Transfusion Donor Research Articles

Blood transfusion induced changes in cell-mediated lympholysis: to immunize or not to immunize.

We have recently observed that the HLA-DR match between recipients and transfusion donors influences the beneficial effect of blood transfusions on allograft survival. To examine the immunologic effects of one-HLA-DR-Ag-matched and completely DR-mismatched transfusions, transfusion-induced changes in cell-mediated lympholysis (CML) were investigated. Blood donor directed CTL activity was measured in vitro before and after blood transfusion in 56 candidates for organ transplantation who received planned HLA-typed blood. We report that blood donor-directed CTL activity increased substantially after a single transfusion mismatched with the recipient for two HLA-DR Ag (p less than 0.0001). A transfusion matched for one HLA-DR Ag did not enhance CTL activity. No correlation was found between CTL reactivity and sharing of HLA class I Ag. The present study supports our previous observation that matching for at least one HLA class II Ag (HLA-DR) between transfusion recipient and blood donor is required if immunization by blood transfusion is to be avoided. These data show that the presence or absence of "autologous" HLA-DR Ag on the leucocytes of the transfusion donor plays a decisive rol whether immunization or immune suppression will ensue.

Suppression of Graft-Versus-Host Reaction in Severe Combined Immunodeficiency with Maternal-Fetal T Cell Engraftment

Simultaneous engraftment of maternal T cells and T cells from unrelated transfusion donor in an infant with severe combined immunodeficiency disease (SCID) was identified by HLA typing blood lymphocytes of the patient, her triplet siblings, parents, and the platelet donor from whom the patient received nonirradiated platelet transfusion. T cell grafts from both the mother and platelet donor were stable for several months, but the graft-versus-host reactions (GvHR) remained mild and immune function was deficient. We hypothesize that immunosuppressive effects of a maternal-fetal GvHR may have modified the expected lethal GvHR from platelet donor's T cells.

Induction of transplantation tolerance by intravenous injection of allogeneic lymphocytes across an H‐2 class II mismatch. Different mechanisms operate in tolerization across an H‐2 class I vs. H‐2 class II disparity

Previously, we have shown that the intravenous (i.v.) injection of allogeneic lymphocytes across an H-2 class I-mutant disparity leads to specific skin allograft tolerance caused by irradiation-sensitive donor T cells, which function as veto cells. In the present study, we show that the i.v. injection of H-2 class II-incompatible spleen cells also results in specific skin allograft tolerance. However, tolerance induction depends on the presence of irradiation-resistant non-T cells in the donor cell inoculum. Thus, different mechanisms operate in tolerance induction across an H-2 class I vs. H-2 class II mismatch. I.v. injection of allogeneic spleen cells across an H-2 class I plus class II disparity does not result in skin allograft tolerance. Finally, our data show that transfusion-induced suppression of the delayed-type hypersensitivity response against alloantigens does not correlate with skin allograft tolerance induced by i.v. injected allogeneic lymphocytes. In conclusion, the type of H-2 mismatch between transfusion donor and recipient not only determines the occurrence of allograft tolerance but also the mechanism leading to tolerization.

SUPPRESSED ANTIDONOR MLC RESPONSES IN RENAL TRANSPLANT CANDIDATES CONDITIONED WITH DONOR-SPECIFIC TRANSFUSIONS THAT CARRY THE RECIPIENTʼS NONINHERITED MATERNAL HLA HAPLOTYPE

Forty-seven patients with end-stage renal disease were entered into a donor-specific transfusion protocol consisting of three infusions of whole blood every two weeks prior to transplantation. Fourteen of the patients became sensitized following transfusion and were not transplanted. Thirty-one patients received a transplant from the DST donor and have an estimated two-year graft survival of 97%, three-year survival of 88%, and four-year survival of 69%. Cells of eleven of the 36 recipients tested in one-way MLC before and two weeks after completion of DST exhibited a significantly decreased antidonor MLC response. Deletion of CD8+ positive lymphocytes from suppressed MLCs resulted in restoration of antidonor MLC reactivity in four of six patients. An analysis of the family HLA profile in patients exhibiting a decreased donor-directed MLC response revealed a significant (P less than 0.02) association between decreased MLC reactivity following DST and the expression of noninherited maternal HLA antigens by cells of the transfusion donor. These alterations in cellular immune responses noted in some patients following DST are consistent with the appearance of specific antidonor T suppressor cells as a result of donor-specific transfusion.

Alloantibody and transferable suppressor activity induced by cyclosporine and blood transfusions in the rat.

The effect of cyclosporine on the alloantibody response to blood transfusion was investigated in inbred strains of rats by IHA and CELISA; recipient animals differed from the donors at the class I (RT1A) or both class I and class II (RT1B) antigens of the major histocompatibility complex. Alloantibody titers stimulated in high responder PVGu/c animals by blood transfusions were attenuated by cyclosporine; this effect was not demonstrated in low responder PVGc rats, as alloantibody titers decreased after further blood transfusions whether or not cyclosporine was given. Cyclosporine not only reduced the initial IgM response but suppressed the subsequent production of IgG. Splenocytes from rats receiving cyclosporine and blood transfusions from donors that differed from the recipients at the class I antigen were effective in suppressing the subsequent antibody response to blood transfusion. When blood transfusions from donors which differed from the recipients at both class I and class II antigenic loci were given after splenocyte transfer, a greater degree of immunosuppression was detected than if the transfusion donor differed only at the class I locus. These data suggest that the sensitization produced by blood transfusions and the persistence or decline of the alloantibody response depend upon the responder status of the recipient. Blood transfusions given with cyclosporine are capable of inducing suppressor activity that is transferable in spleen homogenates. Subsequent alloantibody responses are influenced by the class I and class II disparities of the donor and recipient animals. If these results can be extrapolated to clinical practice, cyclosporine should be given with pretransplant blood transfusions to prevent sensitization, and the transfusion donor should differ from the recipient at both class I and class II antigenic loci.

Donor-specific transfusions. Donor-recipient HLA compatibility, recipient HLA haplotype, and antibody production.

HLA profiles of 25 donor-specific transfusion (DST) kidney donor-recipient pairs were analyzed for HLA antigen compatibility. Serum samples collected during and after DST were tested for cytotoxic antibodies against T and B lymphocytes of the donors and 30 normal individuals. Eleven recipients did not produce cytotoxic antibodies to the antigens of their DST donors, and eight produced cold and/or warm, broadly reactive B-cell antibodies. Six patients (24%) produced HLA-A, B, C, and/or DR antibodies. Three of these individuals produced antibodies after two immunizations, while others required three immunizations. Three of the 11 antibody nonproducers (17%) had not received previous transfusions, as compared to three of the eight antibody producers (43%). Comparison of HLA profiles revealed 22 percent of the HLA-A, B, DR identities between the transfusion donor and recipient in antibody nonproducers as compared to 9 percent of the HLA-A, B, DR identities in antibody producers. The HLA-A2, B40, DR4 haplotype and HLA-DRW6 antigen were more common among antibody producers than among nonproducers, who had an excess of the HLA-B8, DR3 haplotype. These results are consistent with the hypothesis that there may be high- and low-responder HLA haplotypes that control immunologic responsiveness to histocompatibility antigens.

Alanine Aminotransferase and Posttransfusion Hepatitis

To the Editor.— The American Red Cross is initiating the screening of donor blood for high alanine aminotransferase (ALT) levels to reduce the incidence of posttransfusion hepatitis. Non-A, non-B hepatitis, probably caused by several viruses, is the major cause of posttransfusion hepatitis. Thirty percent of patients with posttransfusion hepatitis are symptomatic, and some develop chronic hepatic disease. Hepatitis B has been virtually eliminated as a source of posttransfusion hepatitis by testing donor blood for hepatitis B surface antigen. Aach et al1prospectively followed up 1513 transfusion recipients and found that the incidence of posttransfusion hepatitis was directly related to the ALT concentration in blood donors. The overall attack rate for non-A, non-B hepatitis was 10%, but recipients of donor units with an ALT concentration of 60 U/L or greater had a 45% attack rate. The direct relationship between transfusion hepatitis and donor ALT levels was confirmed by Alter et

Fatal Graft Versus Host Disease Following a Blood Transfusion in a Child with Neuroblastoma

A 2-year-old boy who was receiving intensive chemotherapy for advanced neuroblastoma developed fatal graft versus host disease following administration of a unit of packed red blood cells from an unrelated donor. Graft versus host disease was documented by demonstrating human leukocyte antigen identity between the transfusion donor and the patient's peripheral circulating lymphocytes. Nonirradiated packed red blood cells contain viable lymphocytes and pose a risk to the immunosuppressed cancer patient.

RETROSPECTIVE AND PROSPECTIVE STUDIES ON THE EFFECT OF BLOOD TRANSFUSIONS IN RENAL TRANSPLANTATION IN THE NETHERLANDS

The effect of blood transfusion on cadaveric kidney graft survival was studied retrospectively in 103 Dutch recipients. Seventy-four patients who had never been transfused or pregnant before transplantation had a 1-year graft survival of 32%, whereas 30 patients with only one transfusion had a very superior graft survival of 87% after 1 year (X2 = 22.85 P < 0.00001). Nineteen patients who received prospectively 1 unit of leucocyte-poor blood before transplantation had a 79% graft survival after 240 days. This is not statistically different from the results in the retrospective group with one transfusion. Patients who received prospectively 1 or 3 units leucocyte-free blood before transplantation had a 25% graft survival after 240 days. This does not differ from the graft survival in the never-transfused retrospective group (X2 = 0.31, P = 0.57). HLA matching between kidney donor and recipient could not explain the difference between the two prospectively transfused groups. For both groups the average number of HLA mismatches was equal, namely, 1.7. The average number of HLA mismatches between blood transfusion donors and kidney recipients also did not play a role of importance in both prospectively transfused groups. Even repeated mismatches on the blood transfusion donor as on the kidney donor did not give a worse graft outcome. There was no clear-cut correlation between kidney graft survival and the time of transfusion in the retro- as well as in the prospective groups. The fact that the mean hemodialysis period was longer in the leucocyte-poor prospectively transfused group than in the leucocyte-free transfused group could not explain the better graft survival in the first group, as they received only one transfusion. Finally, there was no beneficial effect seen of peroperative blood transfusions on kidney graft prognosis.

Erythroid colony stimulating and inhibiting cells in peripheral blood of transfused dogs: separation of function by velocity sedimentation

We have previously shown that the addition of normal dog peripheral blood lymphocytes (PBL) to cultures of allogeneic marrow increases the number of marrow-derived erythroid colonies (EC), but that PBL from transfused dogs usually inhibit EC growth from marrow of the transfusion donor. In this study, the cells in normal dog PBL responsible for stimulating EC growth were shown to sediment in a narrow peak at 4.30 mm/hr. A similar population of stimulating cells exists in transfused dogs and can be separated, on the basis of size, from cells that inhibit EC growth. EC-stimulating cells from transfused dog PBL sediment at 3.3--5.0 mm/hr, while cells responsible for inhibition are larger and sediment more rapidly at 5.4--8.1 mm/hr. These data demonstrate that cells capable of stimulating allogeneic EC are present in transfused dogs, but their stimulating ability is masked by the presence of EC-inhibiting cells. Thus, coculture experiments designed to test lymphocyte/marrow cell interactions may miss significant but opposing effects if unfractionated cells are used.

Erythroid Colony Stimulating and Inhibiting Cells in Peripheral Blood of Transfused Dogs: Separation of Function by Velocity Sedimentation

We have previously shown that the addition of normal dog peripheral blood lymphocytes (PBL) to cultures of allogeneic marrow increases the number of marrow-derived erythroid colonies (EC), but that PBL from transfused dogs usually inhibit EC growth from marrow of the transfusion donor. In this study, the cells in normal dog PBL responsible for stimulating EC growth were shown to sediment in a narrow peak at 4.30 mm/hr. A similar population of stimulating cells exists in transfused dogs and can be separated, on the basis of size, from cells that inhibit EC growth. EC-stimulating cells from transfused dog PBL sediment at 3.3-5.0 mm/hr, while cells responsible for inhibition are larger and sediment more rapidly at 5.4-8.1 mm/hr. These data demonstrate that cells capable of stimulating allogeneic EC are present in transfused dogs, but their stimulating ability is masked by the presence of EC-inhibiting cells. Thus, coculture experiments designed to test lymphocyte/marrow cell interactions may miss significant but opposing effects if unfractionated cells are used.

Effect of Six Weekly Transfusions on Canine Marrow Grafts: Tests for Sensitization and Abrogation of Sensitization by Procarbazine and Antithymocyte Serum

We have previously shown that blood transfusions can immunize a dog and lead to rejection of a subsequent marrow graft despite lethal total body irradiation (TBI). Sensitization to histocompatibility antigens induced by two prior transfusions of whole blood could be overcome by a regimen of procarbazine and anti-thymocyte serum (ATS) preceding TBI. The current study investigated a) whether this regimen could abrogate sensitization induced by six weekly transfusions given from days --50 to --15 preceding a marrow graft, and b) whether platelet survival studies and two in vitro tests of immunity could predict marrow graft rejection. All donor-recipient pairs were histoincompatible, unrelated, and of different breed. Twenty-two recipients received platelet concentrate transfusions and eight received whole blood transfusions. Recipients were given 1200 R TBI and a graft of marrow and peripheral blood leukocytes from the transfusion donor on day 0. Three of 15 recipients (20%) given procarbazine, 12.5 mg/kg i.v. on days --8, --6, and --4, and ATS, 0.6 ml/kg subcutaneously on days --7, --5 and --3, Rejected their grafts, whereas 11 of 15 dogs (73%) not given procarbazine and ATS rejected their grafts (p less than 0.01). Serum lymphocytotoxic antibodies, peripheral leukocyte migration inhibition, and in vivo donor platelet recovery and survival were studied in those recipients receiving six weekly transfusions and in 18 other recipients receiving a single donor transfusion 3 months before marrow grafting. No significant correlation was found among these in vitro and in vivo tests of sensitization. Sensitization to marrow grafts was not reliably detected by the presence of cytotoxic antibodies or leukocyte migration inhibition. Platelet survival, however, was positively correlated with the results of marrow grafting in 12 of 15 (80%) evaluable recipients (p approximately 0.15).