Induction of a secondary human anti-HLA alloimmune response in severe combined immunodeficient mice engrafted with human lymphocytes.

BACKGROUND: Experimental manipulation of transfusion-induced alloimmunization is limited in humans by ethical considerations. Conversely, studies of alloimmunization in animal models may not reflect the human immune system closely enough to be of optimal benefit. The development of an in vivo model of human alloimmunization that is amenable to experimental manipulation is thus desirable. STUDY DESIGN AND METHODS: An in vivo model of human alloimmunization was evaluated by using mice with severe combined immunodeficiency (SCID). SCID mice underwent gamma-radiation (200 cGy) and received an intraperitoneal injection of human peripheral blood lymphocytes (PBLs) from donors immunized to HLA antigens by prior pregnancy (reconstitution). These Hu [human]-PBL-SCID mice were then challenged with HLA-mismatched PBLs. Alloantibodies were evaluated by flow cytometry and a standard two-stage microlymphocytotoxicity assay. RESULTS: Hu-PBL-SCID mice (n = 22) that were challenged with PBLs expressing the HLA antigens to which the donors had previously been immunized, made significantly more IgM and IgG alloantibodies than did the unchallenged mice. Responses were measurable by 1 week after reconstitution and challenge. Prior treatment of SCID mice with anti-asialo GM1, which depletes murine natural killer cells and macrophages, further increased the alloantibody response of challenged mice. The human alloantibodies generated were specific to the challenge HLA antigens as assessed by microlymphocytotoxicity assay. CONCLUSION: Hu-PBL-SCID mice are a useful model system in which to study and manipulate the induction of secondary human alloimmune responses against cellular HLA class I antigens. This model will be valuable for testing the in vivo effect of novel immunotherapies on the inhibition of the human alloantibody response.

An investigation of nonhemolytic transfusion reactions.

This study was undertaken to document the incidence of immediate, nonhemolytic transfusion reactions and to identify a technique or set of techniques that would best identify the different causes of these reactions. A variety of tests were employed to detect lymphocyte, granulocyte, platelet and anti-IgA antibodies. During this study 26,318 units of blood components were transfused on 5,030 occasions. 191 immediate, nonhemolytic reactions were experienced giving an incidence per unit of 0.73%. Blood specimens from 101 of these patients were investigated along with serum from 57 patients who showed no reaction to transfusion as controls. We show that standard B cell lymphocytotoxicity testing is the technique with which most antibodies can be detected (64% of reactors positive vs. 30% of controls, p less than 0.001). Additional tests did not significantly increase the level of antibody detection.

Splits of the HLA DR2 antigen.

The reaction of ninety DR2 positive persons with 20 anti-DR2 sera were studied. Four of the panel and members of two families did not react with two of these anti-DR sera and were considered to have the short-split "DR2.2". DR2 positive samples reacting strongly with all anti-DR2 were designated "DR2.1".