, Volume 2, Issue 1, pp 183–197 | Cite as

Association of HL-A antigens andβ2-microglobulin at the cellular and molecular level

  • Ralph A. Reisfeld
  • Ernest D. Sevier
  • Michele A. Pellegrino
  • Soldano Ferrone
  • Miroslav D. Poulik


Surfaces of cultured human lymphoid cells RPMI 1788, RPMI 4098, RPMI 8866, Raji, and WI-L2 were found to contain bothβ2-microglobulin (β2-μ) and HL-A determinants when tested by direct complement-dependent cytotoxicity andquantitative absorption with different cytotoxic antiβ2-μ antisera and specific HL-A alloantisera. The same antigenic specificities were found in 3M KCl extracts of these cultured cells with a sensitiveβ2-μ radioimmunoassay and an HL-A antigen blocking assay. Daudi cells provided a contrast, since noβ2-μ or HL-A determinants were found on their surfaces or in 3 M KCl extracts prepared from them. Results from specific antibody blocking tests suggest a close association betweenβ2-μ and HL-A determinants on plasma membranes of cultured human lymphoid cells. A solid state immunoadsorbent containing antiβ2-μ antibodies effectively removed all detectable HL-A antigenic activity from some 3M KCl extracts of cultured human lymphoid cells as well as from some sera. Adsorption of HL-A antigens to these immunoadsorbents was specific since it was blocked only by prior addition ofβ2-μ. Once on the antiβ2-μ immunoadsorbents, HL-A antigens still reacted specifically with HL-A alloantibodies in quantitative absorption experiments. HL-A antigens andβ2-μ could be eluted from antiβ2-μ immunoadsorbents with a variety of chaotropic reagents and detergents, but thus far potassium bromide and sodium dodecyl sulfate (SDS) appear to be the most effective. SDS-PAGE of these eluates indicated that HL-A antigens were considerably purified by adsorption to antiβ2-μ immunoadsorbents and that two major molecular size fragments were distinguishable, i.e., ∼33,000 for HL-A and ∼ 12,000 forβ2-μ.


Sodium Dodecyl Sulfate Sodium Dodecyl Sulfate Antigenic Specificity Size Fragment Absorption Experiment 
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Copyright information

© Springer-Verlag New York Inc 1975

Authors and Affiliations

  • Ralph A. Reisfeld
    • 3
  • Ernest D. Sevier
    • 3
  • Michele A. Pellegrino
    • 3
  • Soldano Ferrone
    • 3
  • Miroslav D. Poulik
    • 1
    • 2
    • 3
  1. 1.Department of ImmunochemistryWilliam Beaumont HospitalRoyal Oak
  2. 2.Department of Immunology and MicrobiologyWayne State University School of MedicineDetroit
  3. 3.Department of Molecular ImmunologyScripps Clinic and Research FoundationLa Jolla

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