The effect of individual HLA-A and -B mismatches on the generation of cytotoxic T lymphocyte precursors

In order to study the correlation between HLA mismatches and the cytotoxic T lymphocyte precursor frequency, we used a limiting dilution analysis to determine the CTLp frequencies against individual mismatched HLA-A and -B alloantigens in 21 patients waiting for a renal transplant. Altogether, thirty-three mismatched HLA-A antigens and 55 HLA-B antigens were tested. The CTLp frequencies against mismatches of HLA-B locus antigens were found to be significantly higher than those against HLA-A antigens (P less than 0.002). This may explain why matching for HLA-B antigens is more important for a good renal allograft survival than matching for HLA-A antigens.     

Drug-Induced Antibodies: Interaction of the Drug with a Polymorphic Platelet-Antigen

Preincubation of donor platelets with ticarcillin will prevent the reactivity of a platelet antibody against these platelets, whereas no influence was observed on antisera against HLA, 5A, 5b and ZWa. The implications for the mechanism of drug-induced antibodies with restricted specificity will be discussed.     

Differential immune response of congenic mice to ultraviolet-treated major histocompatibility complex class II-incompatible skin grafts

The influence of ultraviolet (UVB) irradiation on the survival of H-2 class II-disparate skin grafts was studied in congenic mouse strains. Isolated skin was UVB irradiated in vitro at a dose of 40 mJ/cm2 from both sides to remove Ia immunogenicity. Immediately after irradiation the skin was transplanted onto the flank of allogeneic mice. When B10.AQR grafts were transplanted onto B10.T(6R) recipients, a significant prolongation of the survival time was observed, while 50% of the UVB-treated grafts were not rejected at all. However, in the opposite direction--i.e., B10.T(6R) grafts onto B10.AQR recipients, no significant prolongation of the survival was observed. To test whether this effect was due to a difference in susceptibility of the donor skin to UVB irradiation or to a different immune response in the recipients, (B10.T(6R) x B10.AQR) grafts were transplanted onto the parent strains. Similar results were obtained, in that UVB-treated grafts did not show a prolonged survival in B10.AQR recipients, whereas a significant prolongation (50% of the grafts survived more than 100 days) was observed in B10.T(6R) recipients. UVB-treated (B10.T(6R) x B10.AQR)F1 grafts were also transplanted onto (B10.T(6R) x C57B1/10)F1, (B10.AQR x C57B1/10)F1, (B10.T(6R) x Balb/c)F1 and (B10.AQR x Balb/c)F1 recipients--but in none of these combinations was a prolonged survival time observed. These data suggest that, in contrast to all in vitro experiments, the abrogation of the immune response by UVB treatment of the stimulator cells is, in vivo, not a general phenomenon. The genetic constitution of the responder mice seems to play an important role in determining whether or not an immune response takes place.     

An increase of donor-specific T helper precursors resulting from blood transfusions

In order to study the effect of blood transfusions on the donor-specific helper T cell repertoire, donor-specific interleukin 2-producing precursors (Th precursors [Thp]) were examined in 10 patients before and after transfusion. These patients were selected to be male, had received no previous transfusions or transplantation, and had no cytotoxic antibodies against MHC antigens in their serum. The limiting dilution curves were linear before as well as after transfusion, showing that only the cell under study was limiting. It was found that the transfusion donor-specific Thp frequencies increased significantly after transplantation in 10 of 10 patients (ratio: 1.5 to 7.1). In addition, the IL-2 production by individual clones tended to be higher after transfusion. In contrast, Thp frequencies against a third-party control showed a small increase in 4 and was unchanged in 6 patients. These data indicate that blood transfusions prime the donor-specific Th compartment.     

Hyperimmunized patients do not need to wait for an HLA identical donor

Hyperimmunized patients tend to accumulate on renal transplant waiting lists because their high level of sensitization leads to positive crossmatches with almost all potential organ donors. The origins of sensitization and the different efforts made to find cross-match negative donors for these patients are discussed. Special emphasis is given to a local strategy based on the determination of HLA-A and -B mismatches, against which the patient did not form allo-antibodies, the so-called acceptable mismatches. Kidney donor selection is based on compatibility with the patient's own HLA-antigens in combination with the acceptable HLA-A and -B antigens, and can be operated from a central organ-sharing office. The acceptable HLA mismatches are often identical with or include the non-inherited HLA class I antigens of the mother (non-inherited maternal antigens: NIMA).     

Variation of the immune response to sheep erythrocytes in several strains of mice and their crosses

Summary There is a continuous variation in the immune response to sheep erythrocytes in inbred and outbred strains of mice. In animals of equal age strain-specific differences are the most important cause of the variation. Female animals are on the average superior to the male ones. Especially in the 7 S response the crossbreds showed positive deviations from the mid-parent values, thus suggesting the existence of non-additive gene effects. In one crossing experiment maternal influences could be shown.

---

Zusammenfassung Die Stärke der Immunantwort nach Injektion einer konstanten Dosis Schaferythrocyten variiert kontinuierlich in verschiedenen Mäusestämmen. Bei gleichaltrigen Tieren stellen stammbedingte Unterschiede eine wichtigere Varianzursache dar als das Geschlecht der Tiere. Trotzdem waren die weiblichen Tiere in der Mehrzahl der durchgeführten Tests den männlichen überlegen. Kreuzungstiere zeigten einen Hybrideffekt, der vor allem die Zahl der indirekten plaquebildenden Zellen in der Milz und den Titer der merkaptoäthanolresistenten Hämagglutinine im Serum betraf. In einem Kreuzungsexperiment konnte ein mütterlicher Effekt beobachtet werden.

---

---

This investigation was supported by the Deutsche Forschungsgemeinschaft.     

The frequency and avidity of committed cytotoxic T lymphocytes (cCTL) for donor HLA class I and class II antigens and their relation with graft vascular disease

Cellular immune processes may trigger the development of graft vascular disease (GVD). CD4 and CD8 cytotoxic T lymphocytes that infiltrate the allograft could play a role in the development of GVD. We studied the presence of in vivo primed or committed CTL (cCTL) and their avidity for donor HLA class I and class II antigens in graft-infiltrating lymphocyte cultures propagated from endomyocardial biopsies derived from patients with and without signs of GVD. The fraction of cCTL with high avidity for HLA class I or class II antigens was estimated by the addition of anti-CD8 or anti-CD4 MoAbs to the cytotoxic phase of the limiting dilution analysis. In the first year after transplantation no difference in the frequency of donor-specific class I cCTL between patients with and without GVD was found. Addition of anti-CD8 MoAb revealed that most cultures predominantly consisted of cCTL with low avidity for donor HLA class I antigens, irrespective of the development of GVD at 1 year after transplantation. However, in patients who did not develop GVD, the frequency of cCTL with donor HLA class II specificity was significantly higher than in patients who did develop GVD. The avidity for donor HLA class II antigens was comparable in both groups. A high frequency of donor-specific cCTL for HLA class II antigens seems to be a protective factor against the development of GVD. These cCTL might be cytotoxic for cells involved in GVD development, e.g. activated endothelium and smooth muscle cells of donor origin.     

Human bone allografts can induce T cells with high affinity for donor antigens

We analysed the cellular immune response in ten transplantations of different massive bone allografts, of which five had a poor clinical outcome. Cytotoxic T lymphocytes (CTL) and T helper lymphocytes (TH) against mismatched donor antigens were found in all patients. More importantly, CTL with a high affinity for donor antigens were found in five cases. High-affinity CTL need no CD8 molecule to stabilise the antigen binding and are strongly associated with rejection of heart and corneal transplants. Even after removal of most of the bone-marrow cells, we found high-affinity CTL and high TH frequencies. This T-cell response could be detected over a period of years. We conclude that frozen bone allografts can induce high-affinity donor-specific CTL. The present assay allows qualification and quantification of the levels of CTL and TH in the blood. This approach may be helpful in studying the effect of the immune response on the outcome of the graft.     

Characterization of high-growth reassortant influenza A viruses generated in MDCK cells cultured in serum-free medium

In the present study reassortant influenza A viruses of both the H1N1 and H3N2 type were generated in Madin Darby Canine Kidney cells grown in the absence of fetal bovine serum (MDCK-SF1 cells). To this end, MDCK-SF1 cells were simultaneously infected with one of the high-growth laboratory strains A/Puerto Rico/8/34 (H1N1) or A/Hong Kong/2/68 (H3N2) and recent H3N2 and H1N1 vaccine strains, respectively. Reassortant viruses obtained from these mixed infections were genetically characterized by RT-PCR and restriction enzyme analysis and their growth properties were compared to those of the corresponding field strains. Reassortant H3N2 viruses inherited the matrix and polymerase pa gene whilst H1N1 reassortant viruses inherited the matrix and polymerase pbl gene of the high-growth parent. Reassortant viruses generally gave higher viral yields, as measured by a haemagglutination assay, than their wild type counterparts. The procedure followed results in the generation of high-growth reassortant viruses in weeks. The use of MDCK-SF1 cells together with these reassortants for generating influenza virus antigens can significantly speed up the vaccine production procedure.