Alternative titles; symbols
HGNC Approved Gene Symbol: CD244
Cytogenetic location: 1q23.3 Genomic coordinates (GRCh38) : 1:160,830,160-160,862,887 (from NCBI)
| Location | Phenotype |
Phenotype MIM number |
Inheritance |
Phenotype mapping key |
|---|---|---|---|---|
| 1q23.3 | {Rheumatoid arthritis, susceptibility to} | 180300 | 3 |
Natural killer (NK) cells express both activating and inhibitory cell surface receptors. Inhibitory signaling receptors all possess cytoplasmic immunoreceptor tyrosine-based inhibitory motifs, or ITIMs, whereas the activating receptors lack ITIMs and associate with DAP12 (TYROBP; 604142), which contains an immunoreceptor tyrosine-based activation motif, or ITAM. Killer cell immunoglobulin (Ig)-like receptors, or KIRs (see KIR2DL1; 604936), and other NK cell receptors interact with major histocompatibility complex (MHC) molecules (see 142800). Members of the CD2 (186990) family adhere to each other instead. The cell surface glycoprotein 2B4 is related to CD2 and is implicated in the regulation of NK- and T-cell function (Boles et al., 1999).
By screening a genomic DNA library and probing with mouse 2b4, followed by screening a human NK-cell cDNA library, Boles et al. (1999) isolated a cDNA encoding human 2B4. Sequence analysis predicted that the 365-amino acid protein, which is 70% similar to the mouse sequence and approximately 45% similar to other members of the CD2 family, has an 18-amino acid leader sequence; an extracellular region of 204 amino acids with 2 Ig-like motifs and 8 potential N-linked glycosylation sites; a 24-amino acid transmembrane domain; and a 120-amino acid cytoplasmic tail with 6 tyrosine residues. Northern blot analysis detected 3- and 5-kb transcripts in T- and NK-cell lines; however, peripheral blood leukocytes, spleen, and lymph node expressed only the 3-kb transcript.
By immunoprecipitation and Western blot analysis, Nakajima et al. (1999) showed that 2B4 was expressed as a 63-kD protein that could be reduced to 42 kD by deglycosylation. Flow cytometric analysis extended the range of cells expressing 2B4 to monocytes as well as a large subset of CD8 (see 186730)-positive cells and a small subset of CD4-positive cells.
By probing a spleen cDNA library with mouse 2b4, Tangye et al. (1999) isolated a cDNA encoding 2B4.
Functional analysis by Boles et al. (1999) demonstrated that engagement of 2B4 with specific antibody activates NK cytolytic activity.
Using recombinant human NK cell-activating ligand 2B4 fused to domains 3 and 4 of rodent Cd4 (186940) and flow cytometric analysis, Brown et al. (1998) demonstrated that CD48 (109530) binds to 2B4. BIAcore surface plasmon resonance analysis showed that the affinity of CD48 for 2B4 is approximately 10-fold higher than that shown for CD48 and CD2.
Nakajima et al. (1999) found that ligation of 2B4 with either antibody or with CD48 led to NK cell-mediated cytolysis but not to T cell-mediated cytolysis.
Binding analysis by Tangye et al. (1999) indicated that phosphorylated 2B4 recruits either SHP2 (176876) or SLAM (603492)-associated protein (SAP, or SH2D1A; 300490).
By studying NK-cell function in patients with X-linked lymphoproliferative disease (XLPD; 308240) and a defect in the SAP gene, Parolini et al. (2000) found that a number of triggering receptors displayed normal function. However, upon 2B4 interaction with CD48, NK-cell function against Epstein Barr virus (EBV)-infected cells, which is primarily mediated via NKp46 (LY94; 604530), was inhibited. Disruption of 2B4-CD48 and/or NK receptor-HLA interaction restored NK cytolytic activity. RT-PCR analysis detected the full-length 2B4 cDNA as well as a 2B4 molecule lacking the Ig C2 domain in both patients and normal individuals. Molecular analysis failed to reveal any differences between normal and patient 2B4 sequences. Immunoblot analysis showed that treatment of normal but not XLPD NK cells with pervanadate led to the association of 2B4 with SAP. Parolini et al. (2000) suggested that anti-2B4 treatment might be of use in XLPD patients awaiting bone marrow transplantation.
By studying an XLPD patient with a SAP deletion, Tangye et al. (2000) found that although XLPD patient NK cells can be active, the absence of SAP selectively cripples the 2B4-mediated activation pathway.
In 2 healthy males carrying the same SAP mutation as 2 deceased relatives who had XLPD, Benoit et al. (2000) found that NK-cell activity could not be enhanced through 2B4 ligation. The authors suggested that this SAP mutation, predicted to disrupt SLAM binding, might interfere with 2B4 binding on the basis of its strong homology to SLAM.
Aoukaty and Tan (2005) found that NK cells from individuals with XLP failed to phosphorylate GSK3A (606784) and GSK3B (605004) after stimulation of 2B4. Lack of GSK3 phosphorylation inactivated GSK3 and prevented accumulation of the transcriptional coactivator beta-catenin (CTNNB1; 116806) in the cytoplasm and its subsequent translocation to the nucleus. Aoukaty and Tan (2005) identified VAV1 (164875), RAC1 (602048), RAF1 (164760), MEK2 (MAP2K2; 601263), ERK1 (MAPK3; 601795), and ERK3 (MAPK6; 602904) as proteins potentially involved in mediating the signaling pathway between 2B4 and GSK3/CTNNB and found that some of these elements were aberrant in XLP NK cells. Aoukaty and Tan (2005) concluded that GSK3 and beta-catenin mediate signaling of 2B4 in NK cells and that dysfunction of some of the elements in the transduction pathway between 2B4 and GSK3/beta-catenin may result in diminished IFNG (147570) secretion and cytotoxic function of NK cells in XLP patients.
Watzl et al. (2000) showed that antibody-mediated cross-linking of 2B4 leads to its rapid tyrosine phosphorylation, which is necessary for 2B4-mediated killer cell activity. However, this activity and 2B4 phosphorylation could be blocked by coligation of 2B4 and an inhibitory receptor, e.g., KIR2DL1 or CD94 (602894).
By Southern blot analysis, Boles et al. (1999) determined that the 2B4 gene spans approximately 25 kb.
Tangye et al. (1999) mapped the 2B4 gene to 1q22, where SLAM, CD48, CD84 (604513), and LY9 are also located.
Suzuki et al. (2008) identified a functional single-nucleotide polymorphism (SNP) in the CD244 gene that contributes to rheumatoid arthritis susceptibility (180300).
Vaidya et al. (2005) generated mice deficient in both isoforms of Cd244. Cd244 -/- mice appeared healthy and developed normally. Female Cd244 -/- mice had a slight increase in immature thymic Cd4-negative/Cd8-negative cells, but the NK cell receptor repertoire was intact in Cd244 -/- mice of both genders. Wildtype mice rejected Cd48-positive melanoma cells poorly compared with Cd48-negative melanoma cells. Male Cd244 -/- mice showed enhanced rejection of Cd48-positive melanoma cells, whereas female Cd244 -/- mice had poor resistance to both Cd48-positive and Cd48-negative melanoma cells. NK cell-depleted Cd244 -/- mice did not show gender distinctions in in vivo and in vitro assays, suggesting that the gender-specific effects may result from interactions of NK cells with other immune cells.
In an analysis of a 1.1-Mb linkage disequilibrium segment associated with susceptibility to rheumatoid arthritis (180300) involving 2 independent rheumatoid arthritis cohorts from Japan, Suzuki et al. (2008) observed the most significant association at rs3766379 (P = 3.23 x 10(-8), odds ratio = 1.31, confidence interval 1.19-1.44) in intron 5 of the CD244 gene. The SNP rs3766379 is located in a heat-shock transcription factor-1 (HSF1)/HSF2/early growth response-2 (EGR2) binding site and an upstream transcription factor-1 (USF1) binding site. The susceptibility allele increased the expression of CD244 in luciferase and allele-specific transcript quantification assays.
Aoukaty, A., Tan, R. Role for glycogen synthase kinase-3 in NK cell cytotoxicity and X-linked lymphoproliferative disease. J. Immun. 174: 4551-4558, 2005. [PubMed: 15814676] [Full Text: https://doi.org/10.4049/jimmunol.174.8.4551]
Benoit, L., Wang, X., Pabst, H. F., Dutz, J., Tan, R. Cutting edge: defective NK cell activation in X-linked lymphoproliferative disease. J. Immun. 165: 3549-3553, 2000. [PubMed: 11034354] [Full Text: https://doi.org/10.4049/jimmunol.165.7.3549]
Boles, K. S., Nakajima, H., Colonna, M., Chuang, S. S., Stepp, S. E., Bennett, M., Kumar, V., Mathew, P. A. Molecular characterization of a novel human natural killer cell receptor homologous to mouse 2B4. Tissue Antigens 54: 27-34, 1999. [PubMed: 10458320] [Full Text: https://doi.org/10.1034/j.1399-0039.1999.540103.x]
Brown, M. H., Boles, K., van der Merwe, P. A., Kumar, V., Mathew, P. A., Barclay, A. N. 2B4, the natural killer and T cell immunoglobulin superfamily surface protein, is a ligand for CD48. J. Exp. Med. 188: 2083-2093, 1998. [PubMed: 9841922] [Full Text: https://doi.org/10.1084/jem.188.11.2083]
Nakajima, H., Cella, M., Langen, H., Friedlein, A., Colonna, M. Activating interactions in human NK cell recognition: the role of 2B4-CD48. Europ. J. Immun. 29: 1676-1683, 1999. [PubMed: 10359122] [Full Text: https://doi.org/10.1002/(SICI)1521-4141(199905)29:05<1676::AID-IMMU1676>3.0.CO;2-Y]
Parolini, S., Bottino, C., Falco, M., Augugliaro, R., Giliani, S., Franceschini, R., Ochs, H. D., Wolf, H., Bonnefoy, J.-Y., Biassoni, R., Moretta, L., Notarangelo, L. D., Moretta, A. X-linked lymphoproliferative disease: 2B4 molecules displaying inhibitory rather than activating function are responsible for the inability of natural killer cells to kill Epstein-Barr virus-infected cells. J. Exp. Med. 192: 337-346, 2000. [PubMed: 10934222] [Full Text: https://doi.org/10.1084/jem.192.3.337]
Suzuki, A., Yamada, R., Kochi, Y., Sawada, T., Okada, Y., Matsuda, K., Kamatani, Y., Mori, M., Shimane, K., Hirabayashi, Y., Takahashi, A., Tsunoda, T., Miyatake, A., Kubo, M., Kamatani, N., Nakamura, Y., Yamamoto, K. Functional SNPs in CD244 increase the risk of rheumatoid arthritis in a Japanese population. Nature Genet. 40: 1224-1229, 2008. [PubMed: 18794858] [Full Text: https://doi.org/10.1038/ng.205]
Tangye, S. G., Lazetic, S., Woollatt, E., Sutherland, G. R., Lanier, L. L., Phillips, J. H. Cutting edge: human 2B4, an activating NK cell receptor, recruits the protein tyrosine phosphatase SHP-2 and the adaptor signaling protein SAP. J. Immun. 162: 6981-6985, 1999. [PubMed: 10358138]
Tangye, S. G., Phillips, J. H., Lanier, L. L., Nichols, K. E. Cutting edge: functional requirement for SAP in 2B4-mediated activation of human natural killer cells as revealed by the X-linked lymphoproliferative syndrome. J. Immun. 165: 2932-2936, 2000. [PubMed: 10975798] [Full Text: https://doi.org/10.4049/jimmunol.165.6.2932]
Vaidya, S. V., Stepp, S. E., McNerney, M. E., Lee, J.-K., Bennett, M., Lee, K.-M., Stewart, C. L., Kumar, V., Mathew, P. A. Targeted disruption of the 2B4 gene in mice reveals an in vivo role of 2B4 (CD244) in the rejection of B16 melanoma cells. J. Immun. 174: 800-807, 2005. [PubMed: 15634901] [Full Text: https://doi.org/10.4049/jimmunol.174.2.800]
Watzl, C., Stebbins, C. C., Long, E. O. Cutting edge: NK cell inhibitory receptors prevent tyrosine phosphorylation of the activation receptor 2B4 (CD244). J. Immun. 165: 3545-3548, 2000. [PubMed: 11034353] [Full Text: https://doi.org/10.4049/jimmunol.165.7.3545]