Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors

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Characterization of a monoclonal antibody directed against mouse macrophage and lymphocyte Fc receptors

JC Unkeless

To investigate the antigenic relationship between the macrophage and lymphocyte Fc receptors (FcR), a monoclonal antibody capable of blocking mouse macrophage Fc receptors was selected. Hybrids were formed by fusing the P3U1 mouse myeloma and spleen cells from a rat immunized with the mouse macrophage-like cell lines J774 and P388D1. The Fab fragment of the monoclonal IgG secreted by clone 2.4G2, inhibited the trypsin-resistant Fc receptor II (FcRII), which is specific for immune aggregates of mouse IgG1 and IgG2b, but had no inhibitory effect on the trypsin-sensitive Fc receptor I (FcRI), which binds monomeric IgG2a and erythrocytes coated with IgG2a. Thus, the monoclonal 2.4G2 IgG appeared to be specific for macrophage FcRII. Further evidence that the 2.4G2 IgG was directed against FcRII came from binding studies of the monoclonal antibody to J774 cells and a series of independently isolated variants which do not express FcRII. These variants of J774 bound 5% as much of the monoclonal antibody as the parent line, which bound 600,000 molecules of 2.4G2 IgG per cell. The antigenic relatedness of mouse lymphocyte FcR to mouse macrophage FcRII was demonstrated by the binding of 2.4G2 IgG to FcR-bearing lymphoid cell lines and the inhibition of the lymphocyte FcR by the monoclonal antibody. Preincubation of spleen cells and peritioneal cells with 2.3G2 IgG likewise inhibited rosette formation with ox erythrocytes coated with rabbit IgG. The ability of the hybridoma IgG to inhibit mouse FcRII was independent of the major histocompatibility complex. The 2.4G2 IgG antigenic determinant was not present on rat, guinea pig, rabbit, or human FcR-bearing cells.
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Smith-Franklin, B. A., Keele, B. F., Tew, J. G., Gartner, S., Szakal, A. K., Estes, J. D., Thacker, T. C., Burton, G. F. (2002). Follicular Dendritic Cells and the Persistence of HIV Infectivity: The Role of Antibodies and Fc{gamma} Receptors. J. Immunol. 168: 2408-2414 [Abstract] [Full Text]
Cashman, J., Clark-Lewis, I., Eaves, A., Eaves, C. (2002). Stromal-derived factor 1 inhibits the cycling of very primitive human hematopoietic cells in vitro and in NOD/SCID mice. Blood 99: 792-799 [Abstract] [Full Text]
Abe, M., Cheng, J., Qi, J., Glaser, R. M., Thall, A. D., Sykes, M., Yang, Y.-G. (2002). Elimination of Porcine Hemopoietic Cells by Macrophages in Mice. J. Immunol. 168: 621-628 [Abstract] [Full Text]
Chung, Y. S., Zhang, W. J., Arentson, E., Kingsley, P. D., Palis, J., Choi, K. (2002). Lineage analysis of the hemangioblast as defined by FLK1 and SCL expression. Development 129: 5511-5520 [Abstract] [Full Text]
Via, C. S., Shustov, A., Rus, V., Lang, T., Nguyen, P., Finkelman, F. D. (2001). In Vivo Neutralization of TNF-{alpha} Promotes Humoral Autoimmunity by Preventing the Induction of CTL. J. Immunol. 167: 6821-6826 [Abstract] [Full Text]
Russell, P. S., Chase, C. M., Sykes, M., Ito, H., Shaffer, J., Colvin, R. B. (2001). Tolerance, Mixed Chimerism, and Chronic Transplant Arteriopathy. J. Immunol. 167: 5731-5740 [Abstract] [Full Text]
Dokun, A. O., Chu, D. T., Yang, L., Bendelac, A. S., Yokoyama, W. M. (2001). Analysis of In Situ NK Cell Responses During Viral Infection. J. Immunol. 167: 5286-5293 [Abstract] [Full Text]
Kubo, S., Matsuoka, K., Taya, C., Kitamura, F., Takai, T., Yonekawa, H., Karasuyama, H. (2001). Drastic Up-Regulation of Fc{epsilon}RI on Mast Cells Is Induced by IgE Binding Through Stabilization and Accumulation of Fc{epsilon}RI on the Cell Surface. J. Immunol. 167: 3427-3434 [Abstract] [Full Text]
Tamura, T., Igarashi, O., Hino, A., Yamane, H., Aizawa, S., Kato, T., Nariuchi, H. (2001). Impairment in the Expression and Activity of Fyn During Differentiation of Naive CD4+ T Cells into the Th2 Subset. J. Immunol. 167: 1962-1969 [Abstract] [Full Text]
Cooney, D. S., Phee, H., Jacob, A., Coggeshall, K. M. (2001). Signal Transduction by Human-Restricted Fc{{gamma}}RIIa Involves Three Distinct Cytoplasmic Kinase Families Leading to Phagocytosis. J. Immunol. 167: 844-854 [Abstract] [Full Text]
Kayaba, H., Dombrowicz, D., Woerly, G., Papin, J.-P., Loiseau, S., Capron, M. (2001). Human Eosinophils and Human High Affinity IgE Receptor Transgenic Mouse Eosinophils Express Low Levels of High Affinity IgE Receptor, but Release IL-10 upon Receptor Activation. J. Immunol. 167: 995-1003 [Abstract] [Full Text]
Doyonnas, R., Kershaw, D. B., Duhme, C., Merkens, H., Chelliah, S., Graf, T., McNagny, K. M. (2001). Anuria, Omphalocele, and Perinatal Lethality in Mice Lacking the Cd34-Related Protein Podocalyxin. JEM 194: 13-28 [Abstract] [Full Text]
Snider, D., Liang, H. (2001). Early Intestinal Th1 Inflammation and Mucosal T Cell Recruitment During Acute Graft-Versus-Host Reaction. J. Immunol. 166: 5991-5999 [Abstract] [Full Text]
Matsumoto, N., Tajima, K., Mitsuki, M., Yamamoto, K. (2001). H-2 allele specificity of the NK cell C-type lectin-like MHC class I receptor Ly49A visualized by soluble Ly49A tetramer. Int Immunol 13: 615-623 [Abstract] [Full Text]