The first EGF-like domain in which the defined polymorphism resides is stippled. b Bone marrow macrophage lysates (left) were separated on an 8% SDS-PAGE under reducing conditions, transferred to PVDF, and probed with 5 μg/ml polyclonal anti-CD93 cytoplasmic tail Ab 1150 (black arrow). Isolated bone marrow B-cell lysate immunoblots (right) were probed with 0.1 μg/ml polyclonal anti-CD93 (anti-C1qR1). The blots were stripped and re-probed for β-actin or β-tubulin to control for similar protein loading. The blots are from one experiment, representative of two, using different animals as a source of bone-marrow-derived macrophages or B cells. c sCD93 is detected in sera from NOD and NZB/W F1 mice. Sera were diluted (1:50) and tested for the presence of sCD93 by sandwich ELISA. Shown is the average of triplicate wells (± SD) from one experiment, representative of two, using different animals as a source of sera

Mentions: Genomic Cd93 DNA from B6, BALB/c, NOD, and NZB/W F1 mice was PCR amplified and sequenced using various primers to walk along the amplified DNA. A hitherto unidentified coding polymorphism at cDNA nucleotide position 790, which converts AAC→CAC, was discovered in NOD and NZB/W F1 mice (Fig. 1a). This mutation causes an amino acid substitution from Asn→His at position 264 in the first EGF-like domain of Cd93 (Fig. 1a).Fig. 1


A novel CD93 polymorphism in non-obese diabetic (NOD) and NZB/W F1 mice is linked to a CD4+ iNKT cell deficient state.

Zekavat G, Mozaffari R, Arias VJ, Rostami SY, Badkerhanian A, Tenner AJ, Nichols KE, Naji A, Noorchashm H - Immunogenetics (2010)

Identification of a coding polymorphism in the first EGF-like domain of Cd93 in NOD and NZB/W F1 mice. a The Cd93 gene was PCR amplified from genomic DNA. Sequencing revealed a point mutation at cDNA nucleotide position 790. The AAC→CAC codon change in NOD and NZB/W F1 mice led to an amino acid substitution in the first EGF-like domain of Cd93 at position 264. The first EGF-like domain in which the defined polymorphism resides is stippled. b Bone marrow macrophage lysates (left) were separated on an 8% SDS-PAGE under reducing conditions, transferred to PVDF, and probed with 5 μg/ml polyclonal anti-CD93 cytoplasmic tail Ab 1150 (black arrow). Isolated bone marrow B-cell lysate immunoblots (right) were probed with 0.1 μg/ml polyclonal anti-CD93 (anti-C1qR1). The blots were stripped and re-probed for β-actin or β-tubulin to control for similar protein loading. The blots are from one experiment, representative of two, using different animals as a source of bone-marrow-derived macrophages or B cells. c sCD93 is detected in sera from NOD and NZB/W F1 mice. Sera were diluted (1:50) and tested for the presence of sCD93 by sandwich ELISA. Shown is the average of triplicate wells (± SD) from one experiment, representative of two, using different animals as a source of sera
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2875467&req=5

Fig1: Identification of a coding polymorphism in the first EGF-like domain of Cd93 in NOD and NZB/W F1 mice. a The Cd93 gene was PCR amplified from genomic DNA. Sequencing revealed a point mutation at cDNA nucleotide position 790. The AAC→CAC codon change in NOD and NZB/W F1 mice led to an amino acid substitution in the first EGF-like domain of Cd93 at position 264. The first EGF-like domain in which the defined polymorphism resides is stippled. b Bone marrow macrophage lysates (left) were separated on an 8% SDS-PAGE under reducing conditions, transferred to PVDF, and probed with 5 μg/ml polyclonal anti-CD93 cytoplasmic tail Ab 1150 (black arrow). Isolated bone marrow B-cell lysate immunoblots (right) were probed with 0.1 μg/ml polyclonal anti-CD93 (anti-C1qR1). The blots were stripped and re-probed for β-actin or β-tubulin to control for similar protein loading. The blots are from one experiment, representative of two, using different animals as a source of bone-marrow-derived macrophages or B cells. c sCD93 is detected in sera from NOD and NZB/W F1 mice. Sera were diluted (1:50) and tested for the presence of sCD93 by sandwich ELISA. Shown is the average of triplicate wells (± SD) from one experiment, representative of two, using different animals as a source of sera
Mentions: Genomic Cd93 DNA from B6, BALB/c, NOD, and NZB/W F1 mice was PCR amplified and sequenced using various primers to walk along the amplified DNA. A hitherto unidentified coding polymorphism at cDNA nucleotide position 790, which converts AAC→CAC, was discovered in NOD and NZB/W F1 mice (Fig. 1a). This mutation causes an amino acid substitution from Asn→His at position 264 in the first EGF-like domain of Cd93 (Fig. 1a).Fig. 1

Bottom Line: This allele carries a coding polymorphism in the first epidermal growth factor-like domain of CD93, which results in an amino acid substitution from Asn-->His at position 264.This polymorphism does not appear to influence protein translation or ecto-domain cleavage, as CD93 is detectable in bone-marrow-derived macrophage and B-cell precursor lysates and in soluble form in the serum.These data suggest that Cd93 may be an autoimmune susceptibility gene residing within the Idd13 locus, which plays a role in regulating absolute numbers of CD4(+) NKT cells.

View Article: PubMed Central - PubMed

Affiliation: Harrison Department of Surgical Research, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.

ABSTRACT
In the present study, we characterize a polymorphism in the CD93 molecule, originally identified as the receptor for the C1q complement component (i.e., C1qRp, or AA4.1) in non-obese diabetic (NOD) mice. This allele carries a coding polymorphism in the first epidermal growth factor-like domain of CD93, which results in an amino acid substitution from Asn-->His at position 264. This polymorphism does not appear to influence protein translation or ecto-domain cleavage, as CD93 is detectable in bone-marrow-derived macrophage and B-cell precursor lysates and in soluble form in the serum. The NOD CD93 isoform causes a phenotypic aberrancy in the early B-cell developmental stages (i.e., pro-, pre-, immature, and transitional), likely related to a conformational variation. Interestingly, the NZB/W F1 strain, which serves as a murine model of Lupus, also expresses an identical CD93 sequence polymorphism. Cd93 is located within the NOD Idd13 locus and is also tightly linked to the NZB/W F1 Wbw1 and Nkt2 disease susceptibility loci, which are thought to regulate natural killer T (NKT) cell homeostasis. Consistent with this genetic linkage, we found B6 CD93(-/-) and B6.NOD(Idd13) mice to be susceptible to a profound CD4(+) NKT cell deficient state. These data suggest that Cd93 may be an autoimmune susceptibility gene residing within the Idd13 locus, which plays a role in regulating absolute numbers of CD4(+) NKT cells.

Show MeSH
Related in: MedlinePlus