Tom Kristensen

Clones of Human Cytotoxic T Lymphocytes Derived from an Allosensitized Individual: HLA Specificity and Cell Surface Markers

By planned immunization of a volunteer, two stable (≥6 months), specific, alloreactive cytolytic T‐cell clones have been established from his peripheral blood lymphocytes. One clone reacts with all serologically defined HLA‐Cw3 cells from our panel, whereas the other defines a split within the serological HLA‐B40 specificity. The two cytotoxic clones are SmIg‐negative, E‐rosette positive, EA and EAC rosette‐negative. HLA‐A, ‐B and ‐C‐positive, and also HLA‐DR‐ or ‘Ia like‐positive. In addition, they present very similar patterns of iodinated cell surface molecules as analysed by sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS‐PAGE), contrasting with that of an EBV cell line derived from the same donor.

Occurrence of lymphocytotoxic lymphocytes and antibodies after corneal transplantation.

Twentyfive recipients of penetrating corneal grafts were investigated for the presence in the peripheral blood of cytotoxic lymphocytes by the Direct Cell Mediated Lympholysis (Direct CML) test as well as for lymphocytotoxic antibodies.

The HLA-D system: At least two loci and four distinct phenotypic traits per haplotype. Introduction to component typing in families and population by primed lymphocyte typing

Using a number of intrafamilial PLTs raised against identical HLA haplotypes it has been possible to construct a model in an informative family defining the HLA-D region as a genetic system. This system consists of at least two regions separated by a recombination between HLA-D and GLO. In relation to the site of recombination, a minimum of one centromeric and three telomeric components can be identified per haplotype.—Fourteen PLTs raised and defined within the family were subsequently tested in a Caucasian population (n=84) and in 13 unrelated, complete families.—It is concluded that the hypothetical model proposed for the HLA-D region as a genetic system of linked loci, coding at the cell surface for associated but distinct components (at least four per haplotype), allows for typing of the components of the HLA-D system of any given haplotype. Serological typing of HLA-D components should, in the near future, provide a more convenient way of establishing component phenotypes than the present use of primed lymphocyte typing reagents. Among the components isolated, some have a high association with the classic alleles defined either by homozygous typing cells or DR serology. Others form the basis of cross-reactivity but their presence does not interfere with standard typing. Others, however, seem by their mere presence to be responsible for false assignments.—The concept of HLA-D as a genetic system clarifies many of the inconsistencies observed with a one-locus system.

Lymphocyte subpopulations in man: suppression of PWM-induced B-cell proliferation by infectious mononucleosis T cells.

The in vitro polyclonal pokewced milogen (PWM)‐induced activation of human B lymphocytes is enhanced by addition of autologous or allogeneic: irrudiated T cells. This model for B/T‐cell cooperation may he used tu define and describe the balance between T helper and T suppressor phenomena. The present study investigates the helper and suppressor capacities of mononuclear cells isolated from peripherial blood of infectious mononucleosis patients during the acute disease and the reconvalescence period. During the acute disease, we found a functional lack of T helper capacity; furthermore, the T cells were able to suppress the PWM and T‐cell‐dependent B‐cell proliferation of healthy donor cells. The suppression was non‐cytotoxic: i.e. not due to destruction of the responder cells. This phenomenon of non‐cytotoxic suppression was found for all seven patients studied and disappeared during the reconvalescence period, indicating that the T lympliocyltosis seen in infectious mononucleosis includes an expansion of T suppressor cells.

Bone marrow transplantation for aplastic anaemia from a HL-A and MLC-identical unrelated donor

SummaryBone-marrow transplantation (BMT) from an unrelated, HL-A-phenotype-identical, MLC-negative donor was performed in a 31 year old woman with severe longlasting aplastic anemia. In vitro assays failed to demonstrate humoral or cellular sensitization of the recipient against donor-type antigens. Following conditioning with cyclophosphamide, prompt but only transient engraftment of the transplant occurred accompanied by signs of mild graft-versus-host-disease (GVHD) of the liver. The results of a second bone marrow transplantation from the same donor cannot be evaluated due to early death of the recipient.It is concluded that bone marrow from unrelated, HL-A- and MLC-identical donors may engraft without severe GVHD. Rejection of the graft in our patient may have been related to greater antigenic differences that can be expected to exist between HL-A-and MLC-identical unrelated individuals than between HL-A-and MLC-identical siblings. However, insufficient preparative immunosuppression with cyclophosphamide due to severe hepatic hemosiderosis appears equally likely as the cause of graft rejection. The possibly increased risk of graft rejection or severe GVHD should not preclude the use of unrelated HL-A-and MLC-identical marrow donors, when histocompatible sibling donors are not available; but more potent immunosuppressive regimens than the cyclophosphamide protocol may be necessary to ensure permanent engraftment.ZusammenfassungBei einer 31 jährigen Patientin wurde wegen schwerer progredienter Panmyelopathie eine Knochenmarktransplantation von einer nichtverwandten, HL-A phänotypisch identischen, MLC-negativen Spenderin durchgeführt. Humorale oder zelluläre Sensibilisierung der Empfängerin gegen zellgebundene Antigene der Spenderin war in vitro nicht nachweisbar. Nach Konditionierung mit Cyclophosphamid waren zeitgerechtes, jedoch nur vorübergehendes Angehen des Transplantates und Symptome einer leichten Transplantat-gegen-Wirt-Reaktion (GVHD) zu beobachten. Der Ausgang einer Retransplantation von Knochenmark derselben Spenderin ist wegen des frühen Todes der Patientin nicht zu beurteilen.Nach Konditionierung mit Cyclophosphamid kann das Knochenmark eines nicht verwandten, HL-A-und MLC-identischen Spenders angehen, ohne daß eine schwere GVHD-Reaktion auftritt. Ob die Abstoßung des Transplantates bei unserer Patientin darauf zurückzuführen ist, daß zwischen Empfänger und HL-A-und MLC-identischem, nichtverwandtem Spender größere, in vitro nicht erkennbare Antigendifferenzen bestehen, als zwischen HL-A-und MLC-identischen Geschwistern, kann nicht entschieden werden. Ungenügende Immunsuppression durch Cyclophosphamid als Folge einer schweren Hämosiderose der Leber mag ebenso zur Transplantatabstoßung geführt haben. Das möglicherweise höhere Risiko einer Transplantatabstoßung oder einer schweren GVHD-Reaktion sollte nicht von der Transplantation des Knochenmarks eines nichtverwandten, HL-A-und MLC-identischen Spenders abhalten, wenn histokompatible Geschwister nicht zur Verfügung stehen; jedoch mag dann stärkere Immunsuppression erforderlich sein, als sie mit Cyclophosphamid zu erzielen ist, um ein permanentes Angehen des transplantierten Knochenmarks zu gewährleisten.

Murine H-2Dd-reactive monoclonal antibodies recognize shared antigenic determinant(s) on human HLA-B7 or HLA-B27 molecules or both

We have evaluated the serological relationships between the murine H-2Dd and human HLA molecules using four H-2Dd-reactive monoclonal antibodies (mAbs) produced in the A.BY (KbIbDb) anti-A.TL (KsIkDd) combination. In the mouse, these reagents exhibited three distinct reactivity patterns: Dd, Ks, and H-2u (mAb 81.L); Dd, H-2p, and H-2u (mAb 81.R); and Dd, Kd, H-2p, H-2u, and H-2v (mAbs 97.G and 97.H). Sequential immunoprecipitation and cross-competitive mAb binding experiments revealed that these mAbs recognized determinants in two spatially distinct polymorphic domains on the H-2Dd molecule of B10.A(5R) cells (defined by mAbs 81.L and 81.R, 97.H, and 97.G, respectively). MAbs 81.R, 97.G, and 97.H, but not 81.L, also defined an HLA-linked polymorphism in the human, the main characteristics of which can be summarized as follows: (i) on B lymphoblastoid cell lines, mAbs 81.R and 97.H bound to cells expressing the HLA-B7, HL-B27 or -Bw40 cross-reacting specificities, (ii) on peripheral blood lymphocyte (PBL) panel mAb 81.R exerted C dependent cytotoxicity to 118 of 400 cells tested, including almost all HLA-B7 or HLA-B27 cells or both (r: 0.952), (iii) the expression of the 81.R. cross-reacting determinant segregated in an informative family with the parental haplotype carrying the HLA-B7 allele, and (iv) mAbs 81.R, 97.G, and 97.H recognized topologically related determinants on the same class I molecule(s) of the human B lymphoblastoid cells JY (HLA-A2,2, -B7,7). These data support the view that some, but not all H-2Dd allotopes have been conserved throughout evolution and are associated in the human with the HLA-B7, -B27 cross-reacting specificities.