Third-party Blood Transfusions Research Articles

Identification of immunodominant donor MHC peptides following rejection and donor strain transfusion-induced tolerance of heart allografts in adult rats.

Pre-graft priming of heart allograft recipients with donor strain blood induces tolerance in 100% of adult rats in the congenic LEW.1W to LEW.1A combination. This tolerant state is specific for donor MHC antigens as third-party blood transfusions fail to induce tolerance, and third-party skin grafts are promptly rejected by tolerant graft recipients. In this study we have characterized the immunodominant donor (RT1u) class I and II allogenic peptides which elicit an in vitro proliferative response to splenocytes from recipients (RT1a) undergoing acute rejection or tolerant to a LEW.1A cardiac allograft. Paradoxically, splenocytes from tolerant animals responded more vigorously to a broader set of donor peptides than splenocytes from rejecting animals. In addition, several of these peptides were observed to be stimulatory only for tolerant splenocytes. These findings suggest that regulatory cells may be involved in tolerance induction or maintenance and are selected by specific motifs, which could be utilized for manipulating the immune system of graft recipients.

Rapamycin, cyclosporine, and perioperative donor-specific transfusions induce prolongation of cardiac allograft survival in the rat.

The effects of a subtherapeutic regimen of rapamycin (RAPA) combined with CsA and donor-specific blood transfusions (DST) were investigated in the ACI to LEW cardiac allograft model. Three protocols were studied. In protocol 1, recipients received a DST (day -1), RAPA intravenously at 0.08 mg/kg/day (day -1 to +5), and CsA intramuscularly at 5 mg/kg/day (day -1 to +1). In protocol 2, the course of CsA was extended to day +5, and in protocol 3 the treatment was further modified to include a second DST on day +3. Control groups received drug treatments alone or combined with DST or third-party blood transfusions. In all protocols, RAPA, CsA, or DSTs alone had little or no effect on graft survival. RAPA-DST or CsA-DST also had no effects beyond the ones induced by the drugs alone. In the RAPA-CsA groups, graft survivals were similar to the ones obtained with CsA alone. In contrast, in each of the protocols, the RAPA-CsA-DST treatment resulted in significant prolongation of graft survival as compared with all controls (P < 0.05). The administration of a second DST (CsA-RAPA-DSTx2) provided no additional benefits over a single DST. Lymph node cells from CsA-RAPA-DST subjects were hyporesponsive in MLR against donor cells and suppressed the proliferation of normal LEW cells in an MLR coculture assay, suggesting the presence of suppressor cells. Furthermore, in the CsA-RAPA-DST group, the anti-donor antibody response was suppressed. These data demonstrate that RAPA, CsA, and DST interact positively by inducing a clear-cut and significant prolongation of allograft survival in the rat model under conditions in which the individual components of the treatment are essentially ineffectual.

Improved survival from potentially lethal graft-vs.-host disease by donor pretreatment with a recipient-specific blood transfusion. I. Requirements for induction and specificity of the effect.

Pretreatment of prospective donors of hemopoietic cells with a single recipient-specific blood transfusion can significantly decrease the morbidity and mortality of graft-vs.-host disease (GVHD) in lethally irradiated, allogeneically reconstituted mice. This beneficial effect of donor pretreatment could be demonstrated in donor-recipient strain combinations that were H-2 + non-H-2, H-2, or only class II-disparate, but not in the class I-disparate C57BL-B6.C-H-2bm1 strain combination. The effect was proportional to the amount of recipient-strain blood used for transfusion. Donor transfusion with a single dose of 1 ml recipient-specific whole blood resulted in minimum GVHD, lower doses being less or not effective. The interval between donor pretreatment and the use of their hemopoietic cells for reconstitution appeared to be important. The best survival was found at an interval of 4 days. Multiple transfusion was not more effective than a single one. We compared the effectiveness of whole blood and irradiated spleen cells for donor pretreatment. Both protocols have been shown previously to suppress anti-recipient delayed-type hypersensitivity. It appeared that the blood transfusion protocol was superior to the spleen cell protocol. The beneficial effect appeared to be recipient specific, since a third-party blood transfusion did not improve GVHD. We found that the beneficial effect of donor blood transfusion was due to suppression of the anti-host immune response. The donor blood transfusion was able to induce bystander suppression to alloantigens that were not used for the induction of suppression, provided they were co-expressed with the specific alloantigens by the recipients. This also indicates that, although the induction of suppression is specific, the ultimate suppressive effect is non-specific.

Allogeneic transplantation of the radial side of the hand in the rhesus monkey: I. Technical aspects.

Since allogeneic transplantation of extremities can only be considered if uneventful long-term survival and functional recovery can be achieved, a series of 12 transplantations of the radial side of the hand were performed in rhesus monkeys so that these factors could be assessed. The transplant incorporated the first ray of the hand in conjunction with the radial forearm flap. Graft survival times varied from 21 to 179 days. Ten of 12 transplants showed rejection. In 2 of the 10, rejection could be reversed. Immunosuppressive therapy consisted of cyclosporin A, prednisone, monoclonal antibodies, and preoperative third-party blood transfusions. Monitoring of the microcirculation of the allograft could not provide a predictive value for transplant rejection. The first clinical signs of sensory and motor function recovery were detected after an average of 42 and 44 days, respectively. Indefinite uneventful allograft survival could not be established. Major complications such as sepsis, shock, and lymphoma development leading to death were encountered. The model, however, is technically feasible, and the results for functional recovery under immunosuppression are promising.

Effect of cyclosporin A on immunoglobulin class in patients receiving blood transfusions

IgG antibodies detected by flow cytometry in sera from potential renal transplant recipients are associated with an increased number of rejection episodes and impaired graft function. Furthermore, cytotoxic antibodies may develop if pre-transplant blood transfusions are given to such patients. We have investigated the effect of cyclosporin A on the development of IgM and IgG antibodies detected by flow cytometry after blood transfusions in 16 previously untransfused dialysis patients. Eight (group 1) received three to five third-party blood transfusions at two weekly intervals and the remaining eight (group 2) received transfusions with concomitant cyclosporin A therapy (10 mg/kg/day). Sera obtained after each transfusion were tested using flow cytometry against lymphocytes from six normal donors. In all 462 serum/cell combinations were tested. Sera from six out of eight patients in group 1 showed IgG antibody activity following blood transfusions and none developed IgM antibodies alone. In contrast IgG antibody activity was detected in one serum sample from only one of the eight patients in group 2 (P less than 0.02); a further three patients developed IgM but not IgG antibody activity during the transfusion protocol. IgG antibodies were found in 25/228 serum/cell combinations in group 1 but in 1/234 in group 2 (P less than 0.001). The patient in group 2 who developed IgG antibodies in one serum/cell combination was known to have red cell autoantibodies. This IgG activity was removed by red cell absorption, suggesting that the autoreactive red cell antibody cross-reacted with lymphocytes. Cytotoxic antibodies were detected in two serum/cell combinations in group 1 alone.(ABSTRACT TRUNCATED AT 250 WORDS)

The Effect of Cyclosporin Administered During a Third-Party Blood Transfusion Protocol on Humoral Immune Responses

Antigen pre-treatment in animals undoubtedly prolongs graft survival. In man, however, routine pre-transplantation blood transfusions have recently become controversial, principally because of the adverse effect of transfusion-induced sensitisation on graft survival rates. We have monitored the effect of cyclosporin administered during a planned programme of third-party blood transfusions on the development of both cytotoxic and anti-idiotypic antibodies. A total of 24 patients were recruited to the study; ten received cyclosporin with blood transfusions (BT) (group 1), 14 received BT alone (group 2). Anti-HLA antibodies developed in 3 of 9 patients in group 1 and 8 of 12 patients in group 2 (P less than 0.05). Anti-idiotypic antibody activity was detectable in 9 of 9 patients in group 1 and 7 of 12 patients in group 2 (P less than 0.006). The mechanism by which cyclosporin can prevent an anti-HLA antibody response and promote an anti-idiotypic response is unclear.

MODERATE EFFECT OF PREOPERATIVE BLOOD TRANSFUSIONS ON PANCREAS ALLOGRAFT SURVIVAL IN RATS AND DOGS

Treatment of type I diabetes by early pancreas transplantation requires the availability of a safe and effective transplantation technique. With the currently available immunosuppressive drugs it is difficult to obtain long-term pancreatic allograft survival. In this respect pancreas grafts compare unfavorably with heart or kidney grafts. Using a relatively simple and safe subcutaneous transplantation technique we investigated the effect of blood transfusions combined with low-dose immunosuppressive drugs in rats and dogs in order to attain an immunosuppressive schedule of low toxicity. Subcutaneous pancreas transplantation appeared to be a feasible technique, with long-term graft survival in syngeneically transplanted rats and autotransplanted dogs. Only a moderate prolongation of pancreatic allograft survival by blood transfusions was demonstrated in both models. In rats one or three preoperative donor-specific blood transfusions significantly prolonged pancreas graft survival to 23 +/- 15 and 29 +/- 15 days, respectively, compared with 12 +/- 2 days in untreated controls. Low-dose cyclosporine (15 mg/kg on the day of operation) led to improved graft survival in nontransfused recipients (17 +/- 4 days), however, this treatment could not further prolong graft survival in transfused animals (34 +/- 20 days). In dogs, treated postoperatively with azathioprine and prednisolone, three preoperative third-party blood transfusions led to 29 +/- 19 days of pancreas graft survival, which was not significantly different from nontransfused controls (17 +/- 5 days). These results indicate that, in rats as well in dogs, pancreatic allografts are less sensitive to the immunomodulating effect of blood transfusions than heart and kidney grafts.

The role of class I major histocompatibility complex antigens in prolonging the survival of hepatic allografts in the rat.

In an attempt to study the role of class I major histocompatibility complex antigens in inducing immunological unresponsiveness, the survival rates of hepatic allografts were compared in rats pretreated with blood taken from various rat strains. A single intravenous injection of 1 ml fresh heparinized whole blood seven days before transplantation significantly prolonged the survival of subsequent donor-specific hepatic allografts in the fully allogeneic ACI(RT1a)-to-LEW(RT1l) rat combination. However, pretreatment with blood taken from the third-party strain BN(RT1n) did not produce suppression of rejection, attesting to the specificity of the pretransplant transfusion effect. Interestingly, pretransplant transfusion of PVG.r1 blood, sharing only the RT1.A MHC region with ACI, significantly prolonged the survival of ACI-to-LEW hepatic allografts. In addition, no lymphocytotoxic antibodies could be detected at 30 or 100 days after transplantation in animals with long-surviving hepatic allografts pretreated with either PVG.r1 or ACI whole blood. On the other hand, pretreatment with PVG(RT1c) blood increased the survival of ACI-to-LEW hepatic allografts only moderately compared with controls. This finding may be consistent with a partial effect of some third-party blood transfusion. The experimental data suggest that the class I MHC antigens can be immunosuppressive in rat hepatic allografts. Adoptive transfer of 5 x 10(7) splenocytes taken from long-term-surviving hepatic allografts pretreated with donor ACI whole blood or PVG.r1 blood into irradiated (750 rads) LEW rats prolonged the survival of donor-type skin grafts, whereas third-party strain (BN) grafts were rejected. This finding suggests the presence of donor-specific suppressor cells.

Failure to prolong pancreatic islet allograft survival in rats with donor-specific blood transfusions and immunosuppression.

The effects on pancreatic islet allograft survival of donor-specific blood transfusions (DST) in combination with pre- and posttransplant immunosuppression were studied. A total of 12 groups of rats (n = 105) with chemically induced diabetes underwent islet allotransplantation. Multiple DST or third-party blood transfusions (TPT) were given prior to transplantation. Pretransplant immunosuppression consisted of azathioprine and prednisolone, and low-dose cyclosporin A was used for posttransplant immunosuppression. TPT, as well as separate or combined pre- and posttransplant immunosuppression without blood transfusions, did not prolong islet allograft survival. DST resulted in either primary nonfunction of the islet allografts or a markedly decreased islet allograft survival. These findings contrast with the beneficial effect of DST on whole-organ allograft survival in rats previously described by others.

Failure to prolong pancreatic islet allograft survival in rats with donor-specific blood transfusions and immunosuppression

Abstract. The effects on pancreatic islet allograft survival of donor-specific blood transfusions (DST) in combination with pre-and posttransplant immunosuppression were studied. A total of 12 groups of rats (n= 105) with chemically induced diabetes underwent islet allotransplantation. Multiple DST or third-party blood transfusions (TPT) were given prior to transplantation. Pretransplant immunosuppression consisted of azathioprine and prednisolone, and low-dose cyclosporin A was used for posttransplant immunosuppression. TPT, as well as separate or combined pre-and posttransplant immunosuppression without blood transfusions, did not prolong islet allograft survival. DST resulted in either primary nonfunction of the islet allografts or a markedly decreased islet allograft survival. These findings contrast with the beneficial effect of DST on whole-organ allograft survival in rats previously described by others.

Prediction of donor-specific transfusion sensitization. I. A linear logistic model.

Using linear logistic regression, six factors were identified as important predictors of risk of DST sensitization in a group of 195 patients. Factors increasing the risk were: percent panel reactive antibody (PRA), previous transplants, and pregnancy; those decreasing the risk were HLA antigens matched, third-party blood transfusions, and Imuran administration. From this analysis, the magnitude of the effect of each factor on the risk of sensitization was obtained. An equation was then obtained that can be used to compute an estimated probability of sensitization (PS) for each patient. As a test of predictive ability of the model, the PS was calculated for 66 patients in an independent patient group. These observations were arranged according to the estimated probability and then divided into intervals of risk. Overall, for each interval, a very high level of agreement was found between the predicted and actual number of sensitized patients. A total of 16.13 patients were predicted to become sensitized and 17 actually did.

Prostaglandin production and suppressor cell induction in transfusion-induced immune suppression.

Two models were used to examine the role of prostaglandin (PGE) in the inductive phase of transfusion-induced suppression. First, it was observed that post-operative allogeneic third-party blood transfusion resulted in prolonged major histocompatibility complex (MHC)-compatible rat heart allograft survival. Additionally, pretransplant antigen-specific allogeneic transfusion decreased graft-versus-host (GVH) reactivity in mice. An inhibitor of PGE synthesis, indomethacin, blocked transfusion-induced suppression when it was administered to either transfused rat heart recipients, or graft-versus-host responder mice. Neutralization of endogenous PGE by anti-PGE antibody also blocked allogeneic blood induced suppression. Allogeneic-blood-induced splenic suppressor cells down-regulated normal GVH responsiveness, and appeared to be generated via a prostaglandin-dependent pathway.

Genetics of the blood transfusion effect on heart allografts in rats.

Working with the recently available recombinant haplotypes of the rat major histocompatibility complex (MHC)--RT1, we investigated the effect of various types of blood transfusion (BT) on allograft prolongation, including blood identical for the whole RT1 haplotype with that of the donor or for only a part of it. One or two milliliters of donor blood significantly prolonged graft survival in the (LEW X BN)F1----LEW or the LEW X 1W----LEW X 1A combination. The optimal regimen consisted of two BTs given 15 and 7 days prior to grafting; BTs given at day -30 were ineffective. A BT given on the day of the operation was effective, but sequential BTs after grafting did not further increase graft survival. In the (LEW X BN)F1----LEW combination, blood from congenic LEW X 1N rats significantly prolonged graft survival, but third-party BTs were ineffective or had only a borderline effect when transfused (1 ml, 8 times) within the three months before transplantation. This showed the major role of the RT1 system as well as the specificity of the model. Although the survival of LEW X 1A heart grafts transplanted into LEW X 1W recipients could not be significantly prolonged by donor blood, with the reverse--and "weaker"--combination (LEW X 1W----LEW X 1A), 2 ml of donor blood led, in all cases, to greater than 100 days graft survival. In this last combination, third-party BT (LEW X 1N) was again totally ineffective. Blood from RT1-recombinant rats was used to test the role of the respective RT1.A, B, and C regions, in the enhancing effect. BTs from LEW X 1AR2 or LEW X 1WR2 recombinants--sharing, respectively, RT1.C and RT1.A with the graft donor--were only moderately effective, as compared with BTs from the graft donor. On the other hand, LEW X 1WR1 BTs--sharing the RT1.A and RT1.B regions with the graft donor--had a much more powerful effect on heart survival. The results strongly suggest that the RT1.B region (coding for Ia-like antigens) must be shared by the graft and blood donor in order to mediate a significant graft prolongation.

Renal Transplantation Update

Various factors affect the outcome of renal transplants in humans. Matching for HLA-A, -B, and -DR histocompatibility antigens improves survival rates for renal allografts from first cadaver donors. Zero-HLA-A- and -B-antigen-mismatched grafts and two-HLA-DR-antigen-matched grafts do better, although results differ depending on the recipient's primary renal disease. Pretransplant third-party blood transfusions significantly improve survival rates of cadaver donor allografts. The mechanism of this beneficial effect has not been identified; however, blood transfusions probably do not "select out" high responders among potential recipients by stimulating the production of lymphocytotoxic antibodies. Cyclosporine has been heralded as a potent, nonspecific immunosuppressive agent that will significantly improve renal allograft survival rates. The selectivity of cyclosporine's effect on T lymphocytes is advantageous; however, its side effects, especially nephrotoxicity, may limit its usefulness. Attention to the potential surgical complications of renal transplantation can significantly reduce morbidity and mortality.

Modification of allograft survival in rats by blood transfusion to the donor.

In the BN/Ro-to-WAG/Ro rat donor-host combination, third-party blood transfusions given to the donor shortly before transplantation can markedly influence heart allograft survival if the immune response of the recipient has been modulated. The WAG recipients were either transfused with BN donor blood, which leads to a permanent graft survival, or postoperatively treated with a single i.m. injection of 15 mg/kg cyclosporine, which leads to a moderate prolongation of graft survival. In the transfused recipients, blood transfusions to the donor had a detrimental influence on graft survival. In the cyclosporine-treated recipients, blood transfusions to the donor had a beneficial effect. There was no significant difference in graft survival after a single transfusion or multiple transfusions to the donor. Recipient-type blood transfusion to the donor did not influence graft survival, nor did irradiated third-party blood or third-party erythrocytes. However, third-party leukocyte suspensions had a significant influence on heart allograft survival. It is concluded that third-party viable leukocytes are responsible for the induction of the donor transfusion phenomenon. It is also postulated that the third-party leukocytes interact with the dendritic cell population of the graft, leading to a reduction in its immunogenicity.

Effets spécifique et aspécifique de la transfusion sanguine sur la survie de la greffe de cœur chez le rat

We investigated the role of various blood-transfusions regimens including donor specific and third-party blood transfusions in rat heart transplantations. One ml donor blood (BN; RT1n) given to the recipients (Lew, RT1(1)) of (Lew.BN)F1 rats 7 days prior to grafting prolonged heart survival (24 +/- 8 vs 9.9 +/- 2.7 in control). Heart graft survival was much less when donor blood was given to recipients 1 month before grafting. There was no complementary effect between BT from donor and from third party blood or vice-versa. In addition, although donor blood prolonged graft survival (19.7 +/- 7.5 vs 9.9 +/- 2.7) when given the day of surgery, post surgery sequential BT from the donor (day 0-3-7 and 14 after transplantation) did not further increased survival (14 +/- 4.6). The effect of donor-BT was RT1a specific since third-party blood (1 ml) was ineffective (9.5 +/- 1) whereas 1 ml of congenic RT1n rat (Lew. 1 N) blood increases heart survival: 14 +/- 2.14 vs 9.9 +/- 2.2 in control animals. Although only 1 ml of donor blood enhanced specifically the heart allografts of the corresponding RT1 phenotype, weekly repeated third-party BT (n = 8) did also prolong graft survival (13.7 +/- 3.4) suggesting that aside from classical and powerful model of active RT1 specific effect of BT, a nonspecific, dose-related mechanism can occur.

Influence of pre- and perioperative blood transfusions on renal allograft survival.

One hundred and seventy-nine patients who received transplants between January 1975 and May 1980 have been studied retrospectively in order to elucidate the effect of pre- and perioperative random third-party blood transfusions on first cadaveric renal allograft survival. The 179 patients were approximately evenly divided into four groups; those who received blood transfusions preoperatively only (group 1, n = 45), received blood both pre- and perioperatively (group 2, n = 48), received blood perioperatively only (group 3, n = 37), or were never transfused before transplantation or perioperatively (group 4, n = 49). Actuarial graft survival for group 1 was 73% at 1 to 4 years, which was significantly better (P less than or equal to 0.02) than each of the other three groups at all four intervals. Groups 2, 3, and 4 were statistically identical to each other and had graft survivals of 52% or less at 1 to 4 years. From our study we conclude that: (1) Preoperative transfusions confer significant benefit on recipients of first cadaveric renal allografts. (2) Perioperative blood transfusions are without significant benefit in previously untransfused patients, and significantly lower allograft survival in previously transfused patients. A rational blood transfusion policy based on these results would be to transfuse renal allograft recipients preoperatively and avoid perioperative blood transfusions as much as possible.