Richard B. Nimberg

Association of anergy with an immunosuppressive peptide fraction in the serum of patients with cancer.

Abstract To study the relation between circulating immunosuppressive factors and anergy in patients with cancer, we tested 53 patients with cancer for hypersensitivity to skin-test antigens and 2,4-dinitrochlorobenzene. Of 41 patients with negative skin tests, 27 (66 per cent) had immunosuppressive serum (≥70 per cent inhibition of lymphocyte stimulation by phytohemagglutinin). None of 12 with positive skin tests had immunosuppressive serum. Thus, anergy and serum immunosuppressive activity were correlated (p<0.05). After ion-exchange chromatography, most of the immunosuppressive activity in cancer serum was in the first peak, fraction I. The same fraction from subjects without cancer contained no detectable immunosuppressive activity. Mean total immunosuppressive activity of cancer serum was 166.0 ± 97.5 (S.D.) units, and that of non-cancer serum was 30.4 ± 9.2 units (p<0.05). Diafiltration of fraction I. from cancer serum yielded a fraction (< 10,000 daltons) that was highly immunosuppressive. Thus, ane...

Correlation Between Anergy and a Circulating Immunosuppressive Factor Following Major Surgical Trauma

In order to clarify the relationship between anergy and immunosuppressive activity in the serum, we studied 46 previously well patients before and at three, five, seven and 28 days after surgery. Delayed hypersensitivity was measured by skin testing with four common recall antigens, and serum immunosuppressive activity was determined by the ability of the patients serum in 10% concentration to suppress by 50% or more the phytohemagglutinin (PHA) stimulation of normal human lymphocytes as compared to pooled normal serum. Prior to surgery, all patients manifested delayed hypersensitivity to one or more antigens, and no patient had immunosuppressive serum. Fifteen patients underwent minor surgery under general anesthesia and did not develop anergy or immunosuppressive serum. Thirty-one patients underwent major cardiovascular surgery. Thirteen of these patients became anergic by day 3 after operation, and 11 of the 13 developed immunosuppressive serum. Eighteen patients maintained delayed hypersensitivity after major surgery, and only three developed immunosuppressive serum. The correlation between anergy and immunosuppressive serum was highly significant (p < 0.001). There was a significant difference in the degree of suppressive activity in the serum of the anergic and reactive patient groups for each postoperative day studied until day 28, when there was recovery of delayed hypersensitivity and lack of immunosuppressive serum. The occurrence of postoperative anergy and immunosuppressive serum was not related to the patients age, sex, number of perioperative blood transfusions or duration of anesthesia but was associated with an increase in postoperative infectious complications (p < 0.05) and in postoperative days in the hospital (p < 0.01). Pooled immunosuppressive serum from anergic patients was fractionated by ion exchange chromatography, gel filtration and preparative high voltage electrophoresis. The majority of the immunosuppressive activity could be accounted for by an electrophoretically homogenous polypeptide-containing fraction not identified in the serum of patients undergoing minor surgery or in normal individuals. We conclude that anergy occurring after major operative trauma is associated with the appearance of a circulating immunosuppressive molecular species and that these events are in turn associated with increased patient morbidity and increased length of hospitalization.

The carbohydrate units of human plasma α1-Acid glycoprotein

Abstract α1-Acid glycoprotein derived from pooled normal human plasma was desialyzed, reduced and carboxymethylated and then digested with chymotrypsin. The resulting glycopeptide mixture, separated from the peptides by gel filtration through Sephadex G-25, was subsequently fractionated by chromatography on CM- and DEAE-celluloses and Dowex 50. The five glycopeptides which were obtained were homogeneous with regard to their electrophoretic properties, amino-terminal amino acids and amino acid compositions, were found to be derived from five specific regions of the polypeptide chain. Each glycopeptide was then subjected to exhaustive treatment with pronase and chromatographed on Dowex-50 at a low ionic strength. Each chromatogram yielded a group of five to eight pronase-glycopeptides, and a total of 26 homogeneous carbohydrate-carrying peptides was obtained. The carbohydrate composition of these compounds indicated that (a) fucose was associated with only the minor glycopeptides of each group of compounds, (b) the heteropolysaccharide units that are linked to each of the five glycosylation sites of the protein are different in size (c) as judged by the composition of the heteroglycans approximately 60% of the carbohydrate units have compositions similar to those of other typical plasma glycoproteins while the remainder of the carbohydrate moiety of this α-globulin is characterized by unusually high galactose contents and high galactose-mannose ratios. Of equal importance, this paper describes a procedure for the isolation and purification of the heteroglycans of each of the carbohydrate attachment sites of a glycoprotein which possesses multiglycosylation sites.

Association of a circulating immunosuppressive polypeptide with operative and accidental trauma.

The serum from 109 traumatized patients was examined for immunosuppressive activity which might explain diminished host immune responsiveness following operative or accidental injury. Twenty-eight fo 31 (90%) severely tralmatized patients, 25 of 60 (42%) moderately traumatized patients, and 0 of 18 minimally traumatized patients developed serum which suppressed the response of normal human lymphocytes to phytohemagglutinin. The degree and duration of serum immunosuppressive activity paralleled the severity of the clinical course but did not correlate with serum cortisol or barbiturate levels. Suppressive sera were not cytotoxic. The immunosuppressive factor(s) was contained in a low molecular weight (less than 10,000 daltons) peptide fraction and was present in 5--10 times the amount recoverable from normal serum. By size and activity the trauma serum factor resembled immunoregulatory alpha globulin, a naturally-occurring serum inhibitor of T-lymphocyte reactions. Thus, depressed immunoreactivity following trauma may be due in part to high concentrations of an endogenous immunosuppressive polypeptide.

The effect of the sialyl residues on the thermodynamic and hydrodynamic properties of α1-acid glycoprotein

Abstract The present study was initiated to define certain hydrodynamic and thermodynamic properties of α 1 - acid glycoprotein and primarily to determine the effect of sialyl residues upon the solution properties of glycoproteins highly hydrated molecule which interacts markedly with the multicomponent system of the solvent. Native α 1 - acid glyco- protein was found to be an asymmetric molecule. In contrast, the desialyzed protein was shown to be spherical and possess a relatively low content of water of hydration. The native and desialyzed glycoproteins were further characterized with respect to several physicochemical properties. Moreover, the molecular weights of these two proteins were determined to be 39 000 and 34 000, respectively.

Three forms of BRP-2 (bone resorptive proteins) from human cancer ascites fluid and their relationship to human serum alpha-2 HS-glycoprotein.

SummaryTwo new forms of BRP-2, a previously described bone resorptive protein, were purified from ascites fluids obtained from patients with hypercalcemia and metastatic bone cancer. The apparent molecular weights of BRP-2 and of these two proteins were 52,000, 48,000, and 46,000, respectively, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The three proteins have essentially the same amino acid compositions but differ with respect to their carbohydrate moieties. The amino-terminal amino acid sequences of the three glycoproteins were identical to each other as well as to human serum α2HS-(human serum) glycoprotein. The relationship of the three forms of BRP-2 to α2HS was also established immunochemically. The ascites proteins, as well as α2HS, on a molar basis, were approximately one-tenth as potent as bovine parathyroid hormone fragment (1–34) in their abilities to stimulate calcium release from bonein vitro. This study describes for the first time a possible function for human serum α2HS.

The prolongation of renal allograft survival by an immunosuppressive peptide isolated from human plasma.

Immunoregulatory α-globulin (IRA), a naturally occurring immunosuppressive peptide isolated from pooled normal human plasma, was administered to Lewis rats before allografting of LBNF1 kidneys. Control Lewis rats received either no treatment or inactive human peptide before kidney allografting. Median survival was 10.0 ± 2.2 (SD) days and 9.0 ± 1.0 days in Lewis rats receiving inactive peptide or no treatment, and 21.0 ± 1.9 days in Lewis recipients treated with IRA, P < 0.05. Three IRA-treated recipients were long-term survivors. At a time when the direct immunosuppressive effect of IRA was long absent, the serum of the long-term survivors could block the killing of LBNF1 target kidney cells by lymphocytes from immune Lewis animals.