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The leucocyte migration test (LMT) was used for the study of cellular responsiveness to nuclear factors, i. e. to calf thymus desoxyribonucleic acid (DNA), calf thymus desoxyribonucleoprotein (DNP), calf liver ribonucleic acid (RNA), in 20 subjects with SLE and in 20 non-immunopathological cases serving as controls. The nucleic acid preparations were found to produce a positive LMT in the SLE-patients as well as in the controls. The proportion of positive responses reached its highest percentage in the cases of active SLE. Two cases were followed up by repeated LMT in the course of immunosuppressive treatment with cytostatics and/or antilymphocyte globulin (ALG) administration indicated by the activity of the process. The LMT proved suitable for assessing the intensity of immunosuppression by cytostatics or by ALG. A critical evaluation of the procedure is given. It is pointed out that inhibition and stimulation of migration are likewise manifestations of cellular immune responsiveness. Both inhibition and stimulation are more marked if serum-free nutrient is used.

A solid-phase, one-step radioimmunoassay was developed for the determination of plasma lactoferrin concentration. The detection limit of the assay is 150 micrograms/l. Leakage of cellular lactoferrin was minimal when EDTA was used as anticoagulant, while results obtained from serum and from heparinized plasma were not reproducible. The plasma lactoferrin concentration of 35 female and 44 male healthy adults was measured in order to determine normal values. The geometric mean of lactoferrin levels in men is about 10% higher than in women: 483 (200-1500) micrograms/l in men and 446 (200-870) micrograms/l in women. Patients with acute and chronic leukaemias were also studied. In 38 patients with chronic myeloid leukaemia plasma lactoferrin levels were increased by three times while the neutrophil count was ten times higher than normal. Normal lactoferrin concentrations were measured in plasma samples from 15 patients with chronic lymphocytic leukaemia in incomplete remission while no detectable lactoferrin was found in samples from those in relapse (10 patients). In the untreated patients or those in relapse (19 cases) of both acute lymphocytic and myeloid leukaemias, plasma lactoferrin concentrations were undetectable while they seemed to return to normal during remission (3 cases). The data obtained indicate that the determination of plasma lactoferrin concentration might play an important role in facilitating the assessment of total blood granulocyte pool (TBGP).

Various lymphocyte subpopulations of peripheral blood of 65 patients with aleukaemic non-Hodgkin lymphoma were studied. The clinically active state and remission in both low and high grade malignancy groups were compared. An elevated lymphocyte count was found in the blood of aleukaemic patients, too. Lymphocytosis in low grade malignancies in the clinically active states was found to be higher than in the corresponding group of the high grade cases. The data showed that the most important factor in the increase of total blood lymphocyte count is the increase in the amount of B-lymphocytes. The TA/TT ratio was significantly reduced in all subgroups. The OKT4/OKT8 ratio was normal in all cases.

Thirty-four children of 12 years mean age having had 3 to 5 BCG vaccinations owing to tuberculin negativity, were subjected to cellular and humoral immune assays (tuberculin skin test, leucocyte migration test, lymphocyte transformation test, estimation of serum IgG, IgA, and IgM levels). It was found that nonreactivity may cease by the age of 11 to 12 years. At the time of study 21 of the 34 children were already tuberculin-positive and only 13 failed to react to 5 TU. The studies confirmed the presence of combined immune deficiencies (negative tuberculin test, negative leucocyte migration test, abnormal immunoglobulin levels) as well as of changes confined to the individual immunoglobulin classes. In the case-history recurrent febrile diseases were found to occur more frequently between the 1st and 6th year of life than between 6 and 12 years of age. It is suggested that nonreactivity to tuberculin results from a congenital partial cellular immune deficiency.

The authors investigated the number and functions of peripheral blood monocytes in patients with mixed connective tissue disease. Moderate monocytopenia was detected in the active stage of the disease. There was a close correlation between the sensitized sheep red blood cell binding capacity of monocytes and the elevated levels of circulating immune complexes. The yeast phagocytosis of monocytes and the chemotactic activity were normal in patients with mixed connective tissue disease. C3b receptor mediated phagocytosis of monocytes decreased in the active and inactive stages of the disease. This study suggests that the decreased function of C3b receptors of monocytes is related to the pathogenesis of this disease.

Haemolytic activities of the classical and alternative complement pathways, and levels of C1, C4, C3, factor B and C1 inhibitor (C1-INH) were measured in 137 serum samples of 69 patients with chronic lymphocytic leukaemia (CLL). In most sera IgG, IgA and IgM concentrations were determined as well. Clinical correlations of these laboratory parameters have been studied. C1 and C4 activities were found to be depressed in almost 50% of the sera tested, and hypogammaglobulinaemia was observed with a similar frequency. Low C1 and C4 levels were found mainly in the early stages of the disease. A strong association between the occurrence of infections and hypogammaglobulinaemia was observed, although low immunoglobulin levels frequently occurred in patients without a history of infections. Low C1 and C4 levels were significantly correlated with the incidence of infections, too, and this correlation was observed mostly in the early stages of the disease. The reason for this is not known. The present results suggest that not only low immunoglobulin levels but low C1 and/or C4 levels may contribute to the increased susceptibility to infections in patients with chronic lymphocytic leukaemia.

In vitro pre-incubation of monocytes derived from patients with Hodgkin's disease with tuftsin (50 micrograms/ml) significantly restored the deficient chemotactic responsiveness of these cells to the complement-derived factor C5a, as demonstrated by a monocyte migration assay based on the Boyden technique. Potentiation of the chemotactic responsiveness of monocytes was most significant after elective splenectomy. The results indicate that the specific receptors required for tuftsin activities may be available on the monocyte membranes in Hodgkin's disease. Since tuftsin is a natural, non-immunogenic tetrapeptide that can also be produced synthetically, it may provide a new therapeutic approach in Hodgkin's disease to at least partial restoration of the defective cellular immunity.

T cells and T-cell subsets were studied with monoclonal antibodies in 15 patients with systemic lupus erythematosus (SLE) during a 6-wk period. During active states of disease, all investigated T-cell subsets decreased, but the reduction of suppressor cells seemed to be more pronounced, therefore the helper/suppressor ratio increased. Less suppressor cells could be detected during clinical impairments and more during improvements.

We have investigated factors which influence HLA-DR expression on thyroid cells. While bTSH (100 mU/ml) did not enhance HLA-DR expression, it increased when brought about by IFN-gamma. Graves' IgG showed a dose-dependent (0.1-2 mg/ml) increase in DR expression and at a concentration of 2 mg/ml prolonged the time for which DR was expressed. The pathway of DR induction by Graves' IgG apparently differs from that by IFN-gamma. The humoral response in Graves' disease, by inducing DR expression, may be instrumental in propagating thyroid specific autoimmunity.

Distribution of the T-lymphocyte subpopulation capable of binding own erythrocytes was examined in patients with Hodgkin's (H) disease. Parallel with the autologous rosette-forming cells (Tar-cells) the absolute lymphocyte count and the proportion of sheep-E-rosette forming T-cells were studied in various stages and histological types of H-disease. A considerable increase in the proportion of Tar-cells was found in untreated or recurrent H-disease, whereas in the periods of remission resulting from therapy no distinct differences between these figures and the mean values of the controls were demonstrable. In the generalized stage of the disease as well as in its mixed-cell and lymphocyte depletion types of poor prognosis, the proportion of Tar-cells was increased. The absolute lymphocyte count and the proportion of T-cells showed a significant fall with the increase of Tar-cells. It is assumed on the grounds of the functional properties of T-cells that the increase in the proportion of Tar-cells in H-disease may be interpreted as a sign of suppressor hyperactivity.

1. IC precipitated by PEG from patients with SLE inhibit in vitro the FcR dependent reaction of normal monocytes with sSRBC, while the C3bR dependent reaction of the cells with sensitized yeast is reduced only by some of them. The monocytes were preincubated with the IC for 30 min at room temperature. 2. When the monocytes were incubated with the IC for 22 hours at 37 degrees C the reaction of FcR with sSRBC increased, while the C3bR dependent reaction did not altered. 3. Simultaneously with the increasing FcR dependent reaction, the secretion of lysosomal beta-glucuronidase of monocytes cultivated with IC is greater than those of the controls.

Forty serum samples of healthy blood donors and 60 sera of SLE patients were tested in parallel by 7 different assays for detecting immune complexes. Significantly higher titres and significant higher incidence of positive results were observed in the patient group than in the control group in 6 tests. No test discriminated between patients in an active stage of the disease and those whose disease was inactive. Significantly higher immune complex levels were found in lupus nephritis than in the non-nephritic patients by the complement consumption test. Significant positive correlation was obtained between the results of the macrophage aggregated IgG uptake inhibition and PEG-precipitation tests and between two tests based on the interaction of the complexes with the complement system. A characteristic 'profile' of the immune complex assay was observed in the course of the repeated testings of the same patients. The results indicate that the different methods detect different types of immune complexes present in the blood of the patients.

The investigation of needle- kidney biopsy from patients with SLE- nephrotic syndrome was carried out by the aid of light- immunofluorescent- and electron microscopy. Membranous-, focal-proliferative-, and diffuse-proliferative forms of the SLE- nephrotic syndrome were distinguished. Clinical characteristics of the different forms are described. Authors believe, that the underlying process of nephrological symptoms of various degree as well as readily distinguishable clinico-pathological pictures is the formation of circulating immunocomplexes of different size, developing due to basic disease itself. Comparison of the clinical symptoms and the morphological picture indicate that the elapse of time between the onset of the underlying disease and the nephrotic syndrome has an important part in the formation of the SLE-nephropathy of focal- and diffuse-proliferative type.

Adenosine reduced the expression of CD4 antigen both in HIV infected and uninfected H9 T cells. The continuous presence of adenosine (1 mumol/ml) resulted in a remarkable delay of HIV expression in the infected cells. A relationship is suggested in the effects of HIV and adenosine on the human T cells.

A micro-ELISA technique has been developed to measure antibodies to native DNA and used in SLE patients. The distribution of antibody to native DNA in the main immunoglobulin classes was studied, using anti-human globulin conjugates labelled with peroxidase. the antigen (double-stranded DNA from calf thymus) used in the assay was adsorbed to the surface of polystyrene plates treated with methylated bovine serum albumin. The standardization of the method was carried out by use of globulin calibration curves.

A 47 year old female presented with a septic clinical picture including fever, abscesses, late cachexia, and unmanageable by disease. Similar characteristics to chronic granulomatosis disease (CGD) seriously decreased intracellular killing activity and chemiluminescence, granulomas in the histology, and the role of genetic factors were found, suggesting that our case is CGD-like disorder, manifested in an adult. Dialysable leukocyte extract (DLE) therapy, complemented with fresh normal plasma, resulted in a striking clinical improvement and there was an increase in the in vitro PMNL intracellular killing activity, too. Although it is generally accepted that DLE derives from monocytes and lymphocytes, it is possible that DLE is a family of DNA-oligopeptide molecules, including factors derived from PMNLs which are capable of influencing PMNL function, transferring information from normal cells. Our results also suggest that it would be worth trying DLE in patients with classic CGD.

The ratio of in vivo activated T helper, T suppressor and B cells in the blood of patients with SLE has been studied through simultaneous subset specific immunofluorescence labelling and the analysis of cellular DNA content with the help of flow fluorimetry. In the active stage of the disease an increase was observed in the proportion of proliferating cells either among the T helper cells or the B cells. Occasionally, however, the rate of proliferation in both subsets grew at the same time. Cases with a high number of activated B cells have proved to be more serious. The percentage of the T suppressor cells in the S-G2-M phase of the cell cycle increased moderately in both the active and the inactive stages of the disease. Our results are also discussed in relation to the pathogenetic mechanism of SLE.

The proportion of T-lymphocytes and monocytes was estimated by the alpha-naphthyl-acetate-esterase-(ANAE)-reaction in blood smears of patients with systemic lupus erythematosus and normal controls. A parallel and significant reduction in the number of T-lymphocytes identified on the basis of both the ANAE and the E-rosette test was found in SLE. In the active periods of SLE more T-cells were identified by the enzyme histochemical study than by the E-rosette test, which points to a functional damage to the T-cells. A marked reduction in the number of the cells considered to be of the suppressor type was demonstrable by both methods in the active period. A distinct reduction in the number of monocytes was also demonstrable by ANAE in the patients with SLE, which may be one of the manifestations of deficient immunoregulation in this disease.

In a collaborative study involving 7 laboratories, sera from 53 patients with lung cancer, 37 primary and 16 secondary tumours, and sera of 40 healthy blood donors were tested by 19 different assays or assay modifications used for detecting immune complexes. In 12 out of 19 assays, significantly higher immune complex levels were found in the cancer patients than in the healthy subjects. Assays based on interactions between immune complexes and Fc receptors of different cells (lymphocytes, macrophages of platelets) discriminated between cancer patients and health subjects and a high percentage (47-87%) of positivity was observed in such assays in patients with lung cancer. In contrast, none of the tests based on immune complex-complement interactions discriminated between cancer patients and health subjects. Immunochemical analyses of the PEG precipitates obtained from the sera tested revealed that the concentrations of IgG, IgA and C3 were significantly higher in the precipitates obtained from patients sera than from control sera, but no significant differences were seen in IgM and C1q concentrations. A 100% correct classification of individuals tested was obtained on discriminant analysis of results with 3 assays: EA rosette inhibition, ADCC inhibition and C3 concentration in PEG precipitates. Correlation between results obtained with individual sera by the different assays was very poor: significant correlation coefficients were found in only 13% of all possible paired comparisons. Our results suggest that Fc receptor-dependent assays are more suitable for detection and measurement of circulating immune complexes in lung cancer than tests based on interactions with complement.

The abnormalities of the humoral immune phenomena in pancreatitis may be summarized on the grounds of published evidence and of the present results as follows: 1. Circulating anti-pancreas antibodies are present in acute pancreatitis as also after its cure, and in chronic pancreatitis. 2. The appearance of soluble immune complexes in pathological amounts was demonstrated in the sera of the patients by radiobioassay in acute pancreatitis of identified aetiology 3 to 4 weeks after onset. On the evidence of follow-up studies over 1 to 8 months the immune complexes were found to persist at increased levels after complete cure of acute pancreatitis. Soluble immune complexes were demonstrable in pathological amounts in the cases of chronic pancreatitis as well. 3. In acute pancreatitis the total serum complement titres decline fast and after a few weeks persistent hypocomplementaemia develops in a number of cases. 4. The possible role of circulating soluble immune complexes and of the complement system in pancreatitis is discussed, and it is suggested on the grounds of the present data that acute pancreatitis is associated with a specific and nonspecific immune sensitization, still demonstrable in a number of cases after a few months.

Antibody-dependent cellular cytotoxicity of peripheral blood mononuclear cells from fourty-six patients with systemic lupus erythematosus was found to be reduced in two independent test systems. The percentage decrease in cytotoxicity was similar in the tests with allogeneic and xenogeneic targets and it was related to the activity of systemic lupus erythematosus.

Searching for the cause of the known immunological abnormalities in systemic lupus erythematosus (SLE), the density of cell surface antigens was measured after immunofluorescent staining in a cell sorter. The densities of CD3, CD4, CD5, CD8 and sIgM lymphocyte antigens were the same on patients' lymphocytes as on lymphocytes from healthy subjects. The intensity of HLA-DR immunofluorescence was found to be decreased on patients' monocytes, while the expression of HLA-DR on lymphocytes of patients with SLE hardly differed from that in healthy subjects. Pretreatment of normal mononuclear cells with patients' sera free from immune complexes decreased the binding of anti-HLA-DR antibody to normal monocytes, but it hardly caused alteration on lymphocytes. After culturing, the expression of HLA-DR antigen on patients' monocytes became the same as on normal cells. A causal role of anti-HLA-DR autoantibodies is suggested and discussed.

It is well known that the serum of SLE patients contain anti-lymphocyte antibodies (ALA). In this paper we present a new rosette-forming assay for investigation of ALA. The method is based on the first step of antibody dependent cellular cytotoxicity (ADCC): the adherence of Fc-receptor bearing effector cells to the lymphocytes coated by antibodies. SLE serum samples were tested simultaneously by the new method (the rosette-test, RT) and the microcytotoxicity test (MCT) based on the complement mediated lysis. The results obtained by use of MCT and RT showed that the latter is suitable for investigation of ALA, indeed its diagnostic parameters are better than those of MCT. Moreover these results suggest that the two tests may differ in the antibody class detected. The new method is simple, time saving and demands no 51Cr-isotope as other ADCC tests do.

Monocyte chemotaxis and its response to levamisole were studied in patients with Hodgkin's disease. Levamisole caused a significant improvement of the deficient monocyte chemotaxis of the patients. By enhancing monocyte functions including chemotaxis, levamisole may contribute to the improvement of cellular immune reactivity, thus increasing the resistance to infections and prolonging the periods of remission.

In vitro and in vivo action of levamisole has been studied in patients with Hodgkin's disease. In vitro levamisole significantly increased the active T cell count of 35 untreated active patients, of those being in complete remission after treatment and of 19 healthy controls; it significantly raised the total T cell count of patients. However, levamisole could not further improve the considerably increased active and total spontaneous rosette formation resulting from in vivo levamisole treatment. Twenty patients with Hodgkin's disease, being in complete remission, were given levamisole for 3 or 6 months, according to 2 different schedules. Levamisole (150 mg) given on 3 consecutive days of every second week for 3 months considerably increased the number of positive skin tests (from 8/60 to 29/60), the numbers of active and total T cells with unchanged absolute lymphocyte count, and decreased the quantity of circulating immune complex. Continuation of treatment for another 3 months resulted in slight, but consistent, decline in all the above parameters; the ratio of active T cells significantly decreased. This decline was even more pronounced if a raised larger dosage of levamisole was administered for 6 months. In the course of the treatment no side-effect or complication was observed. Levamisole is able to improve the weak cellular immune reactivity of patients with Hodgkin's disease, it can, however, result in undesired suppression in the case of too long treatment with high dose.

The solubilization of artificial immune complexes mediated by complements has been well-known since 1975. It is known that the process is bound to the integrity of the alternative complement pathway. The phenomenon of solubilization can be used in the investigation of the function of the complement system. We have studied the solubilization of artificial complexes containing BSA and 125I labelled anti-BSA on the effect of sera of healthy subjects and those of patients suffering from SLE. We have observed that the solubilization capacity of SLE sera is significantly lower than that of healthy persons. The decrease is the most distinct at the time of the activity of the disease.

Lymphocyte surface markers of 100 tissue specimens (67 lymph nodes and 33 spleens) of 40 patients with Hodgkin's disease were studied by means of spontaneous E-rosette formation and direct immunofluorescence method. The occurrence of active and total T cells was signigicantly higher (p less than 0.001) in the tumor-involved lymph nodes and spleens, while the frequency of B cells was practically the same in the histologically negative lymph nodes and spleens. In the tumorous spleen of 6 untreated patients the frequency of active and total T cells was higher in the tumorous tissue specimens than in the surrounding histologically negative tissue specimens. No correlation has been found between the simultaneously investigated peripheral T and B cell counts, clinical stage and the histological type of the tumor.

Antigranulocyte antibodies were studied in patients with systemic lupus erythematosus. The frequency and type of the antibodies were identified with ELISA (Enzyme-Linked Immunosorbent Assay) and MGCT (microgranulocytotoxicity test). To check antibody specificity, a LCT (lymphocytotoxicity test) was used parallel with the above technique. The ELISA proved to be suitable for determining antigranulocyte antibodies. There was a correlation between the serum level of IgG-type antigranulocyte antibodies and granulocytopenia, but the IgM-type antigranulocyte antibodies failed to show a similar correlation. A good parallelism was found between MGCT--a test to be used to determine IgM-type antibodies--and the IgM-type antigranulocyte antibodies determined by ELISA. Of 25 SLE sera, 13 were found positive for antigranulocyte antibodies. LCT was used to examine the presence of HLA antibodies in these sera and 39% of the sera positive for antigranulocyte antibodies appeared to be granulocyte-specific while 61% reacted in the LCT, too.

Presence of different types of autoantibodies is a basic feature of systemic lupus erythematosus (SLE). Though monocytes, macrophages play an important role in cellular immunity, autoantibodies against monocytes have not been sufficiently studied. The authors used automatic fluorochromatic assay to detect monocyte reactive autoantibodies in the sera of SLE patients. Of SLE 35.5% sera showed complement-mediated monocytotoxic activity against healthy monocytes. Monocyte reactive SLE sera as well as monoclonal antibodies against human monocytes inhibited chemotaxis of control monocytes. The results suggest that monocyte reactive autoantibodies may play a role in the decreased monocyte number and defective monocyte functions observed in SLE.

18 patients with progressive systemic sclerosis were investigated. An absolute lymphopenia and a decrease in the number of E-rosettes and early E-rosette forming cells were found. The number of T gamma cells was reduced as compared to the control values. No diminution was found in the number of histamine receptor bearing cells but the number of T lymphocytes capable of recognizing autologous red blood cells was considerably decreased. The number and the ratio of these T cell subpopulations remained relatively stable even after 6 months in the patients with progressive systemic sclerosis. No individual correlation was found between the clinical findings and the ratio of T cell subpopulations.

The authors report their observations gathered by follow-up studies of many years duration on 67 patients suffering from mixed connective tissue disease. The period preceding the overt disease, its duration and its characteristic symptoms were examined. Concerning the severity of the clinical course, the authors distinguished between a less and a more severe form of the disease. Organ specific manifestations occurring in the two types of the disorder, the number of active phases per year, therapeutic tools and the prognosis were analysed. The authors assume that the more severe form of the disease may have some traits resembling those of systemic lupus erythematosus, but in the meantime, in the less severe type of the disease some features shared by rheumatic disorders can be found.

The chemiluminescence (CL) induced by zymosan phagocytosis was tested in mouse peritoneal macrophages infected with three different types of herpes viruses: herpes simplex type-1 (HSV-1), human cytomegalovirus (HCMV) and murine cytomegalovirus (MCMV). The intensity of CL was tested in various intervals of virus infections. In the first eight hours zymosan induced chemiluminescence decreased in all the three systems. By the 24th hour, the macrophages infected with HCMV had almost completely recovered from the early defect, while in the macrophages infected with both HSV-1 and MCMV, the chemiluminescence induced by zymosan remained impaired.

Plasma samples of patients with angioimmunoblastic lymphadenopathy (AIBL) were tested for anti-HTLV antibodies and for interferon content. Out of 12 patients 4 had antibodies to HTL-III. Two of these plasma samples contained antibodies reacting with HTLV-I and HTLV-II, too. Activated interferon (IFN) system was found in patients with clinical remission of AIBL, as it was detected by IFN titration in their plasma samples. Data suggest the etiological role in AIBL of virus(es) related to the HTLV family.