Richard M. Condie

Prevention of cytomegalovirus infection by prophylaxis with an intravenous, hyperimmune, native, unmodified cytomegalovirus globulin: Randomized trial in bone marrow transplant recipients

We have completed a randomized trial to evaluate the safety and effectiveness of hyperimmune cytomegalovirus intravenous human globulin in prevention of cytomegalovirus infection and related problems in bone marrow transplant recipients. Prophylactic intravenous administration of this native, intact, hyperimmune, cytomegalovirus IgG, at a dose of 200 mg/kg 25, 50, and 75 days following transplant resulted in complete protection against cytomegalovirus infection during the 120 days covered by the treatment (p = 0.009). There was no interstitial pneumonia or mortality in the group receiving the hyperimmune IgG. This is significant at the p = 0.014 when compared with the supporting treatment control group. In bone marrow transplant recipients, prophylaxis with a total dosage of 0.6 g/kg of an intravenous hyperimmune cytomegalovirus globulin was safe and afforded effective protection against cytomegalovirus infection and interstitial pneumonia in this high-risk population.

Suppression of plasma cell hepatitis with 6-mercaptopurine

Abstract Four patients with plasma cell hepatitis were treated with 6-mercaptopurine (6-MP). In each case evidence of liver disease, as measured by liver function tests, was suppressed. Studies in human beings on the effect of 6-mercaptopurine on antibody production and on the expression of delayed hypersensitivity showed no effect with the dosages used. 6-Mercuptopurine in a dose of 1.5 mg. per kg. was effective in reducing the inflammatory response in half the patients studied. This antiinflammatory effect was not seen in the three patients with plasma cell hepatitis studied. Evidence from other laboratories indicates that 6-mercaptopurine prevents virus replication. We think that the best explanation of the effect of 6-mercaptopurine on plasma cell hepatitis is that it suppresses a chronic virus infection.

Seven years' experience with antilymphoblast globulin for renal transplantation from cadaver donors.

Antibody of the IgGab type can be isolated from horses immunized with cultured human lymphoblasts plus complete Freunds adjuvant. The essential steps for the production of a safe, potent anti-human lymphoblast globulin (ALG) are: A) the use of early bleedings after immunization to reduce the titer of antibodies which react with red blood cells and platelets; B) careful absorption with human red blood cell stroma and platelets; C) stabilization with non-crystalline silica dioxide; D) chromatography through QAE sephadex to remove pyrogens, microaggregates and possible inhibitors of ALG activity; E) careful safety testing in animals for toxicity and pyrogenicity; and F) testing in vitro for sterility. Such a purified horse ALG (IgGab) can be administered safely intravenously to patients to supplement a standardized immnnosuppressive regimen incorporating azathioprine and prednisone. Under these circumstances, allergic reactions are very rare, antibodies to horse IgG do not develop, skin tests to horse IgG remain negative, and immune elimination of circulating horse IgG from the human circulation cannot be demonstrated. The overall results of ALG patient survival and transplant function after 184 consecutive first cadaver transplants at the University of Minnesota demonstrate a statistically significant improvement in both parameters accompanying increases in ALG dose while rigidly utilizing standardized doses of azathioprine and prednisone. There is a significant reduction in the number of grafts lost to rejection; significant reduction in the number of rejection episodes; significant delay in the onset of rejection episodes; but there is no increase in septic loss of patients or kidneys. These efforts could be seen in the gross data or when subgroups controlling for patient age, tissue typing were analyzed. Excluding patients at high risk did not alter the results. The beneficial effects of ALG were particularly striking in good matches. In the highest doses, ALG may be dangerous for older patients with poor matches who develop an increased incidence of septic loss of kidney and/or life. Thus, ALG appears to be a useful adjunct in the early management of cadaver transplants by reducing the incidence and frequency of rejection episodes. The dose should probably be reduced in the older patients who receive kidneys from badly mismatched donors. One cannot conclude from this study that ALG manufactured in other centers by this or other techniques, will accomplish the same results since the multiplicity of factors involved in the success and failure of transplants must be controlled so that the influence of intravening variables in the assessment of ALG effectiveness can be assessed.

Prolongation of Skin Homograft Survival in Rabbits by 6-Mercaptopurine.

Summary 6-MP, a powerful metabolic antagonist to nucleic acid synthesis, prolongs survival time of skin homografts in rabbits when administered after grafting. Treatment must be given continuously to maintain graft viability. However, toxicity of 6-MP limits the length of time treatment may be given.

Effect of T cell modulation on the translocation of bacteria from the gut and mesenteric lymph node

Although the ability of the gut-associated lymphoid tissue (GALT) to respond to orally ingested foreign antigens has been studied extensively, its function in preventing or limiting escape of resident gut bacteria has not been assessed. The following studies were performed to examine what role cell-mediated immunity (CMI) plays in this process. The ability of suppression of CMI to induce escape of gut bacteria (translocation) to the mescnteric lymph node (MLN) in immunocompctent mice whose gut flora was unaltered was examined. Administration of cyclosporine or anti-L3T4 antibody failed to induce translocation of indigenous gut bacteria after 7 or 14 days of treatment. Antithymocyte globulin (ATG) also failed to induce translocation after 7 days of treatment, despite depletion of all Thy 1, Lyt 1, L3T4, and Lyt 2 positive cells from the spleen, MLN, and intestine as demonstrated by immunofluorescent microscopy. Finally, cultures of the MLN, spleen, liver, and peritoneum of T cell-deficient BALB/c nude mice and their heterozygous T cell-replete littermatcs were also sterile, demonstrating that congenital suppression of T CMI also docs not lead to translocation of indigenous gut bacteria. The role of CMI in limiting systemic spread of bacteria that were already translocating to the MLN was also examined. Translocation of Escherichia coli C25 to the MLN was induced by gastrointestinal (GI) monoassociation, which leads to translocation of E. coli C25 to the MLN in 80–100% of mice. Treatment with ATG during monoassociation failed to induce spread of E. coli C25 to the spleen, liver, or peritoneum, despite the same degree of T cell depletion achieved with ATG in the previous experiment. Monoassociation of conventional T cell-deficient BALB/c nude and hetero/ygous mice and germfree T cell-deficient BALB/c nude and heterozygous mice also did not lead to spread of E. coli C25 beyond the MLN. However, in ATG-treated, conventional nude, and germ-free nude mice, the average number of translocating E. coli C25 per MLN was consistently higher. In separate experiments the ability of stimulation of T cell function to inhibit translocation of E. coli C25 was examined. Rccombinant intcrleukin-2, 25,000 units, was administered intraperitoneally every 8 hours during exposure to E. coli C25. This reduced the incidence of translocation of E. coli C25 from 85% to 51% (p = 0.02). Suppression of CMI, either systemically or within the GALT, has n minimal influence on the mechanisms by which the normal gut flora are translocated to the MLN. However, suppression of CMI promotes increased survival of the bacteria that have translocated to the MLN. Conversely, augmentation of T cell function may reduce the number of bacteria that survive in the node, possibly by enhanced bacterial killing within the local environment of the MLN.

Effect of Meningococcal Endotoxin on the Immune Response.

Summary 1. The enhancement of antibody production against crystalline bovine serum albumin by the simultaneous injection of meningococcal endotoxin is reported. 2. An optimum dosage of endotoxin for production of the adjuvant effect is approximately the same as that required to prepare rabbits for the generalized Shwartzman reaction or to provoke the local Shwartzman phenomenon. 3. Failure of inhibition of antibody production by dosages of endotoxin capable of producing delayed uptake of radio active colloidal gold by the reticuloendothelial system is mentioned. 4. Possible mechanisms responsible for the adjuvant phenomenon are discussed.

INHIBITION BY TRIPARANOL OF CHOLESTEROL FORMATION IN THE BRAIN OF THE NEWBORN MOUSE

TRIPARANOL (MER-29): is a potent inhibitor of cholesterol synthesis (BLOHM and MACKENZIE, 1959). Treatment with this drug leads to the accumulation of desmosterol (A5*W-cholestadicnol) in thc tissucs of thc rat (AVIGAN et al., 1960a, h ; FRANTZ et al., 1960). Experimental evidence available at this time (AVIGAN et a/., 19600, 6 ; FKANTZ e ta [ . , 1960; STEINBERG and AVIGAN, 1960; SCHROEPIER, 1961) is compatible with the hypothesis that Triparanol inhibits the reduction of the AZ1-double bond of cholesterol precursors. We considercd that if Triparanol inhibited the formation of cholcsterol in nervous tissue, the use of this drug might provide a new approach to the study of myelin formation and structure. In the present study we have found that Triparanol inhibits the formation of cholesterol in the brain of the developing mouse. We have studied the sterol composition of the brains and livers of mice treated with this drug during the first month after birth. The isolation of dcsmostcrol from thc brains of these mice is reported. M A T E R I A L S A N D M E T H O D S

Hemoglobin: A Lifesaver and an Oxidant how to tip the Balance

Hemoglobin solutions were assessed in terms of their ability to promote lipid peroxidation, which was quantitated by measuring the formation of thiobarbituric acid reactive substances (TBARS) under specified conditions in murine brain homogenates. Solutions designed for use in acute treatment of hypovolemic shock and trauma should incorporate ingredients specifically aimed at decreasing oxygen and lipid radical mediated injury occurring secondary to ischemia and reperfusion. A number of strategies aimed at decreasing the oxidant effect of hemoglobin solutions and other blood and plasma substitutes have been evaluated. These include use of the naturally occurring anti-oxidants in human plasma, specifically transferrin and ceruloplasmin. Similarly, certain iron chelators, such as deferoxamine (Desferal, Ciba-Geigy), effectively prevent molecular and cellular damage caused by iron catalyzed formation of oxygen derived radicals.

Enzyme therapy II purified human α-galactosidase A: Stabilization to heat and protease degradation by complexing with antibody and by chemical modification

Abstract Methods were investigated for the stabilization of human splenic α-galactosidase A (α- d -galactoside galactohydrolase, EC 3.2.1.22). Anti-α-galactosidase A antiserum was produced in a goat by repeated immunization with highly purified α-galactosidase A. When this antiserum was incubated at 37°C with α-galactosidase A in varying enzyme: antiserum ratios, a significant increase in the thermal (50 °C) stability and in the resistance to protease digestion of these mixtures was observed compared to appropriate controls. The enzyme was also treated with the bifunctional cross-linking reagent, hexamethylene diisocyanate, and the thermal stability and protease resistance of the cross-linked derivative were increased compared to the native enzyme treated with the monofunctional reagent, butyl isocyanate.

Antithymocyte globulin treatment of severe aplastic anaemia

Nineteen patients with severe aplastic anaemia were treated with antithymocyte globulin. Ten patients obtained remissions (transfusion independent, at least 45000 platelets and 2000 PMN/mm3) within 2–3 months and continue in remission 5–35 months after antithymocyte globulin. Ages of responders ranged from 17 to 71. Complications of antithymocyte globulin included arthralgias, rash, serum sickness, angioedema and fever. Two patients died during, two shortly after, and one 10 months after therapy. One patient with a previous remission following antithymocyte globulin relapsed and achieved a second remission with retreatment. Previous androgen therapy did not affect outcome since two of four patients with and eight of 15 patients without previous androgen therapy achieved remission with ATG. Treatment with antithymocyte globulin is a promising alternative to bone marrow transplantation in the treatment of severe aplastic anaemia.