Zeitschrift fur Immunitatsforschung. Immunobiology最新文献

Protein I from the outer membrane of Escherichia coli is a B-lymphocyte mitogen in mice. Polyclonal activation of mouse splenocytes was demonstrated by 3H-thymidine incorporation into DNA, 3H-uridine incorporation into RNA, and by a hemolytic plaque assay in three inbred mouse strains. B-lymphocytes from LPS responder mice (C57Bl/10, STU/nu/nu) and LPS non-responder mice (C3H/HeJ) both responded well to protein I. The presence of serum was not necessary for mitogenicity; bovine serum albumin exhibited a beneficial effect on serum-depleted cultures. Thymocytes of C3H/HeJ mice were not activated by protein I. Protein II* from E. coli was also tested in our systems and showed a weak B-lymphocyte stimulatory activity. Human peripheral blood lymphocytes were not activated.

A new procedure for the isolation of the human complement component C9 is described. This procedure offers the possibility to prepare functionally pure C9 in a one-step procedure with a high recovery of 10-22% of the biological activity. The 125iodinated C9 had a molecular weight of 78,000 daltons and was only contaminated in traces with other proteins. Further purification by absorption with an "anti-impurity" column lead to a C9 preparation which behaved as a homogenous single polypeptide chain in SDS polyacrylamide gel electrophoresis after reduction with mercaptoethanol. It formed a single bell-shaped precipitate in crossed immunoelectrophoresis with antibodies against human serum in the gel of the second dimension. The recovery of the biological activity after the second purification step was in the order of 6-10%. Both preparative steps could be performed within a few hours 450 microgram C9 protein were isolated from 135 ml human serum. A human serum completely defective in C9 was prepared by the extensive absorption of a smaller volume of human serum proteins with the anti-C9 column.

The influence of the Lewis blood group system on transplant survival was studied retrospectively in 161 kidney transplantations. Le (a-b+) recipients had significantly higher graft survival rates than Le (a+b-) or Le (a-b-) recipients. From the known distribution of the Lewis blood groups among the European population, a high percentage of Lewis-compatible transplants would be expected among Le (a-b+) recipients in contrast to the Le (a+b-) and Le (a-b-) recipients. Other factors which are known to influence transplant prognosis such as HLA-match between donor and recipient, ischemic time of the transplants and pretransplant blood transfusions did not differ significantly in any of the three groups studied. Our data again suggest the relevance of the Lewis blood group system for clinical kidney transplantation. The findings should be confirmed by prospective typing of donor and recipient for Lewis antigens.

Rosette formation of human lymphoid cells with mouse erythrocytes has recently been proposed as a marker for a subpopulation of B lymphocytes. In this work we studied the percentage of mouse rosette forming cells (MRFC) in normal and pathological conditions and compared them to the percentage of sheep rosette forming cells (SRFC) a marker for T lymphocytes. Peripheral blood lymphocytes (PBL) from normal donors contained 6.2 +/- 1.1% (mean +/- 1 S.D.) MRFC. High percentages of MRFC were found in CLL patients, and a slight increase was observed in patients with systemic lupus erythematosus. MRFC were absent in Bruton's type agammaglobulinaemia, but were normally present in patients with T cell defects. Cryopreservation of lymphocytes in 10% DMSO did not significantly affect the mean percentages of SRFC and MRFC, though a slight increase of the former and a small reduction of the latter was observed. Double binding experiments on peripheral blood lymphocytes showed a predominant association of MRFC with cells staining for surface IgM and/or IgD. In all samples tested, we also observed a small population of MRFC negative for sIgM or sIgD and a few sIgM or sIgD positive cells that did not rosette with mouse erythrocytes.

While evaluating herpes simplex virus (HSV) typing by indirect immunofluorescence staining, an undesired specific staining pattern turned out to be a reliable marker for herpes simplex type 1. Cells infected with herpes simplex type 1 displayed clear staining with a FITC-conjugated antiglobulin preparation, also in the absence of herpes simplex-specific antibodies. Using the same conjugate, herpes simplex type-2-infected cells exhibited no fluorescence. The particular type of staining observed was influenced by neither the anatomical site of origin of the virus isolate nor the cell type used for virus preparation. Herpes simplex type 1-specific fluorescence was only obtained with the use of FITC-conjugates possessing anti-IgG activity. Both reliability and specificity of this discriminating procedure as a diagnostic tool has been established by typing 282 virus isolates over a period of 4 years.

An adaptation of the nephelometric assay for serum immunoglobulins has been developed for detection and quantitation of extracellular immunoglobulins in cultures of lymphoblastoid cell lines. This assay employs the standard equipment for laser nephelometry and commercial reagents for immunoglobulin quantitation. By adjusting dilutions of controls and sample volumes of culture supernatants, amounts of IgG and IgM below 1 microgram/ml can be detected in culture supernatants. At concentrations between 1 and 4 microgram/ml, day-to-day and within-run variations for IgM assays were 16 and 11% respectively. The possibility of measuring immunoglobulins secreted by cell lines by conventional laser nephelometry opens several areas of application in the study of the functional activity of B cells and of cell-cell interactions.