Zeitschrift fur Immunitatsforschung, experimentelle und klinische Immunologie最新文献

The effect of 3 schedules for tetanus vaccination on the immunity developed by guinea-pigs was investigated, using 1.5 Lf adsorbed tetanus toxoid injected subcutaneously. The following injection schedules were used: a) 5 injections: initially and after 3, 7, 10, and 13 days; b) 3 injections:: initially and after 2 and 4 weeks; c) 2 injections: initially and after 4 weeks. Blood samples were taken after 1, 2, 3, 4, 6, and 8 weeks and the titres of tetanus antitoxin were recorded. No significant difference in the titres was observed within 2 weeks. Immunity was also tested by challenge with different amounts of tetanus toxin after 1 week, 10 days, 2, 3, and 8 weeks. A tendency to a higher immunity with schedule a) was observed after 10 days to 2 weeks; thereafter no acceleration of immunity could be shown. The results indicate that frequent injections over a short period of time do not establish rapid immunity against tetanus.

The leucocyte migration test from caillary tubes was examined for its usefulness as an assay for cell-mediated immunity in melanoma patients. Formalin fixed melanoma cells either obtained from established cell lines or freshly excised tumors were used as antigen source. From a group of 33 melanoma patients 14 reacted positively (42%) when fixed cultured cells were used, whereas, no positive reactions were found in a group of 14 control donors. However, a considerable proportion of patients with chronic inflammatory processes of the skin reacted positively (4/17 = 24%). A smaller proportion of positive reactions were found in melanoma patients when fixed melanoma cells from excised tumors were used (1/16 = 6%). The positive results in the group with chronic inflammatory processes may be explained as reactions against melanoma associated, but not melanoma-specific, antigens. Three possible reasons are discussed for the low frequency of positive reactions with cells from excised tumors: 1. the cells used in this study display only a few of the antigenic determinants typical for malignant melanoma, 2. antigen expression is quantitatively lower. 3. the surface antigens are covered by blocking factors.

The administration of mycobacterial adjuvant produced a stimulation of the reticuloendothelial system (RES) phagocytic function. The degree of such a stimulation was greater in Lewis than in AVN inbred strain of rats. There was no relationship between the degree of RES stimulation and clinical signs of adjuvant-induced arthritis.

In 32 beagles heterotopic renal allotransplantations and bilateral nephrectomies were carried out. Control animals (5 dogs) survived 9.4 +/- 1.6 days. 22 recipients were pretreated with donor-specific semisoluble spleen antigen; 5 recipients pretreated with the antigen alone (780 mg/kg body weight) survived 16.6 +/- 2.3 days. A pretreatment with antigen combined with 5 mg/kg body weight Prednisolone resulted in no significant prolongation of survival time (5 animals), but 5 dogs survived significantly longer after pretreatment with the antigen and 50 mg/kg body weight Prednisolone (27.4 +/- 2,7 days). In 5 recipients after preoperative application of 3 doses Prednisolone alone (50 mg/kg body weight each) no prolongation of survival time could be observed (10.2 +/- 0.7 days). We found no correlation between lymphocytotoxic antibody titre and survival time.

Spontaneous cell-mediated cytotoxicity (SCMC) and antibody-dependent cellular cytotoxicity (ADCC) against 51Cr labeled allogeneic target cells of a human melanoma cell line (IGR3) were determined with purified effector lymphocytes and defibrinated whole blood from 14 melanoma patients and 13 healthy control persons. Peripheral blood lymphocytes were isolated by Ficoll gradient centrifugation (fraction F); subsequently the phagocytic and adherent cells were removed and the supernating cell population (fraction fff) was passaged through IgG anti-IgG columns to obtain a B cell free lymphocyte suspension (fraction fff-c). cells from the 3 lymphocyte fractions and from defibrinated whole blood were simultaneously tested for cytotoxic activity against unsensitized IGR3 target cells (SCMC assay) and IGR3 cells perviously sensitized with a rabbit anti-melanoma IgG (ADCC assay). Dose-response curves were established with all lymphocyte fractions and with whole blood. The following results were obtained. 1. With all lymphocyte fractions tested, ADCC was approximately 15 time higher than SCMC, whereas with whole blood, the difference tended to be less pronounced. 2. Elimination of phagoctic and adherent cells had no significant effect on SCMC and ADCC. 3. Passage over IgG anti-IgG columns drastically reduced cytotoxicity in both assays without, however, completely abolishing it. 4. The only difference seen between lymphocyte cytotoxicity of melanoma patients and control persons was a slight, but non-significant depression of SCMC and ADCC in melanoma patients. The results confirm and extend our previous report that SCMC against an allogeneic tumor cell-line is due to not-specific "Null" or "K" cell-activity rather than to specific T cell cytotoxicity. In one experiment freshly explanted melanoma cells were labeled with 51Cr and reacted wiht autologous blood and purified lymphocyte fractions. It was found that cellular cytotoxicity depending on serum factors (ADCC) was an effective lytic mechanism, whereas T cell-mediated cytotoxicity could not be demonstrated.

In a group of 48 children the following streptococcal antigens were tested by means of the lymphocyte transformation test: group A streptococcal M1- and M19-protein, mucopeptide, polysaccharides from group A and group C streptococci, erythrogenic toxins and streptokinase. Specific transformations were obtained only by M-proteins. Erythrogenic toxines induced an unspecific stimulation comparable to the mitogenic activity of phytohaemagglutinins. No reactions could be found with streptococcal mucopeptide, the group-specific carbohydrates of groups A and C-streptococci and streptokinase, resp. A significant difference was seen in the group of young children (mean age 5 years) between the specific transformability by M1-protein (a common type) and M19-protein (a rare type). No difference was seen with both M-proteins in the group of older children (mean age 11 years). In both groups of children no differences exist in the number of positive cutaneous reactions after injection of M1- and M19-proteins, but the intensity of reactions (diameters of erythema) were more pronounced with the M-protein of the common type 1.

Using DNCB contact sensitization, the primary cellular immune response of 40 normal individuals was investigated. The response to DNCB in the younger age group was higher and stronger than in the older age group, suggesting a decrease of T-cell function in elderly subjects. The simultaneous application of a high (1000 mug) and a low (100 mug) dose of DNCB for quantitation of the reactivity gave good results. However, in several cases - especially in the younger age group - the local reactions reached unexpected intensity followed by long lasting pigmentation. One investigator exhibited contact eczema on both hands after sensitization by merely touching the skin area of a previous DNCB reaction. Because of the potent immunogenicity of this substance and the possibility of cross reactions with numerous similar antigens widely used in the chemical and related industry, this test should only be applied in selected cases.

Sheep erythrocytes were coated with bencylpenicilloyl-(BPO)groups. Different incubation periods resulted in erythrocyte preparations with different hapten density. Complement dependent lysis induced by IgM or IgG antibodies was studied with the cell preparations. The calculation of hapten density on the erythrocyte surface was not possible by direct measurement of coupled radioactive BPO since more than 90% of radioactive material was found in the soluble supernatant after osmotic cell lysis and less than 10% was fixed to the cellular membrane. Measurement of membrane bound immunologically relevant BPO-groups was achieved, therefore, by comparison of the inhibitory capacity of the test cells with that of a standard cell preparation. The latter consisted of tannic acid treated erythrocytes coated with protein complexed radioactive BPO. Surface hapten density of the different target cell preparations varied between 1.9 x 10(5) and 4.8 10(5) BPO-groups per cell depending on the time of incubation. Complement dependent antibody mediated cell lysis was significantly reduced by reduction of haptenic sites per target cell, IgG induced lysis being much more affected than hemolysis induced by IgM antibodies. Statistical calculations led to the conclusion that 18,000 protein islets per cell bearing 4 or more BPO-groups are not sufficient for hemolysis induced by IgG antibodies. 48,000 protein islets with this hapten density are necessary for "optimal" sensitization. IgG antibodies must be apparently bound to the cell surface in bivalent form.

Previous investigations showed that quantitative immunofluorescence using antigens covalently bound to agarose particles represents a reproducible and sensitive assay for antibodies in experimental antisera. In this paper data are presented which show that also thyroglobulin antibodies in patients sera can be demonstrated using the defined antigen substrate spheres (DASS) system with thyroglobulin as antigen. Sera of 45 patients with various thyroid diseases were investigated for the presence of thyroglobulin antibodies using passive haemagglutination and the quantitative immunofluorescence technique. Comparable results were obtained with both techniques. Advantages and disadvantages of both methods are discussed.

Parameters which are of interest for the better performance of the Cr-releasing cytotoxicity tests were examined, such as type of cells used as targets; influence of various media on the viability of the cells, on uptake of radioactive chromium and on nonspecific chromium release; and influence of specific activity of chromium on the extent of labelling of the targets. The test has been applied to follow the course of primary and secondary immune responses to H-2 alloantigens.