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Altered macrophage function after thermal injury is associated with increased production of PGE2 and TNF. However, it is not clear why synthesis of both cellular products remains elevated, as PGE2 is a potent inhibitor of TNF secretion. We studied the relationship between PGE2 and TNF synthesis in a murine model of thermal injury, and examined the effect of prostaglandin blockade on splenic macrophage secretion of these mediators of inflammation. LPS-stimulated production of PGE2 was significantly elevated in burn groups compared with sham-burned controls (pg/ml mean(SEM); sham 151(32): burn 597(147), p < 0.01). TNF production was similarly increased after thermal injury (pg/ml mean(SEM); sham 62(20): burn 928(316), p < 0.01). In vitro culture of macrophages with indomethacin augmented LPS stimulated TNF production in sham-burned controls but did not affect synthesis in burn groups, suggesting a loss of PGE2-dependent regulation of TNF synthesis after thermal injury. Direct measurement of TNF secretion as a function of exogenous PGE2 confirmed this dissociation between PGE2 and TNF synthesis, as burned animals displayed a 5-fold reduction in sensitivity to PGE2-induced inhibition of TNF, when compared with sham-burned controls (ID50 PGE2 molar; sham 1.26 x 10(-8): burn 6.43 x 10(-8), p < 0.05). In vivo pretreatment of burn groups with indomethacin for 5 days before assay partially restored sensitivity to the prostaglandin, and significantly down-regulated synthesis of both TNF and PGE2. These data show that thermal injury is associated with a loss of PGE2-dependent down-regulation of TNF synthesis, which accounts at least in part for increased TNF in these animals. In vivo cyclooxygenase blockade partially restored sensitivity to the prostaglandin and consequently down-regulated synthesis of TNF. These data further support existing evidence that suggests a potential therapeutic role for cyclooxygenase blockade after major thermal injury and trauma.

S-nitrosylation/denitrosylation of critical cysteine residues on proteins serves as a redox switch that regulates the function of a wide array of proteins. A key signaling pathway that is regulated by S-nitrosylation is apoptotic cell death. Here we will review the proteins in apoptotic pathways that are known to be S-nitrosylated by endogenous NO production. The targets and functional consequences of S-nitrosylation during apoptosis are multifaceted, allowing cells to fine tune their response to apoptotic signals.

Interleukin 10 (IL-10) is thought to be protective in injury and sepsis. However, we recently reported that IL-10 antagonism can be beneficial after burn injury. This study used IL-10-deficient (IL-10 [-/-]) mice to further define the role of IL-10 after injury.Wild-type (WT) C57BL/6 or IL-10 (-/-) mice were anesthetized, sham or burn injured, and immunized subcutaneously with a T-cell-dependent protein antigen. Ten days later antigen-specific serum antibody isotype formation was measured by enzyme-linked immunosorbent assay. In addition, antigen-stimulated splenic T-cell proliferation and cytokine production (interleukin 2, interferon gamma, and tumor necrosis factor-alpha) were measured.Burn-injured IL-10 (-/-) mice survival (80%) was equivalent to that of burn-injured WT mice (74%). An injury-dependent loss of T-helper 1 (Th1)-type antibody isotype (IgG2a) formation occurred in both WT and IL-10 (-/-) mice. In vitro studies indicated that burn injury caused reduced antigen-stimulated splenic T-cell proliferation and Th1-type (interleukin 2 and interferon gamma) cytokine production in WT and IL-10 (-/-) mice, whereas burn-injured IL-10 (-/-) mice produced high levels of antigen-stimulated tumor necrosis factor-alpha.IL-10 is not essential for survival after burn injury or for several injury-induced changes in adaptive immune function, including Th1-type antibody isotype formation, T-cell proliferation, and Th1-type cytokine production.

The Committee on Workforce Issues of the Society for Vascular Surgery (SVS) and the North American Chapter, International Society for Cardiovascular Surgery (NA-ISCVS) generated data on the numbers and trends of operations performed and workforce requirements to provide surgical care of patients with vascular disease. Data for analysis were obtained from The National Center for Health Statistics-National Hospital Discharge Survey and questionnaires sent to 2732 vascular surgeons. The data showed that SVS/NA-ISCVS and regional vascular society (RVS) surgeons performed 51% of 583,000 vascular procedures undertaken in the United States in 1992. This represents a 24% increase over the 41% reported in 1985 by similarly defined surgeons. Analysis of 1992 index cases documented that SVS/NA-ISCVS and RVS surgeons accounted for 80% of 31,000 aortoiliofemoral bypasses, 68% of 46,000 aortic aneurysmectomies, 64% of 91,000 carotid endarterectomies, and 72% of 98,000 angioaccess procedures. The mean numbers of vascular operations performed in 1992 by SVS/NA-ISCVS and RVS surgeons were 144 and 100, respectively. These procedures represented 64% and 39% of the total surgical caseload of SVS/NA-ISCVS and RVS surgeons, respectively. American Board of Surgery (ABS)-certified vascular surgeons performed a mean of 171 vascular operations in 1992. Other surgeons, including ABS-certified general surgeons, appear to be performing fewer vascular operations. The latter fact and the increasing incidence of vascular disease in an expanding elderly population supports a continued need for vascular surgery specialists. Evolving technology and new health care delivery systems, however, may lessen the need for surgical care of these patients. Continued assessments of workforce activity will allow better definition of changing vascular surgery needs.

To find out if an infective challenge caused by a burn followed by caecal ligation and puncture in mice caused more abnormalities of the immune response than burn alone or caecal ligation and puncture alone.Laboratory study.University hospital, USA.80 male 7-8 week old A/J mice.Burn followed 10 days later by caecal ligation and puncture (n = 18), caecal ligation and puncture alone (n = 24), burn alone (n = 20), and controls (n = 18). The mice had their spleens removed on day 11 (n = 28; 6, 8, 8, and 6 in the respective groups), day 12 (n = 26; 6, 8, 6, and 6), and day 13 (n = 26; 6, 8, 6, and 6), and splenocytes and adherent cells were harvested for measurement of prostaglandin E2 (PGE2), interleukin 1 (IL-1), interleukin 2 (IL-2), interleukin 6 (IL-6), and tumour necrosis factor alpha (TNF-alpha).Alterations in the production of the cytokines.After the double challenge (burn followed by caecal ligation and puncture) there were significant reductions in production of TNF-alpha and IL-6 compared with caecal ligation and puncture alone (p0.05), burn alone (p0.05), and controls (p0.05). These findings indicate that activation of macrophages was reduced after infection; production of TNF-alpha, IL-1, and IL-6 by splenocytes stimulated by lipopolysaccharide was reduced.The differences do not seem big enough to indicate that mortality would be increased after caecal ligation and puncture alone. Only when there has been a previous injury (which resulted in hyperactivation of macrophages followed by a more pronounced hypoactivation) would mortality increase. In view of clinical trials with antiendotoxin and antiTNF antibodies that failed to improve survival in infected patients, we suggest that the mechanisms of the cellular immune response need further clarification.

As a result of major advances in the care of thermally injured patients over the last two decades, mortality from hypovolemic shock, acute renal failure, and metabolic wasting has fallen dramatically. However, infection remains an important cause of morbidity and mortality in these patients [1]. The prevalence and severity of septic episodes is largely secondary to profound alterations in defence mechanisms which often characterise the host’s response to major burn injury.

Severe thermal injury is associated with major alterations in cell-mediated immunity. Because most B-cell responses are regulated or critically dependent on T-cell help, it is not surprising that many studies have also shown a variety of defects in humoral immunity after thermal injury. However, the nature of the relationship between the in vitro ability to produce antibody and subsequent in vivo responses remains unclear.With a murine model of thermal injury, the primary and secondary humoral immune response to tetanus toxoid (TT) was examined during a 6-week period after sham burn or burn injury. Serum anti-TT titers and the numbers of anti-TT-secreting splenocytes were determined.Splenocytes from burned animals displayed normal or decreased TT-specific immunoglobulin (Ig) M plaque formation. In contrast, however, IgG plaque formation was persistently increased for up to 6 weeks after thermal injury, suggesting a switch from IgM to IgG antibody production. Conversely serum titers of TT-specific IgG antibody were persistently lower in burn, compared with sham groups. Changes in serum immunoglobulin levels did not account for this marked discrepancy between enhanced in vitro IgG plaque formation but impaired in vivo levels of TT antibody.The data suggest that thermal injury is associated with a diminished ability to propagate and maintain a normal IgG antibody response, despite the presence of normal or increased numbers of antigen-specific B cells.

Among the fundamental immunologic abnormalities induced by serious traumatic or thermal injury are alterations in T cell activation, reduced lymphocyte interleukin-2 (IL-2) production, and associated depression of T lymphocyte proliferation. This study attempts to localize the cellular mechanisms underlying abnormal IL-2 production in thermal injury.Following National Institutes of Health guidelines, 150 A/J mice were anesthetized, subjected to a 20% full-thickness scald burn injury or sham burn, and killed at intervals from 4 to 21 days later; splenocytes were harvested for in vitro studies. For measurement of IL-2 production, cells were cultured with either concanavalin A or a combination of the phorbol ester PMA, which directly activates protein kinase C, and the calcium ionophore A23187, which increases intracellular calcium. Cytokine mRNA expression was measured by Northern blot analysis and IL-2 production by bioassay.Both IL-2 production and IL-2 mRNA expression were consistently suppressed in concanavalin A-stimulated cells from burned mice compared with sham burns. This suppression of IL-2 and IL-2 mRNA also occurred when T cells were activated with PMA and A23187, bypassing the earlier stages of the signal transduction mechanism. IL-1 beta and tumor necrosis factor-alpha mRNA expression were consistently increased in burned animals, indicating that decreased IL-2 mRNA expression was specific to IL-2 and not representative of a global decrease in cytokine mRNA expression.These results suggest that the principal cellular abnormalities that result in altered T cell activation and IL-2 production after thermal injury lie downstream of the initiating signal transduction events and before IL-2 gene transcription.

Traumatic injury and burns remain a major cause of morbidity and mortality in the United States and despite modern techniques of resuscitation and intensive care and an ever increasing number of powerful and effective antibiotics nearly 80% of deaths that occur in trauma and burn patients more than 7 days after injury are caused by sepsis [1]. During the past 10 years the longstanding clinical assumption that host defenses against invading microorganisms were impaired following severe injury has become grounded in fact through the study of injured patients and appropriate animal models. Abnormalities in both specific and nonspecific mechanisms of defense against microorganisms have been described by a number of laboratories, including our own. Nonspecific mechanisms include function of polymorphonuclear [2–4] and mononuclear phagocytes [5–6], natural killer cells [7–9], and products of complement activation [10–13]. Specific defense mechanisms include antibody formation by B lymphocytes [14–16], direct microbial killing by T lymphocytes, and T lymphocyte activation of mononuclear phagocytes [17–19].

The purpose of this study was to evaluate the efficacy of angioplasty in the treatment of femoropopliteal arterial disease.From 1980 to 1991, 126 angioplasty procedures were performed in 131 limbs of 106 patients with 175 femoropopliteal lesions (26 common femoral, 118 superficial femoral, and 31 popliteal). Critical ischemia was present in 55 limbs (42%), and claudication was present in 76 (58%). Angioplasty was performed for a single lesion in 87 limbs (66%) and for multiple lesions in 44 (34%). In 13 limbs (10%) the most severe lesion was an occlusion; in 118 (90%) all lesions were stenoses. Distal runoff was good (2 or 3 vessels patent) in 72 limbs (55%) and poor (0 or 1 vessel patent) in 59 (45%).Death within 30 days occurred in 0.8%, nonfatal systemic morbidity in 7.1%, and local morbidity in 1.6% of procedures. Multivariate analysis revealed that indication and age were predictive of increased morbidity and mortality rates. Immediate success was achieved in 95% of limbs treated. Mean follow-up time was 2.0 years. The overall 5-year cumulative primary patency rate was 45% (+/- 5%). In a proportional hazards model indication and lesion type were predictive (p0.01) of long-term failure, with relative risks of 2.0 (1.2 to 3.3) and 2.7 (1.3 to 5.6), respectively. The 5-year primary patency rate after angioplasty for stenoses and claudication was 55% (+/- 7%), for stenoses and critical ischemia it was 29% (+/- 11%), and for occlusions it was 36% (+/- 14%).These results suggest that femoropopliteal angioplasty is a low-risk procedure with acceptable long-term results in patients with claudication and stenoses.

Vein graft stenosis remains an important contributing factor to the failure of infrainguinal arterial reconstruction. Repair of these lesions before graft occlusion provides sustained patency, yet the optimal method of repair has not been established. Percutaneous transluminal balloon angioplasty of these vein graft lesions has been repeatedly advocated as an alternative to surgical revision. Balloon angioplasty was used in 30 patients with 54 stenotic lesions occurring in autogenous vein grafts after infrainguinal reconstruction. The primary 5-year cumulative patency rate was 18% overall, with no significant differences observed among patency rates based on initial indication, length of stenotic lesion, or requirement for preliminary thrombolytic therapy. The 3-year patency rate associated with vein graft lesions requiring only a single angioplasty proved significantly higher (59%) than those requiring repetitive dilations (6%). It is our conclusion that balloon angioplasty for vein graft stenosis has significant limitations in providing sustained secondary patency.

Bypass vein grafts to the infrapopliteal arteries now achieve a 5-year cumulative patency equivalent to that of vein grafts to the popliteal arteries. The technique of in situ vein grafting to the tibial arteries is described and the results are presented. The 5-year cumulative patency for such bypasses and the 5-year limb salvage in the same patients have both been approximately 80%. These results coupled with those reported from other centers that have sizable experience in tibial artery reconstruction suggest that there has been real progress over the past decade in the salvage of lower extremities in patients with far advanced peripheral vascular disease through the use of autogenous venous bypass grafts.

The purpose of the present study was the investigation of the effect of cyclosporin A (CsA) on the induction of alloreactivity in vitro and in vivo. Addition of CsA to mouse mixed lymphocyte cultures (MLC) not only inhibited lymphocyte proliferation but also prevented the generation of alloreactive cytolytic lymphocytes (CL). It was necessary to add CsA within the first 3 days of a 5-day MLC in order to achieve a significant suppressive effect. Lymphocytes, after being cultured in MLC with CsA for 4 days or longer, were incapable of being activated upon re-exposure to the same alloantigens although their responses to unrelated antigens remained intact, indicating antigen specificity of the suppression induced by CsA and its long-lasting effect. Furthermore, lymphocytes from mice treated with CsA after allosensitization failed to manifest primary cytotoxicity and could not be reactivated in a secondary MLC. Finally, CsA had no effect on those CL already generated, suggesting that CsA acts upon the induction of CL rather than the effector phase.

Mice sensitized with alloantigens and treated with cyclosporin A (CsA) were incapable of generating antigen-specific cytolytic lymphocytes (CL). Lymphocytes from these CsA-treated animals could not be reactivated upon exposure to the same alloantigens in mixed lymphocyte culture (MLC), whereas their response to a third-party antigen remained intact, suggesting a long-lasting and specific effect of CsA. After being irradiated, these lymphocytes from CsA-treated animals were added to normal MLC and were shown to prevent normal lymphocytes from becoming cytolytic in a dose-dependent and antigen-nonspecific fashion. These suppressor cells were not detected in mice receiving CsA only, indicating that CsA did not induce but rather permitted the expression of suppressor cells possibly generated by allosensitization. The suppressor cells appeared to be T lymphocytes, because treatment with anti-Thy-1.2 antibody and C abrogated their suppressive activity. The present results suggest that activation and/or sparing of suppressor cells by CsA may account for the long-lasting unresponsiveness seen in CsA-treated animals.

There exists in the serum α-globulins an immunosuppressive factor(s) which was first described by Kamrin (1), and subsequently separated chromatographically by Mowbray (2), and by our laboratories (3, 4). This factor has been demonstrated to prolong skin grafts (3) and to inhibit antibody formation (5, 6). We have also found that this factor inhibits lymphocyte proliferation induced in vitro by both mitogens and antigens (4). We have recently found that the active factor is a polypeptide(s) which migrates electrophoretically with, and may be bound to, an α-globulin protein(s) (7). We have called the active material “immunoregulatory α-globulin” (IRA).4 The mechanism of action of this immunosuppressive factor remains to be determined. Studying mitogen stimulation of lymphocytes in tissue culture, we have observed that the IRA is not suppressive if this agent is added after the lymphocytes have become activated (8). This observation suggests that the IRA acts upon the early events involved in antigen recognition.

The causes of failure of infrainguinal arterial reconstruction vary with the method of reconstruction used. The commonest cause of autogenous vein-graft failure is myointimal hyperplasia of the graft itself. The commonest cause of polytetrafluoroethylene-graft failure, on the other hand, appears to be a myointimal hyperplastic lesion of the artery at, or just beyond, the distal anastomosis of the graft. Strategies for dealing with these two forms of graft failure, including the use of thrombolytic therapy, are discussed.

Suppression of cellular immunity and increased susceptibility to sepsis frequently accompany thermal injury. However, a convincing association between the two has been difficult to establish in human beings. Therefore we chose to investigate the relationship of impaired cell-mediated immunity with susceptibility to sepsis in an animal model. We studied the response to phytohemagglutinin (PHA) and interleukin-2 (IL-2) production by splenocytes from mice subjected to a standard 25% scald burn and killed at intervals of 3, 5, 7, 10, 14, and 25 days after thermal injury. Burned mice were compared in all instances to sham-burn animals (i.e., animals that had been anesthetized and shaved but not burned). We also studied mortality after cecal ligation and puncture (CLP), as a septic challenge, in burned and control animals at the same postburn intervals. We found maximal suppression (50% to 55%) of the PHA response at 10 to 14 days after injury and maximum suppression (68%) of IL-2 production at 7 days. Both of these parameters returned to normal by postinjury day 28. Mortality after CLP increased gradually from control levels after thermal injury up to a maximum of 88% on postburn day 10 and also returned to control levels after 28 days after burn. Significant correlations were found between mortality after CLP in the postburn period and suppression of the PHA response, on the one hand, and the suppression of IL-2 production, on the other (r = 0.89 and 0.91, respectively; p less than 0.05). This result implies a causal relationship between impaired cell-mediated immunity and susceptibility to sepsis after burn injury.

A total of 130 axillofemoral grafts were peformed in 120 patients from 1966 to present; 66 were axillobilateral femoral moral and 64 were axillounilateral femoral grafts. The average patient age was 66 years. The indications for operation were occlusive disease with rest pain or gangrene in 102 patients, severe claudication four, and bypass of a thoracic aneurysm in one. The one month operative mortality rate was 8 percent (ten patients). Four deaths occurred in elderly patients operated upon as emergencies with acute aortic occlusion. In the 126 elective cases, there were six postoperative deaths (4.7 percent). Twenty thrombectomies were performed in 15 axillobilateral femoral grafts of which 15 (75 percent) were successful. In the unilateral grafts there were 25 thrombectomies in 22 grafts, of which only eight (32 percent) were successful. The 5 year patency rate by the life-table method was 74 percent for axillobilateral femoral grafts and 37 percent for axillounilateral femoral grafts. This difference in patency rates is statistically significant (p0.01). Average flow measured by electromagnetic flow meter in six axillobilateral femoral grafts was 621 c.c. per minute and in 11 unilateral femoral grafts was 273 c.c. per minute. We attribute the higher patency rate in axillobilateral femoral grafts to this increased flow. These data indicate that axillobilateral femoral grafts may be performed with anticipation of a 5 year patency rate comparable to that with conventional aortoiliac reconstruction and that axillobilateral femoral grafts should be performed in preference to axillounilateral femoral grafts in poor-risk patients requiring extra-anatomic bypass for aortoiliac occlusive disease.

Six patients with metastatic renal cell carcinoma were treated with five intravenous infusions (every other day) of autologous lymphocytes incubated in vitro with I-RNA extracted from the lymphoid tissue of guinea pigs immunized with the patient's own tumor. No toxicity was evident. One patient showed regression of multiple pulmonary metastases beginning three months after therapy with complete remission by six months. She remained without evidence of disease until 18 months after therapy. Two other patients had more than 50% regression of measurable metastases lasting eight and ten months after therapy. Two patients showed stabilization of previously growing renal cell carcinoma pulmonary metastases. A single patient with renal cell carcinoma metastatic to brain had progressive tumor growth after a single I-RNA treatment. Serial peripheral blood lymphocyte samples obtained from each of the patients during I-RNA therapy demonstrated progressive increase in in vitro cytolysis of allogeneic renal cell carcinoma targets. Boosts in cytolytic effect were shown in all patients during I-RNA treatment regardless of their subsequent clinical course. These results seem to justify a randomized, prospective trial of xenogeneic I-RNA therapy in renal cell carcinoma patients with lesser tumor burden.

We attempt to elucidate the mechanisms of neutrophil (PMN) activation after burn injury. We previously reported prolonged elevations of PMN cell surface complement (C) opsonin receptor levels after burn trauma with a corresponding period of depressed PMN chemotaxis to C5a, which suggests that the C product, C5a, was responsible for PMN activation. However, a lack of direct correlation of C activation with C receptor levels soon after injury raised the possibility of a second PMN-activating substance. We therefore investigated the effect of endotoxin (LPS) on the expression of the C receptors (CR1 and CR3) by normal human PMNs. Concentrations from 0 to 50 ng/ml of LPS 026:B6 caused a dose response increase in the PMN surface expression of CR1 and CR3 as assessed by monoclonal antibody binding and indirect immunofluorescence. The relative CR1-dependent fluorescence rose from a mean of 50 to 385 and CR3 from 50 to 300. Chelation by ethylenediaminetetra acetic acid (EDTA) did not influence this dose response, thus ruling out the possibility of C activation by LPS--an inference supported by the lack of complement activation observed with these concentrations of LPS in normal serum. A similar dose response was obtained in the absence of other cell types or serum, which implies a direct effect that mimicked that of C5a. To determine the mechanism of the later, prolonged C activation after burn injury, we next examined C activation products in 22 patients with burn injuries. Elevations of plasma C3a desArg were present and persisted for 50 days. Elevations were at maximum levels on days 9 through 13 postburn (mean +/- standard error of mean [SEM], 496 +/- 47 ng/ml versus normal 113 +/- 32; p less than 0.01). These were accompanied by elevations of C4a desArg (917 +/- 154 ng/ml versus normal 424 +/- 50; p less than 0.01), which are indicative of classic pathway activation. Finally, we examined PMN function, phagocytosis and percentage killing of Staphylococcus aureus, and found PMN function to be unaltered in the 22 patients. Thus PMN activation after burn injury appears to be caused by LPS soon after injury and by C5a later after injury and affects only selected PMN functions.

Patients who have sustained a large hemispheric stroke are not candidates for early carotid endarterectomy, but there is less agreement regarding the role of carotid endarterectomy in patients with small, fixed neurologic deficits. Accepted practice in many centers is to wait 4 to 6 weeks after the onset of the deficit before proceeding with carotid endarterectomy because of the fear that early revascularization will increase the size of the infarct. Earlier endarterectomy, however, in patients with significant residual ipsilateral carotid territory at risk may prevent repeated infarctions. For the past 5 years our approach to patients with a small stable stroke and significant stenosis (greater than 75%) has been prompt ipsilateral endarterectomy. Of the 337 carotid endarterectomies at our institution since 1979, a subset of 28 patients with hemodynamically significant carotid lesions presented with a small, fixed stroke. The period of time between the appearance of the stroke and carotid endarterectomy averaged 11 days, but 53% of patients were operated on within 7 days of the onset of symptoms. Selective shunting based on intraoperative EEG monitoring was utilized and 40% of the 28 patients required shunts. Operative mortality consisted of one death from a pulmonary embolus, and no patient sustained a new postoperative deficit. Long-term follow-up was available for 96% of patients over a mean of 2 years. During this time two new neurologic events occurred: one fatal stroke and one transient deficit. This experience indicates that patients with small, fixed neurologic deficits who continue to have carotid territory at risk may safely undergo carotid endarterectomy without waiting 4 to 6 weeks.

The immunoreactivity of lymphocytes from mice bearing large tumors was investigated. As compared to normal lymphocytes, lymphocytes from tumor bearers manifested a lessened proliferative response to T-cell mitogens and were less capable of destroying syngeneic tumor target cells. However, the impaired reactivity of these lymphocytes was improved significantly after washing repeatedly in tissue culture medium in vitro. The increase in cytolytic activity was tumor specific. A membrane-associated suppressive substance was detected in the washing medium which inhibited the cytotoxicity of washed lymphocytes. The effect of suppressive substance was tumor specific and sensitive to treatment with protein A and anti-immunoglobulin antibody, indicating that the suppressive substance contained immunoglobulin. The present findings suggest that a membrane-associated suppressive substance(s) may be in part responsible for the diminished immune responsiveness of lymphocytes from animals bearing large tumors.

The suppression of tumor-specific cell-mediated cytotoxicity by human immunoregulatory alpha-globulin (IRA), by a peptide fraction derived from IRA, and by IRA-like peptides from the serum of cancer patients was studied in a syngeneic murine tumor-host system. Splenic lymphocytes from tumor-immunized mice were cytotoxic specific tumor cells in vitro as measured by the [125]-iododeoxyuridine release microcytotoxicity assay. However, this effect was significantly depressed if 1.25 to 5 mg of IRA per ml were added to the cultures. Pooled lyophilized normal human serum protein was inactive. IRA peptide and IRA-like peptide fractions from cancer patients were also highly suppressive of cell-mediated cytotoxicity at much lower concentrations (0.05 to 0.5 mg/ml). Control human serum peptide, which failed to inhibit the induction of hemolytic plaque-forming cells in sheep erythrocyte-injected mice, had no effect on cell-mediated cytotoxicity. IRA and IRA-like peptide fractions were not cytotoxic to the effector lymphocytes or to the target cells at the concentrations used.

An immunosuppressive peptide fraction was isolated by means of gel filtration, membrane partition, and ion-exchange chromatography from the sera of patients hospitalized for cancer. The resulting peptide fraction, which was heterogeneous as judged by high-voltage electrophoresis, was found to suppress both phytohemagglutinin-induced proliferation of lymphocytes in vitro and the in vivo induction of splenic plaque-forming cells in mice. The specific activity of the peptide fraction, which was isolated from the sera of cancer patients, was significantly increased over that of the unfractionated starting material. Moreover, in control experiments, when the sera of normals or non-cancer-bearing hospitalized individuals were subjected to the same chromatographic techniques, no active peptide fraction could be obtained.