Education of human lymphocytes against mouse cells: specific recognition of H‐2 antigens

Abstract

INSERM U 25, HBpital Necker, ParisXeno-sensitization of human peripheral blood lymphocytes (PBL) against mouse lymphoid cells has been studied in vivo in a local graft-vs. -host (GVH) assay and in virro in a mixed lymphocyte culture-cell-mediated lympholysis system. Human PBL were injected into the footpads of mice rendered unresponsive by total-body irradiation, and these were subsequently tested for the presence of cytotoxic cells in the draining popliteal lymph node (LN). In spite of a definite PBL-induced LN proliferation, n o cytotoxic activity was detected against mouse target cells. In contrast with the GVH assay, human PBL collected after 7 days of in vitro culture in the presence of irradiated mouse cells, were strongly cytotoxic against mouse target cells. The antigenic specificities recogniied by in vitro educated cells were primarily those coded for by the H-2 complex. Only target cells with a n H-2 haplotype identical to that of the sensitizing mouse strain or with a t least an H-2D end in common with that strain were killed by xeno-sensitized lymphocytes. Mouse target cells derived from congenic resistant strains remained unaffected. It was verified that in vitro educated PBL depleted of B cells retained their cytotoxic effect, indicating that non-B cells and probably T cells were involved in the response .

Induction of secondary cytotoxic T lymphocytes by purified HLA-A and HLA-B antigens reconstituted into phospholipid vesicles.

“…2) and, in several experiments, the level of cytotoxicity observed at a given effector-to-target ratio (E/T) was equal to or only slightly lower than that observed in response to intact cells. CTLs shown (i) the induction of primary and secondary allogeneic CTL responses with purified plasma membranes (10); (ii) the induction of secondary allogeneic CTL responses with detergent-solubilized (11) and reconstituted (11,12) Although allogeneic and xenogeneic CTLs might be expected to differ with respect to their target antigens, it has been shown genetically (15)(16)(17) and by inhibition with alloantisera (16) that the xenogeneic target antigens are at least partially defined by the major histocompatibility complex. The existence of alloreactive T-cell clones with specificity for major histocompatibility complex antigens in association with minor histocompatibility antigens has been proposed (9).…”

Selective Depression of the Xenogeneic Cell-Mediated Lympholysis in Systemic Lupus Erythematosus

“…It has been shown previously that the cytotoxic cells that are generated in a xenogeneic mixed lymphocyte reaction are of T origin, and recognize specifically H-2 K or H-2 D gene products (10,(16)(17)(18). Furthermore, we have recently observed (a) that the cytotoxic reaction was not inhibited by the presence in the medium of heat-aggregated immunoglobulins, nor by immune complexes and (b) that supernates from xenogeneic mixed leukocyte cultures were unable to induce cytotoxicity either per se or after the addition of naive PBL Xenogeneic Cell-Mediated Lympholysis in Lupus Patients (47) and SLE patients (48).…”

“…All SLE patients were female: 4 presented a major exacerbation at the time of the test, 10 had a mild or moderate activity, and 3 could be considered as inactive. The degree of disease activity graded from one to four integrates both the number of organs involved and the severity of the lesions, and was independently evaluated by two physicians.…”

“…The blood was diluted at 1:3 with Hanks' balanced salt solution (HBSS) (Institut Pasteur, Paris, France). 30-ml aliquots were layered on 10 ml of a Ficoll-Angiocontrix mixture (1,077 specific density): 24 parts of 9% wt/vol Ficoll (Pharmacia, Uppsala, Sweden) and 10 parts of a 30% solution of iothalamate sodium (Angiocontrix, Guerbet, Paris, France) in 50-ml plastic conical tubes (2070, Falcon Labware, Div. Becton, Dickinson & Co., Oxnard, California).…”

“…We have recently shown that human peripheral blood lymphocytes (PBL) can be educated against mouse cells to differentiate into H-2-specific cytotoxic T cells (10). This model offers the possibility to evaluate a well-defined T-cell function, namely the generation of killer cells, under conditions where the HLA haplotype of the responder should supposedly not interfere with the magnitude of the response.…”