Biomedical subjects

P R Odgren

Publications and source records attributed to P R Odgren.

26 records · Page 2Linked to original sources

PubMed1989

Storage of platelets in earth orbit. Effect of gravity and the formulation of plastic storage containers on platelet IgG and C3.

We investigated the effects of gravitational force and three different plastic formulations of the storage bags on the quality of stored platelets by measuring the changes in platelet-associated immunoglobulin G (IgG) and two antigenic markers of the third component of human complement (C3), C3d and C3c. Pooled platelets were stored in parallel at microgravity (MG) and on the ground (1 g) using three different plastic polymers: (1) polyvinylchloride (PVC) plasticized with di-2-ethylhexyl phthalate (DEHP); (2) PVC plasticized with trioctyl trimellitate (TOTM), and (3) unplasticized polyolefin (PO). The IgG and C3 were quantified by an automated antiglobulin consumption test in freeze/thaw disrupted platelets (total IgG or C3). The baseline values for platelet associated IgG and C3, measured after 2.5 days of storage at 1 g just prior to the launch, fell within the normal range. Following an additional 6.5 day storage, platelets stored at MG had accumulated significantly less C3d than those stored at 1 g, suggesting that MG storage was beneficial. Specific plastic formulations also exerted a significant effect on the accumulation of these immunoproteins, the effect being particularly pronounced at MG. The smallest increases of IgG and C3 were seen in platelets stored in TOTM and the largest in those stored in DEHP. It is possible that further studies at MG would permit a clear characterization of the effects of other independent storage variables.

Benzoates↗

PubMed1985

Relationship between the third component of human complement (C3) bound to stored preserved erythrocytes and their viability in vivo.

In this study we evaluated whether the 4 degrees C storage-induced coating of red blood cells (RBC) with molecules of the third component of human complement, C3, affects the viability of the preserved RBC. To this end, we determined whether the amount of C3 bound to preserved RBC correlated with the 24-hour survival value. The % anti-C3c-induced agglutination of stored RBC provided an estimate of the amount of RBC-bound C3. In some cases, the number of RBC-bound C3c-containing molecules was also quantitated. The 24-hour survival of autologous RBC was measured in 114 cases. All units were initially stored at 4 degrees C as RBC concentrates followed in 21 cases by frozen storage and in 75 cases by biochemical rejuvenation and frozen storage. The data showed a significant correlation between % anti-C3c-induced agglutination of the preserved RBC and the length of 4 degrees C storage of the RBC concentrates. Neither freezing nor rejuvenation cleaved the C3c fragment from stored RBC. The 24-hour survival was significantly and negatively correlated with both the storage length of RBC concentrates at 4 degrees C and with the amount of RBC-bound C3 but not with RBC ATP level. These data suggest that the RBC-bound C3 either contributes to or is a marker for the extent of the preservation injury of RBC.

Adenosine Triphosphate↗

PubMed1984

Measurement of fragments of the third component of human complement on erythrocytes by a new immunochemical method.

This paper describes a new antiglobulin consumption method to quantitate the fragments of the third component of human complement (C3) on red blood cell (RBC) membranes. Zymosan-bound C3, which can be stored frozen at -80 degrees C for years, was used as a standard in these tests. Using anti-C3c antibody, zymosan-bound C3 could be calibrated against soluble converted C3 (beta 1A), but not against soluble, native C3 (beta 1C). Calibration with several commercial serum standards yielded virtually identical values. Approximately 79.8 +/- 28.2 C3d molecules (mean +/- 1 SD, n = 50) were detected on normal, freshly collected RBC by this method, whereas no C3c fragments were noted. EC43, prepared by dilution of blood samples with low ionic strength solution, had between 650 and 3,100 C3 molecules/RBC when measured with anti-C3c and between 1,140 and 6,500 C3 molecules when measured with anti-C3d. These data indicated that part of the C3b molecules on EC43 had cleaved to C3d. EC43 are reported to have up to 200,000 C3 molecules when measured by other techniques. To resolve this discrepancy, EC43 were prepared by dialysis of blood samples against low ionic strength solution. About 97.5% of C3 remained in plasma after dialysis supporting the results of our tests. The new assay is an accurate and sensitive method of quantitating C3 fragments bound to RBC in vivo and in vitro.

Binding Sites, Antibody↗

PubMed1981

Complement and Ig uptake by erythrocytes in low ionic strength solutions.

We studied the uptake of immunoglobulin (Ig) and complement from plasma by red blood cells (RBC) under low ionic strength (LIS) conditions using Rh-negative RBC and anti-Rh antibodies. The ionic strength of plasma/RBC mixtures was lowered by dialysis against 5% mannitol, pH 6.0. The studies revealed that bromelin-modified RBC failed to bind either Ig or complement in low ionic strength conditions, but that unmodified RBC bound the majority of anti-D. The bound anti-D (consisting of IgG1, IgG3, IgA, and IgM molecules) could be eluted into 0.9% NaCl yielding a higher recovery of antibody than that obtained by specific absorption-elution techniques. Although uptake of Ig by RBC was independent of complement activation, the subsequent elution of bound Ig into normal ionic strength solutions was not. Small amounts of Ig remained on EC43 and EC4, but not on RBC that did not become complement sensitized during low ionic strength exposure. These findings suggest that Ig remained attached to RBC via a complement-RBC bond.

Agglutination↗

PubMed1980

Red blood cell associated IgG in normal and pathologic states.

We studied the anti-IgG-induced agglutination of both normal and abnormal red blood cells (RBC) using a sensitive, automated antiglobulin test. Normal RBC agglutinated strongly with anti-IgG antibody, indicating that IgG was present on the erythrocyte membrane. Young RBC, recovered by centrifugation from a normal RBC population, agglutinated with anti-IgG less than the old cells, suggesting that immunoglobulin G accumulated gradually on the RBC membrane in vivo. The degree of anti-IgG-induced RBC agglutination correlated negatively with the reticulocyte count and positively with the concentration of plasma IgG. RBC from patients with hypogammaglobulinemia appeared to have a low subnormal quantity of membrane-bound IgG, whereas the reverse was the case in hypergammaglobulinemia. During hemolytic episodes, RBC of patients with hereditary spherocytosis agglutinated poorly with anti-IgG, apparently due to predominance of young RBC. RBC of patients with nonspherocytic. Coombs-negative, nonimmune hemolytic anemia usually also agglutinated poorly with anti-IgG. However, in some cases of active hemolytic anemia, decreased agglutination with anti-IgG was not observed, suggesting that these young RBC had increased amounts of membrane-bound IgG.

Agammaglobulinemia↗

PubMed1979

Studies on the preservation of red blood cells. Attachment of the third component of human complement to erythrocytes during storage at 4 degrees C.

Utilizing an automated antiglobulin test, we have investigated the presence of the third and fourth components of human complement on normal red blood cells (RBCs). Only negligible amounts of the fourth component, C4, could be detected on either freshly collected or stored RBCs. The fragment C3d of the third component, C3, was detectable on both freshly collected and stored normal RBCs. A product derived from C3 and reacting with anti-C3c antibody was only barely detectable on freshly collected normal RBCs. During storage of blood at 4 degrees C, increasing quantities of this material were detected on the RBC membrane. Bromelin treatment rendered stored RBCs completely nonreactive with anit-C3c antibody, whereas only partial loss of reactivity was observed following incubation with heated plasma. In contrast, incubation of EC43 with heated plasma completely abolished their ability to react with anti-C3c antibody. We suggest that the presence of this C3 fragment on stored RBCs may contribute to the development of "preservation injury".

Agglutinins↗

PubMedDate not supplied

Freezing red blood cells prepared for quality control of antiglobulin sera.

A small-aliquot freezing technique was employed to store at -80 C red blood cells (RBC) prepared for quality control of antiglobulin sera. These RBC were used to test the specificity and potency of both polyspecific and monospecific antiglobulin sera on the day of use. Following deglycerolization, about 83 per cent of test RBC were recovered. They were then stored as 5 per cent suspensions in 0.9% NaCl at 4 C for up to two weeks and tested for specific agglutination. EIg (RBC sensitized with immunoglobulins) and EC4 (RBC sensitized with the fourth component of human complement) remained reactive for the two-week period. EC43 (RBC sensitized with both the fourth and third components of human complement) tended to lose reactivity only with anti-C3c antibodies following deglycerolization and storage. EC3d (RBC sensitized with the C3d fragment of the third component of human complement), produced in vivo as a result of Mycoplasma pneumoniae infection, remained reactive for the two-week period, whereas EC3d prepared by the alternative pathway of complement activation was useful only for one week. Use of deglycerolized test RBC improved quality-control procedures by saving materials and technician time as well as by providing a constant supply of uniformly prepared test RBC.

Complement C3↗

PubMedDate not supplied

Quantitation of the third component of complement on stored red cells.

By means of an automated anti-C3c consumption technic, we quantitated the molecules of the third component of human complement (containing the C3c fragment) that accumulate on red cells (RBCs) during liquid whole-blood storage. Calibrated C-3 sensitized zymosan particles ( ZyC3 ) were used as standards for bound C3. Although the dose-response curves of anti-C3c neutralization by ZyC3 and stored RBCs were similar, mathematical analysis showed that the shapes of these two sigmoidal curves were significantly different. This indicated that the large C3 molecules bound to stored RBCs differed antigenically from those bound to ZyC3 . Despite this difference, the former could be quantitated adequately by the automated anti-C3c consumption technic. Red cell-bound large C3 molecules were measured following various periods of storage at 4 degrees C of whole blood samples (n=102). An average of 48 molecules were detected on RBCs stored for 21 days. Statistical analysis of these data indicated that during storage at 4 degrees C there was a continuous accumulation of C3 on RBCs, followed by cleavage of C3c fragments. The degree of agglutination of stored RBCs with anti-C3c was proportional to the number of cell-bound large C3 molecules.

Blood Preservation↗